目的:研究人参中提取的小分子活性物质皂苷Rg1能否促进大鼠骨髓间充质干细胞(marrow—derived mesenchymal stem cells,MSCs)分泌血管内皮生长因子,并进一步探究其作用机制与缺氧诱导因子1α(Hypoxia—induciblefactor,HIF-1)的相关性。方法:用酶联免疫吸附剂测定法(enzyme linked immunosorbent assay,ELISA)观察细胞培养上清中的血管内皮生长因子(vascularen—dothelial growth factor,VEGF)以及蛋白印迹法(western blot)观察细胞中缺氧诱导因子相关蛋白的变化。结果:①通过对照组与人参皂苷Rgl处理组比较发现,1小时前血管内皮生长因子的分泌在两组之间没有明显差异,而在1小时后人参皂苷Rg1处理组相比较于对照组的血管内皮生长因子分泌显著性增加;②进一步实验发现人参皂苷Rgl处理的骨髓问充质干细胞中的HIF—1α在1小时后明显增加,而HIF-1β没有随时间有明显变化。结论:人参皂苷Rgl具有促进骨髓间充质干细胞分泌VEGF的功能,并且发现HIF-1α在这一过程中发挥关键作用,为将来人参皂苷Rgl在临床中的广泛应用提供了重要的理论依据。
Objective: To investigate the promotive effects of Ginsenoside-Rgl on the marrow-derived mesenchymal stem cells producing VEGF via HIF-1α. Methods: Mononuclear cells derived fi'om rat bone marrow were isolated by density-gradient centrifugation and were cultured on fibronectin-coated plates, supplied with bovine pituitary extract. After Ginsenoside-Rg 1 acting on the marrow-derived mesenchymal stem cells, we compared the production of VEGF by ELISA, the protein expression of HIF-1α in different groups by western blot. Results: 0 the production of VEGF had significantly increased with Ginsenoside-Rgl. (2)Rg1 induced a robust accumulation of HIF-1α protein in MSCs under normoxic conditions, which was clearly detectable after 1 h. In contrast, Rgl did not change levels of HIF-1β protein. Conclusion: HIF-1α had crucially effects on the marrow-derived mesenchymal stem cells for VEGF production and it provides important theoretical basis for future ginsenoside Rgl widely used in clinical.