雷公藤红素对体外人肝癌HepG2细胞增殖、凋亡的影响及机制研究  

Study on the Effects and Mechanism of Celastrol on Proliferation and Apoptosis of Human Hepatoma HepG2 Cells in vitro

作　　者：张彦[1] 祝晨蔯[1] 

机构地区：[1]广州中医药大学临床药理研究所,广州510405

出　　处：《中国药房》2017年第10期1342-1345,共4页China Pharmacy

基　　金：广东省普通高校"青年创新人才"资助项目(No.2015KQNCX077)

摘　　要：目的:研究雷公藤红素对体外人肝癌HepG2细胞增殖、凋亡的影响,并探讨其作用机制。方法:采用CCK-8法测定2、5、10μmol/L雷公藤红素分别作用24、48、72 h后的细胞活性,并计算增殖抑制率和半数抑制浓度(IC_(50));采用流式细胞术检测2、5、10μmol/L雷公藤红素分别作用24 h后细胞的凋亡率及周期变化,并以二甲基亚砜(DMSO)为阴性对照;采用罗丹明123染色法测定2、5、10μmol/L雷公藤红素分别作用48 h后的细胞线粒体膜电位,并以DMSO为阴性对照;采用Western blot法检测5μmol/L雷公藤红素作用0、12、24、36 h后细胞中促凋亡相关基因Bax和B淋巴细胞瘤2(Bcl-2)的蛋白表达。结果:2、5、10μmol/L雷公藤红素均可抑制细胞的增殖,IC_(50)为5.834μmol/L。2、5、10μmol/L雷公藤红素均可诱导细胞凋亡。5、10μmol/L雷公藤红素可阻滞细胞于G_0/G_1、S期,并可降低线粒体膜电位,较阴性对照差异均有统计学意义(P<0.05或P<0.01),且以上作用均具有浓度依赖性。5μmol/L雷公藤红素作用12、24、36 h后可上调细胞中Bax蛋白表达、下调Bcl-2蛋白表达,并呈现一定的时间依赖性,较0 h时差异有统计学意义(P<0.05或P<0.01)。结论:雷公藤红素可明显抑制体外人肝癌HepG2细胞的增殖并诱导其凋亡,其机制可能与增强线粒体通透性、促使凋亡诱导因子释放有关。OBJECTIVE：To study the effects of celastrol on the proliferation and apoptosis of human hepatoma HepG2 cells,and investigate its mechanism. METHODS：CCK-8 method was used to determine the cell activity 24,48,72 h after treated by 2,5,10 μmol/L celastrol,and the proliferation inhibition rate and half inhibitory concentration（IC50）were calculated;flow cytometry was conducted to detect the cell apoptosis rate and cycle change 24 h after treated by 2,5,10 μmol/L celastrol,and the DMSO was used as negative control;rhodamine 123 staining method was used to determine the mitochondrial membrane potential 48 h after treated by 2,5,10 μ mol/L celastrol,and the DMSO was used as negative control;Western blot was adopted to detect the pro-apoptotic related genes Bax and B lymphoma 2（Bcl-2）protein expressions 0,12,24,36 h after treated by 5 μmol/L celastrol.RESULTS：2,5,10 μmol/L celastrol can inhibit cell proliferation,IC50 was 5.834 μmol/L. 2,5,10 μmol/L celastrol can induce apoptosis;5,10 μmol/L celastrol can block cell in G0/G1,S phases,compared with negative control group,with significant differences（P〈0.05 or P〈0.01）,and the above effects all showing certain concentration-dependent manner. 5 μmol/L celastrol can increase Bax protein expression and decrease Bcl-2 protein expression after cultured for 12,24,36 h,showing certain time-dependent manner;compared with 0 h,there was significant difference（P〈0.05 or P〈0.01）. CONCLUSIONS：Celastrol can obviously inhibit the proliferation of human hepatoma HepG2 cells and induce their apoptosis,and the mechanism may be related with strengthening mitochondrial permeability and promoting the release of apoptosis-inducing factor.

关 键 词：雷公藤红素 人肝癌HEPG2细胞 增殖 凋亡 线粒体通透性 体外 

分 类 号：R961[医药卫生—药理学]