来源：风湿病与关节炎,2023,12（8）:1-7.

松萝提取物对胶原诱导性关节炎大鼠HMGB1-RAGE自噬通路及关节炎性损伤的影响（基础研究）

谢良山¹，安　阳²，张　军²，黄　颖²，潘晓艺²，徐　晖²

　　【摘　要】目的：探讨松萝提取物对胶原诱导性关节炎（CIA）大鼠高迁移率族蛋白B1（HMGB1）-晚期糖基化终产物受体（RAGE）自噬通路及关节炎性损伤的影响。方法：将54只大鼠随机分为空白对照组，模型对照组，松萝提取物低、中、高剂量组和羟氯喹组，每组9只。除空白对照组外，其余组采用Ⅱ型胶原乳剂建立CIA模型。松萝提取物低、中、高剂量组分别给予2 mg·kg^-1·d^-1、6 mg·kg^-1·d^-1、54 mg·kg^-1·d^-1松萝提取物灌胃，羟氯喹组给予36 mg·kg^-1·d^-1羟氯喹灌胃，空白对照组及模型对照组则予以等量的生理盐水灌胃。观察各组大鼠关节炎评分、肿胀程度和滑膜病理的变化，采用ELISA检测肿瘤坏死因子-α（TNF-α）、基质金属蛋白酶（MMPs）、血管内皮生长因子（VEGF）、HMGB1的含量，PCR、WB检测蛋白HMGB1、RAGE、Beclin1、B淋巴细胞瘤-2基因（Bcl-2）、哺乳动物雷帕霉素靶点（mTOR）、p38丝裂原活化蛋白激酶（p38 MAPK）的蛋白浓度及表达情况。结果：模型大鼠出现关节肿胀、变形，活动量减少，关节炎评分提示造模成功。经松萝提取物治疗后，大鼠的关节肿胀减轻，活动量增加，炎症反应减轻。与空白对照组比较，模型对照组血清TNF-α、MMPs、VEGF、HMGB1含量均增高，且HMGB1、RAGE、Bcl-2、mTOR、p38 MAPK蛋白表达量均上升，Beclin1蛋白表达量下降（P < 0.05）。与模型对照组比较，松萝提取物中、高剂量组及羟氯喹组HMGB1、RAGE、Bcl-2、mTOR、p38 MAPK、TNF-α、MMPs、VEGF含量均降低，Beclin1蛋白表达量上升（P < 0.05）；与羟氯喹组比较，松萝提取物中、高剂量组抑制HMGB1、RAGE、Bcl-2、mTOR、p38 MAPK表达，激活Beclin1的效果相当，差异无统计学意义（P > 0.05）。结论：松萝提取物能减轻CIA大鼠关节肿胀程度，降低血清TNF-α、MMPs、VEGF、HMGB1含量，抑制HMGB1、RAGE、Bcl-2、mTOR、p38 MAPK蛋白的表达，提升Beclin1表达，增强细胞自噬，减少滑膜炎症反应。

　　【关键词】　类风湿关节炎；胶原诱导性关节炎；松萝提取物；HMGB1；RAGE；细胞自噬；炎性损伤；大鼠

Effects of Usnea Extract on HMGB1-RAGE Autophagy Pathway and Arthritis Induced Injury in Collagen Induced Arthritis Rats

XIE Liang-shan,AN Yang,ZHANG Jun,HUANG Ying,PAN Xiao-yi,XU Hui

　　【ABSTRACT】Objective:To investigate the effects of usnea extract on HMGB1-RAGE autophagy pathway and arthritis induced injury in collagen induced arthritis（CIA）rats.Methods:Fifty-four rats were randomly divided into a blank control group,a model control group,low,medium and high dose groups of usnea extract and an Hydroxychloroquine control group,with 9 rats in each group.Except the blank control group,the other groups used type Ⅱcollagen emulsion to establish CIA models.The low,middle and high dose groups were given 2 mg·kg^-1·d^-1,6 mg·kg^-1·d^-1 and 54 mg·kg^-1·d^-1 respectively by gavage,the Hydroxychloroquine group was given 36 mg·kg^-1·d^-1 of Hydroxychloroquine by gavage,and the blank control group and the model control group were given the same amount of normal saline by gavage.Observe the changes in joint score,swelling degree measurement,and synovial pathology of rats in each group,and detect the content of TNF-α,MMPs,VEGF and HMGB1 by ELISA.The protein concentration and expression of HMGB1,RAGE,Beclin1,Bcl-2,mTOR,p38 MAPK were detected by PCR and WB.Results:The model rats showed joint swelling and deformation,reduced activity,and arthritis scores indicating successful modeling.After treatment with usnea extract,the joint swelling of rats decreased,the activity increased,and the inflammatory response decreased.Compared with the blank control group,the concentrations of TNF-α,MMPs,VEGF,HMGB1 increased,and the expression levels of HMGB1,RAGE,Bcl-2,mTOR,and p38 MAPK proteins increased,while the expression levels of Beclin1 protein decreased（P < 0.05）.Compared with the model control group,the content of HMGB1,RAGE,Bcl-2,mTOR,p38 MAPK,TNF-α,MMPs and VEGF in the middle and high dose groups and the Hydroxychloroquine group decreased,while the expression of Beclin1 protein increased（P < 0.05）.Compared with the Hydroxychloroquine group,the middle and high dose groups had similar effects in inhibiting the expression of HMGB1,RAGE,Bcl-2,mTOR,p38 MAPK and activating Beclin1,with no statistically significant difference（P > 0.05）.Conclusion:Usnea extract can reduce the degree of joint swelling and concentrations of TNF-α,MMPs,VEGF,and HMGB1 in CIA rats,inhibit the expression of HMGB1,RAGE,Bcl-2,mTOR,and p38 MAPK proteins,increase the expression of Beclin1,and enhance cell viability.

　　【Keywords】 rheumatoid arthritis;collagen induced arthritis;usnea extract;HMGB1;RAGE;cellular autophagy;inflammatory injury;rats