骨碎补总黄酮对骨关节炎家兔骨髓间充质干细胞软骨定向分化的实验研究  

Experimental study of the proliferation,Chondrocyte differention .Effects of Gusuibu mesenchymal mtem cells from models with knee osteoarthritis

作　　者：尚平[1] 贺宪[1] 安耀武[1] 江永发[1] 杨俊龙[1] 陈晓陇[1] 潘勇泉[1] 赖震[1] 

机构地区：[1]南方医科大学附属花都医院骨科(广州市花都区人民医院骨科),广东广州510800

出　　处：《生物骨科材料与临床研究》2009年第6期10-13,共4页Orthopaedic Biomechanics Materials and Clinical Study

基　　金：广州市卫生局资助课题(No.2006A38)

摘　　要：目的观察骨碎补总黄酮(Gusuibu,GSB)对骨性关节炎(Osteoarthritis,OA)家兔骨髓间充质干细胞(Mesen-chymal mtem cells,MSCs)体外扩增及向软骨细胞分化的影响,深入探讨骨碎补防治OA的作用机理。方法选用6月龄家兔采用Hulth法建立OA模型。运用全骨髓贴壁法和密度梯度离心法分离培养OA家兔MSCs,通过流式细胞仪对其表面标志测定鉴定MSCs。运用MTT法检测不同浓度的GSB对OA家兔MSCs增殖的影响,以确定最佳促增殖添加剂量。运用流式细胞仪对细胞周期的测定来探讨GSB对OA家兔MSCs增殖周期的影响。将OA家兔MSCs分为空白对照组、经典诱导组、GSB组和经典诱导+GSB组。用免疫组织化学方检测Ⅱ型胶原的表达,阿利新蓝比色法测定GAG含量来检测GSB对OA家兔MSCs体外成软骨细胞的影响。结果经MTT法检测GSB对MSCs增殖作用,发现0.2mMGSB组的OD值较其它组明显提高。流式细胞仪对细胞周期的测定显示0.2mMGSB干预的MSCs细胞周期的比率最高(<0.05)。GSB和成软骨诱导液能增加OA家兔MSCs的活性,Ⅱ型胶原的表达,促进软骨的形成(<0.05)。结论GSB能明显促进OA家兔MSCs增殖;促进OA家兔MSCs软骨向分化。Objiective To sdudy the proliferation and chondrocyte differention effect of Gusuibu on MSCs in Osteoarthritis rabbits, and to illuminate the possible mechanisms. Methods The animal model of osteoarthritis was induced by Hulth method in 6-month-old rabitts. MSCs were isolated from the rabitts by means of the density-gradient centrifugation and cell attachment to tissue culure plates.The cell surface antigens were detected by flow cytometry for identiflication of the MSCs. Mtt method was used to investigate the proliferation efects .Gusuibu on MSCs in rabbits, and to determine the optimal concentration of Gusuibu. The third passage MSCs were divided into control group ,induced group and Gusuibu group,to illuminate the effect of Gusuibu on chondrocyte differention of MSCs. The expressions of collagen Ⅱ and content of GAG were to examinate the effect of chondrocyte differention. Results The Gusuibu had the proliferation effects on MSCs in rabbits.The best promotion of proliferation on MSCs was achieved when the concentration of Gusuibu was 0.2 raM. 0.2 mm Gusuibu can increase proliferation index of Osteoarthritis Rabbit MSCs （P〈0.05）. MSCs with cultured medium or Gusuibu could increase the expression of type II collagen.Also there was a significant increase in GAG compared to the control group （P〈0.05） Conclusion Gusuibu powerfully promotes proliferation of MSCs and induces chondrocyte differentiou Of MSCs derived from osteoarthritis rabbits.

关 键 词：骨髓间充质干细胞 骨性关节炎 骨碎补总黄酮 分化 成软骨细胞 

分 类 号：R285.5[医药卫生—中药学]