摘要
目的 研究游离脂肪酸 (FFA)、胰岛素 (Ins)、葡萄糖 (Glu)及其共同作用时对血管内皮细胞 (VEC)的影响。 方法 用MTT法测定经FFA、Ins、Glu处理后脐静脉内皮细胞的活力 ,用绿色荧光蛋白标记的膜联蛋白V/碘化丙啶 (annexinV GFP /PI)双染色法检测细胞凋亡或死亡率。 结果 (1)生理浓度 (10 0 μmol/L)的软脂酸 (PA)作用于细胞 72h ,与空白对照组 (对照组 )相比 ,细胞的存活率降低了 (18 7± 5 8) % ,差异有显著意义 (P <0 0 5 )。 (2 )高浓度的Ins (5 0U/L)可使细胞生存率增加 (36 8± 4 6 ) % ,与对照组相比差异有显著意义 (P <0 0 5 )。 (3) 5 0U/LIns +40 0μmol/LPA组细胞的生存率较 4 0 0 μmol/LPA组增加 (2 8± 3 2 ) % (P <0 0 5 )。 (4 ) 2 2 2mmol/LGlu+40 0 μmol/LPA组细胞生存率较PA组减少了 (2 1± 7 2 ) % (P <0 0 5 )。 (5 ) 2 2 2mmol/LGlu +40 0 μmol/LPA +5 0U/LIns组生存率较 2 2 2mmol/LGlu +40 0 μmol/LPA组增加了 (17 6±2 3) % (P <0 0 5 )。 (6 )Glu致VEC的损伤以引起细胞凋亡为主 ,而FFA以引起细胞非凋亡性的死亡为主。 结论 PA对血管VEC的毒性作用远远大于Glu ,Ins本身对血管VEC有保护作用 ,Glu可加重PA对VEC的毒性作用。
Objective To study influence of saturated fatty acids, high glucose, high insulin and combination of them on cell survival of human umbilical vein endothelial cells (HUVEC) and to explore the degree and manner of vascular endothelial cells injury under the factors. Methods (1) The cell survival rate was assessed using MTT method after the cells were treated with various concentrations of palmitic acid (PA), insulin (Ins), D-glucose (Glu). (2) Apoptosis and necrosis of the cells were determined using annexin V- GFP/PI staining. Results (1)PA of 100 μmol/L markedly suppressed cell survival. (2)When cells were incubated in 50 U/L Ins, the cell survival rate was increased by (36.8±4.6)% comparing with the control group at 72 h.(3) The cell survival rate was increased by (28±3.2)% when cells were incubated with mixture of 400 μmol/L PA plus 50 U/L Ins comparing with 400 μmol/L PA alone (P<0.05). (4) The cell death was accelerated when cells were incubated at mixture of 400 μmol/L PA plus 22.2 mmol/L Glu .(5) The cell survival rate was increased by (17.6±2.3)% when cells were incubated with mixture of 400 μmol/L PA +50 U/L Ins+22.2 mmol/L Glu comparing with 400 μmol/L PA+ 22.2 mmol/L Glu. (6) 22.2 mmol/L of Glu could not cause cell death, but induced apoptosis . (7) PA induced apoptosis at low dose within 4 h, but induced necrosis at high dose and longer time. Conclusions PA has much stronger toxicity than glucose on vascular endothelial cells. High level of insulin has protective effect on HUVEC. High concentration of glucose has no effect on cell survival, but Glu can accelerate the cell death induced by PA.
