苗药五藤膏外敷对膝骨关节炎兔模型的治疗作用及机制  

Therapeutic effect and mechanism of external application of Wuteng ointment on rabbit knee osteoarthritis

作　　者：韩珊 周静 马武开 肖丽娜 姚血明 唐芳 申海艳 陈琳 王颜君 HAN Shan;ZHOU Jing;MA Wukai;XIAO Lina;YAO Xueming;TANG Fang;SHEN Haiyan;CHEN Lin;WANG Yanjun(Department of Rheumatology and Immunology,The Second Affiliated Hospital of Guizhou University of Chinese Medicine,Guiyang 550001,China;不详)

机构地区：[1]贵州中医药大学第二附属医院风湿免疫科,贵阳550001 [2]贵州省骨科医院疼痛科 [3]贵州中医药大学研究生院

出　　处：《山东医药》2023年第7期1-6,共6页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81760884);国家自然科学基金青年项目(81804127);贵州卫生计生委科学技术基金项目(gzwjkj2018-1-028);贵州省中医药(民族医药)治疗风湿病医学平台建设项目(黔科合平台人才[2018]5707);国家重点研发计划中医药现代化研究重点专项(2017YFC1703904);贵州省科技计划项目(黔科合平台人才[2020]2202号)。

摘　　要：目的 探讨苗药五藤膏外敷对膝骨关节炎(KOA)兔模型的治疗作用及机制。方法 采用随机数字表法将96只家兔分为空白对照组、模型组、巴布膏组及五藤膏低、中、高剂量组,每组16只。除空白对照组外,其余组用4%木瓜蛋白酶生理盐水溶液制备KOA模型。造模第6周,空白对照组、模型组每日给予无菌纱布包扎固定右膝关节;巴布膏组给予氟比洛芬巴布膏贴敷于兔右膝关节前方;苗药五藤膏低、中、高剂量组分别给予1、2、4 g五藤膏贴敷于兔右膝关节前方。均用绷带包扎固定4 h,每日给药1次,干预4周。分别于给药1、2、3、4周每组各取4只家兔处死,取右侧股骨内髁顶点钙化层以上软骨。HE染色后观察软骨组织形态变化,用Mankin’s评分评价膝关节软骨组织病理学改变程度;用qRT-PCR法检测膝关节软骨组织中Wnt/β-catenin信号通路下游靶基因(GSK-3β、MMP-3、OPG)及Notch信号通路下游靶基因(Notch1)表达。结果 空白对照组关节软骨表面光滑平整,结构清晰;模型组关节软骨表面粗糙不平整,结构模糊,软骨细胞数目少且分布紊乱;五藤膏高剂量组关节软骨表面较光滑平整,结构较清晰,软骨细胞数目较多且分布较均匀;苗药五藤膏低、中剂量组及巴布膏组伴不同程度的关节肿胀及滑膜充血,层次结构较模糊,软骨细胞数量较少且分布轻度紊乱。与空白对照组比较,模型组膝关节软骨组织Mankin’s评分高(P<0.05);与模型组比较,各治疗组Mankin’s评分低(P均<0.05),且五藤膏高剂量组Mankin’s评分最低(P均<0.05)。与空白对照组比较,模型组膝关节软骨组织中GSK-3β、MMP-3、Notch1 mRNA表达高(P均<0.05),OPG mRNA表达低(P<0.05);与模型组比较,各治疗组GSK-3β、MMP-3、Notch1 mRNA表达低(P均<0.05),OPG mRNA表达高(P均<0.05),且五藤膏高剂量组以上指标变化最明显(P均<0.05)。结论 苗药五藤膏可改善KOA兔膝关节软骨及软骨下病变,Objective To investigate the therapeutic effect and mechanism of the external application of Miao medicine Wuteng ointment on rabbit knee osteoarthritis(KOA). Methods Ninety-six rabbits were divided into the blank control group, model group, babu ointment group, low-dose, medium-dose, high-dose Wuteng ointment groups, with 16 rabbits in each group. Except for the blank control group, rabbit KOA cartilage models were established with 4% papain saline solution in the remaining groups. In the 6th week of modeling, the rabbits in the blank control group and the model group were given sterile gauze bandaging and fixation of the right knee joint every day. The rabbits in the low-dose, medium-dose and high-dose groups of Wuteng ointment were given 1, 2 and 4 g of Wuteng plaster in front of the right knee joint of the rabbit. The rabbits in the babu plaster group were given flurbiprofen and the plaster was applied to the front of the right knee joint. All were fixed with bandages for 4 h, administered once a day, and intervened for 4 weeks. Four rabbits were sacrificed in each group at 1, 2, 3, and 4 weeks of administration, and cartilage above the calcified layer at the apex of the right femoral medial condyle was taken. After HE staining, cartilage tissue morphological changes were observed, and the degree of histopathological changes of knee cartilage was evaluated by Mankin’s score, and the expression levels of Wnt/β-catenin signaling pathway downstream target genes GSK-3β, MMP-3, OPG and Notch signaling pathway downstream target gene Notch1 in the knee cartilage tissues were detected by qRT-PCR. Results The surface of articular cartilage in the blank control group was smooth and flat, and the structure was clear. In the model group, the surface of articular cartilage was rough and uneven, the structure was blurred, the number of chondrocytes was small and the distribution was disordered. The rabbits in the high-dose group of Wuteng ointment had smooth and flat articular cartilage surface, clearer structure, and

关 键 词：膝骨关节炎 苗药五藤膏 外敷法 Wnt/β-catenin信号通路 NOTCH信号通路 兔 

分 类 号：R683.4[医药卫生—骨科学]