[Effects of VEGF-Notch Signaling Pathway on Proliferation and Apoptosis of Bone Marrow MSC in Patients with Aplastic Anemia]

To evaluate the regulation of VEGF-Notch signaling pathway on proliferation and apoptosis of mesenchymal stem cells (MSC) in the patients with aplastic anemia (AA).The bone marrow specimens of AA patients were collected for isolation and identification of BM MSC. Westen blot was used to detect the expression of VEGF-Notch signaling pathway-related proteins (VEGF, VEGFR, Notch 1, Jagged 1, Delta-like 1 and Hes1). The VEGF (100 ng/ml) and DAPT (γ-secretase inhibitor, 10 μmol/L) were respectively added into MSC culture system in oder to activate and inhibit the signaling transduction of VEGF-Notch in BM MSC. The proliferation, apoptosis and cell cycle of MSC in AA patients were detected by CCK8 assay and flow cyfometry. The adipogenic differentiation of BM MSC was detected by oil red O staining.The VEGF-Notch signaling pathway was significantly inhibited in AA BM tissues and AA MSC (P＜0.05) detected by Western blot. The intervention of VEGF and DAPT significantly activated and inhibited VEGF-Notch signaling in AA MSC, respectively. CCK8 assay showed that VEGF intervention significantly promoted the proliferation of MSC in AA patients (P＜0.05). Flow cytometry showed that VEGF significantly inhibited apoptosis of MSCs by blocking S phase cells (P＜0.05).The activation of VEGF-Notch can restore the proliferation function of MSC in AA patients.VEGF-Notch信号通路对再生障碍性贫血患者骨髓MSC增殖和凋亡的作用.评估VEGF-Notch信号通路对再生障碍性贫血（aplastic anemia，AA）患者骨髓间充质干细胞（mesenchymal stem cell, MSC）增殖和凋亡的调节作用.收集再生障碍性贫血患者骨髓标本，进行骨髓MSC分离及鉴定，应用Western blot检测AA患者骨髓（BM）中VEGF-Notch信号通路相关蛋白（VEGF，VEGFR，Notch-1，Jagged1，Delta-like1和hes1）的表达。在MSC培养体系中分别加入VEGF（100 ng/ml）和DAPT（γ-secretase inhibitor）（10μmol/L），以活化和抑制MSC中的VEGF-Notch信号传导。利用细胞计数试剂盒8和流式细胞术评估AA患者MSC的细胞增殖、凋亡和细胞周期；应用油红0染色法检测成脂分化.AA患者MSC中VEGF-Notch信号通路受到明显抑制（P＜0.05）。VEGF和DAPT的干预分别激活和抑制AA患者MSC中的VEGF-Notch信号传导。CCK8检测结果显示，VEGF的干预明显促进MSC的增殖（P＜0.05）。流式细胞仪检测显示，VEGF通过阻断S期细胞显著抑制MSC的凋亡（P＜0.05）.VEGF-Notch信号通路的激活有可能恢复AA患者MSC的增殖功能.