基于AMPK/Sirt1信号通路研究黔北绿茶水提物对过氧化氢诱导的PC12细胞氧化损伤的保护作用  

Protective effect and mechanism of Qianbei green tea water extract on hydrogen peroxide-induced PC12 cell injury based on AMPK/Sirt1 signaling pathway

作　　者：陈娜娜[1] 雷鸣[1] 周清娣[2] 徐应淑[1] 

机构地区：[1]遵义医学院药学院,遵义563000 [2]悉尼大学

出　　处：《中药药理与临床》2017年第6期80-84,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：教育部"春晖计划"合作科研项目(Z2016007);贵州省高层次创新人才"百"层次项目(黔科合平台人才20165684号);贵州省科技创新人才团队项目(黔科合人才团队(2015)4023号)

摘　　要：目的:探讨黔北绿茶水提物通过AMPK/Sirt1信号通路抗过氧化氢诱导的PC12细胞凋亡作用及机制。方法:采用噻唑蓝(MTT)法测定黔北绿茶水提物对过氧化氢诱导的PC12细胞存活率和死亡率的影响,ELISA法检测活性氧(ROS)含量,TUNEL实验检测细胞凋亡情况,罗丹明123荧光染色观察线粒体膜电位的改变,Western-blot法检测相关蛋白表达。结果:不同浓度黔北绿茶水提物(12.5~50μg/ml)能够提高过氧化氢作用的细胞活力并呈浓度依赖性。黔北绿茶水提物(12.5~50μg/ml)作用48 h后能够显著提高细胞线粒体膜电位并上调抗凋亡蛋白Bcl-2蛋白、p-AMPK、Sirt1蛋白表达,下调促凋亡蛋白Bax、caspase 3蛋白表达。结论:黔北绿茶水提物能够显著抑制过氧化氢诱导的PC12细胞凋亡,其作用可能与调控AMPK/Sirt1依赖的细胞凋亡途径相关。Objective： To investigate the protective effect of Qianbei green tea water extract against hydrogen peroxide-induced injury in PC12 cells.Methods： PC12 cells exposed to hydrogen peroxide（ 400 μmol/L） were used as an in vitro model to mimic oxidative stress injury. The protective effect of Qianbei green tea water extract was detected by pre-treatment for 1 hour with Qianbei green tea water extract（ 12. 5,25,50μg/ml） prior to being exposed to hydrogen peroxide. The cell viability and morphology were detected by MTT assay and light microscope,respectively. Contents of reactive oxygen species（ ROS）,malonaldehyde（ MDA） and superoxide dismutase（ SOD） activity were determined by kits,respectively. Cell apotosis and mitochondrial membrane potential（ MMP） were determined by TUNEL assay and rhodamine 123,respectively. Apoptosis related proteins were determined by Western blot. Results： Compared with the hydrogen peroxide case,the cell viability pre-treated with Qianbei green tea water extract（ 12. 5,25,50 μg/ml） was elevated in a concerntration-dependent manner. Qianbei green tea water extract lowered contents of ROS and MDA,and enhanced SOD activity. Moreover,Qianbei green tea water extract supressed oxidative stress-induced cell apoptosis and MMP desfunction,and up-regulated expressions of p-AMPK,Sirt1,Bcl-2,down-regulated expressions of Bax and Caspase 3. Conclusion： Qianbei green tea water extract executs protective effect against hydrogen peroxide-induced PC12 cell injury,which may be contributed to regulating apoptosis-dependent AMPK/Sirt1 signaling pathway.

关 键 词：黔北绿茶 PC12 过氧化氢 凋亡 

分 类 号：R285[医药卫生—中药学]