来源：风湿病与关节炎,2022,11（7）:1-6,17.

基于RAF-MEK1/MEK2信号途径探讨脊蛇祛湿胶囊改善胶原诱导关节炎模型大鼠肠道炎症的作用机制（基础研究）

李湖帆¹，钟　琴²，马武开²，黄　颖²，朱　丹²，张潇东²，陈昌明²

　　【摘　要】目的：探讨脊蛇祛湿胶囊通过RAF-MEK1/MEK2信号途径改善胶原诱导关节炎（CIA）模型大鼠肠道炎症的作用机制。方法：将70只SPF级Wistar雌性大鼠随机分为空白对照组、模型对照组、脊蛇祛湿胶囊低剂量组、脊蛇祛湿胶囊中剂量组、脊蛇祛湿胶囊高剂量组、益生菌组及甲氨蝶呤组，每组10只。喂养1周后，除空白对照组外，其余6组构建CIA模型，连续灌胃4周后处死大鼠，分离大鼠肠道组织，进行HE染色，并观察其病理，采用Western blot法检测各组大鼠肠道组织RAF、MEK1及MEK2蛋白的表达水平，RT-qPCR法检测大鼠肠道组织RAF、MEK1及MEK2 mRNA的表达水平。结果：大肠病理及Western blot、RT-qPCR结果显示，与空白对照组比较，模型对照组肠道组织黏膜层及黏膜下层可见大量细胞增生，并可见黏膜隆起及充血、水肿。模型对照组RAF、MEK1、MEK2蛋白及基因在大肠组织中表达增加，当予以一定脊蛇祛湿胶囊干预后，大鼠肠道组织的病变情况明显改善，且大肠组织中RAF、MEK1、MEK2蛋白及基因表达降低。结论：脊蛇祛湿胶囊可能通过下调RAF-MEK1/MEK2信号途径改善CIA大鼠的肠道炎症。

　　【关键词】　类风湿关节炎；肠道炎症；脊蛇祛湿胶囊；MAPK信号通路；RAF-MEK1/MEK2信号；胶原诱导关节炎；大鼠

Study on the Mechanism of Jishe Qushi Jiaonang（脊蛇祛湿胶囊）in Improving Intestinal Inflammation in Collagen-induced Arthritis Model Rats Based on RAF-MEK1/MEK2 Signal Pathway

LI Hu-fan,ZHONG Qin,MA Wu-kai,HUANG Ying,ZHU Dan,ZHANG Xiao-dong,CHEN Chang-ming

　　【ABSTRACT】Objective:To explore the mechanism of Jishe Qushi Jiaonang（JQJ,脊蛇祛湿胶囊）in improving intestinal inflammation in collagen-induced arthritis（CIA）model rats through RAF-MEK1/MEK2 signaling pathway.Methods:Seventy female Wistar rats of SPF grade were randomly divided into a blank control group,a model control group,a low-dose JQJ group,a medium-dose JQJ group,a high-dose JQJ group,a probiotics group and a methotrexate group,with 10 rats in each group.After fed for one week,except for the blank control group,the other six groups was used to construct CIA models.The rats were killed after continuous gavage for 4 weeks.The intestinal tissues of the rats were separated,HE staining was made,and their pathology was observed.The expression levels of RAF,MEK1 and MEK2 proteins in the intestinal tissues of the rats in each group were detected by Western blot,and the expression levels of RAF,MEK1 and MEK2 mRNA in the intestinal tissues of the rats were detected by RT-qPCR.Results:The results of colorectal pathology,Western blot and RT-qPCR showed that compared with the blank control group,a large number of cell proliferation,mucosal swelling,congestion and edema could be seen in the intestinal mucosa and submucosa of the model control group.In the model control group,the expressions of RAF,MEK1,MEK2 proteins and genes increased in large intestine tissue.When given a certain JQJ,the pathological changes of rat intestinal tissue were significantly improved,and the expression of RAF,MEK1,MEK2 proteins and genes in large intestine tissue decreased.Conclusion:JQJ may improve intestinal inflammation in CIA rats by down regulating RAF-MEK1/MEK2 signal pathway.

　　【Keywords】 rheumatoid arthritis;intestinal inflammation;Jishe Qushi Jiaonang（脊蛇祛湿胶囊）;MAPK signal path;RAF-MEK1/MEK2 signal;collagen-induced arthritis;rats