金茵清热口服液对甲型H1N1流感病毒致细胞病变效应的影响

The effects of Jinyin Qingre Oral Liquid on cytopathic effects induced by A/H1N1 influenza virus

目的:研究金茵清热口服液(试药)抑制甲型H1N1流感病毒的作用及其对喉癌上皮细胞(Hep-2细胞)免疫机能的影响。方法:甲型H1N1流感病毒感染狗肾细胞(MDCK细胞)和Hep-2细胞后,给予不同浓度的试药。显微镜观察细胞病变效应(cytopathic effect,CPE),MTT法检测细胞活性,荧光定量PCR(qRT-PCR)检测试药作用于Hep-2细胞后细胞因子(TNF-α,IFN-1α,IFN-1β,IL-1α和IL-6)mRNA的水平。结果:实验细胞感染病毒后用试药处理,当试药质量浓度为3 mg·m L-1时,MDCK细胞存活率提高了61.6%;试药质量浓度为4 mg·m L-1时,Hep-2细胞存活率提高了40.6%,显著高于药物空白组细胞的存活率(P<0.05)。同时,试药显著上调Hep-2细胞的TNF-α(2.39倍)和IFN-1α(1.98倍)水平,下调炎症因子IL-1α(90%)和IL-6(64%)水平(P<0.05),而IFN-1β水平的差异无统计学意义(P>0.05)。结论:金茵清热口服液可以显著提高染毒细胞的存活率,并能显著上调Hep-2细胞抗病毒免疫分子,同时抑制炎症反应,推测金茵清热口服液调节细胞免疫应答可能是其抗流感病毒作用机制之一。

OBJECTIVE To investigate the effects of Jinyin Qingre Oral Liquid on inhibition on influenza A/H1 N1 influenza virus infection and on regulation of the immune functions of Hep-2 cells. METHODS MDCK and Hep-2 cells were infected with A/H1 N1 influenza virus,and were treated with various concentrations of Jinyin Qingre Oral Liquid. Cell pathogenic effects(CPE) were observed under the inverted microscope,the cell viability rate was determined by MTT assay. In addition,mRNA expression levels of the cellular cytokines(TNF-α,IFN-1α,IL-1α,IL-6 and IFN-1β) in Hep-2 cells treated with the Jinyin Qingre Oral Liquid were determined by quantitative real time PCR(qRT-PCR). RESULTS Compared with the control group,Jinyin Qingre Oral Liquid(3 mg·m L-1 and 4 mg·m L-1) significantly increased the survival rates of MDCK and Hep-2 cells(P < 0. 05) by 61. 60% and 40. 60%,respectively. The mRNA expression levels of TNF-α,IFN-1α of Hep-2 cells were significantly upregulated by 2. 39 and 1. 98 folds(P <0. 05),respectively,while the mRNA expression levels of IL-1α and IL-6 were downregulated by 90% and 64%(P < 0. 05),respectively. CONCLUSION Jinyin Qingre Oral Liquid has obviously protective effects against the influenza A virus infection in cells,which may be one of the mechanism for its stimulatory effects on cellular innate immunity.