低分子肝素和Galectin-3对血管内皮细胞迁移和增殖的影响

发布时间：2019-04-21 15:08

【摘要】：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(b FGF),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Ed U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(P0.05);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(P0.05)。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。
[Abstract]:Aim to investigate the effects of low molecular weight heparin (LMWH) combined with galactose lectin 3 (Galectin-3) on migration and proliferation of (MSCs) derived vascular endothelial cells from bone marrow mesenchymal stem cells (MSCs). Methods SD rat bone marrow MSCs, was isolated and purified by density gradient centrifugation. The third passage was induced with vascular endothelial growth factor (VEGF) (VEGF) and basic fibroblast growth factor (bFGF) (b FGF), for 14 days in vitro. Immunofluorescence staining and electron microscope identification were performed. Four groups of drugs were used for intervention: low molecular weight heparin (LMWH) group: low molecular heparin (LMWH) with 20 mg/L was added to the cells; Galectin-3 group: 5 mg/L Galectin-3; combined group: 20 mg/L low molecular weight heparin (LMWH) group and 5 mg/L Galectin-3; control group (5 mg/L): the same amount of PBS buffer was added to the cells. The proliferation of vascular endothelial cells was detected by Ed U incorporation method, the cell cycle was detected by flow cytometry, and the migration of vascular endothelial cells was detected by scratch test to explore the effects of different conditions on the migration and proliferation of vascular endothelial cells. Results the OD490 values of low molecular weight heparin group, Galectin-3 group, combined group and control group were (0.285 卤0.018), (0.297 卤0.041), (0.351 卤0.016), (0.233 卤0.005), respectively. This indicated that the proliferation of vascular endothelial cells in the combined group was the most significant (P0.05). The migration rate of vascular endothelial cells was (42.02 卤7.62), (45.82 卤3.96), (68.53 卤11.22), (34.21 卤3.99) at 24 h, indicating that the combination group had the highest migration rate (P0.05). Conclusion low molecular weight heparin combined with Galectin-3 can significantly enhance the migration and proliferation of vascular endothelial cells derived from bone marrow mesenchymal stem cells.
【作者单位】：安徽医科大学第二附属医院血管外科
【基金】：安徽省自然科学基金(编号:KJ2013Z115)
【分类号】：R543.5

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