·論著·
曲莉1,白雪松2,張晶瑩3(
1.北華大學(xué)公共衛(wèi)生學(xué)院,吉林 吉林132013;2.吉林醫(yī)藥學(xué)院營(yíng)養(yǎng)教研室,吉林 吉林132013;3.吉林省疾病預(yù)防控制中心,吉林 長(zhǎng)春130062)
摘要:目的研究牛磺酸對(duì)致小鼠腦組織氧化損傷的保護(hù)作用。方法選用健康雄性ICR小鼠40只隨機(jī)均分為正常對(duì)照組、模型組、?;撬岣邉┝拷M、?;撬岬蛣┝拷M。采用跳臺(tái)實(shí)驗(yàn)觀察其對(duì)小鼠學(xué)習(xí)記憶的影響;測(cè)試后處死小鼠,計(jì)算腦臟器系數(shù),測(cè)定腦組織中丙二醛(MDA)、B型單胺氧化酶(MAO-B)含量及超氧化物歧化酶(SOD)活性。結(jié)果跳臺(tái)試驗(yàn)顯示模型組錯(cuò)誤次數(shù)明顯高于正常對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P<0.05);高劑量組錯(cuò)誤次數(shù)明顯低于模型組,差異有統(tǒng)計(jì)學(xué)意義(P<0.05);模型組、各劑量組小鼠重量及臟器系數(shù)均低于正常對(duì)照組,但差異無(wú)統(tǒng)計(jì)學(xué)意義。模型組MDA水平升高與高劑量組相比,有統(tǒng)計(jì)學(xué)意義(P<0.05);模型組SOD活性降低與正常對(duì)照組相比,有統(tǒng)計(jì)學(xué)意義(P<0.05);高劑量組SOD活性升高與模型組相比,有統(tǒng)計(jì)學(xué)意義(P<0.05);模型組MAO-B活性增強(qiáng)與正常對(duì)照組、高、低劑量組相比,有統(tǒng)計(jì)學(xué)意義(P<0.05);高劑量組MAO-B活性明顯低于低劑量組,差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。結(jié)論?;撬釋?duì)所致小鼠腦組織氧化損傷具有保護(hù)作用。
關(guān)鍵詞:牛磺酸;氯化
文章編號(hào):1673-2995(2015)06-0412-04
中圖分類號(hào):R114
基金項(xiàng)目:吉林省衛(wèi)生廳重點(diǎn)實(shí)驗(yàn)室資助項(xiàng)目(2010Z056).
作者簡(jiǎn)介:曲莉(1978-),女(漢族),講師,碩士.
收稿日期:(2015-05-18)
;氧化損傷;腦組織
The protection of taurine on brain tissue oxidative damage induced by cadmium in mice
QU Li1,BAI Xuesong2,ZHANG Jingying3(1.College of Public Health,Beihua University,Jilin City,Jilin Province,132013;2.Department of Nutrition,Jilin Medical University,Jilin City,Jilin Province,132013;3.Disease Control Prevention of Jilin Province,Changchun,Jilin Province,130062,China)
Abstract:ObjectiveTo study the protection of taurine on brain tissue oxidative damage in mice exposed to cadmium chloride. Methods 40 ICR mice were randomized divided into 4 groups:control group,cadmium-exposed group,two administrated groups.All mouse were evaluated for their learning and memory performance by step-down test.After testing,viscera coefficient,the level of MDA and MAO-B,the activity of SOD were detected. ResultsThe results of step-down test indicated that the wrong number of the cadmium-exposed group was higher than those in the control group and the wrong number of high dose taurine treated group was less than those in the cadmium-exposed group,The difference was statistically significant (P<0.05).The results indicated that viscera coefficient of two administrated groups was lower than those in the control group,but there was no statistically significant difference (P>0.05).Compared with the cadmium-exposed group,the MDA levels of high dose taurine treated group were decreased significantly,The difference was statistically significant (P<0.05),while the SOD activities were increased significantly (P<0.05).Compared with the cadmium-exposed group,the MAO-B levels of control group and two administrated groups decreased significantly (P<0.05),and the MAO-B level in the high dose taurine treated group was lower than those in the low dose group. ConclusionTaurine has protective effect on brain tissue oxidative damage
通訊作者:張晶瑩(1978-),女(漢族),醫(yī)師,碩士.
induced by cadmium in mice.
Key words:ttaurine;cadmium chloride;oxidative damage;brain tissue
牛磺酸(taurine,Tau)是一種β-氨基酸,為人體條件性必需氨基酸,是人體內(nèi)含量最豐富的低分子有機(jī)成分,以游離的形式普遍存在于人體的各種組織中,在大腦中的含量顯著高于其他臟器,以小分子二或三的形式存在于中樞神經(jīng)系統(tǒng)。?;撬峋哂芯S持滲透壓平衡,降血壓、降血脂,調(diào)節(jié)鈣在體內(nèi)的平衡等方面的作用,近年來(lái)有研究表明牛磺酸具有廣泛的細(xì)胞保護(hù)作用[1],可以促進(jìn)大腦神經(jīng)細(xì)胞的增殖、分化和發(fā)育,可增強(qiáng)腦細(xì)胞內(nèi)DNA、RNA的合成,能增強(qiáng)腦細(xì)胞對(duì)蛋白質(zhì)的利用率,有研究顯示?;撬徇€可改善學(xué)習(xí)和記憶能力[2]。(cadmium,Cd)是一種有毒重金屬,1994年被國(guó)際癌癥研究所(International Agency for Research Off Cancer,IARC)定為IA級(jí)致癌物,即人類致癌物。對(duì)腎臟、肝臟、腦、肺、睪丸、骨及血液系統(tǒng)均可產(chǎn)生毒性,被美國(guó)毒物管理委員會(huì)(Agency for Toxic Substances and Disease Registry,ATSDR)列為第6位危及人類健康的有毒物質(zhì)。在人體內(nèi)可長(zhǎng)期蓄積,通過(guò)多種途徑進(jìn)入人體的,其半衰期為10~30年,近年研究發(fā)現(xiàn)對(duì)神經(jīng)系統(tǒng)有明顯的毒性作用[3-4],的毒性與氧化損傷密切相關(guān)。本研究主要研究?;撬釋?duì)致小鼠腦組織氧化損傷的保護(hù)作用。
?;撬?C2H7NO3S),分子量125.15,含量≥98.0%(產(chǎn)品批號(hào):F20071022);氯化(CdCl2),分析純,含量≥99.0%(產(chǎn)品批號(hào):F20110222),購(gòu)自國(guó)藥集團(tuán)化學(xué)試劑有限公司;離心機(jī)為北京醫(yī)用離心機(jī)廠產(chǎn)品;組織勻漿機(jī)為江蘇金壇金城國(guó)勝實(shí)驗(yàn)儀器廠產(chǎn)品;722分光光度計(jì)為上海奧普勒儀器有限公司產(chǎn)品;分析天平為北京塞多利斯天平公司產(chǎn)品。
清潔級(jí)健康雄性ICR小鼠,體重18~22 g,購(gòu)自吉林大學(xué)基礎(chǔ)醫(yī)學(xué)院動(dòng)物實(shí)驗(yàn)中心,動(dòng)物許可證號(hào)SCXK-(吉)2011-0003,自由吃食和飲水,室溫(20±2)℃,濕度(55%~65%)。將40只小鼠按體重隨機(jī)分為4組:正常對(duì)照組、模型組、高劑量組、低劑量組,每組10只。除正常對(duì)照組外,其余各組每周二次腹腔注射CdCl22.0 mg/kg,連續(xù)4周,正常對(duì)照組每周2次腹腔注射生理鹽水(相同體積)。同時(shí),高劑量組、低劑量組每日分別灌胃?;撬? g/kg和750 mg/kg,連續(xù)4周,正常對(duì)照組和模型組每日灌胃蒸餾水。
將小鼠置于箱中,自由活動(dòng)5 min,然后置于安全臺(tái)上,箱底通電,觀察其跳臺(tái)的潛伏期并記錄5 min內(nèi)小鼠跳下臺(tái)的次數(shù),此為錯(cuò)誤次數(shù),即學(xué)習(xí)成績(jī)。
測(cè)試結(jié)束后,頸椎脫臼處死小鼠,迅速取腦組織,稱重,計(jì)算腦臟器系數(shù)(腦臟器系數(shù)=腦重量/體重×100%)。
斷髓處死小鼠,立即取全腦,用冰冷的生理鹽水漂洗,除去血液,濾紙拭干,稱取組織重量,制成10%勻漿液。用丙二醛(MDA)、B型單胺氧化酶(monoamine oxidase B type,MAO-B)、超氧化物歧化酶(SOD)試劑盒法分別檢測(cè)腦中MDA、SOD活性及MAO-B含量,同時(shí)用考馬斯亮藍(lán)試劑盒檢測(cè)蛋白質(zhì)含量。
數(shù)據(jù)采用SPSS11.5統(tǒng)計(jì)軟件進(jìn)行單因素方差分析。方差齊性檢驗(yàn)部分,組間比較采用最小顯著差異法檢驗(yàn)(LSD法)。不齊時(shí),各組間兩兩比較采用Tamhane’s方法。
小鼠跳臺(tái)試驗(yàn)學(xué)習(xí)成績(jī)模型組(11.7±3.0)與正常對(duì)照組(4.0±1.7)比較,錯(cuò)誤次數(shù)增加(P<0.05),說(shuō)明模型組小鼠學(xué)習(xí)能力下降;高劑量組(9.5±2.5)與模型組相比,錯(cuò)誤次數(shù)減少(P<0.05),而低劑量組(10.2±2.1)與模型組相比則沒有差別。
表1可見,模型組、各劑量組小鼠重量及臟器系數(shù)均低于正常對(duì)照組,但差異無(wú)統(tǒng)計(jì)學(xué)意義。
表2可見,模型組MDA水平升高與正常對(duì)照組相比,有統(tǒng)計(jì)學(xué)意義(P<0.05);模型組SOD活性降低與正常對(duì)照組相比,有統(tǒng)計(jì)學(xué)意義(P<0.05);高劑量組SOD活性升高與模型組相比,有統(tǒng)計(jì)學(xué)意義(P<0.05);模型組MAO-B活性增強(qiáng)與正常對(duì)照組、高、低劑量組相比,有統(tǒng)計(jì)學(xué)意義(P<0.05);高劑量組MAO-B活性減弱與低劑量組相比,有統(tǒng)計(jì)學(xué)意義(P<0.05)。
表 1 ?;撬釋?duì) 致小鼠腦臟器系數(shù)的影響( ± s, n=10)
表 1 ?;撬釋?duì) 致小鼠腦臟器系數(shù)的影響( ± s, n=10)
組 別重量(g)臟器系數(shù)正常對(duì)照組0.49±0.091.16±0.10模型組0.45±0.041.13±0.05高劑量組0.48±0.061.15±0.05低劑量組0.48±0.071.16±0.08
表 2 ?;撬釋?duì) 致小鼠腦組織生化指標(biāo)影響( ± s, n=10)
表 2 ?;撬釋?duì) 致小鼠腦組織生化指標(biāo)影響( ± s, n=10)
組 別MDA(μmol/g)SOD(μU/g)MAO-B(μU/g)正常組0.78±0.10a81±14a0.54±0.12a模型組1.01±0.0949±101.69±0.14高劑量組0.84±0.11a60±11a1.49±0.20ab低劑量組0.91±0.1351±91.56±0.15a
與模型組比較,aP<0.05;與低劑量組比較,bP<0.05。
參考文獻(xiàn):
[1]Pan Chunliu,Giraldo GS,Prentice H,et al.Taurine protection of PC12cells against endoplasmic reticulum stress induced by oxidative stress[J].J Biomed Sci,2010,17(Suppl 1):S17.
[3]Robinson JF,Yu Xiaozhong,Hong S,et al.Cadmium-Induced differential toxicogenomic response in resistant and sensitive mouse strains undergoing neurulation[J].Toxicol Sci,2009,107(1):206-219.
[4]Borges VC,Santos FW,Rocha JB,et al.Heavy metals modulate glutamatergic system in human platelets[J].Neurochem Res,2007,32(6):953-958.
[5]劉宗平.動(dòng)物中毒病學(xué)[M].北京:中國(guó)農(nóng)業(yè)出版社,2006:394-402.
[6]Liu Jie,Qu Wei,Kadiiska MB.Role of oxidative stress in cadmium toxicity and carcinogenesis[J].Toxicol Appl Pharmacol,2009,238(3):209-214.
[7]Zamora Z,Gonzalez R,Guanche D,et al.Ozonized sunflower oil reduces oxidative damage induced by indomethacin in rat gastric mucosa[J].Inflamm Res,2008,57(1):39-43.
[9]Bar-Am O,Weinreb O,Amit T,et al.The novel cholinesterase-monoamine oxidase inhibitor and antioxidant,ladostigil,confers neuroprotection in neuroblastoma cells and aged rats[J].J Mol Neurosci,2009,37(2):135-145.