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目的:观察淫羊藿苷(ICA)对骨性关节炎(OA)家兔骨髓间充质干细胞(BMSCs)体外增殖及软骨定向分化的影响。方法:选用6月龄家兔采用Hulth法复制OA动物模型,全骨髓贴壁法分离培养OA家兔BMSCs。根据ICA的不同浓度分为25p M、50p M、0.1p M、0.2p M、0.4p M、0.8p M ICA组,并设空白组,运用MTT法检测不同浓度ICA对BMSCs增殖的影响。将BMSCs分为空白组,4μM、1μM、0.25μM ICA组,经典诱导组及1μM ICA+经典诱导组,运用免疫细胞化学方法检测Ⅱ型胶原的表达,运用阿利新蓝比色法测定各组葡萄糖氨基聚糖(GAG)的含量。结果:全骨髓贴壁分离法可获得优质性较好的BMSCs,体外培养下OA家兔BMSCs软骨定向分化能力下降,而脂向分化能力增强。MTT法检测ICA对BMSCs的增殖作用,结果发现0.1p M、0.2p M、0.4p M、0.8p M ICA组的OD值较空白组高(P<0.01)。4μM、1μM、0.25μM ICA组,经典诱导组及1μM ICA+经典诱导组的Ⅱ型胶原表达的阳性率、CAG含量均高于空白组(P<0.05或P<0.01)。结论:ICA能明显促进OA家兔BMSCs的体外增殖及软骨定向分化。
Objective: To observe the effect of icariin (ICA) on proliferation and differentiation of rabbit bone marrow mesenchymal stem cells (BMSCs) induced by osteoarthritis (OA) in vitro. Methods: 6-month-old rabbits were used to copy the animal models by Hulth method. BMSCs were isolated and cultured in osteoblasts of all rabbits. According to the different concentrations of ICA, they were divided into 25p M, 50p M, 0.1p M, 0.2p M, 0.4p M and 0.8p M ICA groups, and blank control group was established. The effects of different concentrations of ICA on the proliferation of BMSCs were detected by MTT assay. BMSCs were divided into blank group, 4μM, 1μM, 0.25μM ICA group, classical induction group and 1μM ICA + classical induction group. The expression of type Ⅱ collagen was detected by immunocytochemical method. Glucosamine Glycan (GAG) content. Results: BMSCs with good quality could be obtained by whole bone marrow adherence separation method. The ability of directional differentiation of cartilage in BMSCs of OA rabbits decreased in vitro and the ability of adipogenic differentiation increased. The proliferation of BMSCs was detected by MTT assay. The results showed that the OD values of 0.1μM, 0.2μM, 0.4μM and 0.8μM ICA groups were higher than that of blank group (P <0.01). The positive rate and CAG content of type Ⅱ collagen in 4μM, 1μM, 0.25μM ICA group, classical induction group and 1μM ICA + classic induction group were higher than those in blank group (P <0.05 or P <0.01). Conclusion: ICA can obviously promote the proliferation and differentiation of BMSCs in OA rabbits in vitro.