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目的:对比研究波动性高浓度胰岛素与恒定性高浓度胰岛素对体外培养的人脐静脉内皮细胞的凋亡的影响,从而探究波动性高浓度胰岛素血症在糖尿病动脉粥样硬化形成中的致病机制。方法:采用体外培养的人脐静脉内皮细胞为对象,研究波动性高胰岛素对血管内皮细胞的凋亡的影响。实验分为4组,即对照组(不含胰岛素)、正常浓度胰岛素组(含胰岛素1nmol/L)、恒定性高胰岛素组(含胰岛素100nmol/L)与波动性高胰岛素组(含胰岛素1nmol/L及含胰岛素100nmol/L两种条件培养液,每8小时轮换一次),每间隔8小时更换新鲜条件培养液一次,一共作用72小时。应用AO-EB染色法与流式细胞仪检测内皮细胞凋亡。结果:1、胰岛素对内皮细胞凋亡的影响1.1经不同浓度的胰岛素作用72小时后,AO-EB检测结果显示:正常浓度胰岛素组的内皮细胞凋亡细胞数明显下降,为对照组的81%(p<0.001),而恒定性高胰岛素组与波动性高胰岛素组的内皮细胞凋亡细胞数明显上升,分别为对照组的1.53倍(p<0.001)与1.75倍(p<0.001),正常浓度胰岛素组的1.88倍(p<0.001)与2.15倍(p<0.001);且波动性高胰岛素组内皮细胞凋亡细胞数显著高于恒定性高胰岛素组(p<0.001)。1.2经不同浓度的胰岛素作用72小时后,流式细胞仪检测结果显示:正常浓度胰岛素组的内皮细胞凋亡率明显下降,为对照组的81%(p<0.001),而恒定性高胰岛素组与波动性高胰岛素组的内皮细胞凋亡率出现明显上升,分别为对照组的1.11倍(p<0.01)与1.39倍(p<0.001),正常浓度胰岛素组的1.37倍(p<0.001)与1.72倍(p<0.001);且波动性高胰岛素组内皮细胞凋亡率显著高于恒定性高胰岛素组(p<0.001)。结论:1、波动性高胰岛素相对于恒定性高胰岛素更能诱导人脐静脉内皮细胞的凋亡。
OBJECTIVE: To compare the effects of high-concentration and high-concentration insulin on the apoptosis of cultured human umbilical vein endothelial cells (HUVECs) in vitro, so as to explore the pathogenesis of high-concentration hyperlipidemia in diabetic atherosclerosis mechanism. Methods: Human umbilical vein endothelial cells cultured in vitro were used to study the effects of hyperinsulinism on the apoptosis of vascular endothelial cells. The experiment was divided into 4 groups: control group (without insulin), normal insulin group (containing insulin 1nmol / L), constant high insulin group (containing insulin 100nmol / L) and high-insulin group L and insulin-containing 100nmol / L two conditioned medium, every 8 hours rotation), every 8 hours to replace the fresh conditioned medium, a total of 72 hours. AO-EB staining and flow cytometry were used to detect the apoptosis of endothelial cells. The effect of insulin on the apoptosis of endothelial cells1.1 After 72 hours of different concentrations of insulin, the results of AO-EB showed that the number of apoptotic cells of endothelial cells in normal insulin group decreased significantly, which was 81% of the control group (p <0.001), while the number of apoptotic cells in the constant high insulin group and the fluctuating high insulin group increased 1.53-fold (p <0.001) and 1.75-fold (p <0.001) (P <0.001) and 2.15-fold (p <0.001), respectively. The number of apoptotic endothelial cells in the high-insulin group was significantly higher than that in the high-insulin group (p <0.001). 1.2 After 72 hours of different concentrations of insulin, the results of flow cytometry showed that the apoptosis rate of endothelial cells in normal insulin group decreased significantly, which was 81% (p <0.001) in control group, while in constant high insulin group (P <0.01) and 1.39-fold (p <0.001), respectively, and 1.37-fold (p <0.001) in the normal insulin group compared with those in the normal insulin group 1.72-fold (p <0.001). The apoptotic rate of endothelial cells in high-insulin group was significantly higher than that in high-insulin group (p <0.001). Conclusions: 1, High blood viscosity insulin can induce apoptosis of human umbilical vein endothelial cells in comparison with constant high insulin.