温阳通络胶囊干预骨关节炎大鼠软骨细胞的炎症反应和血管新生  

Wenyang Tongluo capsule intervenes with inflammation and angiogenesis in rat osteoarthritic chondrocytes

作　　者：李钢[1] 宋芸 梁国光 Li Gang;Song Yun;Liang Guoguang(Guangdong Vocational College of Food and Drug,Guangzhou 510000,Guangdong Province,China;Department of General Practice,School Hospital of South China Normal University,Guangzhou 510000,Guangdong Province,China;Department of Orthopedics,Nanhai Third People’s Hospital,Foshan 528000,Guangdong Province,China)

机构地区：[1]广东食品药品职业学院,广东省广州市510000 [2]华南师范大学校医院全科,广东省广州市510000 [3]佛山市南海区第三人民医院骨科,广东省佛山市528000

出　　处：《中国组织工程研究》2023年第23期3682-3692,共11页Chinese Journal of Tissue Engineering Research

基　　金：广东省中医药局科研项目(20181183),项目负责人:李钢;广东省教育厅科研项目(2021KTSCX235),项目负责人:李钢。

摘　　要：背景:温阳通络胶囊在临床上治疗骨关节炎具有明显的疗效,但其作用机制不明确。目的:探讨温阳通络胶囊对骨关节炎的治疗作用,以及其对软骨细胞中miR-140/血管内皮生长因子信号的调控机制。方法:(1)动物实验:将75只SD大鼠分为5组,每组15只:正常组、模型组、药物组每天左后肢膝关节内注射miR-140-antagomir NC和siRNAVEGF NC,抑制剂组每天左后肢关节内注射mi R-140-antagomir和si RNA-VEGF NC,干扰组每天关节内注射mi R-140-antagomir和si RNA-VEGF,各组均注射2周;2周后,药物组、抑制剂组和干扰组灌胃给予温阳通络胶囊,连续灌胃4周;模型组、药物组、抑制剂组、干扰组实验的1,4,7 d关节腔内注射木瓜蛋白酶复制关节炎模型。给药结束后,进行相关检测。(2)细胞实验:将软骨细胞ATDC5分5组处理:正常组加入含mi R-140-antagomir NC和si RNA-VEGF NC的培养基;模型组加入含白细胞介素1β、mi R-140-antagomir NC和si RNA-VEGF NC的培养基;药物组加入含白细胞介素1β、mi R-140-antagomir NC、si RNA-VEGF NC及温阳通络胶囊的培养基;抑制剂组加入含白细胞介素1β、mi R-140-antagomir及温阳通络胶囊的培养基;干扰组加入含白细胞介素1β、mi R-140-antagomir、si RNA-VEGF及温阳通络胶囊的培养基,进行相关检测。结果与结论:(1)动物实验:与正常大鼠相比,骨关节炎大鼠软骨组织中miR-140 mRNA表达降低(P <0.05),血管内皮生长因子mRNA表达升高(P <0.05)。与模型组相比,药物组、抑制剂组、干扰组大鼠血清中白细胞介素6和肿瘤坏死因子α水平、软骨细胞凋亡、新生血管数量、血管内皮生长因子蛋白表达均明显减少(P <0.05),其中以药物组效果最明显。(2)细胞实验:与正常软骨细胞相比,骨关节炎软骨细胞中miR-140 mRNA表达降低(P <0.05),血管内皮生长因子mRNA表达升高(P <0.05)。与模型组相比,药物组、抑制剂组、干扰组细胞增殖活性提高(P <0.05),细�BACKGROUND:Wenyang Tongluo capsule has obvious clinical effects in the treatment of osteoarthritis,but its mechanism of action is unclear.OBJECTIVE:To investigate the therapeutic effect of Wenyong Tongluo capsule on osteoarthritis and its regulation mechanism on microRNA-140(miR-140)/vascular endothelial growth factor(VEGF) signaling in chondrocytes.METHODS:(1) Animal experiments:A total of 75 Sprague-Dawley rats were randomized into 5 groups(n=15 per group):normal group,model group,drug gro up,inhibitor group,and interference group.In the latter three groups,miR-140-antagomir NC and siRNA-VEGF NC,miR-140-antagomir and siRNA-VEGF NC,and miR-140-antagomir and siRNA-VEGF were injected into the left hindlimb joint daily,res pectively.Administration in each group lasted for 2 weeks.Afte rwards,the rats in the drug,inhibitor,and interference groups were intragastrically given Wenyong Tongluo capsule for 4 continuous wee ks.Animal models of osteoarthritis were prepared via the intraarticular injection of papain on days 1,4,and 7 in the model,drug,inhibitor,and interference groups.After the completion of drug administration,related measurements were performed.(2) Cell assays:ATDC5 chondrocytes were divided into five groups:normal group(cultured in the medium containing miR-140-antagomir NC and siRNA-VEGF NC),model group(cultured in the medium containing interleukin-1β,miR-140-antagomir NC and siRNA-VEGF NC,drug group(cultured in the medium containing interleukin-1β,miR-140-antagomir NC,siRNA-VEGF NC,and Wenyong Tongluo capsule),inhibitor group(cultured in the medium containing interleukin-1β,miR-140-antagomir and Wenyang Tongluo capsule),and interference group(cultured in the medium containing interleukin-1β,miR-140-antagomir,siRNA-VEGF and Wenyang Tongluo capsule).Relevant detections were then performed.RESULTS AND CONCLUSION:(1) Animal expe riments:Compared with normal rats,the expression of miR-140 mRNA in cartilage tissue of osteoarthritic rats was decreased(P <0.05),while the expression of vascular endothelial

关 键 词：骨关节炎 温阳通络胶囊 微小RNA-140 血管内皮生长因子信号 软骨细胞 

分 类 号：R459.9[医药卫生—治疗学] R313[医药卫生—临床医学] R684.3