摘要目的 观察WNT激活剂SB-216763是否具有促进人骨髓基质干细胞(hMSCs)向软骨细胞分化的功能.方法 6例hMSCs,将成软骨细胞诱导分化的hMSCs与SB-216763共同培养,光镜下计数Alcian blue染色阳性的软骨岛的数目、以Alcian blue染色法定量检测由hMSCs分化而来的软骨细胞分泌硫酸化糖胺聚糖的能力,以逆转录-聚合酶链反应(RT-PCR)检测软骨细胞标志基因SOX9、Collagen Ⅱ、Aggrecan的表达变化.结果 在hMSCs成软骨诱导分化过程中,SB-216763可以显著增加软骨岛的数目(133.17±16.58比9.67±1.87,P<0.01),显著增加hMSCs分化的软骨细胞分泌硫酸化糖胺聚糖的量(1.6～2.6倍,P<0.01),并促进SOX9、Collagen Ⅱ和Aggrecan基因的表达.结论 SB-216763可以促进体外hMSCs向软骨细胞的分化和成熟.

abstractsObjective To investigate the effects of a WNT signaling pathway activator, SB-216763,on chondrocytogenesis of human bone marrow stromal cells (hMSCs) in vitro. Methods Fresh bone marrow samples were obtained with institutional review board (IRB) approval as discarded femoral tissue. Low-density mononuclear cells were isolated by density centrifugatian on Ficoll/Histopaque 1077. Adherent hMSCs were expanded 2 passages. Upon near-confluence of hMSCs ,medium was changed to α-MEM with chondrogenic supplements (1% ITS~(+1), 10~(-8) mol/L dexamethasone,50 mg/L ascorbate-2-phosphate, 40 mg/L proline,100 mg/L pyruvate) with or without 5 μmoL/L SB-216763. After 10 days,the cultures were stained with 0.1% alcian blue in 3% acetic acid (pH=1.0) for sulfated glycosaminoglycans and the number of alcian-blue-positive nodules was counted with 4×magnification, hMSCs from five women and one man, aged 57-77 years,wero used to measure content of sulfated glycesaminoglycans by absorbance (A). After 14 days, the effects of SB-216763 on the expression of chondrocyte signature genes were quantified with reverse transeription polymerase chain reaction (RT-PCR). Results During the process of induction differentiation of hMSCs into chondrocytes ,SB-216763 could produce more cartilage nodules (133.17±16.58) than the control group (9.67±1.87 ) ( P<0.01). SB-216763 stimulated the production of sulfated glycosaminogly-cans by 1.6 to 2.6-fold as indicated by alcian blue staining (P<0.01). Consistent with the matrix data, analysis of chondrocyte signature genes showed that the expression of SOX9, collagen Ⅱ, and aggrecan was enhanced by 5 μmol/L SB-216763. Conclusion The WNT signaling pathway activator,SB-216763 stimu-lates in vitro chondrocytogenesis and cartilage matrix production by hMSCs.