三羟基异黄酮对APP695基因转染PC12细胞凋亡的保护作用  

Protective effect of genistein on the apoptosis of PC12 cells transfected with mutant APP695 type

作　　者：王凤斌[1] 龙云[2] 王亚南[1] 丁振禹[3] 赵翠香[1] 

机构地区：[1]潍坊医学院生理学教研室,山东潍坊261053 [2]重庆三峡医药高等专科学校,重庆404120 [3]上海交通大学医学院,上海200025

出　　处：《中国药理学通报》2011年第12期1736-1740,共5页Chinese Pharmacological Bulletin

基　　金：山东省自然科学基金资助项目(No Y2004C29);山东省科技发展计划项目(No 2007GG30002007)

摘　　要：目的探讨植物雌激素三羟基异黄酮(Genistein,GST)对APP695基因转染PC12细胞凋亡的保护作用及机制。方法用pIRES2-EGFP空载体和pIRES2-EGFP/APP695MT表达载体转染正常的PC12细胞,将细胞分为对照组、空载组、APP695转染组、GST干预组。应用流式细胞仪测定细胞凋亡率,免疫细胞化学SABC法和Western blot方法检测凋亡相关蛋白Caspase-3的表达。结果 APP695转染组与空载组比较,PC12细胞凋亡率明显升高(P<0.01),Caspase-3为强阳性表达(P<0.01);GST干预组与APP695转染组比较,PC12细胞凋亡率明显降低(P<0.01),Caspase-3的免疫反应产物明显减弱(P<0.01),且15μmol.L-1和20μmol.L-1 GST处理组与5μmol.L-1和10μmol.L-1 GST处理组比较,Caspase-3的免疫反应产物减弱更明显(P<0.01)。结论 GST能降低APP695基因转染的PC12细胞凋亡率,其机制可能与减少凋亡蛋白Caspase-3的表达有关。Aim To investigate the protective effect of genistein against apoptosis of PC12 cells induced by transfection mutant APP695 plasmid DNA and its mechanisms.Methods PC12 cells were transfected with pIRES2-EGFP plasmid or pIRES2-EGFP/APP695MT expression plasmid,and then were divided into control group,vectortransfected group,APP695 transfected group and GST treatment group.Flow cytometry was applied to detect cell apoptosis.Caspase-3 protein expression in PC12 cells was obseved by Immunocytochemistry SABC and Western blot.Results Compared with vectortransfected group,PC12 cells in APP695 transfected group,apoptotic rate was increased significantly（P0.01）,and the expression of Caspase-3 protein was strongly increased（P0.01）.Compared with APP695 transfected group,PC12 cells in GST treatment group,apoptotic rate was decreased significantly（P0.01）,and the expression of Caspase-3 protein was significantly decreased（P0.01）.Conclusion GST can reduce apoptotic rate in PC12 cells induced by transfection mutant APP695 plasmid DNA,which might be correlated with the expression reduction of Caspase-3 protein.

关 键 词：三羟基异黄酮 APP695 PC12细胞 凋亡 CASPASE-3 保护作用 

分 类 号：R284.1[医药卫生—中药学] R329.25[医药卫生—中医学]