华蟾素对HepG_(2)/ADM细胞耐药的抑制作用及其机制研究  

Inhibitory Effect and Mechanism of Huachansu on Drug Resistance in HepG_(2)/ADM Cells

作　　者：陈婷 李嘉龙 白蕊 史晓燕 段丽芳 李翠娟[1,3] 范妤 CHEN Ting;LI Jialong;BAI Rui;SHI Xiaoyan;DUAN Lifang;LI Cuijuan;FAN Yu(School of Basic Medicine,Shaanxi University of Chinese Medicine,Shaanxi Xianyang 712046,China;School of Medical Technology,Shaanxi University of Chinese Medicine,Shaanxi Xianyang 712046,China;Key Laboratory of Traditional Chinese Medicine System and Disease Prevention and Control in Shaanxi Province,Shaanxi Xianyang 712046,China)

机构地区：[1]陕西中医药大学基础医学院,陕西咸阳712046 [2]陕西中医药大学医学技术学院,陕西咸阳712046 [3]陕西省中医体制与疾病防治研究重点实验室,陕西咸阳712046

出　　处：《现代中医药》2024年第1期89-94,共6页Modern Chinese Medicine

基　　金：国家级大学生创新创业项目(202110716011);陕西中医药大学创新团队项目(2019-YL14);陕西省科技厅项目(2021JM-472);陕西省教育厅2021年度重点科研计划项目(21JS007)。

摘　　要：目的观察华蟾素对人肝癌细胞HepG_(2)/阿霉素(ADM)耐药性的影响并分析其可能的作用机制。方法用不同浓度的华蟾素处理HepG_(2)和HepG_(2)/ADM细胞24 h,MTT法检测细胞增殖抑制率,计算半数抑制浓度(IC 50);倒置显微镜观察细胞形态学变化;药物蓄积实验检测对细胞内药物累积的影响;Western blot实验检测凋亡相关蛋白Bcl-2、Bax、Caspase-9、Caspase-3表达情况。结果华蟾素能对HepG_(2)和HepG_(2)/ADM细胞具有增殖抑制作用,耐药倍数为4.67;随华蟾素处理浓度升高,HepG_(2)/ADM细胞体积变小,细胞皱缩突起减少,细胞间连接减少;与耐药组比较,华蟾素可提高HepG_(2)/ADM细胞对阿霉素的蓄积水平,上调Bax、Caspase-9、Caspase-3蛋白表达(P<0.05),下调Bcl-2蛋白表达(P<0.01),促进细胞凋亡。结论华蟾素能够显著抑制HepG_(2)/ADM细胞增殖,增加ADM在细胞内的蓄积,其作用机制可能与调控Bcl-2/Bax蛋白表达,诱导细胞凋亡有关。Objective To observe the effect of Huachansu on the drug resistance of human liver cancer cells HepG_(2)/doxorubicin(ADM)and its possible mechanism of action.Methods HepG_(2) and HepG_(2)/ADM cells were treated with different concentrations of bufalin for 24 hours,and the cell proliferation inhibition rate was detected by MTT assay.The half inhibitory concentration(IC 50)was calculated;Observing cellular morphological changes under an inverted microscope;The impact of drug accumulation experiments on intracellular drug accumulation;Western blot assay was used to detect the expression of apoptosis related proteins Bcl-2,Bax,Caspase-9,and Caspase-3.Results The results showed that Huachansu can inhibit the proliferation of HepG_(2) and HepG_(2)/ADM cells,with a resistance multiple of 4.67;As the concentration of Huachansu increased,the volume of HepG_(2)/ADM cells decreased,the number of cell wrinkled processes decreased,and the intercellular connections decreased;Compared with the drug resistant group,Huachansu can increase the accumulation level of doxorubicin in HepG_(2)/ADM cells,upregulate the expression of Bax,Caspase-9,and Caspase-3 proteins(P<0.05),and downregulate the expression of Bcl-2 protein(P<0.01),promoting cell apoptosis.Conclusion Huachansu can significantly inhibit the proliferation of HepG_(2)/ADM cells,increase the accumulation of ADM in cells,and its mechanism of action may be related to regulating the expression of Bcl-2/Bax protein and inducing cell apoptosis.

关 键 词：华蟾素 多药耐药 人肝癌细胞HepG_(2)/阿霉素 BCL-2/BAX 凋亡 

分 类 号：R285.5[医药卫生—中药学]