论文部分内容阅读
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: To analyze the mechanism of Huangqi Decoction on liver fibrosis induced by dimethylnitrosamine (DMN) in rats based on differential proteomic analysis of liver tissue. Methods: The rat model of hepatic fibrosis was established by intraperitoneal injection of DM DM for 12 times in 4 weeks. After the model was established, the mice were killed and then randomly divided into model control group and treatment group. The treatment group was given Astragalus soup ig for 2 weeks. Two-dimensional gel electrophoresis -DE) were used to separate the total proteins in liver tissues. Some differentially expressed proteins were identified by matrix-assisted laser desorption ionization time of flight mass spectrometry and peptide mass fingerprinting analysis. Western blotting and biochemical methods were used to detect the expression of Prdx6, Shock Protein 70 (HSP70) and catalase (CAT) activity. RESULTS: Seven of the 18 proteins identified were associated with dyslipidemia and peroxidation damage, three were associated with protein synthesis, processing and degradation, two with biotransformation, and three with energy metabolism. The three protein spots verified (Prdx6, Hsp70, CAT) and two-dimensional electrophoresis analysis results are basically the same. Conclusion: Peroxidation injury is an important pathological process of DMN-induced cirrhosis in rats and enhances the intrinsic antioxidant capacity. Antioxidant stress is one of the main mechanisms of Huangqi Decoction in reversing hepatic cirrhosis in rats.