从TLR2/TLR4介导的炎症因子表达水平探讨麻杏石甘汤抗流感病毒的免疫机理

The Immune Mechanism of Maxing Shigan Decoction(麻杏石甘汤)Against Influenza Virus from the Expression Level of Inflammatory Factor Protein Mediated by TLR2/TLR4

目的:从TLR2/TLR4介导的炎症因子分泌水平探讨麻杏石甘汤抗流感病毒效应机制。方法:传代培养小鼠肺巨噬细胞,实验设正常对照组、TLR2激活剂组、TLR4激活剂组、TLR2/4抑制剂组、病毒对照组、奥司他韦组、抗病毒口服液组、麻杏石甘汤组。各组以相应因素干预24、48 h后用ELISA法检测细胞上清液中IL-1、IL-8、TNF-α含量;用免疫组化法检测干预24 h后各组TLR2、TLR4蛋白表达水平。结果:与正常对照组比较,TLR2激活剂组、TLR4激活剂组、病毒对照组肺巨噬细胞TLR2、TLR4蛋白表达水平明显升高(P<0.01),细胞上清液中IL-1、IL-8、TNF-α含量明显升高(P<0.01);与病毒对照组比较,麻杏石甘汤含药血清干预24 h后肺巨噬细胞TLR2、TLR4蛋白表达水平明显降低(P<0.05),细胞上清液中IL-1、IL-8、TNF-α含量明显降低(P<0.01)。结论:调节流感病毒感染肺巨噬细胞TLR2、TLR4表达水平进而影响炎症因子的分泌水平可能是麻杏石甘汤抗病毒作用的效应机制之一。

Objective:To investigate the anti-influenza effect mechanism of Maxing Shigan Decoction from the secretion of inflammatory factors mediated by TLR2/TLR4.Methods:Mouse lung macrophages were subcultured.The experiment set up normal control group,TLR2 activator group,TLR4 activator group,TLR2/4 inhibitor group,virus control group,oseltamivir group,antiviral oral liquid group and Maxing Shigan Decoction Group.The levels of IL-1,IL-8 and TNF-αin the cell supernatant were detected by ELISA after 24 and 48 hours intervention with corresponding factors;The protein expression levels of TLR2 and TLR4 in each group were examined by immunohistochemistry after 24 hours intervention.Results:Compared with normal control group,the protein expression levels of TLR2 and TLR4 in lung macrophages significantly increased in TLR2 activator group,TLR4 activator group and virus control group(P<0.01),and the contents of IL-1,IL-8 and TNF-αin supernatant significantly increased in TLR2 activator group,TLR4 activator group and virus control group(P<0.01).Compared with the virus control group,the protein expressions of TLR2 and TLR4 in lung macrophages decreased(P<0.05),and the contents of IL-1,IL-8 and TNF-αin the supernatant of lung macrophages significantly decreased(P<0.01)after 24 hours intervention with Maxing Shigan decoction containing serum(P<0.01).Conclusion:Regulating the expression levels of TLR2 and TLR4 in influenza virus-infected lung macrophages to affect the secretion of inflammatory factors may be one of the effective mechanisms of Maxing Shigan Decoction's antiviral effect.