基于肝组织差异蛋白质组解析黄芪汤治疗二甲基亚硝胺大鼠肝纤维化的效应机制  

Changes of Hepatic Proteome in Dimethylnitrusamine-induced Cirrhosis in Rats Treated with Huangqi Tang

作　　者：仝欣[1,2,3,4] 王高强[1,2,3,4] 王磊[1,2,3,4] 刘莺[4] 孙明瑜[1,2,3,4] 刘平[1,2,3,4] 

机构地区：[1]上海中医药大学附属曙光医院 [2]上海市中医药研究院肝病研究所 [3]肝肾疾病病证教育部重点实验室(上海中医药大学)上海高校中医内科学E研究院,上海201203 [4]上海中医药大学,上海201203

出　　处：《中国实验方剂学杂志》2010年第11期111-116,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家重点基础研究发展计划(2006CB504801);国家自然科学基金重大研究计划重点项目(90409020);上海市重点学科建设项目(Y0302);上海市教育委员会E研究院建设计划项目(E03008);上海高校创新团队建设项目资助

摘　　要：目的:基于肝组织差异蛋白质组表达解析黄芪汤治疗二甲基亚硝胺(dimethylnitrosamine,DMN)大鼠肝纤维化的效应机制。方法:4周内给大鼠ip DMN 12次制备肝纤维化模型,成模后停止染毒,随机分为模型对照组与治疗组,治疗组以黄芪汤ig 2周;双向凝胶电泳(2-DE)分离肝组织总蛋白,用基质辅助激光解析离子化飞行时间质谱和肽质量指纹图谱分析鉴定部分差异表达蛋白质;蛋白质免疫印迹法及生化学方法对过氧化还原蛋白6(Prdx6)、热休克蛋白70(HSP70)及过氧化氢酶(CAT)活性进行验证。结果:鉴定的18个蛋白中7个与脂质代谢紊乱及过氧化损伤相关,3个与蛋白合成、加工和降解相关,2个与生物转化相关,3个与能量代谢相关。验证的3个蛋白点(Prdx6,Hsp70,CAT)与双向电泳分析结果基本一致。结论:过氧化损伤是DMN诱导大鼠肝硬化形成的重要病理环节,提高机体内在的抗氧化能力、抗氧化应激效应是黄芪汤逆转大鼠肝硬化的主要机制之一。Objective：Proteomics approaches were performed to identify proteins that may be involved in dimethylnitrusamine（DMN）-induced cirrhosis /fibrosis and the effects of Huangqi Tang（HQT） intervention were observed in rats.Method：Differential protein spots that exhibited changes in HQT group in DMN-induced cirrhosis were selected by two-dimensional gel electrophoresis and identified by matrix-assisted laser desorption /ionization time-of-flight and peptide mass fingerprinting analysis.After that,these protein spots were recognized by bioinformatics information retrieval.Lastly,the results of this proteomic analysis were confirmed by immunoblotting and detection of enzyme activity.Result：Eighteen protein spots were found to be differentially expressed,including 7 spots involved in disorder of lipid metabolism and oxidative damage,3 spots involved in protein metabolism,2spots involved in biotransformation and 3 spots involved in material synthesis and energy metabolism in liver.Among them,3 spots for confirmation（Prdx6,Hsp70,CAT） were consistent with the results of two-dimensional gel electrophoresis.Conclusion：There are significant oxidative stresses occurred during hepatofibrogenesis of DMN-induced cirrhosis/fibrosis in rats and HQT can significantly improve the antioxidant capacity of liver and protect peroxide damage of cells.

关 键 词：肝纤维化 蛋白质组学 氧化应激 黄芪汤 

分 类 号：R285.5[医药卫生—中药学]