杜金梁, 曹麗萍, 賈睿, 劉英娟, 申玉金, 殷國俊,*
(1.中國水產(chǎn)科學(xué)研究院淡水漁業(yè)研究中心,農(nóng)業(yè)部淡水漁業(yè)和種質(zhì)資源利用重點(diǎn)實(shí)驗(yàn)室,江蘇 無錫214081;2.中國水產(chǎn)科學(xué)研究院淡水漁業(yè)研究中心,農(nóng)業(yè)部魚類免疫藥理學(xué)國際聯(lián)合實(shí)驗(yàn)室,江蘇 無錫214081;3.南京農(nóng)業(yè)大學(xué)無錫漁業(yè)學(xué)院,江蘇 無錫214081)
甘草提取物對(duì)2,3,7,8-四氯二苯并-p-二英致建鯉肝損傷的影響
杜金梁1,2, 曹麗萍1,2, 賈睿3, 劉英娟3, 申玉金3, 殷國俊1,2,3*
(1.中國水產(chǎn)科學(xué)研究院淡水漁業(yè)研究中心,農(nóng)業(yè)部淡水漁業(yè)和種質(zhì)資源利用重點(diǎn)實(shí)驗(yàn)室,江蘇 無錫214081;2.中國水產(chǎn)科學(xué)研究院淡水漁業(yè)研究中心,農(nóng)業(yè)部魚類免疫藥理學(xué)國際聯(lián)合實(shí)驗(yàn)室,江蘇 無錫214081;3.南京農(nóng)業(yè)大學(xué)無錫漁業(yè)學(xué)院,江蘇 無錫214081)
為了評(píng)價(jià)甘草提取物是否對(duì)2,3,7,8-四氯二苯并-p-二英(2,3,7,8-tetrachlorodibenzo-p-dioxin,TCDD)引起建鯉肝組織損傷具有保護(hù)作用,本實(shí)驗(yàn)用含甘草提取物的飼料(0.1、0.5和1.0 g/kg)飼喂建鯉60 d,再腹腔注射0.6 μg/kg TCDD,72 h后收集血液和肝組織,檢測血清和肝組織勻漿中谷丙轉(zhuǎn)氨酶(alanine transaminase,ALT)、谷草轉(zhuǎn)氨酶(aspartate transaminase,AST)、乳酸脫氫酶(lactate dehydrogenase,LDH)、超氧化物歧化酶(superoxide dismutase,SOD)、堿性磷酸酶(alkaline phosphatase,AKP)、總蛋白(total protein,TP)、白蛋白(albumin,Alb)、過氧化氫酶(hydrogen peroxidase,CAT)、谷胱甘肽過氧化物酶(glutathione peroxidase,GPx)、總抗氧化能力(total antioxidant capacity, T-AOC)、丙二醛(malondialdehyde,MDA)等生化指標(biāo),同時(shí)制備肝組織病理切片,觀察甘草提取物對(duì)TCDD誘導(dǎo)建鯉肝組織損傷的影響。結(jié)果表明:1.0 g/kg甘草提取物顯著降低血清中ALT、AST、LDH和AKP活性,顯著提高總蛋白(TP)、白蛋白(Alb)質(zhì)量濃度、過氧化氫酶(CAT)、谷胱甘肽過氧化物酶(GPx)、總抗氧化能力(T-AOC)和超氧化物歧化酶(SOD)活性,降低丙二醛(MDA)質(zhì)量摩爾濃度(P<0.05或P<0.01)。病理組織切片觀察結(jié)果顯示,甘草提取物處理組可以明顯減輕TCDD引起肝組織損傷。提示,甘草提取物對(duì)TCDD引起的建鯉肝組織損傷具有較好的保護(hù)作用,而且甘草提取物的保護(hù)作用可能與其抗氧化性能有關(guān)。
甘草提取物; 2,3,7,8-四氯二苯并-p-二英; 肝損傷; 建鯉
中藥的應(yīng)用在我國有著悠久的歷史,隨著人類對(duì)中藥研究的不斷深入,中草藥被廣泛應(yīng)用于一些疾病的治療。甘草,又名烏拉爾甘草,是一種多年生草本植物,屬豆科?,F(xiàn)已證明甘草的功能有很多,如甘草具有抗感染、增強(qiáng)機(jī)體免疫功能、防治病毒性肝炎、抗氧化、清除多種自由基等作用[15]。關(guān)于其在肝損傷治療方面應(yīng)用研究,曹麗萍等[16]在甘草提取物對(duì)叔丁基氫過氧化物誘導(dǎo)的建鯉原代培養(yǎng)肝細(xì)胞損傷的研究中發(fā)現(xiàn),甘草提取物對(duì)叔丁基過氧化氫(tert-butyl hydroperoxide,t-BHP)造成的魚類肝細(xì)胞損傷具有一定的保護(hù)作用;高雪巖等[17]通過研究甘草中總皂苷對(duì)四氯化碳所造成的小鼠急性肝損傷影響發(fā)現(xiàn),甘草總皂苷可以明顯降低四氯化碳造成的肝損傷,且效果顯著。而關(guān)于甘草提取物是否也可以抑制TCDD造成的魚類肝組織損傷,還未見相關(guān)報(bào)道。
本研究以TCDD 誘導(dǎo)鯉肝組織的急性損傷模型為基礎(chǔ),通過測定肝功能生化指標(biāo)和肝組織病理切片的變化來探索甘草提取物是否對(duì)TCDD引起的建鯉急性肝組織損傷具有保護(hù)作用,以期為防治TCDD引起的肝組織損傷提供理論依據(jù)。
1.1 試驗(yàn)用魚
試驗(yàn)用建鯉取自中國水產(chǎn)科學(xué)研究院淡水漁業(yè)研究中心漁場,體質(zhì)健康、無傷,體質(zhì)量為(30.0±1.0) g左右,取回后將建鯉飼養(yǎng)于循環(huán)水系統(tǒng)中暫養(yǎng)1周。
1.2 試劑和儀器
甘草提取物(Glycyrrhizaglabraextracts,GGE)購自西安應(yīng)化生物技術(shù)有限公司;二甲基亞砜 (dimethyl sulfoxide, DMSO)購自美國Sigma公司;2, 3, 7, 8-四氯二苯并-p-二英購自廈門慧嘉生物科技有限公司;谷丙轉(zhuǎn)氨酶、谷草轉(zhuǎn)氨酶等測定試劑盒購自南京建成生物工程研究所;723分光光度計(jì)購自上海欣茂儀器有限公司;酶標(biāo)儀MK3購自美國Thermo公司。
1.3 TCDD誘導(dǎo)建鯉肝損傷模型建立
將大小規(guī)格基本一致的建鯉180條隨機(jī)分為6個(gè)處理組,即1個(gè)對(duì)照組和5個(gè)處理組(TCDD質(zhì)量比為0.1、0.3、 0.6、 1.2和2.4 μg/kg)。5個(gè)處理組按建鯉體質(zhì)量來進(jìn)行腹腔注射染毒,注射劑量為每10 g體質(zhì)量0.05 mL,對(duì)照組僅注射同體積DMSO(0.1%的DMSO,注射劑量為0.05 mL/10 g體質(zhì)量)。
1.4 試驗(yàn)組劃分及處理
選取健康、無傷,體質(zhì)量為(30.0±1.0) g左右的建鯉120尾,隨機(jī)將建鯉設(shè)5個(gè)處理組和1個(gè)對(duì)照組,每組20尾魚,即正常對(duì)照組和TCDD對(duì)照組(僅飼喂基礎(chǔ)飼料);藥物(GGE)對(duì)照組基礎(chǔ)飼料中添加1.0 g/kg甘草提取物;3個(gè)試驗(yàn)組:基礎(chǔ)飼料中分別添加質(zhì)量比為0.1、0.5和1.0 g/kg甘草提取物。連續(xù)飼喂60 d,正常對(duì)照組和GGE對(duì)照組按體質(zhì)量注射同體積DMSO(0.1%的DMSO,注射劑量0.05 mL/10 g體質(zhì)量)。TCDD組和3個(gè)試驗(yàn)組一次性腹腔注射0.6 μg/kg TCDD(用最終不超過體積分?jǐn)?shù)0.1%的DMSO將TCDD助溶),注射量:按照建鯉體質(zhì)量來進(jìn)行注射(0.05 mL/10 g體質(zhì)量)。72 h后采集血液和肝組織。
1.5 血清及肝組織勻漿制備
于建鯉尾靜脈處進(jìn)行采血,用4 ℃低溫離心機(jī),3 500 r/min離心10 min,分離上層血清,將分離好血清置于-20 ℃保存待用。
用0.9%氯化鈉溶液將建鯉肝組織中殘存血液清洗干凈后,用干凈濾紙吸干其表面水分,稱取肝組織0.1 g,然后加入9倍體積的0.9%氯化鈉溶液,用玻璃勻漿器在冰浴上制備10%的肝組織勻漿液。用4 ℃低溫離心機(jī),3 000 r/min離心10 min,收集肝組織勻漿上清液,于-20 ℃保存?zhèn)溆谩?/p>
1.6 生化指標(biāo)測定
參照南京建成生物工程研究所試劑盒說明書測定谷丙轉(zhuǎn)氨酶(alanine transaminase,ALT)、谷草轉(zhuǎn)氨酶(aspartate transaminase,AST)、乳酸脫氫酶(lactate dehydrogenase,LDH)、白蛋白(albumin,Alb)、總蛋白(total protein,TP)、 超氧化物歧化酶(superoxide dismutase,SOD)、堿性磷酸酶(alkaline phosphatase,AKP)、過氧化氫酶(hydrogen peroxidase,CAT)、谷胱甘肽過氧化物酶(glutathione peroxidase,GPx)、總抗氧化能力(total antioxidant capacity,T-AOC)和丙二醛(malondialdehyde,MDA)等各項(xiàng)生化指標(biāo)。
1.7 肝組織切片制備與觀察
將采集好的肝組織放入波恩氏固定液中固定48 h后,制備石蠟組織切片,通過蘇木精-伊紅(hatmatoxylin-eosin,HE) 染色來觀察TCDD對(duì)建鯉肝組織的影響。
1.8 數(shù)據(jù)分析
2.1 不同質(zhì)量比的TCDD對(duì)建鯉血清和肝組織中生化指標(biāo)的影響
由圖1可以看出,腹腔注射TCDD質(zhì)量比≥0.3 μg/kg后,建鯉血清中ALT、AST和LDH的活性在注射72~96 h出現(xiàn)顯著升高(P<0.05或P<0.01)。
n=5; *,**分別表示與對(duì)照組相比在P<0.05和P<0.01水平差異有統(tǒng)計(jì)學(xué)意義。n=5; *,** indicate statistically significant difference from control group at the 0.05 and 0.01 probability levels, respectively.圖1 TCDD致建鯉肝損傷血清和對(duì)肝組織中AST(A)、ALT(B)、LDH(C)、GST(D)和T-AOC(E)活性的影響Fig.1 Effects of TCDD on AST (A), ALT (B), LDH (C), GST (D) and T-AOC (E) activities in serum and liver tissue of Jian carp
建鯉肝組織中谷胱甘肽S-轉(zhuǎn)移酶(glutathioneS-transferase,GST)和T-AOC活性在腹腔注射TCDD質(zhì)量比≥0.3 μg/kg后,48 h和72 h出現(xiàn)顯著降低(P<0.05或P<0.01),綜合結(jié)果,以72 h損傷較為嚴(yán)重。
2.2 甘草提取物對(duì)建鯉血清中ALT、AST、LDH、AKP活性的影響
圖2顯示,建鯉肝組織受到損傷后,血清中ALT、AST、LDH、AKP活性顯著升高(P<0.05或P<0.01)。飼料中添加3種不同質(zhì)量比的甘草提取物飼喂建鯉后,血清中ALT、AST、LDH、AKP活性出現(xiàn)不同程度降低。
2.3 甘草提取物對(duì)建鯉血清中TP和Alb質(zhì)量濃度的影響
由圖3顯示,建鯉肝組織的損傷引起了血清中TP和Alb質(zhì)量濃度顯著降低(P<0.01)。這種降低趨勢在加入甘草提取物后被抑制,以1.0 g/kg甘草提取物效果最好。
n=10; *,**分別表示與TCDD組相比在P<0.05和P<0.01水平差異有統(tǒng)計(jì)學(xué)意義。n=10; *,** indicate statistically significant difference from TCDD group-treated at the 0.05 and 0.01 probability levels, respectively.圖2 甘草提取物對(duì)TCDD致建鯉肝損傷血清中ALT(A)、AST(B)、LDH(C) 和AKP(D)活性的影響Fig.2 Effects of GGE on the serum ALT (A), AST (B), LDH (C) and AKP (D) in TCDD-treated Jian carp
n=10; *,**分別表示與TCDD組相比在P<0.05和P<0.01水平差異有統(tǒng)計(jì)學(xué)意義。n=10; *,** indicate statistically significant difference from TCDD group-treated at the 0.05 and 0.01 probability levels, respectively.圖3 甘草提取物對(duì)TCDD致建鯉肝損傷血清中TP和Alb質(zhì)量濃度的影響Fig.3 Effects of GGE on the serum TP (A) and Alb (B) in TCDD-treated Jian carp
2.4 甘草提取物對(duì)建鯉肝組織中抗氧化指標(biāo)的影響
由圖4顯示,經(jīng)過TCDD處理后的建鯉肝組織中抗氧化指標(biāo)(SOD、GPx、CAT和T-AOC)出現(xiàn)顯著的降低(P<0.05或P<0.01)。飼料中添加甘草提取物后發(fā)現(xiàn),0.5 g/kg和1.0 g/kg甘草提取物處理組效果最好,可以顯著增加肝組織中SOD、CAT和T-AOC活性,GPx在1.0 g/kg甘草提取物處理組也顯著升高,而0.1 g/kg甘草提取物處理組僅顯著增加了SOD活性值。
n=10; *,**分別表示與TCDD組相比在P<0.05和P<0.01水平差異有統(tǒng)計(jì)學(xué)意義。n=10; *,** indicate statistically significant difference from TCDD group-treated at the 0.05 and 0.01 probability levels, respectively.圖4 甘草提取物對(duì)TCDD致建鯉肝損傷組織中SOD(A)、GPx(B)、CAT(C)和T-AOC(D)的影響Fig.4 Effects of GGE on liver SOD (A), GPx (B), CAT (C) and T-AOC (D) in TCDD-treated Jian carp
2.5 甘草提取物對(duì)建鯉肝組織中MDA質(zhì)量摩爾濃度的影響
肝組織的損傷引起MDA的大量生成(P<0.01)(圖5),與TCDD 組相比,0.5和1.0 g/kg甘草提取物有效地抑制了MDA的生成(P<0.01),對(duì)肝組織起到有效的保護(hù)作用。
2.6 病理組織學(xué)觀察
在光學(xué)顯微鏡下觀察發(fā)現(xiàn),正常對(duì)照組肝細(xì)胞核和細(xì)胞質(zhì)未見有異常變化(圖6A,B),細(xì)胞質(zhì)均勻、細(xì)胞界線清晰、細(xì)胞核清晰可見;TCDD 對(duì)照組肝組織結(jié)構(gòu)明顯遭到破壞,肝細(xì)胞體積明顯增大,細(xì)胞界限變得不清楚,肝細(xì)胞核仁大部分消失,細(xì)胞呈蜂窩狀,出現(xiàn)大小不等的空泡化(圖6C);加入3種不同質(zhì)量比的甘草提取物后,肝組織結(jié)構(gòu)發(fā)生了不同變化,加入0.1 g/kg甘草提取物后,肝細(xì)胞病變未出現(xiàn)明顯改變(圖6D);0.5 g/kg的甘草提取物組
n=10;**表示與TCDD組相比在P<0.01水平差異有高度統(tǒng)計(jì)學(xué)意義。 n=10; ** indicate highly statistically significant difference from TCDD group-treated at the 0.01 probability level.圖5 甘草提取物對(duì)TCDD 致建鯉肝組織中MDA質(zhì)量摩爾濃度的影響Fig.5 Effects of GGE on liver MDA contents in TCDD-treated Jian carp
HE染色. A:正常組;B:藥物對(duì)照組;C:TCDD對(duì)照組;D:0.1 g/kg甘草提取物處理組;E: 0.5 g/kg甘草提取物處理組;F:1.0 g/kg甘草提取物處理組. HE stain. A: Control group; B: GGE control group; C: TCDD control group; D: 0.1 g/kg GGE treatment group; E: 0.5 g/kg GGE treatment group; F: 1.0 g/kg GGE treatment group.圖6 甘草提取物對(duì)建鯉肝組織損傷的影響Fig.6 Effects of GGE on liver injury induced by TCDD in Jian carp
肝細(xì)胞損傷程度變小,細(xì)胞數(shù)量增多,細(xì)胞空泡化減少,細(xì)胞輪廓逐漸清晰(圖6E);質(zhì)量比為1.0 g/kg的甘草提取物效果最好,損傷程度明顯降低,細(xì)胞核位于細(xì)胞中心清晰可見 (圖6F)。
當(dāng)前研究表明,TCDD可以對(duì)肝組織產(chǎn)生損害作用,可以導(dǎo)致魚類肝腫大且表面結(jié)節(jié)、脂肪肝、肝萎縮等[18]。關(guān)于TCDD的毒性機(jī)制研究,目前認(rèn)為TCDD發(fā)揮毒性作用主要通過2種途徑,一種是芳香烴受體/芳香烴受體核轉(zhuǎn)運(yùn)蛋白質(zhì)途徑,另一種是芳香烴受體/蛋白質(zhì)磷酸化信號(hào)轉(zhuǎn)導(dǎo)途徑。當(dāng)前研究熱點(diǎn)主要集中在第一種途徑,主要引起機(jī)體一系列氧化應(yīng)激反應(yīng),從而改變酶的活性。能否成功制備TCDD誘導(dǎo)的肝損傷模型是本實(shí)驗(yàn)成功開展的關(guān)鍵所在。本研究根據(jù)前期造模實(shí)驗(yàn)結(jié)果確定以0.6 μg/kg的TCDD腹腔注射來制備肝損傷模型,實(shí)驗(yàn)結(jié)果顯示0.6 μg/kg的TCDD可以造成ALT、AST等肝指標(biāo)的變化,這說明造模實(shí)驗(yàn)是成功的,可以進(jìn)行后續(xù)實(shí)驗(yàn)研究。
肝組織受損傷后,肝細(xì)胞內(nèi)的ALT、AST、AKP、LDH溢出量顯著增高,這4個(gè)指標(biāo)是反映肝細(xì)胞受損的敏感指標(biāo)[19]。在本實(shí)驗(yàn)中,TCDD引起建鯉肝組織損傷,導(dǎo)致4種酶活性顯著升高,飼料中添加不同質(zhì)量比的甘草提取物后發(fā)現(xiàn),ALT、AST、AKP和LDH 4種酶的升高趨勢被抑制,出現(xiàn)顯著性降低。推測其具體保護(hù)作用可能與甘草提取物中甘草酸成分有關(guān),能夠保護(hù)正常肝細(xì)胞免受TCDD損害,對(duì)肝細(xì)胞有修復(fù)作用[20]。類似結(jié)果在小鼠和魚上也有相關(guān)報(bào)道,如陳紅艷等[21]研究發(fā)現(xiàn)甘草提取物對(duì)于四氯化碳誘導(dǎo)急性化學(xué)性肝損傷具有較好的保護(hù)作用。曹麗萍等[16]報(bào)道甘草提取物對(duì)t-BHP引起建鯉原代肝細(xì)胞損傷具有良好保護(hù)作用。
肝在蛋白質(zhì)代謝過程中起著重要作用,血漿內(nèi)主要的蛋白質(zhì)幾乎全部由肝制造,肝合成的蛋白質(zhì)主要為白蛋白(Alb)。血清中總蛋白(TP)和白蛋白(Alb)是反映肝中蛋白質(zhì)丟失情況的重要指標(biāo),當(dāng)肝受到外界物質(zhì)損害時(shí),就會(huì)造成肝合成蛋白質(zhì)出現(xiàn)紊亂,使得白蛋白合成變少[22-23]。在本實(shí)驗(yàn)中,TCDD 誘導(dǎo)肝組織損傷后,導(dǎo)致了血清中TP和Alb質(zhì)量濃度的降低,但飼料中加入甘草提取物可以明顯升高其質(zhì)量濃度,說明甘草提取物可以降低TCDD對(duì)肝造成的損害,促進(jìn)肝恢復(fù)合成蛋白質(zhì)功能。
SOD、GPx、CAT和T-AOC是機(jī)體內(nèi)重要的抗氧化酶指標(biāo)[24],SOD的主要功能是對(duì)抗氧自由基,將氧自由基歧化,防止催化產(chǎn)生的H2O2與O2-結(jié)合形成危害更大的羥基自由基。過氧化氫酶(CAT)是生物體內(nèi)的一種末端氧化酶,主要功能是催化H2O2,防止其含量過高,對(duì)機(jī)體造成損傷[25]。谷胱甘肽過氧化物酶(GPx)的功能較多,不僅可以清除過氧化氫還可以清除脂質(zhì)過氧化物產(chǎn)物??偪寡趸芰-AOC的測定很重要,它可直接反映體內(nèi)抗氧化酶的活力高低,其強(qiáng)弱與生物體健康程度存在密切關(guān)系。在本實(shí)驗(yàn)中,圖4數(shù)據(jù)顯示,TCDD使建鯉肝組織顯著的損傷,引起SOD、GPx、CAT和T-AOC活性的降低,而用甘草提取物飼喂后的建鯉4種酶指標(biāo)被有效地提高,表明甘草提取物能增強(qiáng)建鯉抗氧化能力,抵抗TCDD造成的肝組織損傷。已有研究報(bào)道甘草提取物中總多酚與黃酮類含量與機(jī)體抗氧化能力密切相關(guān)[26],因此可以推斷甘草中多酚和黃酮類物質(zhì)可能是抵抗肝組織氧化損傷的主要因素。
當(dāng)機(jī)體受到自由基攻擊后就會(huì)出現(xiàn)脂質(zhì)過氧化反應(yīng),MDA就是其中一個(gè)判定指標(biāo),其質(zhì)量摩爾濃度高低反映了機(jī)體受損傷程度[27]。TCDD作為一種肝毒物可以引起脂質(zhì)過氧化,使得MDA質(zhì)量摩爾濃度升高,邊芳等[28]研究結(jié)果顯示,甘草提取物可以顯著降低MDA質(zhì)量摩爾濃度,通過抑制自由基和脂質(zhì)過氧化物的產(chǎn)生。在本實(shí)驗(yàn)中,MDA在質(zhì)量比為0.5和1.0 g/kg甘草提取物中被顯著地抑制。這種抑制作用可能和甘草提取物增強(qiáng)自由基清除能力有關(guān)[29]。
綜合以上結(jié)果發(fā)現(xiàn),甘草提取物對(duì)于建鯉肝組織的保護(hù)呈劑量效應(yīng)關(guān)系,以質(zhì)量比為1.0 g/kg甘草提取物保肝效果最好,可以很好地清除氧自由基、羥自由基,具有良好的抗氧化能力,可以明顯抵抗TCDD造成的肝組織損傷。本實(shí)驗(yàn)的順利開展為以后防治TCDD誘導(dǎo)的肝組織損傷奠定了基礎(chǔ)。
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Hepatoprotective and antioxidant effects ofGlycyrrhizaglabraextracts against TCDD-induced hepatocyte damage in Jian carp (Cyprinuscarpiovar. Jian).
Du Jinliang1,2, Cao Liping1,2, Jia Rui3, Liu Yingjuan3, Shen Yujin3, Yin Guojun1,2,3*
(1.KeyLaboratoryofFreshwaterFisheriesandGermplasmResourcesUtilization,MinistryofAgriculture,FreshwaterFisheriesResearchCenter,ChineseAcademyofFisherySciences,Wuxi214081,Jiangsu,China; 2.InternationalJointResearchLaboratoryforFishImmunopharmacology,FreshwaterFisheriesResearchCenter,ChineseAcademyofFisherySciences,Wuxi214081,Jiangsu,China; 3.WuxiFisheriesCollege,NanjingAgriculturalUniversity,Wuxi214081,Jiangsu,China)
In recent years, the mechanism of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) toxicity has been a focal point in research. Generally it enriched in adipose tissue, and gradually amplified through food chains. It is seriously interfered with the normal function of the endocrine system of the organism, and displayed a wide spectrum of toxic effects, including dermal toxicity, reproductive toxicity, immunotoxicity and hepatotoxicity. The current research about the toxic mechanism of TCDD was mainly focused on the oxidative stress reaction, which was considered to be playing an important role in TCDD toxic research. Normally, the production and elimination of activated oxygen through the antioxidant system was in a balanceable and dynamic state. The antioxidant system could prevent excessive reactive oxygen species causing oxidative stress, keep the body balance, maintain the normal structure and function of cells. TCDD combined with aromatic hydrocarbon receptor and entered the nucleus, which changed the mRNA transcription of target gene, caused adverse change in the activity of enzymes and proteins, produced a series of oxidative stress reaction. Chinese medicinal herbs had been widely used in animal disease prevention due to their characteristics of rich bioactive components, free pollution, low side effects and immune enhancement. Many studies had reported that natural antioxidants were efficacious in preventing oxidative stress-related liver pathologies due to particular interactions and synergisms.Glycyrrhizaglabraextracts (GGE) was one of the common traditional Chinese herbal medicine, it could adjust the immunity, and had anti-tumor, anti-virus, antioxidant effects and so on. Although numerous studies had reported its beneficial effects on mammals, there was a lack of reports on its function in aquatic animals.
The aim of this study was to evaluate the hepatoprotective and antioxidant effects of GGE on the TCDD-induced liver injury in Jian carp (Cyprinuscarpiovar. Jian).
Jian carp were fed with diet containing 3 doses of GGE (0.1, 0.5 and 1.0 g/kg diet) for 60 days before a single intraperitoneal injection of TCDD (0.6 μg/kg), 72 h after TCDD injection, blood and liver samples were taken for biochemical analysis.
The results showed that GGE at 1.0 g/kg diet for 60 days prior to TCDD intoxication significantly reduced the elevated activities of ALT, AST, LDH, AKP and increased the reduced levels of total protein (TP), albumin (Alb) in the serum, markedly inhibited the reduction of the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), total antioxidant capacity (T-AOC), and reduced the malondialdehyde (MDA) formation in liver tissue. Meanwhile, histopathological observation also showed that GGE can significantly relieve TCDD induced liver injury.
It is concluded that GGE exhibited protective effect against TCDD-induced hepatotoxicity in fish, which is likely related to its antioxidant activity, and it is suggested that GGE may potentially be used as a hepatoprotective agent for fish liver injury.
Glycyrrhizaglabraextracts; 2,3,7,8-tetrachlorodibenzo-p-dioxin; liver injury;Cyprinuscarpiovar. Jian
Journal of Zhejiang University (Agric. & Life Sci.), 2015,41(5):593-601
國家自然基金青年科學(xué)基金(31202002;31200918);中央級(jí)公益性科研院所基本科研業(yè)務(wù)費(fèi)專項(xiàng)資金 (2013JBFM12);中國水產(chǎn)科學(xué)研究中央級(jí)公益性科研院所基本科研業(yè)務(wù)費(fèi)專項(xiàng)資金項(xiàng)目(2014A08YQ01)。
聯(lián)系方式:杜金梁(http://orcid.org/0000-0002-4924-3527),E-mail:dujl@ffrc.cn
2015-03-06;接受日期(Accepted):2015-06-24;網(wǎng)絡(luò)出版日期(Published online):2015-09-18
X 503.225
A
*通信作者(Corresponding author):殷國俊(http://orcid.org/0000-0001-6261-7506),Tel:+86-510-85558876;E-mail:yingj@ffrc.cn
URL:http://www.cnki.net/kcms/detail/33.1247.s.20150918.1804.020.html