VEGF与Notch信号通路对再生障碍性贫血患者骨髓间充质干细胞的调控机制研究

目的观察血管内皮生长因子(VEGF)通过Notch信号通路对再生障碍性贫血（AA）患者骨髓造血微环境的影响，探讨VEGF与Notch信号通路对骨髓间充质干细胞（MSC）的调控机制。方法选取AA患者与健康志愿者的骨髓MSC，VEGF干预后分别采用CCK-8法、流式细胞仪检测MSC细胞增殖、分化情况，分别采用Westernblot、qRT-PCR法检测MSCVEGF与Notch信号通路相关蛋白、基因表达情况，采用免疫荧光法检测MSC内皮分化能力。结果CCK-8法检测显示，VEGF的干预可明显上调AA患者MSC的增殖能力。流式细胞仪检测显示，VEGF通过阻断S期细胞促进AA患者MSC的增殖，抑制其凋亡。Westernblot、qRT-PCR法检测显示，AA患者MSCVEGF与Notch信号通路相关蛋白、基因表达受抑制，VEGF的干预可促进MSC的Notch信号通路传导。免疫荧光法检测提示，AA患者MSC内皮分化能力弱于正常MSC。结论VEGF或可通过Notch信号通路调控AA患者MSC，从而改善骨髓造血微环境。

Effect of VEGF on bone marrow mesenchymal stem cells from patients with aplastic anemia by Notch signaling pathway

Objective To investigate the effect of vascular endothelial growth factor ( VEGF ) on bone marrow mesenchymal stem cells ( MSC ) from patients with aplastic anemia ( AA ) and its regulating mechanism. Methods The bone marrow MSCs of AA patients and healthy subjects were isolated and cultured. The proliferation and differentiation of MSCs were detected by CCK-8 and flow cytometry . Western blot and qRT-PCR were used to detect VEGF and Notch signaling pathway-related protein and mRNA expression in MSCs, and immunofluorescence assay was applied to measure the MSC endothelial differentiation. Results CCK-8 assay showed that VEGF intervention significantly up-regulated the proliferation of MSCs from AA patients. Flow cytometry showed that VEGF promoted the proliferation of MSCs of AA patients by blocking S phase cells and inhibiting apoptosis. Western blot and qRT-PCR showed that the expression of VEGF and Notch signaling pathway-related proteins and mRNA was inhibited in MSCs of AA patients. The intervention of VEGF promoted the transmission of Notch signaling pathway in MSCs. Immunofluorescence assay showed that the endothelial differentiation ability of MSCs from AA patients was weaker than that of normal MSCs. Conclusion VEGF may regulate MSCs of AA patients through Notch signaling pathway, thereby improving the bone marrow hematopoietic microenvironment.