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-b by the p38 Signal Factor

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

Key words:  epigallocatechin gallate(EGCG)  p38 factor  IFN-β  influenza A virus

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-β mRNA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（0 μg·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（20 μg·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-β mRNA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。

Abstract:

OBJECTIVE To explore the mechanism of epigallocatechin gallate(EGCG) inhibit influenza A virus H1N1 replication through inducing production of IFN-β by the p38 signaling factor. METHODS EGCG was applied to human tubular epithelial cells(BEAS-2B), the expression of IFN-β mRNA 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-β protein and mRNA. IFN-β antibody was used to neutralize the production of IFN-β in 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(0 μg·mL^-1), as the dose of EGCG increased, IFN-β protein and mRNA increased significantly. Compared with 0 h of treatment, EGCG(20 μg·mL^-1) induced significant IFN-β protein 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-β mRNA and protein expression(P<0.01). Compared with EGCG alone in the treatment of H1N1 cells, the co-treatment of EGCG and IFN-β antibody 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-β in BEAS-2B by up-regulating the phosphorylation level of p38 signal factor, thereby inhibiting H1N1 replication.