EGCG通过p38信号因子诱导IFN-β抗甲型流感病毒感染的研究  

Study on EGCG Against Influenza A Virus Through Inducing the Production of IFN-β by the p38 Signal Factor

作　　者：马贵凤 祝洁 曹慧军 陈强 江滟 MA Guifeng;ZHU Jie;CAO Huijun;CHEN Qiang;JIANG Yan(Department of Microbiology and Immunology,Inspection Center,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China;Microbiology Teaching and Research Section,Guizhou Medical University,Guiyang 550004,China;Key Laboratory of Medical Microbiology and Parasitology of Education Department of Guizhou,Guiyang 550004,China)

机构地区：[1]贵州医科大学附属医院临床检验中心微生物免疫科,贵阳550004 [2]贵州医科大学微生物学教研室,贵阳550004 [3]贵州省普通高校病原生物学特色重点实验室,贵阳550004

出　　处：《中国现代应用药学》2021年第6期661-666,共6页Chinese Journal of Modern Applied Pharmacy

基　　金：国家自然科学基金项目(81260249)。

摘　　要：目的探讨表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)通过p38信号因子诱导IFN-β抑制甲型流感病毒H1N1复制作用机制。方法将EGCG作用于人气管上皮细胞(BEAS-2B),通过qRT-PCR和ELISA检测BEAS-2B中IFN-βmRNA和蛋白的表达,Western blotting检测p38和p-p38蛋白表达;使用p38抑制剂抑制p38信号因子的信号传导,检测EGCG作用BEAS-2B细胞诱导的IFN-β蛋白和mRNA表达;用IFN-β抗体中和细胞中IFN-β的产生,通过qRT-PCR和Western blotting检测EGCG作用H1N1后NP蛋白和mRNA的表达。结果与EGCG(0μg·mL^(−1))相比,随着EGCG剂量增加,IFN-β蛋白和mRNA明显增加。与作用0 h相比,EGCG(20μg·mL^(-1))作用细胞12 h时诱导IFN-β蛋白表达显著(P<0.01)。EGCG可促进BEAS-2B中p38磷酸化;使用p38抑制剂抑制p38信号通路后,EGCG诱导IFN-βmRNA和蛋白表达减少(P<0.01)。与EGCG单独治疗感染H1N1的细胞相比,EGCG和IFN-β抗体共同治疗,可明显升高感染细胞中NP蛋白和mRNA表达水平(P<0.05)。结论EGCG可通过上调p38信号因子磷酸化水平诱导BEAS-2B产生IFN-β,从而抑制H1N1复制。OBJECTIVE To explore the mechanism of epigallocatechin gallate(EGCG)inhibit influenza A virus H1N1 replication through inducing production of IFN-βby the p38 signaling factor.METHODS EGCG was applied to human tubular epithelial cells(BEAS-2B),the expression of IFN-βmRNA and protein in BEAS-2B was detected by qRT-PCR and ELISA,and the expression of p38 and p-p38 protein was detected by Western blotting.Used p38 inhibitors to inhibit the signal transduction of p38 signal factors,and detected the effect of EGCG on BEAS-2B cells to induce the expression of IFN-βprotein and mRNA.IFN-βantibody was used to neutralize the production of IFN-βin cells,and the expression of NP protein and mRNA after EGCG treated H1N1 was detected by qRT-PCR and Western blotting.RESULTS Compared with EGCG(0μg·mL^(-1)),as the dose of EGCG increased,IFN-βprotein and mRNA increased significantly.Compared with 0 h of treatment,EGCG(20μg·mL^(−1))induced significant IFN-βprotein expression when cells were treated for 12 h(P<0.01).EGCG could promote the phosphorylation of p38 in BEAS-2B;after the use of p38 inhibitors to inhibit the p38 signaling pathway,EGCG induced a decrease in IFN-βmRNA and protein expression(P<0.01).Compared with EGCG alone in the treatment of H1N1 cells,the co-treatment of EGCG and IFN-βantibody could significantly increase the expression of NP protein and mRNA in the infected cells(P<0.05).CONCLUSION EGCG can induce the production of IFN-βin BEAS-2B by up-regulating the phosphorylation level of p38 signal factor,thereby inhibiting H1N1 replication.

关 键 词：表没食子儿茶素没食子酸酯 p38信号因子 IFN-Β 甲型流感病毒 

分 类 号：R966[医药卫生—药理学]