葛根汤颗粒对甲型流感病毒感染小鼠TNFR1/NF-κB/JNK等炎症和免疫指标的影响

Effects of Gegen Decoction on Inflammatory and Immune Indexes TNFR1,NF-κB,JNK in Mice Infected with Influenza A Virus

目的基于PR8感染所致病毒性肺炎小鼠模型,探寻葛根汤颗粒抗流感病毒性肺炎损伤的作用机制。方法32只雌性SPF级ICR小鼠随机分为空白组、模型组、奥司他韦组、葛根汤颗粒组,每组8只。滴鼻法建立PR8毒株感染所致病毒性肺炎小鼠模型,计算肺指数和肺指数抑制率,应用光镜观察各组肺组织病理变化,运用ELISA和Western blotting法检测各组肺组织炎性细胞因子白细胞介素-6(IL-6)、细胞间黏附分子-1(ICAM1)、血管细胞黏附分子-1(VCAM1)和炎性免疫蛋白人肿瘤坏死因子受体-1(TNFR1)、核转录因子-κB(NF-κB)、Jun氨基末端激酶(JNK1)表达情况。结果与模型组比较,奥司他韦组和葛根汤颗粒组可降低小鼠肺指数(P<0.001),葛根汤颗粒组对肺指数抑制效果优于奥司他韦组。病理提示,与模型组比较,葛根汤颗粒组可显著减轻支气管炎症反应渗出,减轻肺泡、肺间质和红细胞渗出。ELISA结果显示,与模型组比较,奥司他韦组和葛根汤颗粒组可显著降低IL-6、ICAM1、VCAM1的表达(P<0.001);与奥司他韦组相比,葛根汤颗粒组IL-6含量降低(P<0.01)。Western blotting结果显示,与模型组比较,奥司他韦组和葛根汤颗粒组可降低TNFR1、JNK1蛋白的表达(P<0.05或P<0.01),葛根汤颗粒可降低NF-κB蛋白的表达(P<0.05)。结论葛根汤颗粒可以降低甲型流感病毒感染小鼠肺组织中IL-6、ICAM1、VCAM1、TNFR1、NF-κB、JNK等炎性因子和炎性免疫蛋白的表达,调节炎症和免疫紊乱,为仲景经典处方临床应用提供基础理论依据。

Objective:To explore the mechanism of anti-influenza viral pneumonia injury of Gegen Decoction based on the mouse model of viral pneumonia caused by PR8 infection.Methods:A total of 32 female SPF ICR mice were randomly divided into the blank group,model group,oseltamivir group and Gegen Decoction group,with 8 mice in each group.A mouse model of viral pneumonia caused by PR8 strain was established by nasal drops.Lung index and inhibition rate of lung index were calculated,and pathological changes of lung tissues were observed by light microscopy.The expression of inflammatory cytokines IL-6,ICAM1,VCAM1 and inflammatory immune proteins TNFR1,NF-κB and JNK1 were detected by ELISA and Western Blot.Results:Compared with model group,oseltamivir group and Gegen Decoction group could reduce lung index(P<0.001),and the inhibition effect of Gegen Decoction group on lung index was better than oseltamivir group.Pathological results showed that compared with model group,Gegen Decoction group could significantly reduce bronchial inflammatory reaction exudation,alveolar,pulmonary interstitial and red blood cell exudation.ELISA results showed that compared with model group,the expressions of IL-6,ICAM1 and VCAM1 in oseltamivir group and Gegen Decoction group were significantly decreased(P<0.001).Compared with oseltamivir group,the content of IL-6 in Gegen Decoction group was decreased(P<0.01).Western Blot results showed that compared with model group,oseltamivir group and Gegen Decoction group could decrease the expressions of TNFR1 and JNK1 proteins(P<0.05 or P<0.01),Gegen Decoction could reduce the expression of NF-κB protein(P<0.05).Conclusion:Gegen Decoction can reduce the expression of IL-6,ICAM1,VCAM1,TNFR1,NF-κB,JNK and other inflammatory factors and inflammatory immune proteins in the lung tissue of mice infected with influenza A virus,regulate inflammation and immune disorders,and provide basic theoretical basis for the clinical application of Zhang Zhongjing′s classic prescription.