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  • 王世杰,张卫,张龙严,罗正汉,尹昊瓒,唐紫薇,戚中田,赵平.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[J].第二军医大学学报,2017,38(5):548-553    [点击复制]
  • WANG Shi-,ZHANG Wei,ZHANG Long-yan,LUO Zheng-han,YIN Hao-zan,TANG Zi-wei,QI Zhong-tian,ZHAO Ping.Verapamil inhibits hepatitis C virus infection via down-regulating thioredoxin-interacting protein expression[J].Acad J Sec Mil Med Univ,2017,38(5):548-553   [点击复制]
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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
王世杰1△,张卫2△,张龙严1,罗正汉1,尹昊瓒3,唐紫薇1,戚中田1,赵平1*
0
(1. 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, 上海 200433;
2. 第二军医大学海军医学系学员5队, 上海 200433;
3. 第二军医大学学员旅10队, 上海 200433
共同第一作者
*通信作者)
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、400 μmol/L)可下调Huh7.5.1细胞中TXNIP的表达,并呈现浓度依赖性(P<0.05);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(P<0.05)。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(P<0.05)。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。
关键词:  维拉帕米  丙型肝炎病毒  硫氧还蛋白互作蛋白  转录启动子
DOI:10.16781/j.0258-879x.2017.05.0548
投稿时间:2017-02-09修订日期:2017-05-04
基金项目: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).
Verapamil inhibits hepatitis C virus infection via down-regulating thioredoxin-interacting protein expression
WANG Shi-1△,ZHANG Wei2△,ZHANG Long-yan1,LUO Zheng-han1,YIN Hao-zan3,TANG Zi-wei1,QI Zhong-tian1,ZHAO Ping1*
(1. Department of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Faculty of Tropical Medicine and Public Health, Second Military Medical University, Shanghai 200433, China;
2. The 5thTeam of Student Brigade, Faculty of Navy Medicine, Second Military Medical University, Shanghai 200433, China;
3. The 10thTeam of Student Brigade, Second Military Medical University, Shanghai 200433, China
Co-first authors.
* Corresponding author)
Abstract:
Objective To investigate whether antihypertensive agent verapamil can inhibit hepatitis C virus (HCV) infection via reducing the expression of thioredoxin-interacting protein (TXNIP) in hepatocytes of the host. Methods Human hepatocellular carcinoma Huh7.5.1 cells were treated with different concentrations of verapamil, and then the mRNA and protein expressions of TXNIP were detected by qPCR and Western blotting, respectively. The HCV infection level of Huh7.5.1 cells was determined 48 h after treatment with verapamil and cell culture-derived HCV (HCVcc). We observed the effect of verapamil on the Huh7.5.1 cells with TXNIP silenced by siRNA after infected by HCVcc. Huh7.5.1 cells were transfected with expression plasmid of enhanced green fluorescent protein (EGFP) controlled by TXNIP promoter (pTXNIP-EGFP), and then the effect of verapamil on transcriptional activity TXNIP promoter was analyzed. Results Compared with the control group, verapamil (100, 200, 400 μmol/L) significantly inhibited TXNIP expression and HCV infection in Huh7.5.1 cells in a dose-dependent manner (P<0.05). Furthermore, verapamil reduced EGFP expression in Huh7.5.1 cells transfected with pTXNIP-EGFP in comparison with the control group (P<0.05). Conclusion Verapamil can inhibit HCV infection via reducing TXNIP expression, which may be associated with the inhibition of TXNIP promotor transcriptional activity.
Key words:  verapamil  hepatitis C virus  thioredoxin-interacting protein  transcription initiation site