运动对二噁英染毒大鼠肝毒性的影响研究
摘要
目的：通过研究运动对急性2,3,7,8-TCDD暴露后大鼠血清指标、肝组织酶活性及肝脏蛋白表达的影响,探讨运动对预防TCDD肝毒性的有效性,为寻找运动对TCDD毒性影响的生物标志物提供依据。
方法：8周龄雄性Wistar大鼠随机分为五组：对照组1(NC1)、染毒组(NTl、NT2)和运动染毒组(ET1、ET2),染毒组腹腔注射10μg/kg体重的2,3,7,8-TCDD,对照组注射同等剂量的玉米油,NT1染毒后静养四周,ETl组染毒后运动四周,NT2、ET2运动四周后染毒,分别静养、运动四周。运动组尾部负重5%进行游泳,每周游泳6天,每天30min。观察每周体重变化,测定血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆固醇(TC)、磷脂(PL)水平及肝组织芳香烃羟化酶(AHH)、7-乙氧基香豆素-0-脱乙基酶(EROD)和7-乙氧基异吩恶唑酮-0-脱乙基酶(ECOD)活性。采用双向凝胶电泳和质谱分析对肝脏组织进行蛋白组学研究,寻找差异蛋白点。
结果：1、TCDD急性暴露对大鼠一般健康状况无明显影响。TCDD使肝脏重量(P<0.01)、肝湿重/体重(P<0.01)、血清AST、PL、TC水平(P<0.01)及AHH(P<0.05)活性明显升高。2、运动使TCDD大鼠TC水平及ECOD活性显著降低(P<0.01)。3、运动预适应四周后TCDD急性暴露,继续运动可使肝绝对重量及肝湿重/体重显著降低(P<0.01),血清AST、TC水平(P<0.01)及EROD、ECOD、AHH活性(P<0.01)显著升高。4、对49个差异点进行质谱分析,阳性点为24个,其中阳性差异点8个。5、运动能影响TCDD急性暴露后肝脏蛋白表达,上调的蛋白有黄素还原酶和肌动蛋白；下调的蛋白包括细胞色素b5、78kDa葡萄糖调节蛋白前体、3-a烃基类固醇脱氢酶及谷胱甘肽S转移酶a等。5、有运动预适应的大鼠在TCDD急性暴露后的肝脏蛋白表达也发生了变化,生物氧化过程中的酶蛋白表达上调。
结论：TCDD急性暴露后四周可致大鼠肝毒性；运动可有效缓解TCDD诱导的肝毒性；运动预适应对预防大鼠肝毒性的有效性还有待进一步研究。
Objective:The aim of the current study was to explore the possible effect of exercise to hepatotoxicity in2,3,7,8-TCDD induced, and provide the basis for researching the biomarkers.
Methods:fifty animals were assigned randomly into five groups of ten each, including a normal control group (NC), two toxic groups (NT1、NT2) and two exercise toxic groups(ET1、ET2). The rats of toxic groups and exercise toxic groups received10μg/kg BW of2,3,7,8-TCDD, the NC group were administered with equivalent volumes of coin oil only, and exercise groups were swimming with5%weights attach to tails for30min daily. Body weights were recorded weekly, and the blood was collected for serum biochemical analysis, the livers were immediately isolated and weighted, and the lobe of each liver were kept at-80℃for detect the activity of hepatic microsomal enzymes and Proteomics analysis.
Results:There were no effects on general health in all group. Significantly increased of Liver weight, Liver relative weight (P<0.01), the serum levels of aspartate aminotransferase (AST), Phospholipid (PL), Total cholesterol(TC)(P<0.01)and the activity of aryl hydrocarbon hydroxylase(AHH)(P<0.05) were observed in TCI group. Significantly decreased were observed of the serum levels of TC (P<0.01)and activites of7-ethoxycoumarin O-deethylase(ECOD)(P<0.01)in TE1group. Significantly decreased were observed of Liver relative weight (P<0.01), and significantly increased of serum levels of AST、TC(P<0.01) and activities of7-ethoxyresorufin0-deethylase (EROD), ECOD and AHH(P<0.01) in ET2group. Analysing the49points of difference by mass spectrometry, the number of positive points was24, and the number of posoitive points of difference was8. Exercise could affected the expression of liver protein, the protein up-regulated were flavin reductase and actin, while the protein down-regulated were cytochrome b5,78kDa glucose-regulated protein precursor,3alpha-Hydroxysteroid(Slash)dihydrodiol Dehydrogenase and glutathione S-transferase alpha-3. Exercise pre-adaptation could also altered the expression of liver protein. Conclusion:TCDD could induced the hepatotoxicity. Exercise could effective prevent the hepatotoxicity induced TCDD, and whether exercise pre-adaptation could prevent the hepatotoxicity induced TCDD need further research.
方法：8周龄雄性Wistar大鼠随机分为五组：对照组1(NC1)、染毒组(NTl、NT2)和运动染毒组(ET1、ET2),染毒组腹腔注射10μg/kg体重的2,3,7,8-TCDD,对照组注射同等剂量的玉米油,NT1染毒后静养四周,ETl组染毒后运动四周,NT2、ET2运动四周后染毒,分别静养、运动四周。运动组尾部负重5%进行游泳,每周游泳6天,每天30min。观察每周体重变化,测定血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆固醇(TC)、磷脂(PL)水平及肝组织芳香烃羟化酶(AHH)、7-乙氧基香豆素-0-脱乙基酶(EROD)和7-乙氧基异吩恶唑酮-0-脱乙基酶(ECOD)活性。采用双向凝胶电泳和质谱分析对肝脏组织进行蛋白组学研究,寻找差异蛋白点。
结果：1、TCDD急性暴露对大鼠一般健康状况无明显影响。TCDD使肝脏重量(P<0.01)、肝湿重/体重(P<0.01)、血清AST、PL、TC水平(P<0.01)及AHH(P<0.05)活性明显升高。2、运动使TCDD大鼠TC水平及ECOD活性显著降低(P<0.01)。3、运动预适应四周后TCDD急性暴露,继续运动可使肝绝对重量及肝湿重/体重显著降低(P<0.01),血清AST、TC水平(P<0.01)及EROD、ECOD、AHH活性(P<0.01)显著升高。4、对49个差异点进行质谱分析,阳性点为24个,其中阳性差异点8个。5、运动能影响TCDD急性暴露后肝脏蛋白表达,上调的蛋白有黄素还原酶和肌动蛋白；下调的蛋白包括细胞色素b5、78kDa葡萄糖调节蛋白前体、3-a烃基类固醇脱氢酶及谷胱甘肽S转移酶a等。5、有运动预适应的大鼠在TCDD急性暴露后的肝脏蛋白表达也发生了变化,生物氧化过程中的酶蛋白表达上调。
结论：TCDD急性暴露后四周可致大鼠肝毒性；运动可有效缓解TCDD诱导的肝毒性；运动预适应对预防大鼠肝毒性的有效性还有待进一步研究。
Objective:The aim of the current study was to explore the possible effect of exercise to hepatotoxicity in2,3,7,8-TCDD induced, and provide the basis for researching the biomarkers.
Methods:fifty animals were assigned randomly into five groups of ten each, including a normal control group (NC), two toxic groups (NT1、NT2) and two exercise toxic groups(ET1、ET2). The rats of toxic groups and exercise toxic groups received10μg/kg BW of2,3,7,8-TCDD, the NC group were administered with equivalent volumes of coin oil only, and exercise groups were swimming with5%weights attach to tails for30min daily. Body weights were recorded weekly, and the blood was collected for serum biochemical analysis, the livers were immediately isolated and weighted, and the lobe of each liver were kept at-80℃for detect the activity of hepatic microsomal enzymes and Proteomics analysis.
Results:There were no effects on general health in all group. Significantly increased of Liver weight, Liver relative weight (P<0.01), the serum levels of aspartate aminotransferase (AST), Phospholipid (PL), Total cholesterol(TC)(P<0.01)and the activity of aryl hydrocarbon hydroxylase(AHH)(P<0.05) were observed in TCI group. Significantly decreased were observed of the serum levels of TC (P<0.01)and activites of7-ethoxycoumarin O-deethylase(ECOD)(P<0.01)in TE1group. Significantly decreased were observed of Liver relative weight (P<0.01), and significantly increased of serum levels of AST、TC(P<0.01) and activities of7-ethoxyresorufin0-deethylase (EROD), ECOD and AHH(P<0.01) in ET2group. Analysing the49points of difference by mass spectrometry, the number of positive points was24, and the number of posoitive points of difference was8. Exercise could affected the expression of liver protein, the protein up-regulated were flavin reductase and actin, while the protein down-regulated were cytochrome b5,78kDa glucose-regulated protein precursor,3alpha-Hydroxysteroid(Slash)dihydrodiol Dehydrogenase and glutathione S-transferase alpha-3. Exercise pre-adaptation could also altered the expression of liver protein. Conclusion:TCDD could induced the hepatotoxicity. Exercise could effective prevent the hepatotoxicity induced TCDD, and whether exercise pre-adaptation could prevent the hepatotoxicity induced TCDD need further research.
引文
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