银葛双解剂抗甲型H1N1流感病毒的作用机制

Mechanism of Yinge Shuangjie Agent on the anti-influenza A(H1N1) virus

目的:研究银葛双解剂体内外抗甲型H1N1流感病毒作用机制。方法:体外实验以利巴韦林为阳性对照药物,采用流式细胞术检测银葛双解剂对甲型H1N1流感病毒所致狗肾细胞凋亡的抑制作用;体内试验以SPF级小鼠滴鼻建立肺炎模型,随机分为6组,正常对照组、模型对照组、利巴韦林组、银葛双解剂高剂量组、银葛双解剂低剂量组、银葛双解剂预防+治疗组,用流式细胞术检测各组小鼠全血中T细胞亚群分类、放射免疫法测定小鼠血清中白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的表达水平。结果:在体外试验中,银葛双解剂可抑制甲型H1N1流感病毒诱导的MDCK细胞凋亡,药物浓度越高,细胞凋亡抑制率越大,银葛双解剂浓度越高,抑制作用越强;体内实验中,银葛双解剂可通过下调甲型H1N1流感病毒感染小鼠血清中炎性细胞因子TNF-α、IL-6的表达水平,升高甲型H1N1流感病毒感染小鼠全血中CD3+、CD4+T细胞的表达百分比,降低CD8+T细胞的表达,上调CD4+/CD8+比值。结论:银葛双解剂具有良好的抗甲型H1N1流感病毒的作用。

Objective： To research the mechanism of Yinge Shuangjie Agent on the anti-influenza A（H1N1） virus in vivo and in vitro. Methods： In vitro experiment, Ribavirin as a positive control drug, the apoptosis induced by influenza A（H1N1） virus in MDCK（Madindarbycaninekidney） cells were observed by flow cytometry（FCM）, showing the inhibitory effect of Yinge Shuangjie Agent. In vivo experiment, the model of pulmonary inflammation mice were established by nasal intubation drip method, randomly divided into six groups： normal control group, model control group, ribavirin control group, the high and low dose of Yinge Shuangjie groups, the group of Yinge Shuangjie for prevention and treatment. T cell subsets in the whole blood were detected by flow cytometry, the expression levels of IL-6 and TNF-α in serum were determined by radioimmunoassay. Results： In vitro（model of cell）, agent of Yinge Shuangjie inhibit influenza a（H1N1） induced MDCK cell apoptosis, the higher the drug concentration, the stronger the inhibition; In vivo（mice model）, agent of Yinge Shuangjie reduce the levels of IL-6 and TNF-α, regulate the subgroup of T cell. Yinge Shuangjie Agent showed a multi-link effect on anti-influenza A（H1N1） virus, which inhibited the apoptosis of MDCK cells caused by influenza A（H1N1） virus, and down-regulated the expression levels of IL-6, TNF-α, and classification of T cell subsets. Conclusion： Yinge Shuangjie Agent has a good effect on anti-influenza A（H1N1）virus.