小分子药物Kartogenin诱导骨髓间充质干细胞定向分化中基质金属蛋白酶2的表达

Variation of matrix metalloproteinase 2 levels during Kartogenin-induced directional differentiation of human bone marrow mesenchymal stem cells

背景:体外实验发现小分子药物Kartogenin(KGN)能诱导间充质干细胞转变成为软骨细胞,KGN的发现为人们找到了修复软骨的新途径,未来很有希望成为新型骨关节炎治疗药物。目的:通过体外实验观察小分子药物KGN在骨髓间充质干细胞定向分化为软骨细胞的过程中的诱导作用。方法:体外传代培养人骨髓间充质干细胞至对数生长期,分为空白对照组、1μmol/L KGN组和10μmol/L KGN组,3组KGN的终浓度分别为0,1和10μmol/L,培养72 h后通过显微镜检骨髓间充质干细胞的增生和分化情况,用酶联免疫吸附实验检测上清中软骨代谢降解标记物基质金属蛋白酶2水平,用免疫荧光染色技术观察Ⅱ型胶原蛋白表达情况。结果与结论:镜检发现随着KGN药物浓度的增高能明显促进骨髓间充质干细胞的增生和分化;免疫荧光染色结果显示随着KGN浓度的增高Ⅱ型胶原蛋白的表达增加;而随着KGN药物浓度增加,基质金属蛋白酶2的表达随之降低。提示小分子药物KGN能诱导人骨髓间充质干细胞向软骨细胞定向分化,且随着药物浓度的增加,软骨细胞外基质的破坏也随之被抑制。

BACKGROUND：Kartogenin can induce chondrogenic differentiation of mesenchymal stem cel s as reported in in vitro experiments. The discovery of Kartogenin finds a novel path to cartilage repair, and it is expected to develop into a new drug to treat osteoarthritis. OBJECTIVE：To observe the inductive role of Kartogenin in the process of human bone marrow mesenchymal stem cel s differentiating into chondrocytes in vitro. METHODS：In vitro cultured human bone marrow mesenchymal stem cel s were grown to the logarithmic phase, and then divided into control group （0μmol/L Kartogenin）, 1μmol/L Kartogenin group, and 10μmol/L Kartogenin group. After 72 hours of culture, cel proliferation and differentiation were observed microscopical y. Matrix metal oproteinase 2 and type II col agen levels in the cel supernatant were detected by enzyme linked immunosorbent assay and immunofluorescence staining, respectively. RESULTS AND CONCLUSION：Under the microscope, Kartogenin was shown to significantly promote the proliferation and differentiation of human bone marrow mesenchymal stem cel s. With the increase of Kartogenin concentrations, the level of type II col agen was increased, while the level of matrix metal oproteinase 2 was decreased. These findings indicate that Kartogenin can induce human bone marrow mesenchymal stem cel s to differentiate into chondrocytes, and with the increase of Kartogenin concentration, destruction of the cartilage extracel ular matrix may be inhibited.