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目的 观察不同浓度葡萄糖和游离脂肪酸 (FFA)对体外培养的人血管内皮细胞增殖和细胞周期的影响 ;了解葡萄糖和FFA对人血管内皮细胞的损伤的协同效应。方法 将人血管内皮细胞培养于不同浓度的葡萄糖和 /或FFA(棕榈酸和 /或油酸 )中 2 4～ 48h。采用倒置显微镜和电子显微镜观察内皮细胞的形态学变化。噻唑蓝比色 (MTT)法检测内皮细胞增殖受抑制的情况 ,用台盼蓝法判定细胞活力 ,流式细胞仪分析法测定细胞周期。结果 1 5或 3 0mmol/L的葡萄糖、0 2 5或 0 5mmol/L的棕榈酸 ,以及 0 5mmol/L的油酸对内皮细胞产生浓度 效应和时间 效应的增殖抑制作用和加速细胞死亡的作用。细胞暴露于高浓度的葡萄糖和 /或FFA中 ,其G0 /G1期比例增高 ,而S期降低 ,提示高浓度的葡萄糖和 /或FFA主要使内皮细胞停滞在G0 /G1期。在同时具有葡萄糖和FFA的培养基中培养的内皮细胞葡萄糖 3 0mmol/L +棕榈酸 0 2 5mmol/L ,葡萄糖 3 0mmol/L +油酸 0 5mmol/L ,葡萄糖 3 0mmol/L +棕榈酸 0 2 5mmol/L +油酸 0 5mmol/L) ,其增殖的抑制率和细胞的死亡数量明显高于在仅有高浓度葡萄糖或FAA(棕榈酸和 /或油酸 )培养基的内皮细胞。结论 高浓度的葡萄糖和FFA都能抑制内皮细胞的增殖和加速细胞的死亡 ,高浓度的葡萄糖和FFA同时存在?
Objective To observe the effects of different concentrations of glucose and free fatty acid (FFA) on the proliferation and cell cycle of cultured human vascular endothelial cells in vitro and to investigate the synergistic effect of glucose and FFA on the damage of human vascular endothelial cells. Methods Human vascular endothelial cells were cultured for 24-48 h at different concentrations of glucose and / or FFA (palmitic acid and / or oleic acid). Inverted microscope and electron microscope were used to observe the morphological changes of endothelial cells. The inhibition of endothelial cell proliferation was detected by MTT assay. Cell viability was determined by trypan blue assay and cell cycle was assayed by flow cytometry. Results The effects of concentration-effect and time-effect inhibition of proliferation and accelerated cell death were induced by 1 5 or 30 mmol / L glucose, 0 2 5 or 0 5 mmol / L palmitic acid, and 0 5 mmol / L oleic acid . Exposure of cells to high concentrations of glucose and / or FFA increased the proportion of G0 / G1 phases and decreased the S phase, suggesting that high concentrations of glucose and / or FFA predominantly arrested endothelial cells in the G0 / G1 phase. Endothelial cell glucose 30 mmol / L + palmitic acid 25 mmol / L, glucose 30 mmol / L + oleic acid 0 5 mmol / L, glucose 30 mmol / L + palmitate 0 cultured in medium with both glucose and FFA 2 5 mmol / L + oleic acid 0 5 mmol / L), the proliferation inhibition rate and the number of cell death were significantly higher than that in endothelial cells only with high concentration of glucose or FAA (palmitic acid and / or oleic acid) medium. Conclusion High concentrations of glucose and FFA can inhibit the proliferation of endothelial cells and accelerate cell death, high concentrations of glucose and FFA co-exist?