文献类型：期刊文献

中文题名：骨痨愈康丸调控Th17/Treg平衡抑制类风湿关节炎骨破坏机制

英文题名：GulaoYukang Pill inhibits the bone destruction of rheumatoid arthritis by regulating Th17/Treg balance

作者：杨芳[1];王涛[1];王佳[1];漆文霞[2];李平顺[1];魏勇[1]

第一作者：杨芳

机构：[1]甘肃中医药大学附属医院,甘肃兰州730000;[2]甘肃中医药大学,甘肃兰州730000

第一机构：甘肃中医药大学第二附属医院

年份：2025

卷号：31

期号：8

起止页码：1107

中文期刊名：中国骨质疏松杂志

外文期刊名：Chinese Journal of Osteoporosis

收录：;北大核心:【北大核心2023】；

基金：甘肃省科技计划项目(23JRRA1584,24JRRA548);甘肃省中医药科研课题(GZKP-2022-18)。

语种：中文

中文关键词：类风湿关节炎;骨痨愈康丸;Th17/Treg;骨破坏

外文关键词：rheumatoid arthritis;GulaoYukang pill;Th17/Treg;osteoclasia

摘要：目的探讨骨痨愈康丸通过调控Th17/Treg平衡抑制炎性因子治疗类风湿关节炎骨破坏的作用机制。方法建立胶原诱导性关节炎(collagen-induced arthritis,CIA)动物模型,模型评价成功后采用随机数字表法将大鼠分为6组:正常对照(NC)组、模型对照(model)组、骨痨愈康丸低(0.27 g/kg)、中(0.54 g/kg)、高(0.54 g/kg)剂量组及甲氨蝶呤[0.1 mg/(kg·周)]组,每组10只,持续给药6周后取材。通过HE染色观察大鼠踝关节的病理改变;通过TRAP染色观察破骨细胞的生长情况;通过ELISA法检测细胞因子IL-17、TNF-α和IL-22的表达;通过Western blot检测破骨细胞分化相关蛋白NFATcl、CTSK、MMP-9、CTR及Th17和Treg细胞转录基因RORγt、Foxp3蛋白的表达,通过流式细胞学技术检测Th17及Treg细胞的数量。结果(1)与NC组相比,model组中滑膜组织中可见有大量的炎细胞浸润形成血管翳。在软骨表层则有大量的细胞变性、坏死,骨小梁结构紊乱,破骨细胞数显著升高,IL-17、TNF-α和IL-22的表达升高(P<0.05),破骨细胞分化相关蛋白NFATcl、CTSK、MMP-9、CTR表达水平升高(P<0.05),RORγt的表达升高(P<0.05),Foxp3的表达降低(P<0.05),Th17数量增加,Treg细胞数量减少,Th17/Treg的比值升高(P<0.05);(2)与model组相比,GL、GM、GH、MTX组中大鼠踝关节病变程度随着给药剂量的增加而降低,滑膜组织中的炎细胞浸润减少。在软骨表层细胞变性、坏死减少,骨小梁结构整齐,破骨细胞数也随着给药剂量的增加而减少,IL-17、TNF-α和IL-22的表达降低(P<0.05),破骨细胞分化相关蛋白NFATcl、CTSK、MMP-9、CTR表达水平降低(P<0.05),RORγt的表达降低(P<0.05),Foxp3的表达升高(P<0.05),Th17数量减少,Treg细胞数量增多,Th17/Treg的比值降低(P<0.05)。结论骨痨愈康丸能够抑制类风湿关节炎骨破坏,其机制可能与调控Th17/Treg平衡减少炎症因子的表达,降低破骨细分化相关蛋白的水平有关。
Objective To investigate the mechanism of GulaoYukang Pill in the treatment of rheumatoid arthritis by regulating Th17/Treg balance and inhibiting inflammatory factors.Methods CIA animal model was established.After successful evaluation,the rats were divided into 6 groups by random number table method:Normal control(NC)group,model control(model)group,low(0.27 g/kg),medium(0.54 g/kg),high(0.54 g/kg)dose groups and methotrexate group[(0.1 mg/(kg·w)]with 10 ratsin each group were collected after continuous administration for 6 weeks.The pathological changes of ankle joint were observed by HE staining.The growth of osteoclasts was observed by TRAP staining.The expression of IL-17,TNF-αand IL-22 were detected by ELISA.The expressions of osteoclast differentiation-related proteins NFATcl,CTSK,MMP-9,CTR and transcriptional genes RORγt and Foxp3 proteins of Th17 and Treg cells were detected by Western blot.The number of Th17 and Treg cells was detected by flow cytometry.Results①Compared with the NC group,there were a large number of inflammatory cells infiltrated in the synovial tissue of model group to form pannus.On the surface of cartilage,there was a large number of cell degeneration and necrosis,trabecular structure disorder,and the number of osteoclasts increased significantly.The expression of IL-17,TNF-αand IL-22 was increased(P<0.05);The expression levels of osteoclast differentiation related proteins NFATcl,CTSK,MMP-9 and CTR were increased(P<0.05);The expression of RORγt was increased(P<0.05),Foxp3 expression decreased(P<0.05);The number of Th17 increased,the number of Treg cells decreased,and the Th17/Treg ratio increased(P<0.05).②Compared with model group,the degree of ankle joint lesions in GL,GM,GH and MTX groups decreased with the increase of dose,and the inflammatory cell infiltration in synovial tissue decreased.The degeneration and necrosis of cartilage surface cells decreased,the trabecular structure of bone was orderly,and the number of osteoclasts decreased with the increase of dosage.The expression of IL-17,TNF-αand IL-22 decreased(P<0.05);The expression levels of osteoclast differentiation related proteins NFATcl,CTSK,MMP-9 and CTR were decreased(P<0.05);The expression of RORγt decreased(P<0.05),the expression of Foxp3 was increased(P<0.05);The number of Th17 decreased,the number of Treg cells increased,and the Th17/Treg ratio decreased(P<0.05).Conclusion GulaoYukang Pill can inhibit the bone destruction of rheumatoid arthritis,and the mechanism may be related to the regulation of Th17/Treg balance to reduce the expression of inflammatory factors and reduce the level of osteoclast differentiation related proteins.