积雪草酸对人肝癌SMMC-7721细胞体内外增殖和凋亡的影响  

Effect of Asiaticacid on the proliferation and apoptosis of human hepatoma SMMC-7721 cells in vitro and in vivo

作　　者：王献哲 苏棋 梁聪 李媛媛 温燕[2] 郭哲 何萍[1] Wang Xianzhe;Su Qi;Liang Cong;Li Yuanyuan;Wen Yan;Guo Zhe;He Ping(Pharmaceutical College,Guangxi Medical University,Nanning 530021,China;Department of Pharmacy,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China;Emergency Department,The Third Affiliated Hospital of Guangxi Medical University,Nanning 530031,China)

机构地区：[1]广西医科大学药学院,南宁530021 [2]广西医科大学第一附属医院药学部,南宁530021 [3]广西医科大学第三附属医院急诊科,南宁530031

出　　处：《广西医科大学学报》2019年第5期729-733,共5页Journal of Guangxi Medical University

基　　金：广西科技基础条件平台建设课题资助项目(广西实验动物资源共享服务平台建设)(No.15-235-06);2018广西研究生教育创新计划课题资助项目(No.YCSW2018110);南宁市科学研究与技术开发计划课题资助项目(No.20153105);广西中医药管理局自筹课题资助项目(No.GZZC14-55);广西卫生厅自筹课题资助项目(No.Z2014060)

摘　　要：目的:研究积雪草酸(AA)体内外抗肝癌效应及其凋亡机制。方法:将人肝癌SMMC-7721细胞分为溶剂(0.1% DMSO溶剂)对照组和不同浓度(20 μmol/L、30 μmol/L、40 μmol/L、50 μmol/L、60 μmol/L)AA组,作用于SMMC-7721细胞24 h后,采用CCK-8法检测细胞活性,Annexin V-APC/7-AAD双染色流式分析检测细胞凋亡率,JC-1染色检测细胞线粒体膜电位,同时采用Western blotting检测线粒体凋亡相关蛋白的表达。建立SMMC-7721细胞裸鼠皮下移植瘤模型,分为对照组、AA 50 mg/kg组和AA 100 mg/kg组,每组5只,观察AA灌胃给药21 d后移植瘤的生长情况。结果:AA浓度依赖性地抑制SMMC-7721细胞增殖(IC 50 =38.31 μmol/L)并促使细胞产生凋亡样改变。与溶剂对照组相比,AA 20 μmol/L组、AA 40 μmol/L组、AA 60 μmol/L组均可提高SMMC-7721细胞凋亡率,并降低细胞线粒体膜电位(均 P <0.01),具有一定的浓度依赖性;此外,AA 40 μmol/L组明显上调Bax、Cyt-C和Cleaved-Caspase-3蛋白的表达,下调Bcl-2蛋白的表达( P <0.05 或 P <0.01),但对p53蛋白表达无明显影响( P >0.05)。AA 50 mg/kg组和AA 100 mg/kg组给药第14、第21天时的肿瘤体积明显小于对照组,给药第21天时的肿瘤质量小于对照组(均 P <0.01)。结论:AA具有体内外抗肝癌作用,其机制可能与其诱导非p53依赖的线粒体途径的凋亡有关。Objective: To investigate the effect and mechanisms of asiatic acid (AA) on the proliferation and apoptosis in human hepatoma SMMC-7721 cells in vitro and i n vivo . Methods: Human hepatoma SMMC-7721 cells were divided into solvent control group and different concentrations (20~60 μmol/L) of AA groups.After 24 h of treatment,the cell viability,apoptosis rate,and the mitochondrial membrane potential changes were examined by CCK-8 method,Annexin V-APC/7-AAD double staining flow cytometric assay and JC-1staining,respectively.The expression of the mitochondrial apoptosis related proteins were detected by western blotting.After the establishment of subcutaneous xenograft model bearing SMMC-7721 cells,the nude mice were divided into the control group,AA 50 mg/kg group and AA 100 mg/kg group ( n =5 per group).The growth of xenograft was observed after 21 days of treatment. Results: AA inhibited the proliferation of SMMC-7721 cells in a dose-dependent manner with IC 50 of 38.31 μmol/L and induced the apoptosis-like morphological change.Compared with the control group,AA dose-dependently elevated the apoptosis rate and lower the mitochondrial membrane potential ( P <0.01).Additionally,at the concentration of 40 μmol/L,AA significantly up-regulated the protein expression levels of Bax,Cyt-C,and Cleaved-Caspase-3,while down-regulated the expression level of Bcl-2 as compared to the control group( P <0.05 or P <0.01),and it had no influence on the expression of p53( P >0.05).Intragastric administration of AA (50 mg/kg and 100 mg/kg) for 21 d significantly decreased the tumor volume and tumor weigh of nude mice ( P <0.01). Conclusion: AA can inhibit the proliferation of human hepatoma SMMC-7721 cells in vitro and in vivo ,and the mechanisms may be related to the mitochondrial apoptosis in a p53-indepandent manner induced by AA.

关 键 词：积雪草酸 人肝癌SMMC-7721细胞 裸鼠移植瘤 线粒体凋亡 

分 类 号：R285.5[医药卫生—中药学]