低分子肝素和Galectin-3对血管内皮细胞迁移和增殖的影响

目的 探究低分子肝素联合半乳糖凝集素-3(Galectin-3)对骨髓间充质干细胞(MSCs)来源的血管内皮细胞迁移和增殖的影响.方法 密度梯度离心法分离纯化SD大鼠骨髓MSCs,取第3代,加入血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(bFGF),体外诱导14 d,进行免疫组织荧光染色和电镜鉴定,用4组不同组合的药物进行干预:低分子肝素组:在细胞中加入20 mg/L的低分子肝素;Galectin-3组:在细胞中加入5 mg/L的Galectin-3;联合组:在细胞中加入20 mg/L的低分子肝素和5 mg/L的Galectin-3;对照组:在细胞中加入等量PBS缓冲液.使用EdU掺入法检测血管内皮细胞增殖,流式细胞仪检测血管内皮细胞周期,使用划痕实验检测血管内皮细胞迁移,探究在不同条件下对血管内皮细胞迁移和增殖的影响.结果 低分子肝素组、Galectin-3组、联合组、对照组的吸光度(OD490)值分别为(0.285±0.018)、(0.297±0.041)、(0.351±0.016)、(0.233±0.005),这表明联合组的血管内皮细胞增殖情况最为显著(P<0.05);培养24 h的血管内皮细胞迁移率分别为(42.02±7.62)、(45.82±3.96)、(68.53±11.22)、(34.21±3.99),这表明联合组的血管内皮细胞迁移率最大(P<0.05).结论 低分子肝素联合Galectin-3可以显著提升骨髓间充质干细胞来源的血管内皮细胞的迁移和增殖.

The effect of low molecular heparin and Galectin-3on the cell migration and proliferation of vascular endothelial cells

Objective To explore the effect of the association of low molecular heparin and Galectin-3 on the cell migration and cell proliferation of vascular endothelial cell from mesenchymal stem cells.Methods Depending on the administration, this study was divided into four groups: low molecular weight heparin group, adding 20 mg/L low molecular weight heparin into the cells;Galectin-3 group, adding 5 mg/L Galectin-3 into the cells;combination group, adding 20 mg/L low molecular weight heparin and 5 μg/ml of Galectin-3 into the cells;control group, equal volume of PBS buffer into the cells.The proliferation of vascular endothelial cell was to be detected by EdU incorporation,the cell growth cycle of vascular endothelial cell was to be detected by flow cytometry,and the cell migration of vascular endothelial cell was to be detected by scrath test.Then to investigate the effect of different conditions on the migration and proliferation of vascular endothelial cell.Results The OD490 of MWH group, Galectin-3 group, combined group and control group were (0.285±0.018), (0.297±0.041), (0.351±0.016), and (0.233±0.005) respectively, which indicates that the combined group could increase the cell proliferation significantly(P<0.05).Cultured for 24 hours, the cell migration rate was(42.02±7.62), (45.82±3.96), (68.53±11.22),and (34.21±3.99), suggesting that combined group have the largest cell migration(P<0.05).Conclusion The association of low molecular heparin and Galectin-3 could improve the cell migration and cell proliferation of vascular endothelial cell from esenchymal stemcells significantly.