汉黄芩素对胶原诱导性关节炎大鼠关节炎症影响的内质网应激途径  

Effects of wogonin on joint inflammation in collagen-induced arthritis rats via the endoplasmic reticulum stress pathway

作　　者：王瑜茹 李思源 徐烨 张雨蒙 刘杨 郝慧琴[1,2] Wang Yuru;Li Siyuan;Xu Ye;Zhang Yumeng;Liu Yang;Hao Huiqin(Third Hospital of Shanxi Medical University,Shanxi Bethune Hospital,Shanxi Academy of Medical Sciences,Tongji Shanxi Hospital,Taiyuan 030032,Shanxi Province,China;Basic Laboratory of Integrated Chinese and Western Medicine,Shanxi University of Chinese Medicine,Jinzhong 030619,Shanxi Province,China)

机构地区：[1]山西医科大学第三医院(山西白求恩医院,山西医学科学院,同济山西医院),山西省太原市030032 [2]山西中医药大学中西医结合基础实验室,山西省晋中市030619

出　　处：《中国组织工程研究》2025年第5期1026-1035,共10页Chinese Journal of Tissue Engineering Research

基　　金：中西医结合防治风湿免疫病山西省科技创新人才重点团队(202204051002033),项目负责人:郝慧琴;中央引导地方科技发展资金项目(YDZJSX2021A040),项目负责人:郝慧琴;山西中医药大学风湿免疫性疾病中西医结合基础学科建设项目(2023XKJS-03),项目负责人:郝慧琴。

摘　　要：背景:类风湿关节炎是一种炎性疾病,已有多项研究表明汉黄芩素具有较好的抗炎作用,但其对类风湿关节炎的确切疗效和具体作用机制尚待阐明。目的:探讨汉黄芩素通过调节内质网应激途径改善胶原诱导性关节炎大鼠关节炎症的作用机制。方法:①动物水平:雌性Wistar大鼠分为健康对照组、关节炎模型组、汉黄芩素治疗组,通过皮下注射牛Ⅱ型胶原与弗式佐剂构建胶原诱导性关节炎大鼠模型,治疗组在成功造模后给予连续28 d汉黄芩素灌胃治疗;在此期间,每隔7 d对各组大鼠进行体质量、关节炎评分和踝关节肿胀度测量;实验结束后,观察关节病理变化,qRT-PCR和Western Blot、免疫组化检测关节中内质网应激通路GRP78、CHOP mRNA和蛋白表达水平。②细胞水平:CCK-8检测汉黄芩素对胶原诱导性关节炎大鼠成纤维样滑膜细胞的细胞毒作用;不同浓度汉黄芩素处理毒胡萝卜素诱导的成纤维样滑膜细胞,ELISA检测细胞上清白细胞介素1β、肿瘤坏死因子α水平,DCFH-DA探针法测定各组细胞内活性氧簇,qRT-PCR和Western Blot检测GRP78、IRE1α、XBP1s、CHOP mRNA和蛋白水平。结果与结论:①与健康对照组相比,关节炎模型组关节炎指数评分、踝关节肿胀度升高(P<0.01),病理切片可见滑膜增生及炎症细胞浸润、软骨破坏和骨质侵蚀,踝关节GRP78、CHOP mRNA和蛋白表达显著增多(P<0.01),主要定位在滑膜组织和关节面;与关节炎模型组相比,汉黄芩素治疗组关节炎指数评分、踝关节肿胀度降低(P<0.05),滑膜增生及炎症细胞减少,软骨破坏和骨质侵蚀减轻,踝关节GRP78、CHOP mRNA和蛋白表达减少(P<0.05),在滑膜组织和关节面显著降低;3组大鼠体质量无显著差别(P>0.05)。②细胞实验中,200μmol/L汉黄芩素组可明显降低成纤维样滑膜细胞存活率(P<0.01);与空白对照组相比,毒胡萝卜素组细胞上清白细胞介素1β、肿瘤坏死因子α水�BACKGROUND:Rheumatoid arthritis is an inflammatory disease.Many studies have shown that wogonin has a good anti-inflammatory effect on rheumatoid arthritis,but its exact efficacy and specific mechanism of action remain to be clarified.OBJECTIVE:To investigate the mechanism of wogonin ameliorating joint inflammation by regulating endoplasmic reticulum stress pathway in rats with collageninduced arthritis.METHODS:(1)At the animal level:Female Wistar rats were divided into healthy control group,arthritis model group and wogonin treatment group.Rat models of arthritis in the latter two groups were established by subcutaneous injection of bovine type II collagen and adjuvant.In the wogonin group,wogonin was given by gavage for 28 consecutive days after modeling.During this period,the rats in each group were weighed,and arthritis score and ankle swelling were measured every 7 days.After the experiment,the pathological changes of the joint were observed,the mRNA and protein levels of endoplasmic reticulum stress pathway GRP78 and CHOP were detected by qRT-PCR,western blot,and immunohistochemistry.(2)At the cellular level,cell counting kit-8 was used to detect the cytotoxic effect of wogonin on fibroblast-like synoviocytes from rats with collagen-induced arthritis.The fibroblast-like synoviocytes induced by thapsigargin were treated with different concentrations of wogonin.The levels of interleukin-1βand tumor necrosis factor-αin the cell supernatant were detected by ELISA,and the intracellular reactive oxygen species in each group were determined by DCFH-DA probe method.The mRNA and protein levels of GRP78,IRE1α,XBP1s and CHOP were detected by qRT-PCR and western blot,respectively.RESULTS AND CONCLUSION:Compared with the healthy control group,arthritis index score and ankle swelling degree in the arthritis model group were increased(P<0.01),synovial hyperplasia,inflammatory cell infiltration,cartilage destruction and bone erosion were observed in pathological sections,and the mRNA and protein expressions of GRP78 and

关 键 词：汉黄芩素 内质网应激 类风湿关节炎 成纤维样滑膜细胞 炎症反应 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R593.2