基于JAK2-STAT3通路研究流感相关脑病的发病机制与麻杏石甘汤的干预作用

Pathogenesis of influenza associated encephalopathy and intervention effect of Maxing Shigan Decoction based on JAK2-STAT3 pathway

目的 基于Janus激酶2/信号转导和转录激活因子3(Janus kinase 2/signal transducer and activator of transcription 3,JAK2/STAT3)通路介导的细胞因子表达研究流感相关脑病的发病机制,并进一步探讨麻杏石甘汤的干预作用。方法 以A型流感病毒(influenza A virus,IAV)滴鼻感染BALB/c小鼠为研究对象,设置对照组、模型组、奥司他韦组、抗病毒颗粒组和麻杏石甘汤组。给予相应药物治疗3、7 d后,计算小鼠脑指数;苏木素-伊红(HE)染色法观察肺组织、脑组织病理变化;qRT-PCR法检测脑组织中JAK2、STAT3、白细胞介素-1β(interleukin-1β,IL-1β)、IL-10和IL-4 mRNA表达;Western blotting法检测脑组织中JAK2、STAT3蛋白表达;免疫组化法检测脑组织中磷酸化STAT3(phosphorylated STAT3,p-STAT3)蛋白表达;ELISA法检测脑组织中IL-1β、IL-10和IL-4水平。结果 IAV肺部感染的小鼠脑指数增加(P<0.05、0.01),肺组织及大脑皮质出现明显病理变化;脑组织中JAK2、STAT3、IL-1β基因和蛋白表达水平升高(P<0.05、0.01),p-STAT3表达水平升高(P<0.01),IL-10、IL-4基因表达水平降低(P<0.01),IL-1β、IL-10和IL-4分泌水平失调(P<0.05、0.01)。麻杏石甘汤能改善脑组织水肿及肺、脑组织病理变化,下调脑组织中JAK2、STAT3、IL-1β基因和蛋白表达水平(P<0.05、0.01),下调p-STAT3表达水平(P<0.01),上调IL-10、IL-4基因和蛋白的表达水平(P<0.01)。结论 JAK2/STAT3通路介导的细胞因子失衡可能是流感相关脑病的分子机制之一,麻杏石甘汤作为有效的抗流感病毒中药复方,可通过调控JAK2/STAT3通路减轻IAV肺部感染小鼠的脑组织病理损伤。

Objective To study the pathogenesis of influenza associated encephalopathy based on cytokines expressions mediated by Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)pathway,and further explore intervention effect of Maxing Shigan Decoction(麻杏石甘汤).Methods Influenza A virus(IAV)intranasally infected BALB/c mice were used as research objects,control group,model group,oseltamivir group,antiviral granule group and Maxing Shigan Decoction group were set up.After 3 and 7 d of corresponding drug treatment,brain index of mice was calculated;Hematoxylin-eosin(HE)staining was used to observe the pathological changes of lung tissue and brain tissue;qRT-PCR method was used to detect JAK2,STAT3,interleukin-1β(IL-1β),IL-10 and IL-4 mRNA expressions;Western blotting was used to detect the protein expressions of JAK2 and STAT3 in brain tissue;Immunohistochemical method was used to detect phosphorylated STAT3(p-STAT3)protein expression;IL-1β,IL-10 and IL-4 levels in brain tissue were detected by ELISA.Results Brain index of mice infected with IAV lung was increased(P<0.05,0.01),lung tissue and cerebral cortex showed obvious pathological changes;JAK2,STAT3,IL-1βgene and protein expressions in brain tissue were increased(P<0.05,0.01),p-STAT3 expression level was increased(P<0.01),IL-10 and IL-4 gene expressions were decreased(P<0.01),levels of IL-1β,IL-10 and IL-4 were unregulated(P<0.05,0.01).Maxing Shigan Decoction improved brain tissue edema and pathological changes in lung and brain tissue,down-regulated JAK2,STAT3,IL-1βgene and protein expression levels in brain tissue(P<0.05,0.01),down-regulated p-STAT3 expression level(P<0.01),up-regulated IL-10,IL-4 gene and protein expression levels(P<0.01).Conclusion Imbalance of cytokines mediated by JAK2/STAT3 pathway may be one of molecular mechanisms of influenza associated encephalopathy.As an effective anti-influenza virus Chinese medicine compound,Maxing Shigan Decoction can improve brain histopathological damage of mice with IAV lung infection by regulating JAK2/STAT3 pathway.