基于JAK1/STAT1信号通路研究流感“肺脑传变”的分子机制及麻杏石甘汤干预作用  

Molecular Mechanism of"Transmission Between Lung and Brain"of Influenza and Intervention Effect of Maxing Shigantang Based on JAK1/STAT1 Signaling Pathway

作　　者：陈纯静 赵澄 张香港 王平 肖荣 胡珏[1] 匡炜 熊涛 卢芳国[1,3] CHEN Chun-jing;ZHAO Cheng;ZHANG Xiang-gang;WANG Ping;XIAO Rong;HU Jue;KUANG Wei;XIONG Tao;LU Fang-guo(Hunan University of Chinese Medicine,Changsha 410208,China;Changsha Hospital of Traditional Chinese Medicine,Changsha 410002,China;The First Hospital of Hunan University of Chinese Medicine,Changsha 410000,China)

机构地区：[1]湖南中医药大学,长沙410208 [2]长沙市中医医院,长沙410002 [3]湖南中医药大学第一附属医院,长沙410000

出　　处：《中国实验方剂学杂志》2022年第12期12-21,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82074250,81774126,81973670);湖南省自然科学基金项目(2020JJ4063);长沙市杰出创新青年人才培养计划项目(2019-23);湖南中医药大学中医药防治新型冠状病毒肺炎科研专项项目(2020XGXM03);湖南中医药大学一流学科《基础医学》资助项目(No.1);湖南中医药大学研究生创新课题(2020CX01);湖南中医药大学中西医结合一流学科开放基金项目(2020ZXYJH36,2020ZXYJH13,2020ZXYJH18);湖南省研究生优秀教学团队项目(基础医学研究生教学团队,编号[2019]370-118)。

摘　　要：目的基于Janus酪氨酸蛋白激酶1/信号转导和转录激活因子1(JAK1/STAT1)信号通路研究流感“肺脑传变”的分子机制,并进一步探讨麻杏石甘汤(MXSGT)的干预作用。方法SPF级BALB/c小鼠100只,随机分为正常组、模型组、奥司他韦组、抗病毒颗粒组和MXSGT组,每组20只。除正常组常规饲养外,其余4组以A型流感病毒(IAV)滴鼻感染构建小鼠IAV肺炎模型,造模24 h后,各药物治疗组分别予以奥司他韦(21.63 mg·kg^(-1)·d^(-1))、抗病毒颗粒(3.9 g·kg^(-1)·d^(-1))、MXSGT(6.05 g·kg^(-1)·d^(-1))灌胃,正常组和模型组以等量生理盐水灌胃,每天1次,连续给药3、7 d。苏木素-伊红(HE)染色观察肺、脑组织病理变化;实时荧光定量聚合酶链式反应(Real-time PCR)检测肺、脑组织中IAV核蛋白(NP)及JAK1、STAT1 mRNA表达水平;蛋白免疫印迹法(Western blot)检测肺、脑组织中JAK1、STAT1蛋白表达水平;免疫组化法检测肺、脑组织中磷酸化(p)-STAT1表达水平;酶联免疫吸附测定法(ELISA)检测血清中白细胞介素-1β(IL-1β)、白细胞介素-10(IL-10)的水平。结果与正常组比较,模型组小鼠肺组织及大脑皮质出现明显病理变化;肺部IAV NP mRNA相对表达量显著增加(P<0.01);肺组织和脑组织中JAK1、STAT1 mRNA和蛋白表达水平明显增加(P<0.05,P<0.01);肺组织和大脑皮质中p-STAT1表达水平明显增加(P<0.05,P<0.01);血清中IL-1β表达量明显增加(P<0.05)。与模型组比较,MXSGT组小鼠肺组织及大脑皮质病理损伤减轻;肺组织IAV NP mRNA相对表达量显著降低(P<0.01);肺组织和脑组织中JAK1、STAT1 mRNA和蛋白表达水平明显降低(P<0.05,P<0.01);血清中IL-10水平显著增加(P<0.01)。结论JAK1/STAT1信号通路异常活化可能是流感“肺脑传变”的分子机制之一,MXSGT作为有效的抗流感病毒中药复方,可通过调控该通路介导的细胞因子水平的变化缓解IAV肺部感染小鼠的脑组织病理损伤。Objective To explore the molecular mechanism of"transmission between the lung and brain"of influenza based on Janus kinase 1/signal transducer and activator of transcription 1(JAK1/STAT1)signaling pathway and further investigate the intervention effect of Maxing Shigantang(MXSGT).Method A total of 100 SPF BALB/c mice were randomly divided into a normal group,a model group,an oseltamivir group(21.63 mg·kg^(-1)·d^(-1)),an antiviral granules group(3.9 g·kg^(-1)·d^(-1)),and an MXSGT group(6.05 g·kg^(-1)·d^(-1)),with 20 mice in each group.The pneumonia model was induced in mice except for those in the normal group by intranasal infection of influenza A virus(IAV).Twenty-four hours after modeling,mice were treated with corresponding drugs,while those in the normal group and the model group received the same amount of normal saline by gavage,once a day for 3 and 7 days.The pathological changes in the lung and brain were observed by hematoxylin-eosin(HE)staining.The mRNA expression of IAV nucleoprotein(NP),JAK1,and STAT1 in the lung and brain was detected by real-time quantitative polymerase chain reaction(Real-time PCR),and the protein expression of JAK1 and STAT1 in the lung and brain was detected by Western blot.Immunohistochemical method was used to detect the expression of phosphorylated(p)-STAT1 in the lung and brain tissues,and enzyme-linked immunosorbent assay(ELISA)was used to detect the serum levels of interleukin-1β(IL-1β)and interleukin-10(IL-10).Result Compared with the normal group,the model group showed obvious pathological changes in the lung tissues and cerebral cortex,increased relative mRNA expression of IAV NP in the lung(P<0.01),elevated mRNA and protein expression of JAK1 and STAT1 in the lung and brain tissues(P<0.05,P<0.01),up-regulated expression level of p-STAT1 in lung tissues and cerebral cortex(P<0.05,P<0.01),and increased serum level of IL-1β(P<0.05).Compared with the model group,the MXSGT group showed alleviated pathological damage to lung tissues and cerebral cortex,decreased rela

关 键 词：A型流感病毒(IAV) 流感“肺脑传变” 麻杏石甘汤 Janus酪氨酸蛋白激酶1/信号转导和转录激活因子1(JAK1/STAT1)信号通路 细胞因子 

分 类 号：R242[医药卫生—中医临床基础] R285.5[医药卫生—中医学]