苗药五藤膏对膝关节炎兔GSK-3β、Wnt3a、Wnt5a、Wnt7a表达的影响  

Effect of Miao medicine wuteng ointment on GSK-3β,Wnt3a,Wnt5a,Wnt7a expression in rabbits with knee arthritis

作　　者：申海艳 周静 肖丽娜 马武开 姚血明 陈琳 韩珊 SHEN Hai-Yan;ZHOU Jing;XIAO Li-Na(Department of Orthopedics,Guizhou Orthopaedic Hospital,Guiyang 550001,Guizhou,China)

机构地区：[1]贵州省骨科医院骨外科,贵州贵阳550001 [2]贵州中医药大学第二附属医院风湿免疫科 [3]贵州中医药大学

出　　处：《中国老年学杂志》2023年第14期3425-3430,共6页Chinese Journal of Gerontology

基　　金：国家自然科学基金资助项目(81760884);国家自然科学基金委员会青年项目(81804127);贵州卫生计生委科学技术基金项目(gzwjkj2018-1-028);贵州卫生健康委科学技术基金项目(gzwjkj2023-558);贵州省护理学基金项目(GZHLKY202106);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);贵州省中医风湿免疫病临床研究中心(黔科合平台人才[2020]2202号)。

摘　　要：目的基于Wnt/β-连环蛋白(catenin)信号,探讨不同剂量苗药五藤膏贴敷对KOA模型兔软骨组织中糖原合成酶激酶(GSK)-3β蛋白及Wnt3a、Wnt5a、Wnt7a mRNA表达的影响。方法将96只家兔随机分为空白组,模型组,巴布膏对照组,苗药五藤膏低、中、高剂量组;除空白组外均采用木瓜蛋白酶关节腔内注射制备兔KOA模型,空白组正常饲养,不干预;模型组患关节无菌纱布包扎固定,其余各组分别予氟比洛芬巴布膏及不同剂量的苗药五藤膏贴敷干预;给药后第1、2、3、4周各组分别处死动物,取材后观察软骨病理变化及行Mankin评分;采用实时荧光定量逆转录-聚合酶链反应(qRT-PCR)检测关节软细胞中Wnt3a、Wnt5a、Wnt7a mRNA的表达水平;Western印迹检测GSK-3β蛋白的表达水平。结果各组关节软骨病理改变比较,经过4 w干预后,除空白组外,模型组患膝关节面软骨重度退性改变;巴布膏对照组关节面软骨轻度退性改变;苗药五藤膏低、中、高剂量组退变程度与剂量呈正相关;空白组、苗药五藤膏低、中、高剂量组、巴布膏对照组改良版Mankin评分均明显低于模型组(P<0.05),且苗药五藤膏低、中、高剂量组组内改良版Mankin评分比较差异均有统计学意义(P<0.05);与空白组比较,模型组、苗药五藤膏低、中、高剂量组、巴布膏对照组GSK-3β蛋白表达水平明显升高,给药后1、2、3、4 w模型组、苗药五藤膏中、低剂量组、给药后4 w苗药五藤膏高剂量组Wnt3a mRNA明显升高(P<0.05),给药后1、2、3、4w模型组、苗药五藤膏中、低剂量组、给药后1 w苗药五藤膏高剂量组、巴布膏对照组Wnt5a mRNA明显升高,模型组、苗药五藤膏中、低剂量组Wnt7a mRNA表达明显升高(P<0.05,P<0.01);与模型组相比,苗药五藤膏低、中、高剂量组、巴布膏对照组GSK-3β蛋白表达水平明显降低,苗药五藤膏低、中、高剂量组、巴布膏对照组Wnt3a、Wnt5a、Wnt7a mRNA明显降�Objective To investigate the effects of different doses of Miao medicine wuteng ointment on glycogen synthetic kinase(GSK)-3βprotein and expressions of Wnt3a,Wnt5a and Wnt7a mRNA in KOA model rabbit cartilage tissue based on Wnt/β-catenin signal.Methods 96 rabbits were randomly divided into blank group,model group,babu paste control group,low,medium and high dose of Miao medicine wuteng ointment groups.Rabbit KOA model was prepared by intracavascular injection of papain except for the blank group.The blank group was fed normally without intervention.In the model group,the affected joints were fixed with sterile gauze,and the other groups were treated with flurbiprofen cataplasms and different doses of Miao medicine wuteng ointment respectively.Animals in each group were sacrificed at 1,2,3 and 4 w after administration.Pathological changes of cartilage were observed after sampling and Mankin score was performed.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression levels of Wnt3a,Wnt5a and Wnt7a mRNA in joint soft cells.Western blot was used to detect the expression of GSK-3βprotein level.Results Comparison of pathological changes of articular cartilage in each group after 4 w intervention,except for the blank group,the model group had severe degenerative changes in the knee surface cartilage;the articular surface cartilage of babu paste control group was slightly regressive,the degree of degeneration of Miao medicine wuteng ointment in low,medium and high dose groups was positively correlated with dose.The modified version Mankin scores of the blank group,low,medium and high dose of Miao medicine wuteng ointment groups and babu paste control group were significantly lower than those of the model group(P<0.05),there were statistically significant differences in the scores of the modified version of Mankin scores in the low,medium and high dose of Miao medicine wuteng ointment groups(P<0.05).Compared with blank group,the expression levels of GSK-3βprotein were significantly

关 键 词：膝骨关节炎 苗药五藤膏 WNT/Β-CATENIN信号通路 

分 类 号：R285.5[医药卫生—中药学]