苦参碱类生物碱联合恩替卡韦抗耐药HBV的作用效果及机制分析

发布时间：2019-04-15 19:08

【摘要】：目的:研究苦参碱类生物碱联合恩替卡韦抗耐药乙型肝炎病毒(HBV)的作用效果以及对p38 MAPK,钠离子-牛磺胆酸共转运蛋白(NTCP)表达的影响。方法:以解放军第302医院自行构建的恩替卡韦耐药乙型肝炎病毒复制细胞株Hep G2.A64为模型,采用CPE,CCK-8分别考察苦参碱类生物碱氧化苦参碱、苦参碱、槐果碱、槐定碱和核苷类药物恩替卡韦单独用药及两者联合用药的细胞毒性;采用酶联免疫法(ELISA),实时荧光定量聚合酶链式反应(Real-time PCR),蛋白质免疫印迹法(Western blot)分别观察联合用药对Hep G2.A64细胞HBs Ag分泌,HBV DNA复制,p38和NTPC mRNA表达,细胞中p38和p-p38蛋白表达水平的影响。结果:与单独用药比较,氧化苦参碱和恩替卡韦联合用药72 h后能显著抑制HBs Ag分泌,HBV DNA复制(P0.05),同时能显著降低NTCP基因和蛋白的表达水平以及磷酸化p38蛋白水平。结论:氧化苦参碱联合恩替卡韦可能通过下调NTCP的表达同时抑制p38蛋白磷酸化达到抗病毒的效果。氧化苦参碱联合恩替卡韦能显著增强抗耐药HBV病毒的效果。
[Abstract]:Aim: to study the anti-hepatitis B virus (HBV) effect of matrine alkaloids combined with entecavir and its effect on the expression of p38 MAPK, sodium ion-taurocholic acid cotransporter (NTCP). Methods: the anti-hepatitis B virus replication cell line Hep G2.A64, which was established by ourselves in 302 Hospital of PLA, was used as the model. The alkaloid oxymatrine and sophorine were examined by CPE,CCK-8. Cytotoxicity of sophoridine and nucleoside drugs Enticavir alone and the combination of them; Enzyme-linked immunosorbent assay (ELISA),) was used to detect the expression of p38 and NTPC mRNA in Hep G2.A64 cells by real-time fluorescence quantitative polymerase chain reaction (Western blot). The effect of p38 and p-p38 protein expression in cells. Results: the combination of oxymatrine and enecavir for 72 h could significantly inhibit the secretion of, HBV DNA by HBs Ag (P0.05), and significantly reduce the expression of NTCP gene and protein and the level of phosphorylated p38 protein. Conclusion: oxymatrine combined with entecavir may have antiviral effect by down-regulating the expression of NTCP and inhibiting the phosphorylation of p38 protein. Oxymatrine combined with enecavir could significantly enhance the resistance to HBV virus.
【作者单位】：成都中医药大学药学院中药材标准化教育部重点实验室四川省中药资源系统研究与开发利用重点实验室——省部共建国家重点实验室培育基地;解放军第302医院全军中药研究所;解放军第302医院中西医结合肝病诊疗与研究中心;
【基金】：国家自然科学基金项目(8133090) 山东省协同创新项目(XTCX2014B01-06)
【分类号】：R96

【相似文献】