[關(guān)鍵詞]
[摘要]
目的 研究注射用丹參多酚酸(SAFI)對氧糖剝奪/復(fù)氧復(fù)糖(OGD/R)損傷后小鼠腦微血管內(nèi)皮細(xì)胞(BEND3)增殖、遷移、成管能力的影響及機(jī)制。方法 CCK-8法檢測SAFI對正常BEND3細(xì)胞活力的影響,篩選出安全作用濃度。取正常生長的對數(shù)期BEND3細(xì)胞,分為對照組、模型組、SAFI(0.63、1.25、2.50、5.00、10.00 μg·mL-1)組,模型組與SAFI組建立OGD/R模型,缺氧缺糖4 h、復(fù)氧復(fù)糖24 h;CCK-8法檢測BEND3細(xì)胞活力;劃痕實(shí)驗(yàn)檢測BEND3細(xì)胞遷移能力;基質(zhì)膠成管實(shí)驗(yàn)檢測BEND3細(xì)胞成管能力;Western blotting法檢測BEND3細(xì)胞血管內(nèi)皮生長因子A(VEGFA)、血管生成素-1 (Ang-1)、Ang-2蛋白表達(dá)。結(jié)果 與對照組比較,模型組細(xì)胞活力、細(xì)胞遷移率及細(xì)胞成管血管網(wǎng)絡(luò)的交叉點(diǎn)數(shù)、節(jié)點(diǎn)數(shù)、分支點(diǎn)數(shù)、血管總長度均顯著降低(P<0.05、0.01);與模型組比較,SAFI濃度為2.50、5.00、10.00 μg·mL-1時(shí)細(xì)胞活力、細(xì)胞遷移率及細(xì)胞成管血管網(wǎng)絡(luò)的交叉點(diǎn)數(shù)、節(jié)點(diǎn)數(shù)、分支點(diǎn)數(shù)、血管總長度、網(wǎng)眼總面積和VEGFA和Ang-1、Ang-2蛋白表達(dá)量均顯著上升(P<0.05、0.01),且作用呈濃度相關(guān)性。結(jié)論 SAFI可提高BEND3細(xì)胞OGD/R損傷后的增殖能力、遷移能力、成管能力,機(jī)制可能與上調(diào)VEGFA和Ang-1、Ang-2蛋白表達(dá)相關(guān)。
[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.
[中圖分類號]
R285.5
[基金項(xiàng)目]