丹参酮ⅡA通过调节线粒体转位蛋白诱导HepG2细胞凋亡  

Mediation of mitochondrial translocator protein by tanshinoneⅡA in apoptosis of HepG2 cells

作　　者：张谊 欧玉龙 王惠霞 黄晓佳 ZHANG Yi;OU Yu-long;WANG Hui-xia;HUANG Xiao-jia(Affiliated Danyang Hospital of Nantong University,Danyang Jiangsu 212300,China;Institute of Biomedical Engineering and Health Sciences,Changzhou University,Changzhou Jiangsu 213164,China)

机构地区：[1]南通大学附属丹阳医院江苏省丹阳市人民医院,江苏丹阳212300 [2]常州大学生物医学工程与健康科学研究院,江苏常州213164

出　　处：《中国药理学通报》2023年第1期101-107,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No81300059);镇江市社会发展指导性项目(NoFZ2019005);镇江“金山英才”高层次领军人才培养计划(第六期“169工程”)。

摘　　要：目的探讨丹参酮ⅡA(tanshinoneⅡA,TanⅡA)通过调节线粒体转位蛋白(translocator protein,TSPO)诱导HepG2细胞凋亡中的作用及可能存在的作用机制。方法采用MTT比色法检测TanⅡA对HepG2细胞增殖的影响,Annexin V-PE/7-氨基-放线菌素D双染色法和Hochest染色法观察细胞凋亡率,荧光素酶发光法检测HepG2细胞中ATP的含量,Clark氧电极法测定细胞耗氧率,JC-1荧光探针法检测细胞线粒体膜电位变化,免疫荧光染色法观察转位蛋白在细胞内的表达及定位,免疫印迹法检测线粒体内细胞色素C、半胱氨酰天冬氨酸特异性蛋白酶-3以及半胱氨酰天冬氨酸特异性蛋白酶-9的表达。结果TanⅡA能够剂量依赖性地抑制肝癌细胞HepG2增殖,诱导细胞凋亡、ATP生成减少、耗氧量降低以及线粒体膜电位的下降。同时,TanⅡA可诱导HepG2细胞TSPO表达上调并聚集于核内。结论TanⅡA可诱导HepG2细胞凋亡,其机制与促进细胞转位蛋白TSPO表达及细胞核转位,进而引起线粒体功能障碍有关。Aim To investigate the role of mitochondrial translocator protein(TSPO)in the apoptosis of HepG2 cells induced by tanshinoneⅡA(TanⅡA)and the involved mechanism.Methods Following the HepG2 cells treated with TanⅡA at 2.5,5 and 10μmol·L-1,the cell viability was determined by MTT assay,and intracellular ATP content was determined by luciferin-luciferase method.Oxygen utilization was measured polarographically with a Clark oxygen electrode.Cell apoptosis was determined by Hoechst 33342 staining and flow cytometry.The mitochondrial membrane potential was assessed with JC-1 staining.The intracellular distribution of TSPO was examined by TSPO immunostaining,and the expressions of TSPO,Cyto C,caspase-3,caspase-9 were determined by immunoblotting analysis.Results TanⅡA inhibited the proliferation of HepG2 cells in a dose-and time-dependent manner.The treatment with TanⅡA inhibited ATP production and oxygen utilization of mitochondria.In addition,TanⅡA enhanced TSPO expression and accumulation in nuclei and up-regulated the expression of Cyto C,caspase-3 and caspase-9.Conclusions TanⅡA induces the apoptosis of HepG2 cells,which may be related to the TSPO-mediated mitochondrial dysfunction.

关 键 词：丹参酮ⅡA HEPG2细胞 细胞凋亡 转位蛋白 线粒体 细胞能量代谢 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学] R329.25R341R735.7