維拉帕米通過下調(diào)硫氧還蛋白互作蛋白的表達(dá)抑制丙型肝炎病毒感染
發(fā)布時(shí)間:2018-05-15 03:14
本文選題:維拉帕米 + 丙型肝炎病毒 ; 參考:《第二軍醫(yī)大學(xué)學(xué)報(bào)》2017年05期
【摘要】:目的探討抗高血壓藥維拉帕米是否可通過下調(diào)宿主硫氧還蛋白互作蛋白(TXNIP)的表達(dá)而抑制丙型肝炎病毒(HCV)的感染。方法用不同濃度梯度的維拉帕米處理人肝癌細(xì)胞系Huh7.5.1,用qPCR和蛋白質(zhì)印跡法檢測TXNIP的表達(dá)。用維拉帕米處理Huh7.5.1細(xì)胞,同時(shí)加入細(xì)胞培養(yǎng)產(chǎn)生的HCV(HCVcc),48 h后檢測HCVcc感染情況。用TXNIP siRNA轉(zhuǎn)染Huh7.5.1細(xì)胞,檢測下調(diào)TXNIP表達(dá)后,維拉帕米對HCVcc感染的影響;用TXNIP啟動(dòng)子調(diào)控的增強(qiáng)型綠色熒光蛋白(EGFP)報(bào)告基因表達(dá)質(zhì)粒(pTXNIP-EGFP)轉(zhuǎn)染Huh7.5.1細(xì)胞,分析維拉帕米對TXNIP啟動(dòng)子轉(zhuǎn)錄活性的影響。結(jié)果與對照孔相比,維拉帕米(100、200、400μmol/L)可下調(diào)Huh7.5.1細(xì)胞中TXNIP的表達(dá),并呈現(xiàn)濃度依賴性(P0.05);并可抑制HCVcc對Huh7.5.1細(xì)胞的感染,且濃度越高抑制作用越明顯(P0.05)。進(jìn)一步研究結(jié)果顯示,與對照組相比,維拉帕米能夠抑制pTXNIP-EGFP轉(zhuǎn)染細(xì)胞中EGFP的表達(dá)水平(P0.05)。結(jié)論維拉帕米可下調(diào)TXNIP的表達(dá)從而抑制HCV感染,這可能是通過抑制TXNIP啟動(dòng)子的轉(zhuǎn)錄活性來實(shí)現(xiàn)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.
【作者單位】: 第二軍醫(yī)大學(xué)熱帶醫(yī)學(xué)與公共衛(wèi)生學(xué)系微生物學(xué)教研室上海市醫(yī)學(xué)生物防護(hù)重點(diǎn)實(shí)驗(yàn)室;第二軍醫(yī)大學(xué)海軍醫(yī)學(xué)系學(xué)員5隊(duì);第二軍醫(yī)大學(xué)學(xué)員旅10隊(duì);
【基金】:國家自然科學(xué)基金(31570170) 上海市公共衛(wèi)生三年行動(dòng)計(jì)劃重點(diǎn)學(xué)科建設(shè)項(xiàng)目(15GWZK0103)~~
【分類號(hào)】:R512.63
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