[关键词]
[摘要]
目的 研究注射用丹参多酚酸(SAFI)对氧糖剥夺/复氧复糖(OGD/R)损伤后小鼠脑微血管内皮细胞(BEND3)增殖、迁移、成管能力的影响及机制。方法 CCK-8法检测SAFI对正常BEND3细胞活力的影响,筛选出安全作用浓度。取正常生长的对数期BEND3细胞,分为对照组、模型组、SAFI(0.63、1.25、2.50、5.00、10.00 μg·mL-1)组,模型组与SAFI组建立OGD/R模型,缺氧缺糖4 h、复氧复糖24 h;CCK-8法检测BEND3细胞活力;划痕实验检测BEND3细胞迁移能力;基质胶成管实验检测BEND3细胞成管能力;Western blotting法检测BEND3细胞血管内皮生长因子A(VEGFA)、血管生成素-1 (Ang-1)、Ang-2蛋白表达。结果 与对照组比较,模型组细胞活力、细胞迁移率及细胞成管血管网络的交叉点数、节点数、分支点数、血管总长度均显著降低(P<0.05、0.01);与模型组比较,SAFI浓度为2.50、5.00、10.00 μg·mL-1时细胞活力、细胞迁移率及细胞成管血管网络的交叉点数、节点数、分支点数、血管总长度、网眼总面积和VEGFA和Ang-1、Ang-2蛋白表达量均显著上升(P<0.05、0.01),且作用呈浓度相关性。结论 SAFI可提高BEND3细胞OGD/R损伤后的增殖能力、迁移能力、成管能力,机制可能与上调VEGFA和Ang-1、Ang-2蛋白表达相关。
[Key word]
[Abstract]
Objective To study the effects of Salvianolic Acid for Injection (SAFI) on proliferation, migration and tubulation of BEND3 after hypoxia injury and its mechanism. Methods The CCK-8 method was used to detect the effect of SAFI on the activity of normal BEND3 cells and screen for safe concentrations. Normal growth of logarithmic BEND3 were divided into control group, model group and SAFI (0.63, 1.25, 2.50, 5.00, 10.00 μg·mL-1) group. Model group and SAFI administration group established oxygen-glucose deprivation/reoxygenation/reglycation (OGD/R) model, hypoxia and glucose deficiency for 4 h, reoxygenation/ reglycation for 24 h. CCK-8 method was used to detect the viability of BEND3 cells. The effect of SAFI on the migration ability of BEND3 cells injured by hypoxia was detected by scratch test. Matrix colloidal tube test was used to detect the effect of SAFI on tube formation ability of BEND3 cells injured by hypoxia. The effect of SAFI on the expression of vascular endothelial growth factor A (VEGFA), angiopoietin-1 (Ang-1) and Ang-2 proteins after hypoxia injury was detected by Western blotting. Results Compared with control group, the cell viability, cell mobility, the number of crossings, nodes, branches, and total length of vessels in the model group were significantly lower than those in the control group (P<0.05 and 0.01). Compared with the model group, the cell viability, cell mobility, the number of crossings, nodes, branches, total length of vessels,total area of meshes and the expression of VEGFA, Ang-1, and Ang-2 proteins increased when the concentration of SAFI was 2.50, 5.00, and 10.00 μg·mL-1, (P<0.05 and 0.01), and the effect was concentration dependent. Conclusion SAFI can improve the proliferation, migration and tubation of BEND3 cells after hypoxia injury, and the mechanism may be related to upregulating the expression of VEGFA and Ang-1 and Ang-2 proteins.
[中图分类号]
R285.5
[基金项目]