摘要
目的构建小鼠酒精性肝纤维化(ALF)模型,探讨补充外源性甲状腺激素T3对小鼠肝脏氧化应激的影响。方法80只小鼠随机分为6组:正常对照组、酒精性肝纤维化模型组、低浓度T3干预组(25μg/kg)、中浓度T3干预组(50μg/kg)、高浓度T3干预组(100μg/kg)及T3对照组(T3浓度为100μg/kg)。采用酒精液体饲料(TP4060A)喂养,联合31.5%乙醇溶液灌胃构建酒精性肝纤维化模型,从第6周起腹腔注射T3,干预持续3周。T3对照组及正常对照组采用对照液体饲料(TP4060C)喂养。通过天狼星红染色检测胶原纤维含量,免疫印迹法检测肝组织α平滑肌肌动蛋白(α-SMA)和Ⅰ型胶原蛋白(collagenⅠ)蛋白表达,以及血清转氨酶活性检测分析小鼠肝脏损伤及纤维化程度;采用ELISA法检测肝组织中超氧化物歧化酶(SOD)活性、谷胱甘肽(GSH)和丙二醛(MDA)含量;免疫组织化学法和免疫印迹法检测肝脏自噬相关蛋白微管相关蛋白轻链3-Ⅱ(LC3-Ⅱ)和p62的表达水平。结果模型组小鼠肝脏结构与功能损伤严重,自噬被抑制,氧化应激反应较对照组明显增强;与模型组相比,T3干预组小鼠肝脏出现不同程度的功能和结构的恢复,自噬功能恢复,低、中浓度T3干预组肝脏中SOD活性和GSH含量升高,MDA含量明显减少;高浓度T3干预组则表现为SOD活性升高,MDA含量明显减少,GSH含量仍明显低于正常对照组,与模型组无明显差异。结论适当补充T3能够通过恢复肝脏自噬功能抑制氧化应激反应,进而影响酒精性肝纤维化的发生发展。
Objective To construct a mouse model of alcoholic liver fibrosis and explore the effect of supplementing exogenous thyroid hormone T3 on oxidative stress in liver.Methods Eighty mice were randomly divided into 6 groups,normal control group,alcoholic liver fibrosis(ALF)model group,and low concentration T3 intervention group(25μg/kg),medium concentration T3 intervention group(50μg/kg),high concentration T3 intervention group(100μg/kg)and T3 control group(the concentration of T3 is 100μg/kg).A model of mice alcoholic liver fibrosis was established by using alcoholic liquid feed combined with 31.5%ethanol gavage.From the sixth week,mice in the T3 intervention and T3 control group were injected with corresponding concentrations of T3 intraperitoneally for three weeks.Mice in the control and T3 control groups were fed with control liquid feed.The degree of mice liver injury and fibrosis was evaluated through the sirius red staining,Western blotting,and serum biochemical testing.The activity of superoxide dismutase(SOD),the content of glutathione(GSH)and malondialdehyde(MDA)in liver tissue were detected by ELISA,and the protein expressions of microtubule⁃associated protein light chain 3⁃Ⅱ(LC3⁃Ⅱ)and p62 were detected by immunohistochemistry and Western blotting.Results The liver structure and function in the ALF group were severely damaged,autophagy was inhibited,and the oxidative stress response was significantly enhanced compared with the control group.Compared with the ALF group,the recovery of liver functional and structure and autophagy were showed in the T3 intervention group,and SOD activity and GSH content in the liver increased in the low and medium concentrations of T3 intervention groups,while MDA content significantly decreased.In the high concentration T3 intervention group,it showed the same increase in SOD activity,a significant decrease in MDA content,while the content of GSH was lower than that in the control group,which was not different with the ALF group.Conclusion Appropriate supplement
作者
罗仁利
李三强
冯家阳
张凯杰
卢杉
吴俊菲
LUO Ren-li;LI San-qiang;FENG Jia-yang;ZHANG Kai-jie;LU Shan;WU Jun-fei(Molecular Medicine Key Laboratory of Liver Injury and Repair,College of Basic Medicine and Forensic Medicine,He’nan University of Science and Technology,He’nan Luoyang 471000,China;He’nan Center for Engineering and Technology Research on Prevention and Treatment of Liver Diseases,He’nan Luoyang 471000,China)
出处
《解剖学报》
CAS
CSCD
2024年第6期753-760,共8页
Acta Anatomica Sinica
基金
国家自然科学基金(82170606)
河南省高等学校重点科研项目计划基础研究专项(23ZX006)。
