川芎嗪对肝星状细胞凋亡的影响及机制研究

目的 研究川芎嗪对大鼠肝星状细胞(HSC)凋亡的影响及可能机制,为肝纤维化的治疗提供新的方法.方法 以血小板衍生生长因子(PDGF-BB)刺激大鼠肝星状细胞系HSC-T6,建立体外肝纤维化细胞模型,在此基础上,给予川芎嗪30μmol/L刺激HSC-T6,作用24 h后,Western blot法检测相关蛋白的表达情况,流式细胞术检测各组细胞凋亡率,Hoechst 33258染色法观察各组HSC的凋亡形态.结果 Western blot法结果表明,与PDGF组比较,川芎嗪组ERS标志性蛋白GRP78表达增多,并且ERS特异性凋亡蛋白CHOP及C-Caspase12表达增强,Bcl-2/Bax比例明显下调,p-ERK/ERK水平降低.流式细胞仪检测结果表明川芎嗪组较PDGF组HSC凋亡率增强,同时Hoechst 33258染色结果表明川芎嗪组HSC呈现典型的核固缩等凋亡形态变化.结论 川芎嗪可以诱导PDGF活化的HSC发生凋亡,其可能与ERS特异性凋亡蛋白CHOP及C-Caspase12的表达上调、ERK1/2磷酸化水平降低,进而下调Bcl-2/Bax比例,发生相关凋亡.

Effect of tetramethylpyrazine on hepatic stellate cell apoptosis and its possible mechanisms

Objective To provide new method for the treatment of liver fibrosis by studying the effect of tetramethylpyrazine on hepatic stellate cells (HSCs) and possible mechanisms.Methods Platelet-derived growth factor (PDGF-BB) was used on HSC to establish liver fibrosis model in vitro,based on this,tetramethylpyrazine 30 μmol/L was given to stimulate HSC-T6,and 1 μmol/L ERS inducer-thapsigargin(TG),was used as ERS positive control,after 24 h,Western blot was used to detect related protein expression,flow cytometry was used to detect each group cells apoptosis rate,Hoechst 33258 staining was used to observe each group of HSC apoptosis morphology.Results Western blot results showed that the expression of ERS iconic protein GRP78,ERS specificity apoptosis protein CHOP and C-Caspase12 of tetramethylpyrazine group were increased compared with PDGF group,and the Bcl-2/Bax ratio of tetramethylpyrazine group was significantly decreased,as well as p-ERK/ERK ratio.Flow cytometry test results showed that apoptosis rate of tetramethylpyrazine group was increased compared with PDGF group,at the same time Hoechst 33258 staining showed that tetramethylpyrazine group HSC present typical apoptotic morphological changes such as nuclear pyknosis.Conclusion Tetramethylpyrazine can induce HSC activated by PDGF apoptosis which may be related to increased expression of ERS specific apoptosis protein CHOP and CCaspase12,as well as decreased phosphorylation level of ERK1/2 and decreased Bcl-2/Bax ratio.