游离脂肪酸对猴胰岛微血管内皮细胞脂毒性的影响

摘要

目的 观察游离脂肪酸(free fatty acid,FFA)对恒河猴胰岛微血管内皮细胞(islet microvascular endothelial cells,IMECs)增殖和凋亡的影响及FFA介导的恒河猴IMECs氧化应激损伤.方法 分离纯化恒河猴IMECs,分别用含有FFA(FFA组)和不含有FFA(对照组)的DMEM培养,MTT实验测定细胞增殖率(成活率),流式细胞仪检测细胞凋亡率,活性氧簇(ROS)酶联免疫分析(ELISA)测定IMECs中ROS的浓度.结果 经24 h培养后,FFA组IMECs增殖率较对照组显著降低(P＜0.01);Annexin V-FITC/PI双染色后FFA组于荧光显微镜下可见明显的凋亡细胞;与对照组凋亡率比较,1.0 mmol/L FFA作用适当时间可显著升高IMECs凋亡率(P＜0.01);FFA组IMECs中ROS水平较对照组显著升高(P＜0.01).结论 高水平FFA可显著降低恒河猴IMECs增殖率,促进细胞发生凋亡,其凋亡可能与FFA介导的氧化应激损伤有关.%Objective To investigate the effect of free fatty acid (FFA) on growth and apoptosis of rhesus monkey islet microvascular endothelial cells (IMECs) , and the FFA - mediated oxidative stress injury. Methods IMECs were isolated and purified from rhesus monkeys, and cultured in the DMEM with and without FFA. Cell growth rate, apoptosis rate and the reactive oxygen species (ROS) concentration were assessed with MTT assay, flow cytometry and enzyme -linked immunoassay (ELISA) , respectively. Results After 24 h incubation, IMECs proliferation rate in FFA group was significantly lower than that in control group ( P＜0. 01) . Obviously apoptotic cells were observed under fluorescence microscopy with Annexin V - FITC/PI double staining in FFA group. Furthermore, the IMEC apoptosis rate and the ROS concentration in FFA group was significantly higher than those in control group ( P ＜0. 01) . Conclusion High concentration FFA greatly reduces the proliferation rate and promotes the apoptosis of IMECs, which may be involved with the FFA - mediated oxidative stress damage.