四逆汤在过氧化氢引起的心肌细胞氧化应激性损伤中的保护效应  

Protective Effects of Sini Decoction on Cardiomyocytes in OxidativeStress Damage Induced by Peroxide

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

机构地区：[1]中山大学中山医学院病理生理学教研室,广州510080

出　　处：《中药材》2005年第5期395-399,共5页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金资助项目(编号: 39770936)

摘　　要：目的:观察四逆汤对乳鼠心肌细胞在过氧化氢损伤中的保护效应,探讨Bcl- xL/xSmRNA在四逆汤抗过氧化氢损伤保护心肌细胞中的作用。方法:采用过氧化氢诱导损伤制备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DMME培养基培养细胞,应用流式细胞仪检测培养细胞的存活率,筛选合适的四逆汤含药血清培养细胞的浓度;用1mmol/L过氧化氢处理心肌细胞,分别在处理0. 5h, 1h, 2h, 3h, 4h, 8h后检测细胞内丙二醛(MDA)和过氧化物歧化酶(SOD)的活性,并用流式细胞仪检测不同作用时间的细胞凋亡率和坏死率;在过氧化氢处理心肌细胞1h和8h后,收获细胞,检测细胞胞浆Bcl- xL和Bcl- xSmRNA的表达情况。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;过氧化氢对心肌细胞的损伤程度与处理时间成正相关;与单纯过氧化氢处理组相比,四逆汤含药血清组细胞内MDA的含量降低,SOD的活性增加,细胞的凋亡率和坏死率降低,Bcl- xLmRNA表达增加,Bcl- xSmRNA表达减少。结论:四逆汤含药血清有较好的抗氧化应激性损伤保护心肌细胞的作用,这种保护作用可能通过增加Bcl- xLmRNA的表达,降低Bcl- xSmRNA的表达来实现。Objective:To investigate protective effects of Sini decoction on cardiomyocytes from oxidative stress damage induced by peroxide (H 2O 2).Methods:Ventricular cardiomyocytes 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 rats serum containing Sini decoction, respectively.Viablity percentage of rat cardiomyocytes was detected by flow cytometry using Annexin V-FITC to screen the appropriate SD rat serum concentration containing Sini decoction in DMEM culture media.Cardiomyocytes were exposed to 1 mmol/L H 2O 2 for 0.5h,1h,2h,4h and 8h respectively.MDA and SOD in cardiomyocytes were measured.Apoptosis and necrosis percentage of cardiomyocytes were measured by flow cytometry using Annexin V-FITC as well.Bcl-xL and Bcl-xS mRNA in cardiomyocytes were detected by polymerase chain reaction.Result:15% SD rat serum with Sini decoction in DMEM culture midia was the appropriate concentrate for cell growth.MDA and SOD in cardiomyocytes were changed with time elapse of cardiomyocytes exposure to H 2O 2.Sini decoction could inhibit rat cardiomyocytes from oxidative stress damage induced by H 2O 2.Conclusion:To some extend,Sini decoction inhibit rat cardiomyocytes from oxidative stress damage induced by peroxide through promoting expression of Bcl-xL mRNA and decreasing expression of Bcl-xS mRNA in cardiomyocytes.

关 键 词：应激性损伤 保护效应 细胞氧化 四逆汤 Bcl-xL 流式细胞仪检测 BCL-XS 丙二醛(MDA) 过氧化氢损伤 保护心肌细胞 过氧化物歧化酶 DMEM培养基 含药血清 培养细胞 心肌细胞生长 mRNA表达 mol/L 细胞凋亡率 损伤模型 不同浓度 

分 类 号：R285.5[医药卫生—中药学] R743.31[医药卫生—中医学]