阿司匹林对心肌微血管内皮细胞血管新生功能的影响

Effect of aspirin on myocardial microvascular endothelial cells and its angiogenesis functions

目的：探讨不同剂量阿司匹林对大鼠心肌微血管内皮细胞（CMECs）血管新生功能的影响和可能的机制。方法：消化法分离大鼠CMECs，用不同浓度（1、10、100、1000和5000μmol／L）的阿司匹林处理体外培养的大鼠CMECs后，分别用CCK-8比色法检测细胞增殖、TUNEL法检测细胞凋亡、划痕试验及Transwell小室评价细胞迁移能力、成管实验评价血管新生能力、Westernblot法检测蛋白激酶B（AKT）磷酸化水平，ELISA法测定纤维连接蛋白（FN）表达。结果：①1、10和100p,mol／L的阿司匹林对，CMECs的增殖、凋亡和AKT磷酸化水平无明显影响，1000、5000p,mol／L的阿司匹林对组显著抑制CMECs增殖、下调AKT磷酸化水平、诱导细胞凋亡；②10、100ixmol／L的阿司匹林促进CMECs迁移、成管；③FN表达随阿司匹林浓度升高而升高。结论：①低浓度阿司匹林（1～100μmoL／L）对CMECs的存活无明显影响，而高浓度的阿司匹林（1000、5000μmol／L）显著抑制CMECs增殖、下调AKT磷酸化水平、细胞凋亡增多。②低浓度阿司匹林（1～100μmol／L）作用下，阿司匹林促进CMECs迁移、成管能力，可能与阿司匹林作为环氧化酶（COX）抑制剂，减少前列环素（PGI2）生成引起FN表达上调有关。

AIM： To investigate the effects of different doses of aspirin on angiogenic functions of rat myocardial microvascular endothelial cells （CMECs） and the possible mechanism. METHODS： CMECs were isolated from rats with collagenase digestion and cultured with different doses of aspirin （1, 10, 100, 1 000, 5 000 μmol/L） in vitro. The proliferation of CMECs was analyzed by CCK-8 assay. Apoptotic cells of CMECs were detected by TUNEL. Migration of CMECs was assessed by migration assays. The angiogenic activity of CMECs was examined by tube formation experiment, pAKT protein expression was measured by Western blotting analysis, and FN protein expression was measured by ELISA analysis. RESULTS： Aspirin enhanced CMEC proliferation and pAKT protein expression and induced cell apoptosis at concentrations between 1 000 and 5 000 μmol/L. Aspirin promoted CMEC migration and angiogenesis at concentrations between 10 and 100 μmol/L, and FN protein expression increased with the increasing concentrations of aspirin. CONCLUSION： Low concentrations of aspirin （ 1 - 100 μmol/L ） exert no obvious effect on the survival of CMECs, whereas high concentrations of aspirin （1 000 -5 000 μmol/L） inhibit the proliferation of CMECs, reduce AKT phosphorylation levels and increase apoptosis. At low concentrations of aspirin （ 1 - 100 μmol/L）, CMEC migration and tube formation are promoted possibly because aspirin, as a ring oxidase （COX） inhibitor, reduces the generation of PGI2 and increases FN expression.