麻杏石甘汤和银翘散体外抗流感病毒的作用机制分析

Analysis of the Mechanism of Maxing Shigan Decoction and Yinqiao Powder against Influenza Virus in Vitro

目的分析麻杏石甘汤和银翘散体外抗流感病毒的作用机制。方法购买石家庄以岭药业股份有限公司提供的麻杏石甘汤、银翘散药物粉剂,将Sigma公司生产的奥司他韦强酸盐(批号:1717-003A9)设定为阳性对照药。测定药物对细胞半数毒性浓度(TC50)、药物对病毒半数有效浓度(IC50),运用qPCR法测定药物对病毒诱导细胞因子及趋化因子基因调节作用,运用ELISA法测定药物对病毒诱导IL-6、IP-10蛋白调节作用,分析麻杏石甘汤、银翘散体外对流感病毒的药效,统计分析麻杏石甘汤、银翘散体外对细胞因子基因表达的作用,并对麻杏石甘汤、银翘散体外对IP-10、IL-6蛋白的调节作用进行统计分析。结果麻杏石甘汤对H1N1、H1N1、B/Lee/1940、H6N2、H9N2的IC50分别为(0.142±0.040)、(0.380±0.081)、(0.432±0.015)、(0.395±0.051)、(0.724±0.204),对5种流感病毒的IC50在0.1~1 mg/mL之间,体外在一定程度上抑制流感病毒;银翘散体外不直接抑制流感病毒。麻杏石甘汤对IP-10、IL-6的基因表达下调作用显著高于病毒组,差异有统计学意义(t=4.303、3.182,P<0.05),麻杏石甘汤、银翘散均将CCL5、IP-10、TNF-α、IL-6、IL-8的基因表达下调,差异有统计学意义(F=6.965、4.541,P<0.05)。银翘散能够将MIG的基因表达下调,差异有统计学意义(P<0.05),麻杏石甘汤能够将COX-2的基因表达下调,差异有统计学意义(P<0.05)。麻杏石甘汤、银翘散均能够将IP-10、IL-6的蛋白表达下调,差异有统计学意义(P<0.05)。结论麻杏石甘汤能够直接抗病毒、对病毒介导炎症进行抑制,银翘散只有抗炎药效。

Objective To analyze the mechanism of Maxing Shigan Decoction and Yinqiao San against influenza virus in vitro.Methods Purchase Maxing Shigan Decoction and Yinqiao Powder provided by Shijiazhuang Yiling Pharmaceutical Co.,Ltd.,and set the oseltamivir strong acid salt(batch number:1717-003A9)produced by Sigma as the positive control drug.Determine the half-toxic concentration of the drug to cells(TC50),the half-effective concentration of the drug to the virus(IC50),use qPCR to determine the regulatory effect of drugs on virus-induced cytokines and chemokine genes,and use ELISA method to determine the drug-induced IL-6 on viruses,IP-10 protein regulation,analysis of the efficacy of Maxing Shigan Decoction and Yinqiao San on influenza virus in vitro,statistical analysis of the effect of Maxing Shigan Decoction and Yinqiao San on the expression of cytokine genes in vitro,the effects of Shigan Decoction and Yinqiao Powder on the regulation of IP-10 and IL-6 proteins in vitro were statistically analyzed.Results The IC50 of Maxing Shigan Decoction for H1N1,H1N1,B/Lee/1940,H6N2,H9N2 were(0.142±0.040),(0.380±0.081),(0.432±0.015),(0.395±0.051),(0.724±0.204),the IC50 for 5 kinds of influenza viruses was between 0.1~1 mg/mL,which inhibited influenza virus to a certain extent in vitro;Yinqiao San does not directly inhibit influenza virus in vitro.Maxingshigan decoction had a significantly higher down-regulation effect on the gene expression of IP-10 and IL-6 than the virus group,the difference was statistically significant(t=4.303,3.182,P<0.05).Maxingshigan decoction and Yinqiao powder both reduced the gene expression of CCL5 and IP-10,TNF-α,IL-6 and IL-8 was down-regulated,the difference was statistically significant(F=6.965,4.541,P<0.05).Yinqiaosan can down-regulate the gene expression of MIG,the difference was statistically significant(P<0.05),and Maxing Shigan Decoction can down-regulate the gene expression of COX-2,the difference was statistically significant(P<0.05).Both Maxing Shigan Decoction and Yinqiao Powder could down-regulate the protein expression of IP-10 and IL-6,the difference was statistically significant(P<0.05).Conclusion Maxing Shigan Decoction can directly resist viruses and inhibit virus-mediated inflammation,while Yinqiao Powder has only anti-inflammatory effects.