Yuan Qin, Kan Wei-Bing, Song Peng-Fei, Zhao Jin, Yu Wei-Guo, Wang Yong-Jun
Department of Orthopaedics, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, China.
Zhongguo Gu Shang. 2012 Sep;25(9):761-5.
To observe beta-catenin expression of Wnt signaling pathway in rats with knee osteoarthritis, and influence of Bushen Huoxue decoction on beta-catenin and MMP-7 expression of synoviocytes in rats with knee osteoarthritis (OA).
Rats model with knee osteoarthritis were established by Hulth method. Primary synoviocytes and OA synoviocytes were cultured with collagenase digestion method. The cultured synoviocytes were divided into normal group, OA model group and Bushen Huoxue decoction group. Western blotting method was used to detect beta-catenin, MMP-7 protein expression of synoviocytes after acting by Bushen Huoxue decoction for 48 h; ELISA method was used to detect MMP-7 expression of synovial supernatant.
OA synoviocytes were cultured successfully. Western blotting showed that beta-catenin, MMP-7 expression in OA synoviocytes was significantly higher than normal group, Bushen Huoxue decoction could significantly reduce beta-catenin, MMP-7 expression; ELISA results showed that MMP-7 expression of OA synovial supernatant was significantly higher than normal synoviocytes supernatant, Bushen Huoxue decoction significantly regulated the level MMP-7 down.
(1) High expression of beta-catenin in OA synoviocytes indicates that Wnt classical signal pathway is activated in rat with knee osteoarthritis; (2) High expression of MMP-7 expression in OA synoviocytes confirms the MMP-7 is downstream genes of Wnt/beta-catenin signal pathway; (3) Activation of Wnt signal pathway and increase of MMP-7 may cause degradation of articular cartilage, and promote the formation of osteoarthritis; (4) Bushen Huoxue decoction can reduce expression of MMP-7, and promote cartilage repair, which may be one of mechanisms of osteoarthritis.
观察Wnt信号通路中β-连环蛋白在膝骨关节炎大鼠中的表达，以及补肾活血汤对膝骨关节炎（OA）大鼠滑膜细胞中β-连环蛋白和基质金属蛋白酶-7（MMP-7）表达的影响。
采用 Hulth 法建立大鼠膝骨关节炎模型。用胶原酶消化法培养原代滑膜细胞和OA滑膜细胞。将培养的滑膜细胞分为正常组、OA模型组和补肾活血汤组。采用蛋白质免疫印迹法检测补肾活血汤作用48 h后滑膜细胞中β-连环蛋白、MMP-7蛋白表达；采用酶联免疫吸附测定法检测滑膜上清液中MMP-7表达。
成功培养出OA滑膜细胞。蛋白质免疫印迹法显示，OA滑膜细胞中β-连环蛋白、MMP-7表达明显高于正常组，补肾活血汤可显著降低β-连环蛋白、MMP-7表达；酶联免疫吸附测定结果显示，OA滑膜上清液中MMP-7表达明显高于正常滑膜细胞上清液，补肾活血汤可显著下调MMP-7水平。
（1）OA滑膜细胞中β-连环蛋白高表达表明膝骨关节炎大鼠Wnt经典信号通路被激活；（2）OA滑膜细胞中MMP-7高表达证实MMP-7是Wnt/β-连环蛋白信号通路的下游基因；（3）Wnt信号通路激活及MMP-7增加可能导致关节软骨降解，促进骨关节炎形成；（4）补肾活血汤可降低MMP-7表达，促进软骨修复，这可能是其治疗骨关节炎的机制之一。
