骨痹散通过调节TGF-β/Smad信号通路减轻兔膝骨关节炎损伤  

Gubi powder alleviates knee osteoarthritis injury by regulating transforming growth factor-β/Smad signaling pathway in rabbits

作　　者：李东东 李晓明 吴雪莲 易露 陈光友 LI Dongdong;LI Xiaoming;WU Xuelian;YI Lu;CHEN Guangyou(the Affiliated Traditional Chinese Medicine Hospital of Southwesst Medical University,Luzhou 646000,China.;Department of Rehabilitation Medicine,Southwest Medical University,Luzhou 646000)

机构地区：[1]西南医科大学附属中医医院,四川泸州646000 [2]西南医科大学康复医学系,四川泸州646000

出　　处：《中国比较医学杂志》2023年第7期100-107,共8页Chinese Journal of Comparative Medicine

基　　金：大学生创业国家级课题(202110632059);西南医科大学附属中医医院课题(2018XYLH-019)。

摘　　要：目的探讨骨痹散对骨关节炎的治疗效果及其机制。方法将新西兰大白兔分为空白组、模型组、阳性组(双氯芬酸钾凝胶)与骨痹散组,每组6只,使用改良Hulth法构建兔膝骨关节炎模型,造模8周后给药4周。对软骨组织进行甲苯胺蓝染色后测量软骨厚度,番红-O染色后进行Mankin评分;使用Tunel荧光染色法检测软骨组织中软骨细胞的凋亡情况;使用免疫组织化学法检测软骨组织中ColⅡ与Col X蛋白的表达水平;使用Western blot法检测软骨组织中Bax、cleaved-caspase-3、p-ERK、p-p38、p-Smad2、p-Smad3、TGF-β1的蛋白表达水平。结果与空白组相比,模型组软骨组织厚度减少,Mankin评分升高,软骨细胞凋亡率升高,ColⅡ、p-Smad2、p-Smad3、TGF-β1蛋白表达水平减低,Col X、Bax、cleaved-caspase-3、p-ERK、p-p38蛋白表达升高。与模型组相比,阳性组与骨痹散组软骨组织厚度增加,Mankin评分降低,软骨细胞凋亡率降低,ColⅡ、p-Smad2、p-Smad3、TGF-β1蛋白表达水平升高,Col X、Bax、cleaved-caspase-3、p-ERK、p-p38蛋白表达降低。结论骨痹散对骨关节炎具有良好的治疗效果,能通过减少软骨细胞凋亡,稳定软骨组织结构,促进软骨组织损伤后再生与修复,其机制可能与抑制ERK/P38通路、激活TGF-β/Smad通路有关。Objective To investigate the therapeutic effect and mechanism of Gubi powder on osteoarthritis.Methods New Zealand white rabbits were divided into a blank group,model group,positive group(diclofenac potassium gel),and Gubi powder group(n=6 rabbits per group).A rabbit knee osteoarthritis model was constructed using the modified Hulth method,and the treatments were administered for 4 weeks after 8 weeks of modeling.Cartilage thickness was measured by toluidine blue staining and the Mankin score was derived after Muscovite O staining of cartilage tissue.Terminal deoxynucleotidyl transferase dUTP nick end labeling fluorescence staining was used to detect the apoptosis of chondrocytes in cartilage tissue.Collagen(Col)Ⅱand collagen X protein expression levels in cartilage tissue were detected by immunohistochemistry and expression levels of Bax,cleaved-caspase-3,phospho(p)extracellular signal-regulated kinase(ERK),p-p38,p-Smad2,p-Smad3,and transforming growth factor(TGF)-β1 in cartilage tissues were detected by Western blot.Results Cartilage thickness was decreased,the Mankin score and chondrocyte apoptosis rate were increased,ColⅡ,p-Smad2,p-Smad3,and TGF-β1 protein expression levels were decreased,and Col X,Bax,cleaved-caspase-3,p-ERK,and p-p38 levels were increased in the model group compared with the control group.Cartilage thickness was increased,the Mankin score and chondrocyte apoptosis rate were decreased,ColⅡ,p-Smad2,p-Smad3,and TGF-β1 expression levels were increased,and Col X,Bax,cleaved-caspase-3,p-ERK,and p-p38 protein expression were decreased in the positive and Gubi powder groups compared with the model group.Conclusions Gubi powder showed good therapeutic efficacy against osteoarthritis in rabbits,by reducing chondrocyte apoptosis,stabilizing cartilage tissue structure,and promoting the regeneration and repair of cartilage tissue after injury.Its mechanism may be related to inhibition of the ERK/P38 pathway and activation of the TGF-β/Smad pathway.

关 键 词：骨关节炎 骨痹散 软骨组织 ERK/P38通路 TGF-β/Smad通路 

分 类 号：R-33[医药卫生]