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目的观察不同浓度血管内皮生长因子(vascular endothelial growth factor,VEGF)对少突胶质前体细胞(oli-godendrocyte precursor cells,OPCs)增殖的影响,探索VEGF促进OPCs体外增殖与Notch信号通道上相关蛋白的关系。方法分离纯化培养OPCs,MTT法检测不同浓度VEGF(50、100、200 ng/ml)促进OPCs增殖的作用以及加入Notch通路γ-分泌酶抑制剂DAPT(VEGF 100 ng/ml+DAPT50μmol/ml)后OPCs增殖变化情况,RT-PCR、Western blot技术检测VEGF处理OPCs后Notch信号通道上Notch-1、Hes-1的基因表达和蛋白表达情况。结果 50、100、200 ng/ml VEGF处理组的细胞增殖率分别为(107±2)%、(124±2)%、(142±7)%,与空白对照组相比差异具有统计学意义(P<0.05);VEGF+DAPT处理组的增殖率下降至(103±4)%(P<0.05)。RT-PCR及Western blot分析显示VEGF处理OPCs后,Notch-1、Hes-1蛋白在基因水平和蛋白水平表达均增加(P<0.05),γ-分泌酶抑制剂DAPT能抑制其表达(P<0.05)。结论 VEGF对OPCs具有促进增殖的作用,其促增殖作用与Nocth信号通路相关蛋白有关。
Objective To observe the effects of different concentrations of vascular endothelial growth factor (VEGF) on the proliferation of oligodendrocyte precursor cells (OPCs) and to explore the correlation between the effect of VEGF on promoting the proliferation of OPCs and the expression of related proteins on Notch signaling pathway Relationship. Methods The proliferation of OPCs was isolated and purified. The proliferation of OPCs was stimulated with different concentrations of VEGF (50, 100, 200 ng / ml) by MTT assay and the effects of DAPT (100 ng / ml VEGF + DAPT 50 μmol / ml) The expression of Notch-1 and Hes-1 mRNA and protein in Notch signaling pathway were detected by RT-PCR and Western blot. Results The cell proliferation rates were (107 ± 2)%, (124 ± 2)% and (142 ± 7)% in 50, 100 and 200 ng / ml VEGF groups, respectively, which were significantly different from those in blank control group (P <0.05). The proliferation rate of VEGF + DAPT group decreased to (103 ± 4)% (P <0.05). RT-PCR and Western blot analysis showed that the expression of Notch-1 and Hes-1 protein at the gene and protein levels increased (P <0.05), and the γ-secretase inhibitor DAPT inhibited the expression of OPCs (P < 0.05). Conclusion VEGF can promote the proliferation of OPCs, and its role in promoting proliferation is related to the Nocth signaling pathway related proteins.