苗药“四大血”通过IL17A-p38MAPK-NF-κB信号通路对类风湿性关节炎因子MMPs和Cyr61的影响  

Effect of Guizhou Miao medicine formula"Sidaxue"on rheumatoid arthritis factors MMPs and Cyr61 via IL17A-p38MAPK-NF-κB signaling pathway

作　　者：陈燕子 郭婉玲 张振鹏 张丹 罗盛岚 杨永馨 吴宁 吴遵秋 CHEN Yanzi;GUO Wanling;ZHANG Zhenpeng;ZHANG Dan;LUO Shenglan;YANG Yongxin;WU Ning;WU Zunqiu(School of Basic Medicine Science,Guizhou Medical University,Guiyang 550004,Guizhou,China;School of Pediatrics,Guizhou Medical University,Guiyang 550004,Guizhou,China;School of Clinical Medicine,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学基础医学院,贵州贵阳550004 [2]贵州医科大学儿科学院,贵州贵阳550004 [3]贵州医科大学临床医学院,贵州贵阳550004

出　　处：《贵州医科大学学报》2025年第2期199-204,211,共7页Journal of Guizhou Medical University

基　　金：国家自然科学基金项目(82260829);贵州省中药管理局项目(QZYY-2022-014);贵州省大学生创新创业计划训练项目(S202110660038)。

摘　　要：目的通过研究IL17A-p38MAPK-NF-κB信号通路对类风湿性关节炎因子基质金属蛋白酶(matrix metalloproteinase,MMPs)和半胱氨酸蛋白61(cysteine-rich 61,Cyr61)的影响,探讨贵州苗医验方“四大血”(SiDaXue,SX)缓解类风湿性关节炎症状的机制。方法将42只SD大鼠随机均分为6组,除7只作为空白对照组(Nor组)外,余35只SD大鼠均进行胶原诱导的关节炎(collagen induced arthritis,CIA)动物模型造模;造模成功后,再均分为SX高、中、低剂量组(40、20、10 g/kg SX)、动物模型对照组(Mod组)及雷公藤多苷片(triptery-gium glycosides,GWT)阳性对照组(GTW组,40 mg/kg GWT);用HE染色法观察不同组别中大鼠滑膜血管的组织病理学特征,RT-PCR、Western bolt法检测膝踝关节滑膜组织中p38丝裂原活化蛋白激酶(P38 mitogen-actived protein kinase,p38MAPK)、Cyr61、MMP-13及激活蛋白-1(activator protein-1,AP-1)mRNA、蛋白表达水平,ELISA法检测血清中MMP-9的变化。结果光镜下大鼠滑膜血管的组织病理学切片显示,与Mod组相比,SX中、低剂量组的滑膜组织增生和炎症细胞浸润较少,SX高剂量组和GTW组滑膜组织中炎症细胞浸润、滑膜细胞增生、软骨和骨质破坏等现象明显减少;与Mod组比较,GTW组和SX高剂量组滑膜组织中的p38 MAPK、Cyr61、MMP13、AP-1 mRNA及其蛋白表达水平均明显下降(P<0.01),大鼠血清中的MMP-9的表达量也明显降低(P<0.01)。结论贵州苗医验方SX能拮抗类风湿性关节炎病情的发展,其作用机制可能与抑制IL17A-p38MAPK-NF-κB信号通路的激活,从而下调基质金属蛋白酶MMP-9、MMP-13和细胞外基质蛋白Cyr61以及AP-1的表达有关。Objective To investigate the mechanism by which"Sidaxue"(SX),a Guizhou Miao medicine formula alleviates the symptoms of rheumatoid arthritis(RA)by studying the effects of the IL17A-p38MAPK-NF-κB signaling pathway on matrix metalloproteinases(MMPs)and cysteine-rich 61(Cyr61)in rheumatoid arthritis.Methods Forty-two SD rats were randomly divided into six groups.Seven rats served as blank control group(Nor group),and the remaining 35 rats were used for collagen-induced arthritis(CIA).After successful modeling,the mice were divided into high-,medium-,and low-dose SX groups(40,20,and 10 g/kg SX,respectively),model control group(Mod group)and Tripterygium wilfordii glycosides positi ve control group(GTW group,40 mg/kg GWT).HE staining was used to observe histopathological changes of rat synovial vessels.RT-PCR and Western blot were used to detect the mRNA and protein expression levels of p38 mitogen-activated protein kinase(p38MAPK),Cyr61,MMP-13,and activator protein-1(AP-1)in the synovial tissues of the knees and ankle joints,respectively.ELISA was used to measure serum MMP-9 change.Results Histopathological examination of rat synovial tissues under a light microscope showed that medium-and low-dose SX groups exhibited less synovial hyperplasia and inflammatory cell infiltration than those in Mod group.High-dose SX and GTW groups showed significantly reduced inflammatory cell infiltration,synovial cell hyperplasia and cartilage and bone destruction,etc.When compared to Mod group,GTW,and high-dose SX groups had significantly decreased mRNA and protein expression levels of p38MAPK,Cyr61,MMP-13,and AP-1(P<0.01)and reduced serum MMP-9 levels(P<0.01).Conclusion Guizhou Miao herbal medicine formula SX can counteract the progression of rheumatoid arthritis.Its mechanism of action may be related to inhibiting the activation of IL17A-p38MAPK-NF-κB signaling pathway,thereby downregulating the expressions of MMP-9,MMP-13,extracellular matrix protein Cyr61and AP-1.

关 键 词：类风湿性关节炎 苗医方“四大血” 基质金属酶 半胱氨酸蛋白61 激活蛋白-1 

分 类 号：R593.22[医药卫生—内科学] R364.5[医药卫生—临床医学]