温针灸对膝骨关节炎大鼠软骨组织ROCK/LIMK1/Cofilin通路的影响  

Warm acupuncture improves arthritic injury by down-regulating expression of skeleton proteins in rats with knee osteoarthritis

作　　者：彭锐 李静[2] 李佳 李柏村 蔡国伟[2] PENG Rui;LI Jing;LI Jia;LI Bo-cun;CAI Guo-wei(College of Acupuncture and Orthopedics,Hubei University of Chinese Medicine/Hubei Provincial Colla borative Innovation Center of Preventive Treatment by Acupuncture and Moxibustion,Wuhan 430061.China;Department of Acupuncture,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430020)

机构地区：[1]湖北中医药大学针灸骨伤学院/针灸治未病湖北省协同创新中心,武汉430061 [2]华中科技大学同济医学院附属协和医院针灸科,武汉430020

出　　处：《针刺研究》2020年第2期105-110,共6页Acupuncture Research

基　　金：国家自然科学基金项目(No.81774410);湖北省卫健委项目(No.ZY2019M041)

摘　　要：目的:观察温针灸对膝骨关节炎(KOA)大鼠软骨细胞骨架蛋白Rho相关蛋白激酶(ROCK)/单丝氨酸蛋白激酶1(LIMK1)/丝切蛋白(Cofilin)的影响,探讨针灸防治KOA的作用机制。方法:SD大鼠随机分为正常组、模型组、针刺组、艾灸组和温针灸组,每组24只。采用右膝关节腔注射4%木瓜蛋白酶制备KOA大鼠模型。针刺组、艾灸组和温针灸组于造模第1天开始,取右侧"内膝眼""外膝眼"和"足三里"穴,分别采用针刺、艾灸和温针灸治疗,20 min/次,1次/d,共治疗21 d。采用游标卡尺测量右膝关节宽度,排水法检测大鼠右侧足部体积,采用蛋白免疫印迹法检测右侧膝关节软骨组织ROCK、LIMK1和磷酸化LIMK1(p-LIMK1)、Cofilin、磷酸化Cofilin(p-Cofilin)蛋白表达水平。结果:与正常组比较,模型组大鼠右膝关节宽度增加(P<0.05),第6天起右侧足部体积增大(P<0.01),Markin评分升高(P<0.05),右侧膝关节软骨组织ROCK、p-LIMK1和p-Cofilin蛋白表达水平升高(P<0.01)。与模型组比较,针刺组、艾灸组和温针灸组大鼠右膝关节宽度降低(P<0.05),第12天起右侧足部体积减小(P<0.05),Markin评分降低(P<0.05),右侧膝关节软骨组织ROCK、p-LIMK1和p-Cofilin蛋白表达水平降低(P<0.05)。与针刺组和艾灸组比较,温针灸组大鼠右膝关节宽度降低(P<0.05),第15天起右侧足部体积减小(P<0.05),Markin评分降低(P<0.05),右侧膝关节软骨组织ROCK、p-LIMK1和p-Cofilin蛋白表达水平降低更明显(P<0.05)。结论:相对于针刺和艾灸,温针灸更能有效调节KOA大鼠关节炎性损伤状况,其机制可能与降低软骨组织骨架蛋白ROCK、p-LIMK1和p-Cofilin蛋白表达有关。Objective To investigate the effect of warm acupuncture on chondrocyte cytoskeleton protein Rho associa-ted protein kinase(ROCK)/monopherine domain kinase 1(LIMK1)/Cofilin signaling of synovial tissue of the knee-joint in knee osteoarthritis(KOA) rats, so as to explore its mechanisms underlying improvement of KOA. Methods One hundred-twenty SD rats(half male and half female) were randomly divided into 5 groups: normal control, model, acupuncture, moxibustion and warm acupuncture, with 24 rats in each group. The KOA model was established by injection of 4% Papain(0.25 mL/kg) into the right knee cavity on day 1, 3 and 7. Rats in the acupuncture, moxibustion and warm acupuncture groups were treated with manual acupuncture, moxibustion and warm acupuncture stimulation of "Neixiyan"(EX-LE4), "Waixiyan"(EX-LE5) and "Zusanli"(ST36), respectively for 20 min, once a day for 21 days. The volume of the right knee-joint was measured by using drainage method and its width measured using a vernier caliper. The histopathological changes of the right knee cartilage were observed after H.E. stain, and scored(0 to 14 points) with reference to Markin’s methods. The expression levels of ROCK, Cofilin, phospho-Cofilin, LIMK1 and phospho-LIMK1 proteins of the right knee synovial tissue were detected by Western blot. Results After modeling, the width and the volume since day 6 of the right knee-joint and Markin score of the cartilage, as well as the expression levels of ROCK, phospho-Cofilin, and phospho-LIMK1 proteins were significantly increased in the model group in contrast to the normal control group(P<0.05,P<0.01). Following the interventions, the width and the volume since day 12 of the right knee-joint and Markin score of the cartilage, as well as the expression levels of ROCK, phospho-Cofilin, and phospho-LIMK1 proteins were reversed in the three treatment groups(P<0.05). The effect of warm acupuncture was significantly superior to that of both simple acupuncture and simple moxibustion in decreasing the width and the volume

关 键 词：温针灸 针刺 艾灸 骨关节炎 软骨退变 骨架蛋白 

分 类 号：R245.31+5[医药卫生—针灸推拿学]