四逆汤抗氧化应激性损伤保护心肌细胞机制的探讨  

Protective mechanism on Sini Decoction on cardiac myocytes from oxidative stress damage

作　　者：聂咏梅[1] 吴伟康[1] 刘颖[2] 段新芬[1] 赵明奇[1] 赵丹阳[1] 

机构地区：[1]中山大学医学院病理生理学教研室,广东广州510074 [2]广东药学院病理生理学教研室,广东广州510224

出　　处：《中成药》2005年第6期697-702,共6页Chinese Traditional Patent Medicine

基　　金：本课题受国家自然科学基金资助(项目编号:39770936)

摘　　要：目的:观察四逆汤在乳鼠心肌细胞氧化应激性损伤过程中的保护效应,探讨四逆汤抗氧化应激性损伤保护心肌细胞的机制。方法:采用H2O2诱导损伤制备SD乳鼠心肌细胞氧化应激性损伤模型,分别用含不同浓度的四逆汤含药血清DMEM培养基培养细胞,应用流式细胞仪检测培养细胞的存活率,用1mmol/LH2O2处理心肌细胞,分别在处理0.5h,1h,2h,4h和8h后检测细胞的凋亡率和坏死率,同时检测细胞培养液中的乳酸脱氢酶(LDH)和细胞内丙二醛(MDA);对H2O2处理不同时间的心肌细胞线粒体膜电位进行检测。结果:采用含15四逆汤含药血清的DMEM培养基是心肌细胞生长的合适浓度;四逆汤含药血清能降低细胞培养液中的LDH活性以及细胞内MDA的含量,减少细胞的凋亡率和坏死率;降低心肌细胞线粒体膜电位的下降速度。结论:四逆汤对于H2O2造成的心肌细胞损伤具有保护作用,这种作用与保护线粒体膜电位的稳定有关。AIM: To investigate the protection of Sini Decoction for SD rat's cardiac myocytes against oxidative stress damage. METHODS: In this study, cardiac ventricular myocytes were prepared from 1-to-2-day-old Sprague-Dawley rats and cultured with DMEM culture media containing 15% neonatal bovine serum. 4 days later, culture media were replaced with neonatal bovine serum-free DMEM with 15%, 1.5% and 0.15% SD rat's serum containing sini decoction respectively. Cell survival rate was detected by flow cytometry using Annexin V-FITC. Cardiomyocytes were exposed to 1 mmol/L H_2O_2 for 0.5 h, 1 h, 2 h, 4 h and 8 h, respectively. Apoptosis and necrosis rate of myocytes were measured by flow cytometry using Annexin V-FITC . Mitochondrial membrane potential of cardiomyocytes was measured by flow cytometry using J-C-1. RESULTS: 15% SD rat serum with Sini Decoction in DMEM culture media was the appropriate concentration for cell growth.Sini Decoction could protect rat's myocytes against oxidative stress induced by H_2O_2. CONCLUSION: To some extent Sini Decoction could protect rat's myocytes from oxidative stress damage induced by H_2O_2 through keeping mitochondrial membrane potential of cardiomyocytes faily stable. nec

关 键 词：四逆汤 心肌细胞 凋亡 线粒体膜电位 

分 类 号：R285.5[医药卫生—中药学]