糖皮质激素影响间充质干细胞成骨分化的实验研究.

Objective To investigate the exact role of Notch signaling pathway in the inhibition of osteogenic differentiation of mesenchymal stem cells(MSCs) by glucocorticoids(GCs). Methods MSCs were cultured. GCs were added to the medium. Cell apoptosis was determined using multi-mode microplate reader by 530 nm. The mRNA expression of Hes1 and osteoblastic markers(ALP, Collagen 1) was determined by real-time PCR. The protein of Hes1 and osteoblastic markers(ALP, Collagen 1) was determined by Western blot. The mineralization was detected by Alizarin red staining. Results After GCs treatment, the number of apoptotic cells was increased [(80±5.789)vs Control(60±7.132), P<0.05]. The mRNA and protein of osteogenic markers ALP and Collagen 1 was decreased compared with the control(P<0.05). The Alizarin staining density was decreased compared with the control(P<0.05). The expression of Notch target gene Hes1 was decreased significantly compared with the control(P<0.05) and the expression of Notch signaling pathway was decreased by GCs significantly. After activation of Notch signaling, the number of apoptotic cells was decreased to 72±2.169(compared with the GCs group, P<0.05). The mRNA and protein of osteogenic markers ALP and Collagen 1 was increased compared with the GCs group(P<0.05). The Alizarin staining density was increased in GCs+JAG1 group compared with the GCs group(P<0.05). Conclusion GCs demolish osteogenic differentiation of MSCs via Notch signaling pathway inhibition. [ABSTRACT FROM AUTHOR]