Résumé
Objective: To investigate the effect of KR-12 analog, KR-12-a5, on osteogenic differentiation of human bone marrow mesenchymal stem cells (HBMSCs). Methods: Osteogenic differentiation-related experiments were performed using HBMSCs. KR-12-a5 was used as additional stimulation under osteogenesis inducing environment. By alkaline phosphatase (ALP) staining assay, ALP quantitative assay, alizarin red staining and elution quantitative analysis, and real-time quantitative polymerase chain reaction (RT-qPCR), the effect of KR-12-a5 on osteogenic differentiation of HBMSCs was verified. Results: As the concentration of KR-12-a5 increased, the staining intensity of ALP and alizarin red also increased. The strongest staining effect was exhibited at the highest concentration (50 μg/mL) of KR-12-a5. Quantitative tests showed similar results. After 3 days of stimulation with KR-12-a5, the mRNA level of RUNX2 increased in a dose-dependent manner. On the 7th day, the expression levels of ALP, COL1A1, BSP and BMP2 in the KR-12-a5-treated group showed significant changes. On the 14th day, OSX, OCN and OPN levels increased significantly. Conclusion: KR-12-a5 promoted the osteogenic differentiation of HBMSCs in a dose-dependent manner.
Résumé
Objective To investigate the differences in eNOS gene expression,activity and its metabolites before and after human bone marrow mesenchymal stem cells (hBMSCs) are induced into vascular endothelial cells.Methods hBMSCs were induced into vascular endothelial cells.The morphological changes of the cells were observed under inverted microscope.Transwell assay was used to detect the cells' migration ability.The protein expression of eNOS was detected by immunofluorescence and Western blot.The activity of eNOS was detected by ELISA and the content of NO in cell culture supernatant was determined by nitrate reduction method.Results Compared with those in the undifferentiated group,the morphological changes of the differentiated cells were obvious.Cell migration ability increased by 238.10％ (73.000±7.002 vs.21.000±4.359,P＜0.05).The expression of eNOS protein increased by 114.72％ (0.423±0.011 vs.0.197±0.079,P＜0.05).The activity of eNOS was enhanced by 157.49％ (4.967±0.073 vs.1.929±±0.103,P＜0.05).The synthesis and release of NO increased by 155.67％ (184.909±1.853 vs.72.323±0.426,P＜0.05).Conclusion After hBMSCs are induced into endothelial cells,the expression of eNOS gene increases,their activities increase,synthesis and release of the metabolite NO increase.It may provide a basis for prevention and treatment of cardiovascular diseases with stem cells.