来源：风湿病与关节炎,2019,8（2）:5-9.

跳骨片调控Wnt/β-catenin信号通路延缓骨关节炎关节软骨退变的机制研究（基础研究）

何晓娟¹，郑文伟¹，贾良良²，李　慧¹，许丽梅²，曾建伟²，许惠凤²，郑春松²，叶锦霞²，吴广文²，李西海²，叶蕻芝²

　　【摘　要】目的：探讨跳骨片调控Wnt/β-连环蛋白（β-catenin）信号通路保护骨关节炎关节软骨的机制。方法：将30只2月龄雄性SD大鼠随机分为空白对照组、模型对照组、跳骨片组，每组10只。采用改良Hulth法建立骨关节炎动物模型。造模成功后跳骨片组给予跳骨片0.32 g·kg^-1·d^-1灌胃，空白对照组和模型对照组给予等量生理盐水灌胃，干预12周后取大鼠关节软骨。Real-time PCR检测关节软骨β-catenin、Frizzled-2、GSK-3β、CKIε mRNA表达，Western blot法检测关节软骨CKIε、CollagenⅡ、PGS1蛋白表达，Massion染色法观察3组关节软骨胶原的表达及形态变化。结果：Real-time PCR结果显示，与空白对照组比较，模型对照组关节软骨β-catenin、Frizzled-2、CKIε mRNA表达明显上升（P < 0.05，P > 0.05，P < 0.05），而GSK-3βmRNA表达明显下降（P < 0.05）；与模型对照组比较，跳骨片组关节软骨β-catenin、Frizzled-2、CKIε mRNA表达明显下降（P < 0.05，P > 0.05，P < 0.05），而GSK-3β mRNA表达明显上升（P < 0.05）。Western blot结果显示，与空白对照组比较，模型对照组关节软骨CKIε蛋白表达明显升高（P < 0.05），而CollagenⅡ、PGS1蛋白表达明显降低（P < 0.05，P < 0.01）；与模型对照组比较，跳骨片组关节软骨CKI-ε蛋白表达明显降低（P < 0.05），CollagenⅡ、PGS1蛋白表达明显升高（P < 0.01，P > 0.05）。Massion染色结果显示，空白对照组和跳骨片组关节软骨结构清晰，软骨层染色明显，软骨组织呈淡蓝色，胶原含量较高；模型对照组软骨层结构紊乱，染色较浅，胶原含量低。结论：跳骨片能调控Wnt/β-catenin信号通路抑制骨关节炎炎症反应，促进细胞外基质的合成，保护关节软骨。

　　【关键词】　骨关节炎；关节软骨；跳骨片；Wnt/β-catenin信号通路；大鼠

Study on the Mechanism of Tiaogu Tablet（跳骨片）Regulating Wnt/β-catenin Signaling Pathway to Delay Articular Cartilage Degeneration in Osteoarthritis

HE Xiao-juan,ZHENG Wen-wei,JIA Liang-liang,LI Hui,XU Li-mei,ZENG Jian-wei,XU Hui-feng,ZHENG Chun-song,YE Jin-xia,WU Guang-wen,LI Xi-hai,YE Hong-zhi

　　【ABSTRACT】Objective:To investigate the mechanism of Tiaogu Tablet（跳骨片）regulating Wnt/β-cat-enin signaling pathway to protect articular cartilage in osteoarthritis.Methods:Thirty two-month-old male SD rats were randomly divided into a blank control group,a model control group and a Tiaogu Tablet group,with 10 rats in each group.Animal models of osteoarthritis were established by modified Hulth method.The Tiaogu Tablet group was given Tiaogu tablet 0.32 g·kg^-1·d^-1 after successful modeling.The blank control group and the model control group were given the same amount of saline by gastric lavage.After 12 weeks of intervention,the articular cartilage of rats was taken.Real-time PCR was used to detect the expressions of β-catenin,Frizzled-2,GSK-3β and CKIε mRNA in articular cartilage.Western blot was used to detect the expressions of CKIε,CollagenⅡand PGS1 in articular cartilage.Massion staining method was used to observe the expressions and morphological changes of collagen in articular cartilage.Results:Real-time PCR showed that the expressions of β-catenin,Frizzled-2 and CKIε mRNA in articular cartilage of the model control group were significantly higher than those of the blank control group（P < 0.05,P > 0.05,P < 0.05）,while the expression of GSK-3β mRNA was significantly lower（P < 0.05）.Compared with the model control group,the expressions of β-catenin,Frizzled-2 and CKIε mRNA in articular cartilage of the Tiaogu Tablet group were significantly lower（P < 0.05,P > 0.05,P < 0.05）,while the expression of GSK-3β mRNA significantly increased（P < 0.05）.Western blot results showed that CKIε protein expression in articular cartilage of the model control group was significantly higher than that of the blank control group（P < 0.05）,while the expressions of CollagenⅡand PGS1 protein were significantly lower（P < 0.05,P < 0.01）.Compared with model control group,CKI-ε protein expression in articular cartilage of the Tiaogu Tablet group was significantly lower（P < 0.05）,and the expressions of CollagenⅡand PGS1 protein were significantly higher（P < 0.01,P > 0.05）.Massion results showed that the structure of articular cartilage in the blank control group and the Tiaogu Tablet tablet group was clear,the cartilage layer stained was obvious,the cartilage tissue was light blue,and the collagen content was high;while the structure of cartilage layer in the model control group was disordered,the staining was light,and the collagen content was low.Conclusion:Tiaogu Tablet can regulate Wnt/β-catenin signaling pathway to inhibit osteoarthritis inflammation,promote the synthesis of extracellular matrix and protect articular cartilage.

　　【Keywords】 osteoarthritis;articular cartilage;Tiaogu Tablet（跳骨片）;Wnt/β-catenin signaling pathway;rats