维拉帕米通过下调硫氧还蛋白互作蛋白的表达抑制丙型肝炎病毒感染

发布时间：2018-05-15 03:14

本文选题：维拉帕米 + 丙型肝炎病毒　；参考：《第二军医大学学报》2017年05期

【摘要】：目的探讨抗高血压药维拉帕米是否可通过下调宿主硫氧还蛋白互作蛋白(TXNIP)的表达而抑制丙型肝炎病毒(HCV)的感染。方法用不同浓度梯度的维拉帕米处理人肝癌细胞系Huh7.5.1,用qPCR和蛋白质印迹法检测TXNIP的表达。用维拉帕米处理Huh7.5.1细胞,同时加入细胞培养产生的HCV(HCVcc),48 h后检测HCVcc感染情况。用TXNIP siRNA转染Huh7.5.1细胞,检测下调TXNIP表达后,维拉帕米对HCVcc感染的影响;用TXNIP启动子调控的增强型绿色荧光蛋白(EGFP)报告基因表达质粒(pTXNIP-EGFP)转染Huh7.5.1细胞,分析维拉帕米对TXNIP启动子转录活性的影响。结果与对照孔相比,维拉帕米(100、200、400μmol/L)可下调Huh7.5.1细胞中TXNIP的表达,并呈现浓度依赖性(P0.05);并可抑制HCVcc对Huh7.5.1细胞的感染,且浓度越高抑制作用越明显(P0.05)。进一步研究结果显示,与对照组相比,维拉帕米能够抑制pTXNIP-EGFP转染细胞中EGFP的表达水平(P0.05)。结论维拉帕米可下调TXNIP的表达从而抑制HCV感染,这可能是通过抑制TXNIP启动子的转录活性来实现的。
[Abstract]:Objective to investigate whether verapamil can inhibit hepatitis C virus (HCV) infection by down-regulating the expression of host thioredoxin interaction protein (TXNIPP). Methods Human hepatoma cell line Huh 7.5.1 was treated with verapamil at different concentrations and the expression of TXNIP was detected by qPCR and Western blotting. Huh7.5.1 cells were treated with verapamil and the HCVcc infection was detected after 48 h. The effect of verapamil on HCVcc infection was detected after down-regulation of TXNIP expression in Huh7.5.1 cells transfected with TXNIP siRNA, and pTXNIP-EGFP) was transfected into Huh7.5.1 cells by TXNIP promoter regulated enhanced green fluorescent protein (EGFP) reporter gene expression plasmid pTXNIP-EGFP. To analyze the effect of verapamil on the transcriptional activity of TXNIP promoter. Results verapamil 100200400 渭 mol / L could down-regulate the expression of TXNIP in Huh7.5.1 cells in a concentration-dependent manner, and inhibit the infection of Huh7.5.1 cells by HCVcc. The higher the concentration of verapamil was, the more obvious the inhibitory effect was on Huh7.5.1 cells. Further studies showed that verapamil could inhibit the expression of EGFP in pTXNIP-EGFP transfected cells compared with the control group (P 0.05). Conclusion verapamil can down-regulate the expression of TXNIP and inhibit HCV infection, which may be achieved by inhibiting the transcriptional activity of TXNIP promoter.
【作者单位】：第二军医大学热带医学与公共卫生学系微生物学教研室上海市医学生物防护重点实验室;第二军医大学海军医学系学员5队;第二军医大学学员旅10队;
【基金】：国家自然科学基金(31570170) 上海市公共卫生三年行动计划重点学科建设项目(15GWZK0103)~~
【分类号】：R512.63

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