專題2:神經(jīng)藥理學(xué)

T2-1

毒品的神經(jīng)免疫藥理學(xué)及創(chuàng)新藥物發(fā)現(xiàn)

王曉輝

（中國科學(xué)院長春應(yīng)用化學(xué)研究所，化學(xué)生物學(xué)實驗室，吉林長春 130022）

關(guān)于鴉片等精神活性物質(zhì)的研究主要圍繞神經(jīng)元，當(dāng)前的戒毒藥物作用于神經(jīng)元的阿片受體或離子通道受體，然而其戒毒效果非常有限。神經(jīng)元不是中樞神經(jīng)系統(tǒng)中調(diào)節(jié)神經(jīng)信號轉(zhuǎn)導(dǎo)的唯一組分，神經(jīng)小膠質(zhì)細(xì)胞占中樞神經(jīng)系統(tǒng)的10%～15%，然而其作用和功能在很長一段時間被忽視。我們和合作者研究發(fā)現(xiàn)鴉片、可卡因、冰毒及其他精神活性物質(zhì)激活Toll樣受體4（TLR4），活化小膠質(zhì)細(xì)胞，產(chǎn)生大量炎癥因子，從而調(diào)節(jié)獎賞信號通路，增加神經(jīng)元的興奮性，導(dǎo)致藥物依賴和成癮，因而TLR4是開發(fā)新型通用戒毒藥物的靶點(diǎn)。以TLR4作為靶點(diǎn)，設(shè)計和發(fā)現(xiàn)一系列TLR4小分子抑制劑，用于治療毒品成癮，其中（+）-naltrexone已轉(zhuǎn)讓給美國Xlaud公司，將進(jìn)行臨床研究新藥（Investiga?tional New Drug，IND）的PhaseⅠ/Ⅱa研究。

鴉片；藥物成癮；疼痛；Toll樣受體4；膠質(zhì)細(xì)胞

T2-2

質(zhì)量標(biāo)準(zhǔn)化的紅花黃酮類提取物抗帕金森病的藥效學(xué)評價

雷 慧1，2，任汝通1，2，努爾·艾買提江·阿布來提1，2，蒲小平1，2

（北京大學(xué)1.藥學(xué)院分子與細(xì)胞藥理學(xué)系，2.天然藥物與仿生藥物國家重點(diǎn)實驗室，北京 100191）

摘要：紅花是一類廣泛用于心腦血管及婦科疾病治療的傳統(tǒng)中藥，主要化學(xué)成分為黃酮類、多糖類、脂肪酸類、生物堿類等。其中黃酮類化合物是紅花發(fā)揮治療作用的主要成分，其主要藥理活性包括抗氧化、抗凝血、抗炎及神經(jīng)保護(hù)作用等。我們的實驗研究表明，質(zhì)量標(biāo)準(zhǔn)化的紅花黃酮類提取物能夠改善MPTP、6-OHDA、魚藤酮誘導(dǎo)的PD動物模型的行為學(xué)、病理學(xué)、腦微環(huán)境等的改變。其指標(biāo)成分紅花黃色素B和山奈酚-3-O-蕓香糖苷，夠抑制H2O2、魚藤酮誘導(dǎo)的細(xì)胞損傷，本文就質(zhì)量標(biāo)準(zhǔn)化的紅花黃酮類提取物的抗帕金森病的藥效學(xué)及其機(jī)制進(jìn)行較為系統(tǒng)的綜述，以期為進(jìn)一步臨床應(yīng)用提供科學(xué)依據(jù)。

關(guān)鍵詞：紅花黃酮類提取物；帕金森??；神經(jīng)保護(hù)；6-OH?DA；腦微環(huán)境

基金項目：科技部重大新藥創(chuàng)制項目（2012ZX09103201-042）；國家重大科學(xué)儀器設(shè)備開發(fā)專項（2013YQ03065106）

通訊作者：蒲小平，E-mail：pxp123@bjmu.edu.cn，Tel：（010）82802431

T2-3

間苯三酚類衍生物WYD1-8抑制神經(jīng)炎癥作用機(jī)制的研究

臧彩霞，鮑秀琦，鄭遠(yuǎn)鵬，張 丹

（中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院藥物研究所，天然藥物活性物質(zhì)與功能國家重點(diǎn)實驗室，北京100050）

摘要：目的帕金森病（PD）是以靜止性震顫、肌強(qiáng)直、運(yùn)動遲緩為特征的神經(jīng)退行性疾病。其病理學(xué)特征主要為黑質(zhì)致密部多巴能神經(jīng)元的選擇性丟失以及路易小體的形成。近年來的研究表明，神經(jīng)炎癥與PD發(fā)病關(guān)系密切，在PD的發(fā)生和發(fā)展過程中發(fā)揮重要的作用?；衔颳YD1-8為間苯三酚類衍生物，前期結(jié)果顯示W(wǎng)YD1-8在體外可通過抑制神經(jīng)炎癥反應(yīng)而保護(hù)神經(jīng)元。在多種PD動物模型中，WYD1-8均可明顯改善小鼠的運(yùn)動行為學(xué)障礙，抑制腦內(nèi)炎癥和保護(hù)多巴胺能神經(jīng)元，機(jī)制研究表明，WYD1-8的神經(jīng)保護(hù)作用與其抑制神經(jīng)炎癥密切相關(guān)，但其抑制神經(jīng)炎癥的機(jī)制尚未明確。本研究在前期研究工作的基礎(chǔ)上，在BV2小膠質(zhì)細(xì)胞水平對其抑制神經(jīng)炎癥作用機(jī)制進(jìn)行初步探討，明確WYD1-8作用機(jī)制。方法采用LPS刺激小膠質(zhì)細(xì)胞BV2，ELISA法測定WYD1-8對LPS誘發(fā)的TNF-α和PGE2釋放量的影響，Western blotting法檢測WYD1-8對炎癥蛋白COX-2、iNOS表達(dá)及JAK/STAT信號通路和NADPH氧化酶亞基膜轉(zhuǎn)位的影響；流式細(xì)胞儀檢測WYD1-8對活性氧ROS產(chǎn)生的影響。結(jié)果WYD1-8能明顯抑制LPS誘發(fā)的BV2小膠質(zhì)細(xì)胞炎癥因子TNF-α和PGE2的釋放，同時能抑制炎癥蛋白COX-2和iNOS的表達(dá)并抑制JAK/STAT信號通路的激活；流式細(xì)胞儀檢測ROS，發(fā)現(xiàn)WYD1-8可明顯抑制ROS的產(chǎn)生；Western blotting結(jié)果發(fā)現(xiàn)WYD1-8能抑制上游的NADPH氧化酶亞基P47的膜轉(zhuǎn)位，從而抑制NADPH氧化酶的激活。結(jié)論實驗結(jié)果表明，間苯三酚類衍生物WYD1-8對神經(jīng)炎癥有明顯的抑制作用。WYD1-8可能通過抑制上游的NADPH氧化酶的激活，進(jìn)而抑制JAK/ STAT炎癥信號通路活化發(fā)揮抑制神經(jīng)炎癥的作用，但其具體的作用靶點(diǎn)仍需進(jìn)一步研究。

關(guān)鍵詞：間苯三酚類衍生物WYD1-8；神經(jīng)炎癥

T2-4

百可利對魚藤酮致帕金森病模型的神經(jīng)保護(hù)作用

張 雪1，杜立達(dá)2，楊玉林3，杜冠華1

（1.中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院藥物研究所，北京市藥物靶點(diǎn)研究與新藥篩選重點(diǎn)實驗室，北京100050；2.香港中文大學(xué)生物醫(yī)學(xué)學(xué)院，香港；3.廣東藥科大學(xué)，廣州廣東528458）

摘要：目的研究百可利對魚藤酮致帕金森?。≒D）模型的神經(jīng)保護(hù)作用。方法SD大鼠腹腔注射魚藤酮（2.5 mg-1·kg·d-1）制備PD動物模型，利用TUNEL染色以及cleaved-caspase 3表達(dá)變化檢測大鼠黑質(zhì)神經(jīng)元凋亡。1 μmol·L-1魚藤酮損傷SH-SY5Y建立PD細(xì)胞模型。MTT法評價細(xì)胞活力；Bcl-2/Bax、細(xì)胞色素c的檢測反映線粒體凋亡途徑。結(jié)果與魚藤酮模型大鼠相比，百可利（400 mg·kg-1）能有效地降低黑質(zhì)神經(jīng)元凋亡指數(shù)（8.9%vs39.6%，P＜0.01），抑制caspase-3的活化（109.5%vs387.0%，P＜0.01）。在魚藤酮損傷SH-SY5Y細(xì)胞模型上，與模型組相比，百可利（10 μmol·L-1）可提高細(xì)胞活力（85.5%vs73.0%，P＜0.01）、抑制細(xì)胞凋亡（194.4%vs225.2%，P＜0.01）；升高Bcl-2/Bax比值，抑制細(xì)胞色素c釋放，從而抑制線粒體凋亡途經(jīng)。結(jié)論百可利通過抑制魚藤酮誘導(dǎo)的線粒體凋亡途徑，緩解魚藤酮引起的黑質(zhì)神經(jīng)元凋亡，從而發(fā)揮對魚藤酮致帕金森病模型的神經(jīng)保護(hù)作用。

關(guān)鍵詞：百可利；帕金森??；線粒體凋亡途徑；魚藤酮

基金項目：北京協(xié)和醫(yī)學(xué)院研究生創(chuàng)新基金項目（2015-1007-05）

通訊作者：杜冠華，E-mail：dugh@imm.ac.cn，Tel：（010）63165184

T2-5

百可利對魚藤酮誘導(dǎo)的帕金森病模型大鼠腦組織抗氧化能力的影響

楊玉林1，張 雪2，杜冠華1，2

（1.廣東藥科大學(xué)，廣東廣州 510006；2.中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院藥物研究所，北京市藥物靶點(diǎn)研究與新藥篩選重點(diǎn)實驗室，北京100050）

摘要：目的考察百可利對魚藤酮誘導(dǎo)的帕金森病模型大鼠腦組織抗氧化能力的影響并探討相關(guān)機(jī)制，為百可利治療帕金森病提供理論依據(jù)。方法SD大鼠腹腔注射魚藤酮（2.5 mg·kg-1·d-1）制備PD動物模型，測定大鼠腦組織組織中超氧化物歧化酶（SOD）、谷胱甘肽過氧化物酶（GPx）和丙二醛（MDA）水平。1 μmol·L-1魚藤酮損傷SH-SY5Y建立PD細(xì)胞模型研究百可利抗氧化機(jī)制。利用DCFH-DA探針反映細(xì)胞內(nèi)ROS水平，并檢測氧化應(yīng)激重要通路NRF-2/HO-1的活化情況。結(jié)果與魚藤酮模型大鼠相比，百可利能顯著地上調(diào)抗氧化酶SOD（200 mg·kg-1·d-1，124.0%vs57.5%，P＜0.05），下調(diào)腦組織中MDA（400 mg·kg-1·d-1，96.9%vs473.8%，P＜0.01）。在魚藤酮損傷SH-SY5Y細(xì)胞模型上，與模型組相比，百可利（10 μmol.L-1）能明顯抑制細(xì)胞內(nèi)ROS水平（159.7%vs88.9%，P＜0.01），促進(jìn)NRF-2的核轉(zhuǎn)位（76.1%vs69.6%，P＜0.01），并增加HO-1的表達(dá)（74.4%vs60.2%，P＜0.01），發(fā)揮抗氧化作用。結(jié)論百可利通過激活NRF-2/HO-1抗氧化通路，調(diào)節(jié)魚藤酮誘導(dǎo)的體內(nèi)體外氧化應(yīng)激失衡，從而發(fā)揮治療帕金森病作用。

關(guān)鍵詞：百可利；帕金森??；氧化應(yīng)激；魚藤酮

通訊作者：杜冠華，E-mail：dugh@imm.ac.cn，Tel：（010）63165184

T2-6

MPTP致慢性帕金森病小鼠模型的潛在生物標(biāo)記物及藥物靶點(diǎn)的研究

崔丹丹，王曉良，彭 英

（中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院藥物研究所，北京100050）

摘要：目的我們前期在帕金森?。≒D）患者和健康人血清蛋白組學(xué)研究中發(fā)現(xiàn)了一些差異性蛋白，本研究將采用PD動物模型對差異蛋白進(jìn)行驗證，以期發(fā)現(xiàn)PD潛在的生物標(biāo)記物。在神經(jīng)毒素致慢性小鼠模型上驗證PD患者血漿來源的差異蛋白。方法構(gòu)建（1-甲基-4-苯基-1，2，3，6-四氫吡啶（MPTP）慢性PD模型。分別于造模結(jié)束后第1周、8周、18周進(jìn)行神經(jīng)行為學(xué)評價、取材，利用高效液相方法檢測神經(jīng)遞質(zhì)，提取黑質(zhì)、紋狀體區(qū)蛋白，進(jìn)行Western Blot差異蛋白驗證。結(jié)果①行為學(xué)評價：經(jīng)MPTP誘導(dǎo)，模型組轉(zhuǎn)棒實驗潛伏期明顯縮短（P＜0.05），掉落次數(shù)增加（P＜0.05）；自主活動次數(shù)減少（P＜0.05）；爬桿實驗爬下時間延長。第18周時，模型組小鼠體重明顯低于同齡對照組（P＜0.05）。②紋狀體區(qū)神經(jīng)遞質(zhì)檢測：模型組的DA，DOPAC，HVA，5-HT水平明顯下降（P＜0.01，n=5），三個時間點(diǎn)的DA含量平均下降70.8%。③Western Blot驗證：進(jìn)行了ITIH4，α-synu?clein，DJ-1，apoA-Ⅳ，apoA-Ⅰ，plasma Kallikrein，MMP-7和HSC-70等蛋白驗證，發(fā)現(xiàn)模型組黑質(zhì)、紋狀體區(qū)上述蛋白的表達(dá)無變化，但血漿中ITIH4 35KD和α-synuclein蛋白含量明顯下降。結(jié)論ITIH4和α-synuclein在PD動物模型中有明顯改變，有望成為PD診斷和治療的新型的生物標(biāo)記物。

關(guān)鍵詞：帕金森??；MPTP模型；生物標(biāo)記物

基金項目：國家科技重大專項（2012ZX09301002-004）；北京市重點(diǎn)實驗室（BZ0150）

通訊作者：王曉良，E-mail：wangxl@imm.ac.cn，Tel：（010）63165330；彭 英，E-mail：ypeng@imm.ac.cn，Tel：（010）63165173

T2-7

曹麗霞，何靜波，鄭延澤，張浩楠，田彩云，武海軍，楊玉梅

（包頭醫(yī)學(xué)院心血管研究室，內(nèi)蒙古包頭 014040）

摘要：目的探討腦缺血損傷后不同時相腦組織和血清中髓過氧化物酶（MPO）的活性變化以及蒙藥嘎日迪-13對其的影響。方法取雄性SD大鼠360只，將大鼠隨機(jī)分為假手術(shù)組、腦缺血模型組、嘎日迪-13大劑量組、嘎日迪-13中劑量組、嘎日迪-13小劑量五個組，每組又隨機(jī)分為缺血1 h組、6 h組、12 h組、24 h組、72 h組與120 h組6個時相組，嘎日迪-13大、中、小給藥組分別以1.2 g/100 mL，0.6 g/ 100 mL和0.3 g/100 mL濃度2 mL/100 g灌胃，1次/d，連續(xù)27 d后，采用改良Zea Longa線栓法制備大鼠大腦中動脈栓塞（MCAO）模型，進(jìn)行神經(jīng)功能評分，在各時相規(guī)定時間點(diǎn)麻醉大鼠后從腹主動脈采血并取腦組織，采用分光光度法檢測MPO活性。結(jié)果與假手術(shù)組比較，腦缺血模型組各時相血清和腦組織中MPO活性都明顯升高（P<0.05）；與腦缺血模型組比較，嘎日迪-13大、中、小劑量組各時相血清和腦組織中MPO活性都明顯降低（P<0.05），且12和24 h時作用尤為明顯（P<0.01）。結(jié)論嘎日迪-13可能是通過減少腦缺血損傷后各時相MPO活性，調(diào)節(jié)各時相炎癥因子相互之間的作用，減輕腦組織損傷，從而發(fā)揮對腦缺血損傷的保護(hù)作用。

關(guān)鍵詞：嘎日迪-13；腦缺血；腦組織；血清；髓過氧化物酶

基金項目：國家自然科學(xué)基金（81160560）

通訊作者：楊玉梅，E-mail：yym457@aliyun.com，Tel：13847230457

T2-8

丁苯酞的抗焦慮作用及機(jī)制研究

大都市區(qū)域產(chǎn)業(yè)發(fā)展的模式是通過單核心區(qū)與多條產(chǎn)業(yè)通道相結(jié)合的布局方式來實現(xiàn)的，以大都市區(qū)核心區(qū)為核心，向外延伸形成了各具不同產(chǎn)業(yè)發(fā)展特色的五條產(chǎn)業(yè)廊道。大都市區(qū)的核心為大都市區(qū)核心增長極，其中昌宜產(chǎn)業(yè)廊道就是依托奉新工業(yè)園，形成了南昌西面產(chǎn)業(yè)發(fā)展廊道。

付治鳳，陳春林

（宜春學(xué)院化學(xué)與生物工程學(xué)院，江西宜春 336000）

摘要：目的觀察丁苯酞在高架十字迷宮實驗、孔板實驗中的抗焦慮作用，并探究其作用機(jī)制。方法將50只雄性ICR小鼠隨機(jī)分為5組，分別為空白對照組（生理鹽水）、地西泮組（2.5 mg·kg-1）、丁苯酞（100，30和10 mg·kg-1）。連續(xù)灌胃給藥14 d，于末次給藥1 h后進(jìn)行行為學(xué)測試。用酶聯(lián)免疫法測定小鼠血清皮質(zhì)酮的含量，化學(xué)顯色法測定小鼠皮層組織中丙二醛（MDA）、超氧化物歧化酶（SOD）、一氧化氮（NO）的含量。結(jié)果在小鼠高架十字迷宮實驗中，與空白對照組相比，丁苯酞高劑量組能夠明顯增加小鼠進(jìn)入開放臂的次數(shù)（OE%，P＜0.05）和進(jìn)入開放臂的時間（OT%，P＜0.01）；在小鼠孔板實驗中，丁苯酞高劑量組能夠顯著提高小鼠探頭次數(shù)和持續(xù)時間以及直立次數(shù)和持續(xù)時間（P＜0.01）；丁苯酞30和100 mg·kg-1能明顯拮抗印防己毒素誘導(dǎo)的驚厥實驗；與空白對照組相比，丁苯酞高劑量組能夠明顯降低小鼠皮層組織中一氧化氮（NO）水平（P＜0.05），但是丁苯酞對小鼠血清皮質(zhì)酮、皮層組織中的丙二醛（MDA）、超氧化物歧化酶（SOD）水平?jīng)]有影響。結(jié)論丁苯酞具有明顯的抗焦慮作用，其作用機(jī)制可能與γ-氨基丁酸（GABA）受體和降低腦內(nèi)一氧化氮（NO）的水平有關(guān)，與小鼠血清皮質(zhì)酮、皮層組織中丙二醛（MDA）、超氧化物歧化酶（SOD）的水平無關(guān)。

關(guān)鍵詞：丁苯酞；抗焦慮；高架十字迷宮實驗；γ-氨基丁酸受體；一氧化氮

T2-9

癲癇發(fā)作和抗癲癇藥物誘導(dǎo)共同導(dǎo)致耐藥性癲癇腦中P-糖蛋白過表達(dá)

劉曉東

（中國藥科大學(xué)藥物科學(xué)研究院，江蘇南京 210009）

摘要：癲癇是最常見的神經(jīng)紊亂癥狀之一。越來越多證據(jù)證實20%～30%癲癇病人是耐藥的。臨床和動物實驗顯示血腦屏障（BBB）上P-糖蛋白過表達(dá)可能是其癲癇耐藥的原因之一。BBB限制藥物進(jìn)入腦內(nèi)，導(dǎo)致腦組織對多種藥物耐受。P-GP廣泛表達(dá)于腦血管內(nèi)皮細(xì)胞腔側(cè)面，在BBB功能中發(fā)揮重要作用。本文總結(jié)我們前期研究成果。研究顯示，在用閾下劑量戊四氮唑（PTZ）連續(xù)28 d誘導(dǎo)頑固性癲癇模型大鼠腦中P-GP的功能與表達(dá)顯著增加，伴隨腦內(nèi)抗癲癇藥物苯巴比妥濃度顯著降低。合用P-GP抑制劑環(huán)胞菌素A在增加腦內(nèi)苯巴比妥濃度的同時，顯著增加苯巴比妥的抗癲癇作用。在體外大鼠腦微血管內(nèi)皮細(xì)胞（rBMEC）模型也證實了苯巴比妥在BBB上轉(zhuǎn)運(yùn)主要是P-GP介導(dǎo)的，提示癲癇大鼠腦內(nèi)苯巴比妥濃度降低可能與P-GP功能與表達(dá)上調(diào)有關(guān)。一般認(rèn)為癲癇往往伴隨短暫的腦缺血/缺氧。為此，我們用rBMEC研究短暫缺氧對P-GP功能與表達(dá)的影響。結(jié)果顯示,每天短暫缺氧15 min，連續(xù)8 d，rBMEC中P-GP蛋白表達(dá)增加約1.6倍，伴隨細(xì)胞對羅丹明123和苯巴比妥的攝取顯著降低，提示缺氧可能是引起癲癇鼠BBB上P-GP功能與表達(dá)的原因之一。進(jìn)一步在體和離體（rBMEC）水平研究幾種常用抗癲癇藥物對BBB上P-GP功能與表達(dá)的影響，結(jié)果證實苯巴比妥，苯妥因，卡馬西平和丙戊酸均可顯著誘導(dǎo)BBB上P-GP的功能與表達(dá)。上述結(jié)果提示頑固性癲癇病人腦內(nèi)P-GP功能與表達(dá)上調(diào)可能歸功于癲癇發(fā)作和藥物誘導(dǎo)的共同作用。進(jìn)一步以苯巴比妥為模型藥物，采用PTZ誘導(dǎo)的癲癇大鼠模型驗證這一推論。癲癇大鼠用苯巴比妥（45 mg·kg-1·d-1）治療40 d，同時考察腦內(nèi)P-GP的功能與表達(dá)及其腦內(nèi)苯巴比妥濃度。與我們預(yù)期結(jié)果一致，苯巴比妥短期治療顯示良好地抗癲癇活性，治療14 d效果最好。同時，腦內(nèi)被癲癇誘導(dǎo)的P-GP功能與表達(dá)被逆轉(zhuǎn)。然而繼續(xù)治療，苯巴比妥的抗癲癇活性逐漸降低，治療40 d后，苯巴比妥不再顯示其抗癲癇作用。此時，腦內(nèi)P-GP功能與表達(dá)又進(jìn)一步加強(qiáng)，伴隨腦內(nèi)苯巴比妥濃度顯著降低，與苯巴比妥抗癲癇作用降低一致。上述結(jié)果提示可以得出這樣的推論：頑固性癲癇病人腦內(nèi)P-GP過表達(dá)可能部分歸功于癲癇發(fā)作和藥物誘導(dǎo)作用。早期主要是癲癇發(fā)作，后期則是由癲癇發(fā)作和藥物誘導(dǎo)共同作用。

關(guān)鍵詞：抗癲癇藥物；P-糖蛋白

基金項目：國家自然科學(xué)基金（81373482）

T2-10

以Nampt/SIRT1為靶點(diǎn)探索通絡(luò)醒腦泡騰片抗氧化應(yīng)激作用

付文君1，2，魏江平1，2，陳 歡1.2，徐世軍1，2

（1.四川省中藥資源系統(tǒng)研究與開發(fā)利用國家重點(diǎn)實驗室培育基地，四川成都 611137；2.成都中醫(yī)藥大學(xué)藥學(xué)院，四川成都 611137）

摘要：目的考察通絡(luò)醒腦泡騰片對SAMP8小鼠學(xué)習(xí)記憶的影響，探究其抗AD作用是否與調(diào)控Nampt/SIRT1有關(guān)。方法SAMP8小鼠給通絡(luò)醒腦泡騰片（4.05g生藥·kg-1）干預(yù)60 d；采用Morris水迷宮評價其認(rèn)知變化，隨后免疫組化法測定海馬中Nampt和SIRT1的平均光密度，ELISA法測定腦組織NAD+/NADH表達(dá)。結(jié)果與模型對照組比較，通絡(luò)醒腦泡騰片使SAMP8小鼠在隱藏平臺實驗中第5天的逃避潛伏期明顯縮短（24.46±16.69）%，平臺象限所占時間百分比顯著增加（54.91±33.28）%，在空間探索實驗中進(jìn)入目標(biāo)象限次數(shù)增加（49.61±13.68）%；海馬中Nampt多分布于細(xì)胞漿，陽性表達(dá)產(chǎn)物為為棕黃色顆粒狀，給藥組Nampt陽性表達(dá)產(chǎn)物顯著增多，著色較淺，平均光密度增加（3.37± 0.23）%，SIRT1主要表達(dá)在細(xì)胞核，陽性染色呈棕黃色，給藥組SIRT1陽性表達(dá)增多，著色較深，平均光密度增加（5.05±0.58）%，且腦組織中NAD+/NADH比值提高（46.04± 5.42）%。結(jié)論通絡(luò)醒腦泡騰片可上調(diào)Nampt表達(dá)而提高NAD+的比率進(jìn)而促進(jìn)SIRT1的表達(dá)，因此通絡(luò)醒腦泡騰片改善SAMP8小鼠認(rèn)知功能與Nampt/SIRT1抗氧化應(yīng)激途徑有關(guān)。

關(guān)鍵詞：阿爾茨海默??；通絡(luò)醒腦泡騰片；SAMP8小鼠；Nampt；SIRT1

基金項目：國家自然科學(xué)基金（81273900）；四川省學(xué)術(shù)和技術(shù)帶頭人培養(yǎng)資金資助項目（2015058）；中藥藥理四川省青年科技創(chuàng)新研究團(tuán)隊（2014TD0007）

通訊作者：徐世軍，E-mail：docxu@126.com，Tel：（028）61800231

T2-11

桃紅四物湯對魚藤酮誘導(dǎo)帕金森細(xì)胞模型保護(hù)作用

李 雙，郭春燕，劉豐熙

（河北北方學(xué)院藥學(xué)系，河北張家口 075000）

摘要：目的研究桃紅四物湯對魚藤酮致帕金森細(xì)胞模型的保護(hù)作用及對細(xì)胞氨基酸代謝與抗氧化活性的影響。方法將細(xì)胞分為0.025%二甲基亞砜（DMSO）溶媒組，魚藤酮0.15 μmol·L-1模型組和桃紅四物湯（TSD）給藥組（10，50和100 μg·mL-1）。魚藤酮作用SH-SY5Y細(xì)胞24 h體外模擬帕金森模型。TSD與魚藤酮同時加入，研究其保護(hù)作用。MTT法檢測細(xì)胞活力；DAPI熒光染色觀察細(xì)胞核型；West?ern蛋白印跡法檢測細(xì)胞中α-突觸核蛋白（α-synuclein）的表達(dá)；高效液相色譜柱前衍生化法測定細(xì)胞氨基酸含量；酶生化法檢測超氧化物歧化酶（SOD）活性。結(jié)果模型組較DMSO對照組，細(xì)胞存活率顯著降低（P＜0.01），細(xì)胞形態(tài)與核型發(fā)生改變，α-synuclein表達(dá)顯著升高（P＜0.01）；胞內(nèi)10種氨基酸（谷氨酸、谷氨酰胺、絲氨酸、組氨酸、甘氨酸、脯氨酸、苯丙氨酸、異亮氨酸、亮氨酸、賴氨酸）含量顯著降低（P＜0.01），興奮性神經(jīng)遞質(zhì)谷氨酸與抑制性神經(jīng)遞質(zhì)谷氨酰胺平衡失調(diào)；SOD活性顯著下降（P＜0.01）。TSD組與模型組相比，劑量依賴地逆轉(zhuǎn)上述變化。結(jié)論TSD對魚藤酮造成的帕金森病模型具有保護(hù)作用，與TSD調(diào)整細(xì)胞氨基酸代謝的紊亂及增強(qiáng)抗氧化酶SOD活力有關(guān)。

關(guān)鍵詞：桃紅四物湯；魚藤酮；SH-SY5Y；帕金森??；α-突觸核蛋白；氨基酸；高效液相色譜法

基金項目：河北省教育廳重大項目（ZD2014075）；河北北方學(xué)院博士科研啟動基金項目資助

通訊作者：郭春燕，E-mail：guochy0311@163.com

T2-12

紅景天苷促大鼠腦缺血再灌注腦組織的神經(jīng)再生作用

韓 靜，張繼州，肖 慶，胡 娟

（福建省中醫(yī)藥研究院，福建福州）

摘要：目的探討紅景天苷對大鼠腦缺血再灌注損傷后腦組織神經(jīng)再生的作用。方法SD大鼠隨機(jī)分為假手術(shù)組、模型組和紅景天苷組，采用線栓法制作大鼠腦缺血再灌注模型，紅景天苷組于造模后給予紅景天苷（40 mg·kg，ip）每日1次，連續(xù)7 d。給藥結(jié)束，采用改良神經(jīng)功能缺損評分（mNSS）、平衡木平衡試驗（Beam Balance）、足失誤試驗評價動物神經(jīng)功能恢復(fù)狀況；采用免疫熒光化學(xué)方法檢測側(cè)腦室下區(qū)（SVZ）和海馬齒狀回（DG）細(xì)胞增殖標(biāo)記BrdU和新生神經(jīng)細(xì)胞標(biāo)記DCX，免疫組織化學(xué)法半定量檢測缺血側(cè)皮質(zhì)和紋狀體NGF和BDNF的表達(dá)。結(jié)果紅景天苷可顯著降低腦缺血損傷大鼠的mNSS評分、降低Beam Balance評分，減小足失誤比率（P＜0.05）。紅景天苷組SVZ區(qū)BrdU和DCX的表達(dá)較模型組顯著升高（P＜0.05），在海馬DG區(qū)，紅景天苷組DCX的表達(dá)較模型組顯著升高（P＜0.05）；在缺血側(cè)大腦皮質(zhì)和紋狀體，紅景天苷組BDNF的免疫陽性反應(yīng)強(qiáng)度顯著高于模型組。結(jié)論紅景天苷可促進(jìn)腦組織內(nèi)源性神經(jīng)再生，改善腦缺血損傷的神經(jīng)行為學(xué)癥狀，該作用可能與紅景天苷增加腦內(nèi)神經(jīng)營養(yǎng)因子的表達(dá)相關(guān)。

關(guān)鍵詞：紅景天苷；腦缺血；BrdU；DCX；BDNF

基金項目：福建省自然科學(xué)基金項目（2014J01340）；福建省衛(wèi)計委中青年骨干人才項目（2014-ZQN-JC-32）

通訊作者：胡 娟，E-mail：huj@fjtcm.edu.cn，Tel：（0591）83570397

T2-13

丹酚酸A對大鼠腦缺血再灌注損傷的保護(hù)機(jī)制

宋俊科，張 雯，何國榮，閆 蓉，杜冠華

（中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院藥物研究所，北京市藥物靶點(diǎn)研究與新藥篩選重點(diǎn)實驗室，北京100050）

摘要：目的研究丹酚酸A（SAA）對大鼠腦缺血再灌注損傷的保護(hù)機(jī)制。方法SD大鼠隨機(jī)分組，線栓法制備大鼠大腦中動脈閉塞/再灌注（MCAO/R）模型，缺血1.5 h，再灌注24 h，對神經(jīng)行為學(xué)缺損程度進(jìn)行評分，TTC法測定大鼠腦梗死體積，Western蛋白印跡法法測定腦組織胞核、胞漿Nrf2和全細(xì)胞HO-1蛋白含量。結(jié)果大鼠MCAO手術(shù)1.5 h，再灌注24 h可造成明顯的神經(jīng)功能障礙，多數(shù)動物出現(xiàn)追尾或向一側(cè)傾倒。SAA（10 mg·kg-1）可改善MCAO/R引起的神經(jīng)功能損傷癥狀，顯著降低神經(jīng)行為學(xué)缺損評分。MCAO手術(shù)1.5 h，再灌注24 h，大鼠缺血腦半球出現(xiàn)梗死，而SAA（10 mg·kg-1）可顯著減小MCAO/R大鼠腦梗死體積，減輕腦水腫程度。同時SAA（10 mg·kg-1）能夠促進(jìn)Nrf2蛋白的核轉(zhuǎn)位，使HO-1蛋白表達(dá)增加。結(jié)論SAA具有抗腦缺血再灌注損傷的作用，其機(jī)制可能與激活Nrf2/HO-1信號途徑，促進(jìn)Nrf2的核轉(zhuǎn)位，從而促進(jìn)下游抗氧化蛋白HO-1的表達(dá)有關(guān)。

關(guān)鍵詞：丹酚酸A；腦缺血再灌注；Nrf2；HO-1

通訊作者：杜冠華，E-mail：dugh@imm.ac.cn，Tel：（010）63165184

T2-14

DL0410改善APP/PS1轉(zhuǎn)基因小鼠學(xué)習(xí)記憶的作用機(jī)制

周 圍，劉艾林，杜冠華

（中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院藥物研究所，北京100050）

摘要：目的探討DL0410改善APP/PS1轉(zhuǎn)基因小鼠學(xué)習(xí)記憶能力的作用機(jī)制。方法將9月齡的APP/PS1雙轉(zhuǎn)基因小鼠隨機(jī)分為模型組、多奈哌齊（3 mg·kg-1）治療組、美金剛（2 mg·kg-1）治療組、DL0410（3，10和30 mg·kg-1）劑量組，并以同背景陰性的C57小鼠作為正常對照組。灌胃給藥2個月后采用筑巢實驗、新物體識別實驗以及Morris水迷宮實驗檢測轉(zhuǎn)基因小鼠的學(xué)習(xí)記憶能力，行為學(xué)測試結(jié)束后，處死小鼠，用Western蛋白印跡法檢測皮質(zhì)和海馬組織中淀粉樣前體蛋白（APP）糖原合成酶激酶-3（GSK-3β），蛋白激酶B（AKT），腦源性神經(jīng)生長因子（BDNF），促凋亡蛋白（Bax）以及抗凋亡蛋白（Bcl-2）的表達(dá)。結(jié)果多項行為學(xué)實驗結(jié)果均表明，DL0410可明顯改善APP/PS1轉(zhuǎn)基因小鼠的學(xué)習(xí)記憶能力。病理染色結(jié)果表明，DL0410不僅可以減輕模型小鼠神經(jīng)元的腫脹，增加神經(jīng)元的存活數(shù)量，還可以有效清除沉積于皮質(zhì)和海馬區(qū)的老年斑及Aβ。同時，DL0410能夠增加小鼠皮質(zhì)和海馬腦組織中AKT、BDNF以及bcl-2的表達(dá)，減少GSK-3β和Bax的表達(dá)。結(jié)論DL0410通過AKT-GSK-3β信號通路改善APP/PS1小鼠的學(xué)習(xí)記憶能力。

關(guān)鍵詞：阿爾茨海默?。籄PP/PS1轉(zhuǎn)基因小鼠；行為學(xué)實驗；GSK-3β；AKT

基金項目：“十二五”科技重大專項（2014ZX09507003-002）

通訊作者：劉艾林，E-mail：liuailin@imm.ac.cn，Tel：（010）63165184；杜冠華，E-mail：dugh@imm.ac.cn，Tel：（010）63165184

T2-15

銀杏二萜內(nèi)酯對缺血再灌注大鼠腦組織中神經(jīng)遞質(zhì)的影響

張 雯1，宋俊科1，何國榮1，張 雪1，周啟蒙1，王振中2，蕭 偉2，杜冠華1

（1.中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院藥物研究所，北京市藥物靶點(diǎn)研究與新藥篩選重點(diǎn)實驗室，北京 100050；2.中藥制藥過程新技術(shù)國家重點(diǎn)實驗室，江蘇康緣藥業(yè)股份有限公司，江蘇連云港 222001）

摘要：目的探討銀杏二萜內(nèi)酯葡胺注射液（DGMI）對缺血再灌注（MCAO/R）大鼠腦內(nèi)氨基酸類和單胺類神經(jīng)遞質(zhì)的影響。方法采用線栓法制備大鼠腦缺血再灌注模型，缺血1.5 h，再灌注24 h，尾靜脈給予不同劑量DGMI后，測定腦內(nèi)氨基酸類和單胺類神經(jīng)遞質(zhì)含量。結(jié)果10 mg·kg-1DGMI治療組能夠顯著降低MCAO/R引起的大鼠神經(jīng)行為學(xué)缺損評分。大鼠腦缺血1.5 h，再灌注24 h，缺血側(cè)半球出現(xiàn)大面積梗死，DGMI 3和10 mg·kg-1治療組均可減小腦梗死體積，與模型組相比，分別降低22%和32%；DGMI還可以劑量依賴的減輕MCAO/R引起的腦水腫，與模型組比較，三個劑量組的水腫程度分別降低9%，10%和12%。DMGI可以抑制缺血再灌注導(dǎo)致的腦內(nèi)天冬氨酸、谷氨酸、甘氨酸和γ-氨基丁酸的升高；降低缺血腦區(qū)皮質(zhì)和海馬內(nèi)去甲腎上腺素、二羥苯乙酸、5-羥色胺、5-羥吲哚乙酸的濃度，抑制皮質(zhì)內(nèi)腎上腺素的減少。結(jié)論DGMI可以通過調(diào)節(jié)腦內(nèi)氨基酸類和單胺類神經(jīng)遞質(zhì)的水平，對大鼠MCAO/R引起的損傷發(fā)揮治療作用。

關(guān)鍵詞：銀杏二萜內(nèi)酯；腦缺血再灌注；氨基酸類遞質(zhì)；單胺類遞質(zhì)

基金項目：“重大新藥創(chuàng)制”國家科技重大專項資助項目（2013ZX09402203）；抗毒藥物與毒理學(xué)國家重點(diǎn)實驗室開放課題（TMC201510）；國家自然科學(xué)青年基金（81603100）；北京協(xié)和醫(yī)學(xué)院研究生創(chuàng)新基金（2015-1007-07）

通訊作者：杜冠華，E-mail：dugh@imm.ac.cn，Tel：（010）63165184

T2-16

DL0410對APP/PS1轉(zhuǎn)基因小鼠的神經(jīng)保護(hù)作用

閆 蓉，劉艾林，杜冠華

（中國醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院藥物研究所，北京市藥物靶點(diǎn)研究與新藥篩選重點(diǎn)實驗室，北京 100050）

摘要：目的研究DL0410對APP/PS1轉(zhuǎn)基因動物模型的神經(jīng)保護(hù)作用。方法本研究選用9月齡APP/PS1轉(zhuǎn)基因小鼠，灌胃給予DL0410（10和30 mg·kg-1）8周后，進(jìn)行被動回避（跳臺）、新物體識別和水迷宮等行為學(xué)實驗，考察DL0410對APP/PS1轉(zhuǎn)基因動物模型學(xué)習(xí)記憶功能的作用。并觀察DL0410對離體海馬腦片CA3區(qū)schaffer側(cè)枝-CA1區(qū)LTP的影響。結(jié)果DL0410在APP/PS1轉(zhuǎn)基因小鼠模型上，在小鼠定向航行第4天與模型組比較給予化合物DL0410（10和30 mg·kg-1）后對小鼠尋找站臺的潛伏期有明顯的縮短（P＜0.05）；被動回避能力試驗結(jié)果顯示，與模型組比較給予DL0410（30 mg·kg-1）對其跳臺潛伏期明顯提高（P＜0.05）。Control組與模型組比較HFS后各段時間點(diǎn)的EPSP斜率值均降低（P＜0.05），而DL0410組各個時間段的EPSP幅明顯高于模型組（P＜0.01）。結(jié)論DL0410通過改善突觸可塑性及空間學(xué)習(xí)記憶功能障礙，發(fā)揮對APP/PS1轉(zhuǎn)基因動物的神經(jīng)保護(hù)作用。

關(guān)鍵詞：DL0410；APP/PS1轉(zhuǎn)基因動物模型；學(xué)習(xí)記憶

基金項目：國家重大新藥創(chuàng)制科技重大專項（2014ZX09507003-002，2013ZX09508104001002）；醫(yī)科院博士后科學(xué)基金（500101234）

通訊作者：杜冠華，E-mail：dugh@imm.ac.cn，Tel：（010）63165184

T2-17

兩種線粒體復(fù)合體抑制劑擬PD動物模型的建立及行為學(xué)檢測

張如意，張 麗，李 林

（首都醫(yī)科大學(xué)宣武醫(yī)院藥物研究室，北京市老年病醫(yī)療研究中心，北京 100000）

摘要：目的比較兩種線粒體復(fù)合體抑制劑所致動物模型的行為學(xué)異同，為實驗動物模型的選擇提供科學(xué)依據(jù)。方法Lewis大鼠經(jīng)魚藤酮（RT）皮下注射建立擬帕金森?。≒D）大鼠模型，采用一般行為學(xué)觀察，曠場實驗，斜板實驗檢測RT大鼠行為學(xué)。C57BL小鼠經(jīng)MPTP 30 mg·kg-1連續(xù)腹腔注射5 d建立擬PD模型，經(jīng)爬桿實驗、轉(zhuǎn)棒實驗和自主活動實驗測試MPTP小鼠行為學(xué)變化。結(jié)果RT模型組大鼠出現(xiàn)行動遲緩，身體向一側(cè)偏斜，毛色發(fā)暗。隨著造模時間延長，模型大鼠癥狀持續(xù)加重，出現(xiàn)偶發(fā)緩慢旋轉(zhuǎn)，僵直。造模2周后，模型組大鼠體重明顯降低。以曠場實驗檢測各組大鼠運(yùn)動功能，記錄各組大鼠3 min內(nèi)的跨格次數(shù)和站立次數(shù)。與對照組相比，模型組大鼠跨格次數(shù)和站立次數(shù)均顯著減少。以改良的Rivlin斜板實驗測定各組大鼠的肌肉協(xié)調(diào)性。與對照組相比，模型組大鼠在斜板的停留角度顯著減小，表明模型組大鼠肌肉協(xié)調(diào)性顯著降低。MPTP模型組小鼠的爬桿時間較對照組顯著延長，在棒時間明顯縮短，自主活動次數(shù)減少。結(jié)論RT模型動物造價高，造模周期長，大鼠行為學(xué)改變明顯，MPTP小鼠模型造模簡單、快速，適宜作為初篩的動物模型。

關(guān)鍵詞：帕金森病；線粒體；動物模型；魚藤酮；MPTP

基金項目：國家自然科學(xué)基金（81273498）；首都衛(wèi)生發(fā)展科研專項（2011-1001-04）

通訊作者：李 林，E-mail：linlixw@126.com；張如意，zhan?gruyi@xwh.ccmu.edu.cn，Tel：83198881

T2-18

匹諾塞林對大鼠腦梗死后rtPA誘導(dǎo)的出血性轉(zhuǎn)化的預(yù)防作用及機(jī)制

馬寅仲1，2，李 莉2，孔令雷2，張 雯2，宋俊科2，杜冠華2

（1.天津藥物研究院新藥評價有限公司，天津 300301；2.中國醫(yī)學(xué)科學(xué)院藥物研究所，北京 100050）

摘要：目的通過不同方式將匹諾塞林與rtPA聯(lián)合使用治療血栓性腦梗死，以期最大限度地降低延遲溶栓造成的出血性轉(zhuǎn)化，進(jìn)而延長rtPA的有效溶栓治療時間窗。方法本實驗設(shè)計了3種匹諾賽林與rtPA的聯(lián)合用藥方式：給予rtPA前5 min靜脈注射匹諾塞林（P+T）；二者注射前預(yù)先混合后再行注射（MIX）；給予rtPA后立即注射匹諾塞林（T+P）。通過本課題組建立大鼠血管內(nèi)成栓腦梗死模型，在造模2，4，6和8 h后，分別進(jìn)行不同的聯(lián)合用藥治療，并對大鼠的神經(jīng)行為學(xué)，腦出血，腦水腫，腦血流量等指標(biāo)進(jìn)行檢測，評價最佳聯(lián)用方式。進(jìn)而對給藥后腦脊液的實時變化進(jìn)行了觀察，結(jié)合腦組織中與血腦屏障相關(guān)的蛋白表達(dá)情況，分析匹諾賽林對出血預(yù)防作用的潛在機(jī)制。結(jié)果與單純進(jìn)行溶栓治療相比，三種聯(lián)合用藥方式均可以在造模4 h內(nèi)顯著降低伊文藍(lán)在腦組織中的滲漏，但P+T效果最強(qiáng)，T+P效果最弱，這與匹諾賽林在大鼠腦脊液的實時血藥濃度正相關(guān)。此外，僅P+T組可以在6 h內(nèi)顯著改善腦部供血，緩解神經(jīng)功能與運(yùn)動功能失調(diào)，降低腦水腫。免疫印跡法證明缺血腦組織中的基質(zhì)金屬蛋白酶受到匹諾塞林的調(diào)節(jié)而表達(dá)下降。結(jié)論匹諾塞林可以顯著改善rtPA對腦栓死大鼠血腦屏障的損傷，緩解出血性轉(zhuǎn)化，延長溶栓治療窗。其作用機(jī)制可能與匹諾賽林對血腦屏障的保護(hù)作用有關(guān)。

關(guān)鍵詞：血管內(nèi)造栓腦梗死模型；rtPA；rPA；匹諾塞林；出血性轉(zhuǎn)化

通訊作者：杜冠華，E-mail：dugh@imm.ac.cn，Tel：（010）63165184

T2-19

復(fù)方柴歸方抗抑郁藥效學(xué)特點(diǎn)

張 濤，令狐婷，張 瀟，田俊生，秦雪梅

（山西大學(xué)中醫(yī)藥現(xiàn)代研究中心，山西太原 030006）

摘要：抑郁癥屬于情感性心境障礙，以顯著而持久的情緒低落、快感缺失為核心癥狀，具有發(fā)病率高、復(fù)發(fā)率髙、致殘率高的“三高”特點(diǎn)。同時還會伴發(fā)意志減弱、思維遲緩、認(rèn)知功能損害、睡眠障礙、性功能障礙、便秘或腹瀉等癥狀。前期經(jīng)過藥效篩選和化學(xué)成分指紋圖譜分析，得到了逍遙散抗抑郁活性部位，并通過藥效成分歸屬和原方湯劑臨床觀察進(jìn)行處方化裁，得到了優(yōu)選復(fù)方柴歸方。針對其抗抑郁藥效特點(diǎn)研究結(jié)果如下：復(fù)方柴歸方各劑量組可以顯著降低懸尾實驗（TST）和強(qiáng)迫游泳實驗（FST）模型小鼠的不動時間；可以顯著改善利血平誘導(dǎo)的小鼠體溫降低和眼瞼下垂等癥狀；對慢性溫和不可預(yù)知應(yīng)激（CUMS）模型大鼠伴發(fā)的焦慮癥狀有顯著的改善作用；可以顯著改善CUMS模型大鼠伴發(fā)的認(rèn)知功能障礙，而文拉法辛對這一癥狀沒有明顯改善作用；可以顯著降低大鼠的排便數(shù)目，調(diào)節(jié)大鼠糞便含水量趨近于空白組水平，表明復(fù)方柴歸方對CUMS大鼠胃腸功能有較好的調(diào)節(jié)作用。之外，復(fù)方柴歸方給與CUMS大鼠停藥后無明顯反彈現(xiàn)象，表明其具有一定程度的防治抑郁復(fù)發(fā)作用。

關(guān)鍵詞：柴歸顆粒，抗抑郁，藥效學(xué)，作用特點(diǎn)

通訊作者：秦雪梅，Email：qinxm@sxu.edu.cn，Tel：（0351）7011501

T2-20

培元解郁方對急慢性應(yīng)激BALb/c小鼠的抗抑郁作用及TRY-KYN代謝途徑的調(diào)節(jié)

暢洪昇，王萍萍，劉保秀

（北京中醫(yī)藥大學(xué)中藥學(xué)院，北京100102）

摘要：目的研究四逆散、天絲飲及其合方培元解郁方的抗抑郁作用及其對血清皮質(zhì)酮含量和色氨酸（TRP）-犬尿氨酸（KYN）途徑關(guān)鍵代謝酶色氨酸2，3-雙加氧酶（TDO）的影響。方法使用強(qiáng)迫游泳和束縛加噪聲分別造成小鼠急慢性抑郁模型，采用Noldus EthoVision XT9系統(tǒng)采集分析小鼠強(qiáng)迫游泳不動時間，采用HPLC-MS/MS技術(shù)檢測血清色氨酸（TRP）、犬尿氨酸（KYN）、犬尿烯酸（KA）、喹啉酸（Q）、皮質(zhì)酮（C）含量，實時熒光定量PCR檢測肝組織中TDO的mRNA表達(dá)水平。結(jié)果急性應(yīng)激后，與模型組比較，天絲飲、培元解郁方能縮短小鼠不動時間（P＜0.01），減少血清皮質(zhì)酮的含量（P＜0.01，P＜0.05），降低TDOmRNA的表達(dá)（P＜0.05）。其中，天絲飲能提高KYN/Q比值（P＜0.05），表現(xiàn)出抑制喹啉酸作用。慢性應(yīng)激后，四逆散、天絲飲、培元解郁方均能縮短慢性應(yīng)激小鼠游泳不動時間（P＜0.05），降低血清皮質(zhì)酮含量（P＜0.01，P＜0.01，P＜0.05），增加KYN/TRP比值（P＜0.01），KYN/Q比值（P＜0.01）。同時，培元解郁方降低KYN/KA比值（P＜0.01）。三方對對慢性應(yīng)激TDO mRNA表達(dá)無明顯影響。結(jié)論天絲飲、培元解郁方具有較明顯的抗抑郁藥物樣作用，這一作用與對抗皮質(zhì)酮和抑制色氨酸-犬尿氨酸通路神經(jīng)毒性代謝產(chǎn)物產(chǎn)生，提高神經(jīng)保護(hù)作用有關(guān)。

關(guān)鍵詞：四逆散；天絲飲；培元解郁方；抗抑郁；色氨酸-犬尿氨酸代謝

基金項目：國家自然科學(xué)基金（81373584）

通訊作者：暢洪昇，E-mail：chs1971@sina.com，Tel：18910848375

T2-21

斯皮諾素誘導(dǎo)小鼠非快動眼睡眠

王 娟1，倪 健1，曲衛(wèi)敏1，2

（復(fù)旦大學(xué)，1.基礎(chǔ)醫(yī)學(xué)院藥理學(xué)系，2.腦科學(xué)研究院協(xié)同創(chuàng)新中心，上海 200032）

摘要：目的研究酸棗仁有效成分斯皮諾素對小鼠睡眠-覺醒的影響。方法C57BL/6J小鼠頭部埋置電極，恢復(fù)10 d后實驗。小鼠在19：00分別腹腔注射斯皮諾素5，10和20 mg·kg-1，記錄腦電和肌電，觀察睡眠-覺醒的變化。為探索斯皮諾素作用機(jī)制，在斯皮諾素（20 mg·kg-1）給藥前15 min，給與GABAA受體苯二氮卓艸類結(jié)合位點(diǎn)拮抗劑氟馬西尼，以及5-HT1A受體拮抗劑WAY-100635。結(jié)果小鼠活躍期給與斯皮諾素20 mg·kg-1，能明顯增加非快動眼（non-rapideye movement，NREM）睡眠量（P<0.01），作用持續(xù)3 h；并顯著縮短小鼠睡眠潛伏期（P<0.05）。累計3 h的睡眠-覺醒量，斯皮諾素10和20 mg·kg-1劑量依賴地分別增加NREM睡眠2.5和3.1倍；減少覺醒量35.9%和54.6%；但REM睡眠無顯著差異。斯皮諾素5 mg·kg-1不影響動物的睡眠-覺醒量。斯皮諾素可增加小鼠覺醒向睡眠的轉(zhuǎn)換次數(shù)（P＜0.01），減少平均覺醒持續(xù)時間（P＜0.01），但不改變NREM睡眠腦波的delta power。進(jìn)一步發(fā)現(xiàn)，氟馬西尼不能阻斷斯皮諾素的促眠作用，而WAY-100635與小劑量斯皮諾素（5 mg·kg-1）聯(lián)合使用，能誘導(dǎo)動物睡眠。結(jié)論斯皮諾素促進(jìn)小鼠NREM睡眠，可能是通過拮抗5-HT1A受體。提示斯皮諾素可能作為新型促眠藥物用于失眠癥的治療。

關(guān)鍵詞：斯皮諾素；睡眠；氟馬西尼；5-HT1A受體

基金項目：國家自然科學(xué)基金（31471064）

通訊作者：曲衛(wèi)敏，E-mail：quweimin@fudan.edu.cn，Tel：（021）54237103

T2-22

白藜蘆醇抑制星形膠質(zhì)細(xì)胞氧化應(yīng)激的細(xì)胞內(nèi)作用機(jī)制

李雪粉1，2，3，陳文武1，郭安臣3，4，5，王擁軍2，3，4，5，王 群2，3，4，5

（1.河南大學(xué)第一附屬醫(yī)院神經(jīng)內(nèi)科，河南開封 475001；2.首都醫(yī)科大學(xué)附屬北京天壇醫(yī)院神經(jīng)病學(xué)中心，北京100050；3.腦血管病轉(zhuǎn)化醫(yī)學(xué)北京市重點(diǎn)實驗室，北京100050；4.北京腦重大疾病研究院，北京100050；5.國家神經(jīng)系統(tǒng)疾病臨床醫(yī)學(xué)研究中心，北京100050）

摘要：中樞神經(jīng)系統(tǒng)損傷以及退行性疾病的病理過程復(fù)雜多樣，但均伴有不同程度的星形膠質(zhì)細(xì)胞氧化應(yīng)激。在各種不同神經(jīng)疾病的病理過程中，活性氧（ROS）生產(chǎn)過剩是星形膠質(zhì)細(xì)胞活化的一個重要的潛在因素。各種內(nèi)源性及外源性因素可直接或間接地通過NADPH氧化酶刺激ROS的產(chǎn)生，ROS激活MAPK家族，通過NF-κB使sPLA2的表達(dá)增加。近年來研究發(fā)現(xiàn)，多酚化合物白藜蘆醇（RES）具有抗氧化、抗炎、抗細(xì)胞增殖等多種功能。RES和Apocynin均屬于植物多酚化合物，其分子結(jié)構(gòu)有相似性，具有抗氧化、抗炎作用。Apocynin作為NADPH氧化酶特異性抑制劑已經(jīng)被作為干預(yù)工具廣泛用于藥理學(xué)研究。因此可以推斷RES可能有類似于Apocynin的NADPH氧化酶抑制特性。拓展RES針對抑制星形膠質(zhì)細(xì)胞NADPH oxidase-ROSMAPKs-NF-κB-sPLA2通路防治神經(jīng)系統(tǒng)疾病的藥理干預(yù)，具有一定的藥理價值，并為RES防治中風(fēng)和癲癇發(fā)作所致神經(jīng)元損傷的臨床前研究向臨床轉(zhuǎn)化奠定基礎(chǔ)，具有重要的臨床應(yīng)用前景。

關(guān)鍵詞：白藜蘆醇；星形膠質(zhì)細(xì)胞；氧化應(yīng)激

基金項目：國家自然科學(xué)基金（81171097，81271312）

通訊作者：王 群，E-mail：wangq@ccmu.edu.cn，Tel：13146254818

T2-23

BKCa通道在適度酒精預(yù)適應(yīng)保護(hù)腦缺血再灌注損傷中的作用及ROS的調(diào)控機(jī)制

蘇 芳1，2，3，4，5， 朱雨嵐2， 郭安臣3，4，5， 王擁軍1，3，4，5，王 群1，3，4，5

（1.首都醫(yī)科大學(xué)附屬北京天壇醫(yī)院神經(jīng)病學(xué)中心，北京100050；2.哈爾濱醫(yī)科大學(xué)附屬第二醫(yī)院神經(jīng)內(nèi)科，黑龍江哈爾濱 150001；3.腦血管病轉(zhuǎn)化醫(yī)學(xué)北京市重點(diǎn)實驗室，北京100050；4.北京腦重大疾病研究院，北京100069；5.國家神經(jīng)系統(tǒng)疾病臨床醫(yī)學(xué)研究中心，北京100050）

摘要：目的探討B(tài)KCa通道在酒精預(yù)適應(yīng)（EtOH-PC）保護(hù)腦缺血再灌注（I/R）損傷中的作用及調(diào)控機(jī)制。方法1.建立原代皮質(zhì)神經(jīng)元OGD/R模型，CCK-8檢測細(xì)胞活性、流式及TUNEL法檢測凋亡、Fluo4孵育檢測細(xì)胞內(nèi)鈣、West?ern檢測凋亡相關(guān)蛋白及BKCa-α亞基表達(dá)，觀察BKCa在EtOH-PC中的作用。2.膜片鉗單通道記錄觀察酒精對BKCa通道的作用。3.建立大鼠MCAO模型，給予Mn-TBAP藥理干預(yù)，再灌注24 h后行為學(xué)評分，TTC染色。結(jié)果1.與OGD/R組相比，10 mmol·L-1EtOH-PC可提高細(xì)胞存活率，降低凋亡細(xì)胞比例；EtOH-PC前給予Pax，神經(jīng)元存活率較酒精組下降；與OGD/R組相比，EtOH-PC降低OGD/R誘導(dǎo)的細(xì)胞內(nèi)鈣升高水平及凋亡細(xì)胞比例，上調(diào)Bcl-2表達(dá)，下調(diào)Bax表達(dá)，而Pax拮抗酒精的作用；OGD/R組BKCa-α亞基蛋白表達(dá)水平較對照組升高。2.酒精可電壓依賴性的增加皮質(zhì)神經(jīng)元BKCa通道電流及開放概率，激活BKCa通道。3．EtOH-PC與BKCa通道激動劑NS11021預(yù)適應(yīng)組大鼠神經(jīng)功能缺損評分及腦梗死體積顯著低于I/R組，預(yù)適應(yīng)前給予Mn-TBAP腹腔注射，神經(jīng)功能評分及腦梗死體積增高。結(jié)論EtOH-PC通過激活BKCa通道降低細(xì)胞內(nèi)鈣、抑制凋亡而發(fā)揮對OGD/R誘導(dǎo)神經(jīng)元損傷的保護(hù)作用；OGD/R誘導(dǎo)BKCa通道α亞基表達(dá)上調(diào)，可能參與了腦缺血再灌注損傷的內(nèi)源性防御機(jī)制。ROS調(diào)控了EtOH-PC激活BKCa通道發(fā)揮對I/R損傷的神經(jīng)保護(hù)作用。

關(guān)鍵詞：腦缺血再灌注；酒精預(yù)適應(yīng)；神經(jīng)保護(hù)；BKCa通道；ROS

基金項目：國家自然科學(xué)基金（81171097；81271312）

通訊作者：王 群，E-mail：wangq@ccmu.edu.cn，Tel：（010）67098544

T2-24

以炎癥為靶點(diǎn)治療慢性缺血后的腦白質(zhì)損傷

胡薇薇，周怡亭，張 菁，馬 婧，陳 忠

（浙江大學(xué)藥醫(yī)藥學(xué)部理系神經(jīng)科學(xué)研究中心，浙江杭州310058）

摘要：大腦長期慢性缺血將可能導(dǎo)致腦白質(zhì)損傷及認(rèn)知功能損害，造成皮質(zhì)下缺血性血管性癡呆。目前臨床上缺乏針對性的有效藥物，因此急需針對其發(fā)病機(jī)制尋找減輕髓鞘丟失或促進(jìn)髓鞘再生修復(fù)的治療新靶點(diǎn)。課題組研究發(fā)現(xiàn)，慢性缺血后髓鞘再生過程受到抑制，主要是因為腦室下區(qū)增殖的前體細(xì)胞OPC向白質(zhì)區(qū)遷移過程受到阻礙，因而OPC遷移障礙是慢性缺血后髓鞘再生和白質(zhì)修復(fù)的關(guān)鍵限速步驟。進(jìn)一步發(fā)現(xiàn)，慢性缺血早期膠質(zhì)細(xì)胞高表達(dá)炎癥因子IL-1β，其受體拮抗劑（IL-1Ra）或基因敲除其受體（IL-1R1 KO）可以促進(jìn)腦室下區(qū)的OPC（逆轉(zhuǎn)錄病毒標(biāo)記）向腦白質(zhì)區(qū)遷移的過程，但對OPC的增殖沒有影響；而在IL-1R1 KO小鼠上，通過腺病毒介導(dǎo)的IL-1R1過表達(dá)可以逆轉(zhuǎn)OPC遷移的增多。并且發(fā)現(xiàn)阻斷IL-1β受體IL-1R1后可以促進(jìn)髓鞘再生、神經(jīng)傳導(dǎo)功能恢復(fù)及改善認(rèn)知功能。該工作提示控制炎癥反應(yīng)、促進(jìn)OPC的遷移可能是減輕慢性缺血引起的白質(zhì)損傷的重要途徑，而IL-1β及其受體IL-1R1是重要的藥物干預(yù)靶點(diǎn)。同時IL-1β的多肽類似物KdPT可以在慢性缺血后進(jìn)入腦內(nèi)促進(jìn)OPC的遷移，從而促進(jìn)白質(zhì)修復(fù)和改善認(rèn)知功能。此外，小膠質(zhì)細(xì)胞抑制劑米諾環(huán)素在慢性缺血早期應(yīng)用可顯著減輕白質(zhì)損傷和認(rèn)知功能損害，其機(jī)制主要是通過減少成熟少突膠質(zhì)細(xì)胞凋亡、促進(jìn)腦室下區(qū)OPC的增殖從而促進(jìn)髓鞘再生過程，為二次開發(fā)“老藥”米諾環(huán)素提供了重要的實驗依據(jù)。以上研究通過針對膠質(zhì)細(xì)胞介導(dǎo)的炎癥反應(yīng)，發(fā)現(xiàn)多個促進(jìn)慢性缺血后白質(zhì)修復(fù)的新靶點(diǎn)，為皮層下缺血性血管性癡呆治療提供了新途徑。

關(guān)鍵詞：慢性缺血；白質(zhì)損傷；少突膠質(zhì)前體細(xì)胞；髓鞘再生；炎癥

基金項目：國家自然科學(xué)基金（81273490，81273506，81473186，81221003）；中央高校基本科研業(yè)務(wù)費(fèi)項目（2016FZA7014）

通訊作者：胡薇薇，E-mail：huww@zju.edu.cn，Tel：（0571）88208227

T2-26

The neuroprotective effect of Rhodiolae on dementia rat induced by ibotenic acid

WEI Jiang-ping1，2，F(xiàn)U Wen-jun1，2，CHEN Huan1，2，ZHANG Zhan-jun3，XU Shi-jun1，2

（1.Key Laboratory of Systematic Research and Exploita?tion of TCM Resources in Sichuan Province，Chengdu 611137，China；2.The College of Pharmacy，Chengdu University of TCM，Chengdu 611137，China；3.Nation?al Key Laboratory of Cognitive Neuroscience and Learn?ing，Beijing Normal University，Beijing 100875，China）

Abstract：OBJECTIVETo research the effect of Rhodio?lae on cognitive function on dementia rats and to investi?gate its potential mechanism.METHODSEstablished dementia model via injecting ibotenic acid into basal nuclei，and randomly divided the model rats into model group，Aricept group（1.4 mg·kg-1）and Rhodiolae group（0.14 mg·kg-1），moreover the sham group was built by injecting normal saline.All animals received homologous drugs after modeling 1 week，and the sham group and model group were gave isometric normal saline（10 mL·kg-1），1/d. Morris water maze was used to evaluate the change of cognitive function after the rats were treated 40 d，and pathomorphism in CA1 and cerebral cortex were ob?served by Nissl staining and the numbers of nerve cell in these areas were counted.The concentration of MDA，H2O2，CAT，SOD and GR in brain tissue were detected by spectrophotography.RESULTSCompared to model group，the escape latency of Rhodiolae groups〔（31.10± 20.97）s compare to（51.07±13.04）s〕was shortened but it hadn′t statistical difference，and the total swimming dis?tance〔（8469.88±4724.74）cm compare to（15862.17± 1315.69）cm〕was shortened obviously，furthermore the times entered the target quadrant〔（9.50±1.87）times compare to（6.50±1.38）times〕and the percentage in target quadrant〔（33.87±4.47）%compare to（26.17± 2.58）%〕were evidently increased.Moderate shrinkage of nucleus of neurons in cerebral cortex of model rats，mild hyperplasia of glial cells and loosely neuron was also observed in CA1 area，but those lesions were dis?tinctly ameliorated in Rhodiolae groups.What′s more，the numbers of nerve cells in CA1〔（70.40±4.00）compare to（45.50±4.95）〕and cerebral cortex〔（63.00± 5.29）compare to（42.40±11.87）〕of Rhodiolae groups were obviously increased.In addition，compared to model group，the content of MDA〔（14.72±4.42）compare to（19.99±4.70）g protein·L-1〕and H2O2〔（60.07±6.38）mmol·L-1compare to（76.37±6.06）mmol·L-1〕were evidently decreased， the activity of GSH〔（41.30±14.81）kU·g-1protein compare to（15.93± 3.59）kU·g-1protein〕was visibly increased，and the activity of SOD〔（5.74±3.76）g protein·L-1compare to（2.72±1.09）g protein·L-1〕and CAT〔（210.45± 60.11）kU·g-1protein compare to（171.48±56.50）kU·g-1protein〕were also increased but without statistical differ?ence.CONCLUSIONRhodiolae improve learning and remembering function，ameliorate pathological changes and protect neuronal loss which respond to decrease the oxi?dative products and enhance the activity of antioxidase.

Key words：Rhodiolae；ibotenic acid；antioxidase

Foundation item：The project supported by Key Proj?ects of National Natural Science Foundation of China（81430100）；Academic and technical leaders of Sich?uan Province to Raise Funds for Funding Projects（2015058）；and Sichuan Province Youth Science and technology Innovation Research Team（2014TD0007）

Corresponding author：XU Shi-jun，E-mail：docxu@126. com，Tel：（028）61800231

T2-27

Lentivirus-mediated knockdown of phosphodiester?ase 4D improved depression-like behaviors in miceZHANG Cong，CHENG Yu-fang，NIU Bo，XU Jiang-ping

（Guangdong Provincial Key Laboratory of New Drug Screening，SchoolofPharmaceuticalSciences，SouthernMedicalUniversity，Guangzhou510515，China）

Abstract：OBJECTIVETo examine whether long-form phosphodiesterase-4（PDE4）knockdown by lentiviral RNA construct containing a specifc microRNA/miRNA-mir hairpin structure reversed depression-like symptoms caused by chronic unpredictable mild stress（CUMS）in mice.METHODSIn this research，the study was per?formed on adult male C57 mice，weighing（25±5）g，kept in a controlled environment.CUMS animal model was used recapitulate a multiple of behavioral character?istics and biochemical states of depression in human. The forced swimming test（FST）and the tail suspension test（TST）were used to detectthe state of depression. Western blotting analysis was used to assess protein lev?els of cAMP response element binding protein（CREB，unphosphorylated and phosphorylated［pCREB］）to ex?plore the neurochemical mechanisms.RESULTSCUMS decreased cAMP levels（P＜0.01）and produced depres?sion-like symptoms in FST（P＜0.01）and TST（P＜0.01）.Microinfusions of lentiviruses reversed CUMS-in?duced cAMP decline（P＜0.05）and depression-like symptoms.Moreover，CUMS caused a significant reduc?tion in protein kinase A and CREB phosphorylation，and brain-derived neurotrophic factor transcription，both of which were partially attenuated by lentivirus-mediated knockdown of PDE4D.Also，the phosphorylation of ex?tracellular signal-regulated kinase 1/2 was reduced in CUMS-exposed mice，which was reversed by 4DmiRNA treatment.Taken together，this study demonstrated that PDE4DmiRNA improved the CUMS-induced depressionlike symptoms that might be related to the increase in hip?pocampal cAMP and pCREB expression.CONCLUSIONHence，PDE4D inhibitors can serve as potential antide?pressants，and their antidepressant activity is partially mediated by the activation of cAMP signaling pathway in the hippocampus.In other words，long-form PDE4D knockdown may offer a promising treatment for major de?pression disorder.

Key words：antidepressant；CREB；long-form PDE4D；major depression disorder

Foundation item：The project supported by National Natural Science Foundation of China（81301099，81373384）；Natural Science Foundation of Guangdong Province（S2013040014202）；China Postdoctoral Science Foun?dation（2013M542192）；and National Science and Tech?nology Major Projects for“Major New Drugs Innovation and Development”（2012ZX09J1211003C）.

Corresponding author：XU Jiang-ping

T2-28

Melatoninalleviateslipopolysaccharide-compromised integrity of blood brain barrier by activation of AMP-activated protein kinase in old mice

WANGXiao-na1， SHU Hui1， WANGMeng-wei1，SUN Yan-yun1，LIU Chun-Feng1，2，LIU Wen-lan3，LIU Jie4，JIN Xin-chun1

（1.Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and Institute of Neuroscience，The Second Affiliated Hospital of Sooch?ow University，Soochow University，Suzhou215004，China；2.Department of Neurology，Jiangsu Key Labo?ratory of Translational Research and Therapy for Neuro-Psycho-Diseases，The Second Affiliated Hospital of So?ochow University，Soochow University，Suzhou 215004，China；3.The Central Laboratory，Shenzhen Second People′s Hospital，Shenzhen University Hospital，Shenzhen 518035，China；4.Translational Center for Stem Cell Research，Tongji Hospital，Stem Cell Research Center，TongjiUniversitySchoolofMedicine，Shanghai 200065，China）

Abstract：The integrity and function dysfunction of the blood brain barrier（BBB）is considered to be an early event in the pathogenesis of a variety of neurological dis?eases in old patients and dysfunction of BBB could be in?duced bycommon stress that old people often face，such as sepsis during which lipopolysaccharide（LPS）is re?leased into circulation and BBB is damaged.Since de?creased melatonin level had been shown in the serum and brain of old people and mice，we aim to investigate whether supplement with melatonin could alleviate LPS-induced BBB damage in old mice.Mice（24-28 months old）received one week melatonin（10 mg·kg-1·d-1，intra?peritoneally，ip）or saline before challenge with LPS（1 mg·kg-1，ip）.Evan′s blue（EB）and immunoglobulin G（IgG）leakage were used to assess the BBB permea?bility.Immunofluoresence and Western blotting were used to detect the protein expression and distribution. Our results showed that LPS significantly increased BBB permeability in old mice accompanied by tight junction protein occludin and claudin-5 degradation，inhibition of AMP-activated protein kinase（AMPK）activation and increase of gp91phox protein expression.Interestingly，one week of melatonin treatment significantly decreased LPS-induced BBB hyperpermeability，activated AMPK and inhibited gp91phox expression upregualtion.More?over，activation of AMPK by metformin significantly inhib?ited LPS-induced gp91phox expression upregualtion in endothelial cells.Taken together，our findings demon?strate that melatonin alleviates LPS-impaired integrity of BBB by activation of AMPK and inhibition of gp91phox expression in old mice.

Key words：melatonin；blood brain barrier

Corresponding author：JIN Xin-chun，Tel：（0512）65883557，E-mail：xinchunjin@gmail.com

T2-29

Propane-2-sulfonic acid octadec-9-enyl-amide，a nov?el PPARα//γdual agonist，protects against ischaemiainduced brain damage in mice by inhibiting inflamma?tory responses

HUANG Jun-xiong，ZENG Kai-yue，XU Lan-xi，JIN Xin，YANG Li-chao

（Faculty of Basic Medicine，Medical College，Xiamen University，Xiamen，China）

Abstract：OBJECTIVEPropane-2-sulfonic acid octadec-9-enyl-amide（N15），an analogue of oleoylethanolamide（OEA），is a novel PPARα/γ dual agonist.Our previous studies verified the positive effects of OEA on the acute and delayed stages of cerebral ischaemia.However，it is not clear whether N15 is effective against ischaemic cere?bral injury.In the present study，male Kunming mice were subjected to middle cerebralartery occlusion（MCAO）.METHODSTo evaluate its preventive effects，N15（50，100 or 200 mg·kg-1，ip）was administered for 3 d before ischaemia.To evaluate its therapeutic effects，N15（200 mg·kg-1，ip）was administered 1 h before reperfusion or 0，1，2 or 4 h after reperfusion.Neurologi?cal deficit scores，infarct volume and the degree of brain oedema were determined at 24 h after reperfusion. Blood brain barrier（BBB）disruption was evaluated by Evans blue（EB）leakage at 6 h after reperfusion.The activation/inflammatory responses ofmicroglia were detected using immunohistochemistry andWestern blotting.RESULTSN15 pretreatment improved neuro?logical dysfunction，reduced infarct volume and alleviat?ed brain oedema in a dose-dependent manner；the most effective dose was 200 mg·kg-1.The therapeutic time window was within 2 h after reperfusion.Moreover，N15 was more potent in the treatment of cerebral ischaemia injury than OEA.N15 treatment preserved the BBB integ?rity and suppressed the activation of microglia.N15 inhib?ited inflammatory cytokine expression not only in MCAO mice but also in lipopolysaccharide（LPS）-stimulated BV-2 microglial cells.Moreover，N15 decreased the phosphor?ylation levels of NF-κBp65，STAT3，and ERK1/2 bothin vivoandin vitro.CONCLUSIONOur findings demon?strated that N15 exerts neuroprotective effects and was more potent than OEA.Additionally，the neuroprotective effects of N15 on cerebral ischaemia may be attributed to its anti-inflammatory properties，at least in part，by en?hancing PPARα/γ dual signalling and inhibiting the acti?vation of the NF-κB，STAT3，and ERK1/2 signalling pathways.These findings suggest that N15 may be a po?tential therapeutic choice for the prevention and treat?ment of ischaemic stroke.

Key words：propane-2-sulfonic acid octadec-9-enyl-am?ide；mice；focal cerebral ischaemia；activated microglia；inflammation

Foundation item：The project supported by National Natural Sciences Foundation of China（81373407）；and the NaturalScience Foundation ofFujian Province（2016D018）

Corresponding author：YANG Li-chao，E-mail：yan?glc116@xmu.edu.cn

T2-30

The effects of 5-lipoxygenase inhibitor and cysteinyl leukotriene receptor 1 antagonist on 1-methyl-4-phe?nylpyridine-induced neurotoxicity

ZHANG Xiao-yan1，2，LI Chen-tan1，WANG Yan-fang1，ZHAO Jian-bo1，WEI Er-qing3，ZHANG Li-hui1

（1.Department of Pharmacology，Hangzhou Key Labo?ratory of Medical Neurobiology，School of Medicine，Hangzhou Normal University，Hangzhou 310036，China；2.Biotherapy Center，The General Hospital of Beijing Military Command，Beijing 100700，China；3.Depart?ment of Pharmacology，School of Medicine，Zhejiang University，Hangzhou 310058，China）

Abstract：OBJECTIVEPreviously we demonstrated the neuroprotective effect of 5-lipoxygenase（5-LOX）inhibi?tor as well as cysteinyl leukotriene receptor 1（CysLT1）antagoniston rotenone-induced microglial activation and neuronal death.In this study，we determined the effects of 5-LOX inhibitor zileuton and CysLT1 antagonist monte?lukast on neurotoxicity induced by 1-methyl-4-phenylpyri?dine（MPP+）in anin vitromodel of Parkinson disease（PD）.METHODSThe neurotoxicity of MPP+，a neurotox?in relevant to PD，on the PC12 cells was measured by MTT assay，lactate dehydrogenase（LDH）release and double fluorescence staining with Hoechst/propidiumio?dide（PI）.The protective effects of 5-LOX inhibitor zileuton and CysLT1 antagonist montelukast were investigated by the above methods.RESULTSWe found that expo?sure of PC12 cells to MPP+led to a reduced cell viability and an increased level of LDH in a concentration-depen?dent manner.Pretreatment with zileuton and montelukast significantly attenuated viability loss and LDH release in MPP+-treated PC12 cells.Furthermore，MPP+increased?necrotic cell death in PC12 cells.Administration of monte?lukast significantly decreased MPP+-induced cell necro?sis in PC12 cells.CONCLUSIONThe 5-LOX inhibitor zileuton and CysLT1 antagonist montelukast have a neu?roprotective effects on MPP+-induced neurotoxicity in PC12 cells.The 5-LOX inhibitor and CysLT1 antagonist might raise a possibility as potential therapeutic agent for PD and other inflammation-related the central nervous system disorders.

Key words：MPP+；neurotoxicity；PC12 cell；5-lipoxy?genase；cysteinyl leukotriene receptor

Foundation item：The project supported National Natu?ral Science Foundation of China（81273491）；and the Zhejiang Provincial Natural Science Foundation（LY12H31010）

Corresponding author：ZHANG Li-hui，Tel：（0571）28861603，E-mail：lihuizhang002@sina.com

T2-31

Attenuation ofScutellaria barbataflavonoids on corti?cal cytoplasm apoptotic factors disorders induced by complex Aβ25-35in rats

SHANG Ya-zhen，GUO Ke，CUI Ya-di，ZHAO Hong-xiang，ZHANG Shu-feng

（Hebei Province Key Laboratory of Traditional Chinese Medicine Research and Development/Institute of Tradi?tional Chinese Medicine，Chengde Medical College，Chengde 067000，China）

Abstract：OBJECTIVETo investigate the effect of SBF on corticalcytoplasm apoptoticfactors disturbances induced by complex Aβ25-35in rats.METHODSThe cere?bral injuries model was established by rats received intra?cere-broventricular injection of RHTGF-β1，Aβ25-35and AlCl3and then accepted SBF treatment.All the rats were sacrificed by decapitation for indicators detection at last the drug treatment.Western blotting method was for cas?pase-3 protein expression and RT-PCR method detected cytochrome c，apoptotic protease activating factor-1（Apaf-1），caspase-9 mRNA expression in cortical cyto?plasm.RESULTSThe protein expression of caspase-3 in cortical cytoplasm of rats was assayed by Western blotting.The results indicated that compared with the sham group，the caspase-3 protein expression of corti?cal cytoplasm in Aβ group was significantly increased（P＜0.01）.However，the increased expression can be obviously reversed by SBF at doses of 35，70 and 140 mg·kg-1，as compared with model group（P＜0.01）.The Cyt-C，Apaf-1 and caspase-9 mRNA expressions in corti?cal cytoplasm of rats were determined by RT-PCR.The results indicated that compared with the sham group，the Cyt-C Apaf-1 and caspase-9 mRNA relative expres?sions of cortical cytoplasm in Aβ group was significantly increased（P＜0.01）.However，these increased expres?sions can be differently reversed by SBF at doses of 35，70 and 140 mg·kg-1，as compared with model group（P＜0.01）.CONCLUSIONSBF can definitely improve rats′cortical cytoplasm apoptotic factors disorders in?duced by complex Aβ25-35，which maybe benefit for treat?ment of degenerative disease.

Key words：Scutellaria barbataflavonoids；apoptosis

Foundation item：The project supported by Hebei Pro? vincialNaturalScience Foundation（C2009001007，H2014406048）；Hebei Provincial Administration of Tradi?tional Chinese Medicine of China（05027，2014062）；and the Key Discipline Construction in Institution of High Education in Hebei Province

T2-32

Moderate alcohol preconditioning activates BKCa channels to protect brain damage-induced by cere?bral ischemia and reperfusion

ZHAO Yi-Long1，2，3，4，GUO An-Chen2，3，4，WANG Yong-Jun1，2，3，4，WANG Qun1，2，3，4

（1.Department of Neurology，Beijing Tiantan Hospital，Capital Medical University，Beijing 100050，China；2. Beijing Key Laboratory of Translational Medicine for Car?diovascular Diseases，Beijing 100050，China；3.Beijing Institute for Brain Disorder，Beijing 100069，China；4. National Research Center for Neurological Diseases，Beijing 100050，China）

Abstract：OBJCETIVEEpidemiologic studies have dem?onstrated that consumption of moderate amounts of red wine is associated with significant reductions in incidenc?esofcardiovascularandcerebrovasculardiseases，which may be related to alcohol in red wine.Our previ?ous study demonstrated that ethanol ingestion 24 h prior to induction of cerebral ischemic/reperfusion（I/R）re?duced delayed neuronal death（DND）.Our most recent results supported a role for big Ca2+-sensitive K+channel（BKCa channel）activation in the neuroprotective effects of ethanol preconditioning（EtOH-PC）in global cerebral I/R.Therefore，we hypothesis that moderate EtOH-PC activates BKCa channel to protect brain damage induced by focal cerebral I/R.This project will utilize focal cere?bral I/R animal model to explore the function of BKCa channel in EtOH-PC protectionin vivolevels by means of pharmacological intervention such as BKCa channel opener（NS11021，NS）and blocker（paxilline，PX）.The re?sults will provide theoretical evidence for neuroprotective effect of moderate alcohol preconditioning against isch?emic stroke，and the conclusion will also bring to a con?cept that extrinsic moderate ethanol preconditioning may activate intrinsic protective mechanism in the brain.METHODSThe SD rat were randomly divided into the following six groups（n=10）：sham，I/R，EtOH-PC+I/R，NS11021-PC+I/R，paxilline+EtOH-PC+I/R，Paxilline+ NS11021-PC+I/R.Both EtOH-PC and NS11021-PC（0.1mg·kg-1；ip）were induced 24 h before I/R.The vol?ume of 95%ethanol to be instilled（in μL）was calculated as follows：〔body weight（g）×0.6〕+0.3.This volume of ethanol was mixed in 0.3 mL of sterile distilled water just before administration to the animals by gavage.The Pax?illine（2.5 mg·kg-1；ip）was administered 10min beforeEtOH-PC and NS11021-PC.The right middle cerebral artery occlusion（MCAO）was produced by inversion of a 4-0-nylon filament.The filament was withdrawn 2 h after onset of MCAO and then reperfused.Neurological deficits and infarct volume were measured 24 h after I/R. Another 36 rats were randomly divided into 6 groups as above，6 in each group.DWI were performed 2h after ischemic and T2WI MRI were performed 24 h after I/R to observe the infarct volume of brain and the penumbra volume of brain in each group.Then rats were killed and detected the apoptotic cell death and degeneration of neurons.RESULTSCompared to I/R group，the neuro?logical score（P＜0.01），the infarct volume of brain（P＜0.01），the infarct volume of ischemic penumbra（P＜0.01），the percentage of apoptotic cell death（P＜0.01）and the percentage of degenerative neurons（P＜0.01）were significantly decreased after ethanol precondition?ing，while these changes were reversed by paxilline（P＜0.05）；compared to I/R group，the neurological score（P＜0.01），the infarct volume of brain（P＜0.01），the in?farct volume of ischemic penumbra（P＜0.01），the per?centage of apoptotic cell death（P＜0.01）and the per?centage of degenerative neurons（P＜0.01）were signifi?cantly decreased after NS11021 preconditioning，while these changes were reversed by paxilline（P＜0.05）.CONCLUSIONOur results show that moderate alcohol preconditioning activates BKCa channels to protect brain damage induced by focal cerebral I/R.

Key words：ethanol preconditioning；neuroprotection；cerebral ischemia/reperfusion；BKCa channels

Foundation item：The project supported by NSFC（81171079，81271312）

Corresponding author：WANG Qun，E-mail：wangq@ ccmu.edu.cn；Tel：13146254818

T2-33

Isoliquiritigenin suppresses microglial activation and neuroinflammation in primary microglia

FU Yan，YANG Pin，ZHAO Yang，XU Ying

（Department of Physiology，Shanghai University of Tra?ditional Chinese Medicine，Shanghai 201203，China）

Abstract：OBJECTIVEMicroglial activation contributes to neuroinflammation and neuronal damages in neurode?generative disorders including Alzheimer and Parkinson diseases.It has been suggested that neurodegenerative disorders may be improved if neuroinflammation can be controlled.Isoliquiritigenin（ISL）isolated from Glycyrrhi?za glabra possess potent anti-inflammatory capability，we thus investigate the inhibitory effects of ISL on LPS-in?duced microglial activation and neuroinflammation and the roles of neuroprotection on neurodegenerative disor?ders.Moveover，we would explore the mechanism of its associated inflammatory signaling passway.METHODSBy cultivating，isolating and purifying，we got the prima?ry microglia of the rat.The form of those cells was ob?served under an optical microscope，the purity was iden?tified by the CD11b immunefluorescence staining.Firstly，the effects of ISL on microglial viability were explored by the MTT assay.The microglial cells were pretreated by ISL（5 μmol·L-1）and then stimulated by LPS（100 ng·mL-1），the production of NO was detected by Griess reagent，the change of microglial morphology was observed by im?munefluorescence staining.The production of IL-1β and TNF-α in culture medium were tested by ELISA，and the mRNA expression of pro-inflammatory factors，including IL-1β，TNF-α，iNOS and COX-2 was detected by real time PCR.The protein expression of iNOS and COX-2 was detected by Western blotting.RESULTS（1）After purifying，microglial showed the typical morphological characteristics.The cell body was small，protruding out?stretched in the branch rod，spindle.After LPS stimula?tion，the shape was changed to amoeba-like，and the protrusions retracted，cell rounding.In the CD11b im?munefluorescence staining，the purity of the microglial was over 98%，and showed powerful phagocytic activity.（2）The results show that ISL（0.16-20 μmol·L-1）concen?tration has no effect on primary microglia′s vitality.And at the concentration of 5 μmol·L-1ISL has the significant?ly inhibitory effect on NO production induced by LPS.（3）The results shows that ISL pretreatment can signifi?cantly inhibit LPS-induced microglial activation，and can significantly reduce the production of NO in a dose-de?pendent manner（P＜0.05）.When stimulated by LPS for 12 h，the expression of TNF-α and IL-1β mRNA in the cul?ture medium are reduced by ISL pre-treatment（P＜0.05）.After LPS stimulation for 24 h，the expression of COX2 and iNOS mRNA and protein can be reduced（P＜0.05）.After stimulated by LPS from 0.5 to 2 h，p-ERK ex?pression has be decreased.CONCLUSIONWe successful?ly obtain primary microglia with high-purity by culturing in vitro.We find that ISL can significantly reduce the levels of proinflammatory facters，which indicate ISL can inhibit microglia activation and neuroinflammation by blocking ERK1/2 signal pathway.

Key words：isoliquiritigenin；primary microglia；inflam?matory factors；neuroinflammation

Foundation item：The project supported by National Natural Science Foundation of China（81274119）

Corresponding author：XU Ying，Tel：（201）51322453，E-mail：ying6122003@aliyun.com

T2-34

Inhibition of chemokine-like factor 1 improves bloodbrain barrier dysfunction in rats following focal cere?bral ischemiaKONG Ling-lei1，2，HU Jin-feng1，YUAN Yu-he1，CHEN Nai-hong1，DU Guan-hua1，2

（1.State Key Laboratory of Bioactive Substances and Functions of Natural Medicines，2.Beijing Key Laborato?ry of Drug Target and Screening Research，Institute of Materia Medica，Chinese Academy of Medical Sciences andPekingUnionMedicalCollege，Beijing100050，China）

Abstract：OBJECTIVETo investigate the role of che?mokine-like factor 1（CKLF1），a novel C-C chemokine，on brain-blood barrier（BBB）integrity in rat focal cerebral ischemia and reperfusion model.METHODSAntibodies against CKLF1 was applied to the rightcerebral ventricle immediately after transient middle cerebral artery occlu?sion.Brain water content，Evans blue leakage and the ex?pression of aquaporin-4（AQP-4），matrix metalloprotein?ase-9（MMP-9），zonula occludens-1（ZO-1）and occlu?din were measured.RESULTSAfter treatment with anti-CKLF1 antibody，brain water content and Evans blue leakage in ipsilateral hemisphere were decreased in a dose-dependent manner at 24 h after reperfusion，but not changed in contralateral hemisphere.Anti-CKLF1 an?tibody reduced the expression of AQP-4 and MMP-9，and upregulated the expression of ZO-1 and Occludin. These results suggest that CKLF1 is involved in BBB dis?ruption after reperfusion.CONCLUSIONInhibition of CKLF1 protects against cerebral ischemia by maintaining BBB integrity，possibly via inhibiting the expression of AQP-4 and MMP-9，and increasing the expression of tight junction protein.

Key words：chemokine-like factor 1；cerebral ischemia；brain-blood barrier

Foundation item：The project supported by National Natural Science Foundation of China（81302760）；and the Chinese Postdoctoral Science Foundation Project（2013M542510）

Corresponding authors：CHEN Nai-hong， E-mail：chennh@imm.ac.cn；DU Guan-hua，E-mail：dugh@imm.ac.cn

T2-35

Hippocampal PPARδdownregulation contri butes to depressive phenotype with decreased neurogenesisYUXu-ben，F(xiàn)ANGShun-chang，CHENFang，MEI Zhen-Lin，LIN Jing-ran，HU Mei，TANG Su-su，LONG Yan，SUN Hong-bin，KONG Ling-yi，HONG Hao

（Department of Pharmacology，Jiangsu Key Laboratory of Drug Discovery for Metabolic Diseases，and State Key Laboratory of Natural Medicines，China Pharmaceu?tical University，Nanjing 210009，China）

Abstract：Our previous study showed that up-regulating hippocampal peroxisome proliferator-activated receptor δ（PPARδ）displays an antidepressive effect and enhanc? es hippocampal neurogenesis in the context of chronic stress.Here，the changes in depressive behaviors and hippocampalneurogenesis were investigated after PPARδ knockdown by microinfusion of the lentiviral vec?tor，expressing short hairpin RNA（shRNA）complemen?tary to the coding exon of PPARδ，into the bilateral den?tate gyri of the hippocampus or PPARδ blockade by re?peated systemic administration of PPARδ antagonist，GSK0660（1 or 3mg·kg-1，ip，for 21 d）.We found that hippocampal PPARδ knockdown or blockade induced de?pressive-like behaviors and increased vulnerability to stress，which is involved in decreased hippocampal neu?rogenesis and neuronal differentiation.Down-regulating hippocampal PPARδ also induced significant decreases in phosphorylation cAMP response element-binding pro?tein（CREB）and BDNF level in the hippocampus.The in vitro study that PPARδ knockdown or blockade inhibited proliferation and differentiation of neural stem cells.Tak?en together，our results suggest that PPARδ could be a novel and promising target for developing new PPARδ agonists for the treatment of depressive disorders.

Key words：depression；peroxisome proliferator-activat?ed receptor δ；hippocampal neurogenesis

Foundation item：The project supported Natural Sci?ence Foundation of Jiangsu Province（SBK201320969）；and NationalNaturalScience Foundation ofChina（81573413 and 81273497）

Corresponding author：HONG Hao，E-mail：honshao@ cpu.edu.cn

T2-36

Adenosine A2A receptor-expressing neurons in the striatum regulate sleep behaviors

YUAN Xiang-shan1，WANG Lu1，DONG Hui1，QU Wei-min1，LI Rui-xi2，HUANG Zhi-li1，3

（1.Department of Pharmacology，2.Department of Anat?omy，Histology and Embryology，3.State Key Laborato?ry of Medical Neurobiology，F(xiàn)udan University，Shanghai 200032，China）

Abstract：OBJECTIVEThe high prevalence of sleep disturbance has been found in patients with striatum-re?lated neurodegenerative disorders.In the striatum，there are abundant adenosine A2A receptors（A2ARs）which?havebeen reported to mediatesleepbehavior for adenos?ine.We hypothesized that the A2AR-expressing neurons in the striatum are involved in sleep-wake regulation.METHODSWe employed a chemogenetic technique，designer receptor exclusively activated by designer drug（DREADD），to specifically and non-invasively manipu?late the neuron activity based on the principle of Cre/ LoxP recombination，EEG/electromyogram recording for sleep-wake behaviors，the neural tracing approach toselectively visualize the perikarya of A2AR-expressing neurons and their axons by adeno-associated virus（AAV）encoding humanized Renilla green fluorescent（hrGFP）as a tracerin A2AR-Cre mice.In addition，we used immunoelectron microscopy，patch-clamp tech?nique，and optogenetics in A2AR-Cre mice to selectively characterize the synapse and functional connectivity between the A2AR-expressing neurons and the neuron of their downstream targetsin vitro.RESULTSThe acti?vation of A2AR-expressing neurons in rostral，centrome?dial and centrolateral striatum increased non-rapid eye movement（non-REM，NREM）sleep，concomitant with a reduction in wakefulness，whereas the activation of A2AR-expressing neurons in caudal striatum didn′t alter sleep-wake profiles at all.Topographical projections in the sagittal section showed that the axons of A2AR-expressing neurons from rostral striatum distributed in the rostral external globuspallidus（GPe）with a discoidal region paralleled to the striato-pallidal border，while the axons of the A2AR-expressing neurons from the central striatum not only distributed in the rostral GPe，but also in the caudal GPe with a similar distributing pattern as did in rostral neurons.However，the axons of A2AR-expressing neurons from caudal striatum just scattered in the caudal GPe.Based on our anatomical findings and patch-clamp technique combining with optogenetics，we found that A2AR neurons in the rostral striatum preferen?tially formed inhibitory synapses with parvalbumin（PV）-positive neurons in the rostral GPe，while A2AR neurons in the caudal striatum preferentially formed inhibitory syn?apses with PV-negative neurons in the caudal GPe.CONCLUSIONThe present results indicated that the A2AR-expressing neurons in rostral and central striatum are involved in sleep-wake regulation，probably via inner?vating PV-positive neurons in the GPe.

Key words：striatum；A2AR neuron；sleep；topographi?cal projection；DREADD

Foundation item：The project supported by National Basic Research Program of China（2015CB856401）

Corresponding author：HUANG Zhi-li，Tel：（021）54237043，E-mail：huangzl@fudan.edu.cn

T2-37

Naturalproductcompound3camelioratesbrain inflammation and brain ischemic stroke via AMPK-mediated microglia polarization

WANG Yun-jie， RUAN Wen-chen， XU Ya-zhou，ZHANG Lu-yong，PANG Tao

（Jiangsu Key Laboratory of Drug Screening，China Phar?maceutical University，Nanjing 210009，China）

Abstract：OBJECTIVEBrain inflammation plays an im?portant role in the pathophysiology of brain ischemic stroke，psychiatric and neurological diseases.During brain inflammation，microglia cells are activated and show pro-inflammatory M1 and anti-inflammatory M2 phe?notypes，producing neurotoxic molecules and neuro?trophic factors，respectively.We have previously discov?ered a novel natural product compound 3c exhibiting antiinflammatory effects in microglia cells，but the underlying mechanisms and its beneficial effects on brain inflamma?tion and brain ischemia are unknown.METHODSThe gene expression of M1 markers and M2 markers was measured by RT-PCR.The AMPK phosphorylation level and M2 marker CD206 protein expression were deter?mined by Western blotting.TNFα release was measured by ELISA.The gene knowdown was performed by the siRNA transfection experiment.The LPS-induced brain inflammation mouse model and transient middle cerebral artery occlusion（tMCAO） stroke model were used.RESULTSWe found that compound 3c inhibited M1 polarization and promoted M2 polarization in LPS-stimu?lated BV2 and primary microglia cells，and these effects are mediated by CaMKKβ/AMPK/JNK signaling pathway. Furthermore， compound 3c prevented M1 gene expression and enhanced M2 gene expression in a mouse model of LPS-induced neuroinflammation，and reduced the LPS-induced sickness behavior.In addition，compound 3c significantly reduced infarctvolume，improved the neurological deficit，and reduced neuroin?flammation in rats with acute focal cerebral ischemia.CONCLUSIONOur results indicate that natural product compound 3c suppresses microglia activation by promot?ing M2 polarization and may provide a novel therapeutic approach to treat brain ischemic stroke associated with enhanced brain inflammation.

Key words：natural product；microglia polarization；AMP-activated protein kinase；neuroinflammation；brain ischemia

Corresponding authors：PANG Tao，E-mail：tpang@ cpu.edu.cn；ZHANG Lu-yong，E-mail：lyzhang@cpu. edu.cn

T2-38

Modulation of abnormal neuronal circuit in temporal lobe epilepsy

WANG Yi1，YING Xiao-ying2，CHEN Bin1，XU Ceng-lin1，CHEN Zhong1

（1.Department of Pharmacology，Key Laboratory of Medical Neurobiology of the Ministry of Health of China，2.Institute of Pharmaceutics，College of Pharmaceutical Sciences，SchoolofMedicine，ZhejiangUniversity，Hangzhou，China）

Abstract：OBJECTIVETemporal lobe epilepsy（TLE）is one of the most common types of human epilepsy，and theyare often resistantto currenttreatments.METHODSBy using optogenetic，electrophysiological，imaging and pharmacology strategies，we aimed toinves?tigate the underlying circuit mechanism of TLE and tried to developthe novel and efficient approach to control epilepsy.RESULTS（1） Using microPET and multichannel EEG recording，we found an abnormal neural network，characterized by early hypometabolism and after discharge spread，during the epileptogenensis of TLE.（2）Deep brain stimulation，especially low frequen?cy stimulation，targeted the epileptic focus and the areas outside of the focus（critical regions for seizure spread），such as the piriform cortex，cerebellum，entorhinal cortex or subiculum，reduced seizure severity in TLE. Its anti-epileptic effect is time-window dependent and polarity dependent，which shows a promising strategy for treating epileptic seizures.（3）Using an optogenetic strategy，we demonstrated that excitatory projection from entorhinal cortex to hippocampus instructs the brain-stimulation treatments of epilepsy.（4）Our data from both the clinical and experimental studies further demonstrated that a disinhibitory GABAergic neuronmediated microcircuit in the subiculum contributes to secondary generalized seizures in TLE.（5）Finally，based on abnormal synchronization of the electrical activity in epileptic circuit， we developed electroresponsive hydrogel nanoparticles modified with angiopep-2 to facilitate the delivery of the antiepileptic drug phenytoin sodium，which greatly improves the therapeuticindex.CONCLUSIONOurfindingsmay update the current view of epileptic neuronal networks and suggestpossible promising ways forepilepsy treatment.

Key words：epilepsy；neural circuits；brain stimulation；optogenetics；electro-responsive

Foundation item：The project supportedp by National NaturalScience Foundation ofChina（91332202，81221003）

Corresponding author：CHEN Zhong，Tel：（0571）88208228，E-mail：chenzhong@zju.edu.cn

T2-39

The basic helix-loop-helix（bHLH）transcription fac?tor，DEC1，provides neuroprotection from apoptosis induced by MPP+through PI3K/Akt pathway in SHSY5Y cells

ZHU Zhu，WANG Yu-wen，YANG Jian

（Nanjing Medical University，Nanjing 210029，China）

Abstract：OBJECTIVETo determine the role of the ba?sic helix-loop-helix（bHLH）transcription factor，differenti?ated embryonic chondrocyte gene 1（DEC1），in the apoptosis induced by 1-methyl-4-phenylpyridiniumion（MPP+）in SH-SY5Y cells.METHODSSH-SY5Y cells were treated with different concentrations of MPP+for 24 or 48 h.The cell inhibition and apoptosis were measured by MTT and DAPI staining.DEC1，the apoptosis-related proteins and PI3K/Akt/GSK3β/β-catenin signaling were determined by Western blotting.The expression of DEC1 was regulated by overexpression and shRNA.RESULTSMPP+induces apoptosis along with decreasing of DEC1 expression in SH-SY5Y cells.Overexpression or knock?down of DEC1 can alleviate or enhance the cell inhibition induced by MPP+.And overexpression of DEC1 can alle?viate the increased cleaved caspase 3/caspase 3 but not alleviate Bax/Bcl-2 induced by MPP+.Meanwhile，MPP+represses PI3Kp110α，p-Akt/Akt，p-GSK-3β/GSK-3β and β-catenin expression，which is accompanied by decreasing DEC1 expressions.It is confirmed that the activator or inhibitor of PI3K/Akt/GSK-3β pathway can alleviate or enhance the repression of PI3K/Akt/GSK3β/ β-catenin signaling cascade induced by MPP+.Further study，we find that overexpression of DEC1 alone can increase PI3Kp110α，p-Akt/Akt，p-GSK-3β/GSK-3β，and β-catenin expression.More importantly，overexpres?sion of DEC1 significantly alleviates the decreased levels of PI3Kp110α，p-Akt/Akt，p-GSK-3β/GSK-3β，and βcatenin induced by MPP+.CONCLUSIONDEC1 pro?vides neuroprotection from apoptosis induced by MPP+through PI3K/Akt pathway in SH-SY5Y cells.Promisingly，DEC1 is a candidate gene that may provide a novel therapeutic approach for the treatment of Parkinson disease.

Key words：differentiated embryonic chondrocyte gene 1 1-methyl-4-phenylpyridiniumion；neuroprotection；PI3K/Akt pathway

Foundation item：The project supported by National NaturalScience Foundation ofChina（81573503，81373443）；and by the Major Project of Jiangsu Provin?cial Department of Education（13KJA310003）

Corresponding author：YANG Jian，E-mail：jianyang@ njmu.edu.cn

T2-40

Mechanical allodynia and affective behavior are im?proved by INI-0602，a gap junction hemichannel in?hibitor，in a rat model of neuropathic pain induced by sciatic nerve injury

ZHANG Xiao-min，F(xiàn)AN Li-xia，PENG Yue-xia，SONG Qi，XIANG Yu-ke，WU Wei-li，WANG Qin，TAO Liang

（Department of Pharmacology，Zhongshan School of Medicine，Sun Yat-Sen University，Guangzhou 510080，China）

Abstract：OBJECTIVETo investigated the effects of INI-0602 on nociceptive reflex，depression-associated andanxiety-related behaviors caused by neuropathic pain in sciatic nerve injury rats.METHODSMale rat were sub?jected to sciatic nerve injury（SNI）or sham surgery.Rat received daily treatment with INI-0602 intrathecally，at a dose of 0.25 μg/10 μL.The response frequency to mechanical allodynia in animals was measured with von Frey hairs on day 1，3，5，7，14，21.Rats were evaluated in the forced swimming test（FST）test，tail suspension test（TST）， sucrose preference test（SPT） for depression-like behavior.We performed open field test（OFT）and elevated plus-maze test（EPM）to evaluate anxiety-associated behaviors.Besides，we investigated the alterations of NMDA receptor and the brain-derived neurotrophic factor（BDNF）and also the expression of connexin43 and connexin32，structure protein of gap junction channel，on the protein level and the number of activated astrocyte showed by immunohistochemical.RESULTSThe SNI procedure produced mechanical al?lodynia and accompanied with depressive-like and anxi?ety-like behavior.Treatment with INI-0602 produced a significant analgesic effect in SNI rats at day 7（model+ NS：11.017±1.506 g；model+INI-0602：31.157±1.532 g，P＜0.01），and still obviously on the 21th day（31.067± 1.787，P＜0.01）.INI-0602 could also improve the perfor?mance of sciatic nerve injury rats among program behav?ior tests related to depression and anxiety.In parallel with relief of pain，the alterations of NMDA receptor and the brain-derived neurotrophic factor（BDNF），involved in central sensitization and synaptic plasticity，were in?vestigated.INI-0602 not only could inhibited spared nerve injury induced up-regulated of NR2B and phos?phorylation NR2B in early and late neuropathic pain（early phase：Nr2b：2.897±0.228，P＜0.01；p-Nr2b：2.984±0.236，P＜0.01；late phase：Nr2b：2.594±0.187，P＜0.01；p-Nr2b：3.124±0.330，P＜0.01），but also could inhibit the increased of BDNF in the early（model+ NS：3.637±0.381，model+INI-0602：1.148±0.372，P＜0.01）and upregulate the BDNF in late stage（model+ NS：0.438±0.103，model+INI-0602：1.222±0.092，P＜0.01）.Meanwhile，INI-0602 significantly decreased the expression of connexin43 and connexin32，structure pro?tein of gap junction channel，on the protein level and the number of activated astrocyte showed by immunohisto?chemical.CONCLUSIONINI-0602 blocked behavioral changes induced by neuropathic pain，suggesting that it might be a promising pharmacological approach of painemotion diseases.

Key words：INI-0602；gap junction hemichannel inhibi?tor；neuropathic pain；depression；anxiety

Foundation item：The project supported by National NaturalScience Foundation ofChina（81473234，U1303221）

Corresponding author：TAO Liang，E-mail：Taol@mail. sysu.edu.cn

T2-41

Synergistic use of geniposide and baicalin on BV2 cell activation damage caused by LPS

SHEN Tian，SONG Ya-gang，ZHANG Huan-huan，LIU Han，LI Min，WANG Bin

（Shaanxi University of Traditional Chinese Medicine，Xianyang 712046，China）

Abstract：OBJECTIVETo explore the synergistic effect of baicalin and geniposide（BG）on BV2 cell activation damage caused by lipopolysaccharide（LPS）.METHODSBV2 murine microglial cell line was culturedin vitro，LPS（final concentration 500 ng·mL-1）and various concentra?tionof Baicalin and Geniposide（BG）（final concentration 12.5，25 and 50 μg·mL-1）were added tointerven，the negative control was establised.MTT method was used to value the effect of LPS on the viability of BV2 cell line. The accumulated nitrite was assayed utilizing the Griess reaction method.RESULTS（1）Morphological observa?tion：The common marphological of quesient microglia is circle，cell bodies smaller and synaptic slender.The en?largement of microglial cell bodies and an amoeboid mor?phology with retraction of extensions are generally in?duced by LPS.BG markedly suppressed the LPS-activat?ed BV2 microglia morphological variations，meanwhile the dose-dependent was dramaticaly performed.（2）MTT test showed that LPS-stimulated BV2 cells viability was significantly decreased compared to the control group；compared to LPS treated cells，drug group（LPS+ BG）effectively improves the LPS-stimulated BV2 cells vi?ability.（3）The Griess reaction method indicated that LPS could obviously promoted the BV2 cells′NO genera?tion contrasted to control group；while the drug group（LPS+BG）can effectively inhibited the generation of NO which activated by LPS.CONCLUSIONThe treatment group could significantly enhance survival rate of LPS-stimulated BV2 cells，while，the level of NO was marked?ly decreased in BV2 induced by LPS.These findings sug?gest that combination of BG could attenuate BV2 microg?lial cells activation and injury which induced by LPS，pos?sessed the capacity of neuroprotective.

Key words：lipopolysaccharide；baicalin；geniposide；microglia；nerve inflammation

Foundation item：The project suppored by National Nat?ural Science Foundation of China（81473385）；Shaanxi Province Education Department Project（13JS029）；and by Shaanxi Province Administration of Traditional Chinese Medicine（13-ZY016）

Corresponding author：WANG Bin，E-mail：wangbin812 @126.com

T2-42

Neuroprotective effects of kaempferol against 2VO-induced chronic cerebral ischemia in rats

ZHANG Jun1，2，CHENG Xiao1，YANG Huan1，YANG Yin-lin1，ZHAO Ting-kun3， WANG Qi2， WANG Yue-hua1，DU Guan-hua1

（1.Beijing Key Laboratory of Drug Target Identification andDrugScreening，InstituteofMateriaMedica，Chinese Academy of Medical Sciences&Peking Union Medical College，Beijing 100050，China；2.Guangzhou University of Chinese Medicine，Guangzhou 510405，China；3.WeifangMedicalUniversity，Weifang 261053，China）

Abstract：OBJECTIVETo investigate the effects of kaempferol（KAE）on chronic cerebral ischemia in rats.METHODSChronic cerebral ischemia was induced in rats by permanent occlusion of bilateral common carotid arteries（2VO）.Then，the rats with chronic cerebral isch?emia were randomly divied into three groups：model group，KAE 10 and 30 mg·kg-1group.Another group rats without occlusion of common carotid arteries were used as the sham-operation group.Memory behavior was investigated by Morris water maze test.Prehensile ability was investigated by prehensile traction test.The structure of hippocampus and cortex neurons was observed with Nissel staining.In addition，the SOD activity and MDA content in brain tissue were determined.The DJ-1 protein level was determined by Western blotting.RESULTSKAE 10 and 30 mg·kg-1could significantly improve cogni?tive impairment and prehensile traction ability（P＜0.01）induced by chronic cerebral ischemia in rats.The results of the pathological analysis also suggested that KAE could ameliorate the pathological damage induced by chronic cerebral ischemia.In addition，KAE 30 mg·kg-1significantly increased the activity of SOD（P＜0.05），but had no effect on the content of MDA in rat brain tissue.Western-blotting confirmed that KAE 10 and 30 mg·kg-1could increase the expression of anti-oxida?tion proteins DJ-1 in hippocampus（P＜0.01）.CONCLU?SIONKAE may attenuate the chronic cerebral ischemic injury in rats.

Key words：kaempferol；chronic cerebral ischemia；occlusion of bilateral common carotid arteries

Foundation item：The project supported by National NaturalScience Foundation ofChina（81473383，81573645）

Corresponding authors：WANG Yue-hua，E-mail：wall?ing@imm.ac.cn；DU Guan-hua，E-mail：dugh@imm.ac. cn

T2-43

Xiao-xu-ming decoction inhibits lipopolysaccharide induced neuroinflammation in mice

CHENG Xiao1，YANG Huan1，ZHANG Jun2，YANG Yin-lin1， WANG Yue-hua1，DU Guan-hua1

（1.Beijing Key Laboratory of Drug Target Identification andDrugScreening，InstituteofMateriaMedica，Chinese Academy of Medical Sciences&Peking Union Medical College，Beijing 100050，China；2.Guangzhou University of Chinese Medicine，Guangzhou 510006，China）

Abstract：OBJECTIVEXiao-xu-ming decoction（XXMD），a well-known traditional Chinese herbal prescription，has been widely used to treat stroke.It is recorded in″Bei Ji Qian Jin Yao Fang″written by Si-miao Sun of the Chinese ancient Tang Dynasty.In our previous study，the active fraction of XXMD（XXM）against cerebral ischemia has been prepared by modern separation and purification techniques.This study was to investigate XXM against lipopolysaccaride（LPS）-induced neuroinflammation in mice.METHODSLPS is an endotoxin from the outer membrane ofGram-negative bacteria thatactivates inflammation.XXM was pre-treated in BALB/C mice fol?lowed by injected intraperitoneally with LPS（5 mg·kg-1）. The effects of XXM on LPS-induced pro-inflammatory factors and proteins were measured by ELISA，Western blot，and immunofluorescencein vivo.RESULTSMice treated with XXM showed significantly decreased proinflammatory factors level，including IL-1β（P＜0.01），IL-6（P＜0.01），TNF-α（P＜0.05），and MCP-1（P＜0.01）. Furthermore，XXM also significantly inhibited the inflam?matory pathway proteins expression induced by LPS，including TLR4，MyD88，and COX-2.CONCLUSIONXXM possesses anti-neuroinflammation in mice and might be a promising therapeutic agent for stroke.

Key words：Xiao-xu-ming decoction；lipopolysaccharide；neuroinflammation

Foundation item：The project supported by National NaturalScience Foundation ofChina（81473383，81573645）

Corresponding authors：WANG Yue-hua，E-mail：wall?ing@imm.ac.cn；DUGuan-hua，E-mail：dugh@imm.ac.cn

T2-44

The extract ofRamulus Cinnamomattenuates inflam?matory injury induced by LPS via regulation of TLR4/ MyD88 signaling pathway in BV2 cells

YANG Huan1，CHENG Xiao1，ZHANG Jun2，YANG Yin-lin1，WANG Yue-hua1，DU Guan-hua1

（1.Beijing Key Laboratory of Drug Target Identification and Drug Screening，Institute of Materia Medica，Chi?nese Academy of Medical Sciences&Peking Union Med?ical College，Beijing 100050，China；2.Guangzhou Uni?versity of Chinese Medicine，Guangzhou 510006，China）

Abstract：OBJECTIVETo investigate the effects of ex?tract of Ramulus Cinnamom（RC）against LPS-induced inflammation in microglia.METHODSActivated microg?lia releases various pro-inflammatory cytokines to induce neuroinflammation in stroke.Lipopolysaccaride（LPS）is an endotoxin from the outer membrane of Gram-negative bacteria that activates microglia.MTT assay was used to observe the cell viability.The content of NO in superna?tant was measured by Griess reagent.The levels of IL-1β，IL-6 and TNF-α in supernatant were detected by ELISA kits.The intracellular COX-2，TLR4，and MyD88 expression was assayed by Western blotting.RESULTSRC extract 30 and 100 μg·mL-1significantly decreased the production of related inflammatory factors such as NO（P＜0.05，P＜0.01），IL-1β（P＜0.01，P＜0.01），IL-6（P＜0.05，P＜0.01）and TNF-α（P＜0.01，P＜0.01）. Furthermore，RC extract significantly inhibited the COX-2，TLR4，and MyD88 expression induced by LPS in BV2 cells.CONCLUSIONRC extract may have therapeutic potential for the improvement of neuroinflammation，and the mechanism may be involved in down-regulation of TLR4/MyD88 inflammation pathway.

Key words：Ramulus Cinnamom；lipopolysaccharide；BV2 cells；neuroinflammation

Foundation item：The project supported by National NaturalScience Foundation ofChina（81473383，81573645）

Corresponding authors：WANG Yue-hua，E-mail：wall?ing@imm.ac.cn；DUGuan-hua，E-mail：dugh@imm.ac.cn

T2-45

Ginkgo biloba extract-761 improves the motor func?tions in permanent middle cerebral artery occlusion rats

ZENG Gui-rong1，2，3，ZHOU Shi-da1，2，SHAO Ya-jie1，2，ZHANG Miao-hong1，2，SUN Mei-zhen1，2，Mudassar Azhar4，Ayaz Aham4，LIU Xin-ming3，JIANG De-jian1，2

（1.Hunan Key Laboratory of Chinese Materia Medica Powder and Innovative Drugs Established by Provincial and Ministry，Changsha 410331，China；2.Hunan Pro?vincial Research Center for Safety Evaluation of Drugs，Changsha 410331，China；3.Institute of Medicinal Plant Development，Chinese Academy of Medical Sciences and Peking Union Medical College，Beijing 100193，Chi?na；4.Dr.Panjwani Center for Molecular Medicine and Drug Research，International Center for Chemical and Bi?ological Sciences，University of Karachi，Karachi，Paki?stan）

Abstract：OBJECTIVETo study the role of Ginkgo bi?loba extract-761（EGb-761）in the recovery of gait abnor?mality and its neuroprotective effect against the brain inju?ry induced by permanent middle cerebral artery occlu? sion in rats.METHODSMale Sprague Dawley rats（n= 200，240-305 g）were anesthetized with 0.2%pentobar?bital sodium diluted in physiological saline（2.0 mL·kg-1，ip）.Then a monofilament coated with poly-L-lysine，was used to occlude the origin of the middle cerebral artery.It was inserted into the internal carotid artery lumen until it met mild resistance，approximately 20mm beyond the common carotid arterybifurcation.The suture was secured with a ligature and maintained in place until sac?rifice.The same surgical procedure was conducted in sham-operated rats in which the middle cerebral artery was not occluded.Motor and behavioral changes were assessed after surgery using a five point scale.The rats securing the point scale above 2 were included in the study.The rats were randomly divided into control，and treated groups：EGb-761（20，50，and 100 mg·kg-1）. The treated groups were orally administered（10 mL·kg-1）for 28 d.On 7th，14th，21st，and 28th day the neurologi?cal scores，rotar rod test and gait assessment（the auto?mated computer-assisted method）were performed.The brains were collected for TTC staining and histopathologi?cal analysis.RESULTS1）Weight：On 28th day，EGb-761（20 mg·kg-1，）significantly increased the weight of the rat by~8%as compared to control（~300 g）.However，at 50 mg·kg-1，and 100 mg·kg-1，a significant increase of~7-7.6%（control：~232 g），and~7.3-7%，respectively from 14 to 28 days was noted.2）Neurological scores：On 28thday，EGb-761（20，50，and 100 mg·kg-1）signifi?cantly decreased the neurological scores by~18%，~22%，~21%，respectively as compared to control（~2.07）. 3）Rotar rod test：On 28thday，EGb-761（50，and 100 mg·kg-1）significantly increased by~69.1%，~74.1%，respectively as compared to control（~28.2）.4）Gait assessment：On 7th，14th，21st，and 28thday，EGb-761（20，50，and 100 m·kg-1）significantly reduced the aver?age body angle，on 7th，14th，21st，and 28thday，EGb-761（100 mg·kg-1）significantly increased the walk speed and reduced the average walking cycle，EGb-761（50，and 100 mg·kg-1）significantly the area of the left brain/right brain area percentage and reduced tissue pathologic neuron injury.CONCLUSIONGinkgo biloba extract EGb-761 has obvious improve behavior disor?ders，and has a protective neuroprotective effect against the brain injury induced by permanent middle cerebral artery occlusion.

Key words：permanent middle cerebral artery occlusion

Foundation item：The project supported by Hunan prov?ince Science and Technology Plan Projects of China（2015DK3010）

Corresponding authors：JIANG De-jian，Tel（0731）83285157，E-mail：jiangdejian@hnse.org；LIU Xin-ming，E-mail：liuxinminuae@yahoo.com.cn，Tel：（010）6281 2595/ 9108

T2-46

Chemogenetic activation of sublaterodorsal tegmental glutamatergic neurons alleviates rapid eye move?ment sleep behavior disorder symptoms in a chronic rat model of Parkinson disease

DU Li-da1，XU Lin-hao1，LIANg Tuo1，Yun-Kwok WING2，KE Ya1，Wing-Ho YUNG1

（1.School of Biomedical Sciences，2.Department of Psychiatry，F(xiàn)aculty of Medicine，The Chinese University of Hong Kong，Hong Kong，China）

Abstract：Rapid eye movement（REM）sleep behavior disorder（RBD）is a parasomnia that is featured by ele?vated motor behaviors and dream enactments during REM sleep.Clinical observations show that RBD bears significant relevance with several synucleinopathies such as Lewy body dementia and Parkinson disease（PD），and often develops prior to their diagnosis.Being a potential biomarker of PD，investigating the relationship of RBD symptoms and their emergence in developing PD would provide insight intoits pathogenesis.Here，in a chronic model of PD，rats with daily rotenone treatment exhibited key RBD features，including elevated sleep muscle tone，sleep fragmentation and EEG slowing at different time points.Based on detectedearly alpha synu?clein aggregation and neural apoptosis in the sublat?erodorsal tegmental nucleus（SLD），an area known to promote REM sleep and maintain sleep muscle atonia，the possible involvement of SLD glutamatergic neurons was interrogated.Via chemogenetic activation of SLD glutamatergic neurons，key RBD symptoms and EEG slowing in REM sleep were alleviated.These results are consistent with a progressive degeneration in REM sleep promoting pathways.Our findings provide a foundation for further studies into RBD and its relationship to neuro?degenerative diseases.

Key words：sublaterodorsal tegmental glutamatergic neurons；rapid eye movement；sleep behavior disorder

Foundation item：The project supported by the HKGRCGRF grant（14111715）

T2-25
Neuroprotective effect of Angiopep-2 peptide modi?fied scutellarin-loaded PEGylated PAMAM dendrimer nanoparticles on ischemic stroke by modulating the Toll-like receptors-dependent MyD88/IKK/NF-κB sig?naling pathway

LIU Xin， LI Yu-tao， LIU Wei， ZHANG Feng-ming，CHEN Zeng-zhen， ZENG Zhi-yong， XU Meng-shu，SUN Xiao-jun

（Department of Pharmaceutical Engineering，School of Chemical and Environmental Engineering，Harbin Univer?sity of Science And Technology，Harbin 150040，China）

OBJECTIVEThe greatest challenge in che?motherapy of ischemic stroke is the construction a suit?able delivery system to overcome the poor physicochemi?cal properties of drug and its low permeability across the blood brain barrier（BBB）.METHODSIn the present study，dendrimer，polyamidoamine（PAMAM），was syn?thesized as the nano-drug carriers.Angiopep-2，which has been proved excellent ability to cross the BBB，was exploited as the targeting ligand to conjugate PAMAM via bifunctional polyethylene glycol（PEG）.Then scutella?rin（STA） was encapsulated into the functionalized nanoparticles（NPs）to formulate Angiopep-2 modified STA-loaded PEG-PAMAM NPs.Ischemic stroke model was established to evaluate the treatment efficacy and protective mechanism of Angiopep-2-STA-PEG-PAMAM NPs.RESULTSThe pharmacokinetics and biodistribu?tion demonstrated that Angiopep-2-STA-PEG-PAMAM NPs exhibited significantly higher plasma concentration from 1 h to 10 h after intravenous administration and im?prove accumulation in brain（4.7-fold）compared with STA solution.Moreover，prolonged elimination half-life（4.8-fold）and lower clearance（3.4-fold）were observed. The brain uptake study of 6-coumarin confirmed that An?giopep-2-PEG-PAMAM NPs possessed better brain tar?geting efficacy（3.2-fold）than PEG-PAMAM NPs.Angio?pep-2-STA-PEG-PAMAM NPs obviously ameliorated in?farct volume，neurological deficit，histopathological se?verity and neuronal apoptosis.In addition，Angiopep-2-STA-PEG-PAMAM NPs markedly inhibited the calcium content and the levels of IL-12p40，IL-13，IL-17 and IL-23.Furthermore，Angiopep-2-STA-PEG-PAMAM NPs significantly decreased the mRNA and protein expres?sions of HMGB1，TLR2，TLR4，TLR5，MyD88，TRIF，TRAM，IRAK-4，TRAF6，IкBα，IKKβ and NF-кBp65.CONCLUSIONThe results suggested that Angiopep-2 modified scutellarin-loaded PEG-PAMAM nanocarriers possessed remarkable neuroprotective effects on ischemic stroke through modulation of inflammatory cascades and HMGB1/TLRs/MyD88-induced NF-κB activation pathways.

scutellarin；cerebral ischemia；Angiopep-2 modified PEG-PAMAM nanoparticles；brain targeting；HMGB1/TLR/MyD88/IKK/NF-κB pathways；neuroprotection

s：LIU Xin，E-mail：xinliu98@ 126.com；SUN Xiao-jun，sunxiaojun@hrbust.edu.cn，Tel：（0451）86392728

國家重點(diǎn)研發(fā)計劃（2016YFC0800900）；中國科學(xué)院百人計劃和吉林省自然科學(xué)基金（20160101211JC）

王曉輝，E-mail：xiaohui.wang@ciac.ac.cn

Foundation item：The project supported by National Natural Science Foundation of China（NSFC 21476054）；and the Natural Science Foundation of Heilongjiang Prov?ince（B201407）