固脾消积饮对人肝癌细胞HepG2线粒体结构功能的调节作用  

Effect of Gupi Xiaoji Decoction on mitochondrial structure and function of human hepatoma cell line HepG2

作　　者：柳卓 田雪飞[3] 郜文辉[4] 谭小宁 俞淑娴[1] 翦慧颖 李可心 张振 曾普华 LIU Zhuo;TIAN Xue-fei;GAO Wen-hui;TAN Xiao-ning;YU Shu-xian;JIAN Hui-ying;LI Ke-xin;ZHANG Zhen;ZENG Pu-hua(Hunan Academy of Traditional Chinese Medicine,Changsha 410006,China;Affiliated Hospital of Hunan Academy of Traditional Chinese Medicine,Changsha 410006,China;College of Integrated Traditional Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;College of Traditional Chinese Medicine,Hunan University of Traditional Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南省中医研究院,湖南长沙410006 [2]湖南省中医药研究院附属医院,湖南长沙410006 [3]湖南中医药大学中西结合学院,湖南长沙410208 [4]湖南中医药大学中医学院,湖南长沙410208

出　　处：《中国药理学通报》2023年第4期781-786,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 82074425);湖南省中医药管理局青年项目(No 2021178);全国青年岐黄学者人才项目;湖南省重点研发项目(No 2021SK2006);湖南省中医药院级联合青年项目;湖南省自然基金项目(No 2021JJ30417);抗肿瘤中药创制技术湖南省工程研究中心项目。

摘　　要：目的 观察固脾消积饮(Gupi Xiaoji Decoction, GPXJY)对人肝癌细胞HepG2细胞线粒体结构功能的影响,并探讨其可能机制。方法 CCK-8检测细胞增殖,Mito-Tracker Green荧光染色观察线粒体结构,流式细胞术检测膜电位,ELISA检测ATP含量,透视电镜观察微观结构变化,高内涵检测相关蛋白。结果 荧光染色发现GPXJY对HepG2细胞线粒体造成损伤并且ATP含量降低;流式结果显示GPXJY能使HepG2细胞线粒体膜电位减少;电镜结果发现GPXJY使癌细胞线粒体肿胀等;高内涵检测发现,固脾消积饮对Hep G2细胞内Bcl-2平均荧光强度值明显降低,凋亡促进蛋白Bax、cytochrome-c、caspase-3、cleaved-caspase-3平均荧光强度值明显增加,差异有统计学意义。结论 GPXJY对Hep G2细胞线粒体结构功能有调节作用。Aim To observe the effect of Gupi Xiaoji Decoction(GPXJY)on the structure and function of mitochondria of human hepatoma cell HepG2 cells and explore its possible mechanism.Methods CCK8 was used to detect cell proliferation,Mito-Tracker Green fluorescence staining was used to observe the mitochondrial structure,flow cytometry was used to detect the membrane potential,Elisa was used to detect the ATP content,fluoroscopic electron microscopy was used to observe the microstructure changes,and high-content screening(HCS)was used to detect the related proteins.Results Fluorescence staining showed that GPXJY damaged the mitochondria of HepG2 cells and decreased the content of ATP.The results of flow cytometry showed that GPXJY could reduce the mitochondrial membrane potential of HepG2 cells.The results of electron microscope showed that GPXJY made the mitochondria of cancer cells swell and so on.HCS found that GPXJY significantly reduced the average fluorescence intensity of Bcl-2 in HepG2 cells,and significantly increased the average fluorescence intensity of apoptosis promoting proteins Bax,cytochrome-c,caspase-3 and cleaved-caspase-3,which was statistically significant.Conclusion GPXJY can regulate the structure and function of mitochondria in HepG2 cells.

关 键 词：固脾消积饮 线粒体 肝癌 流式细胞术 高内涵 透射电镜 

分 类 号：R-332[医药卫生] R256.4R329.24R446.8R735.7