The Effects Of HbA_1C On Production Of ENOS, NO, E-selectin, SICAM-1, T-PA And PAI-1 In Endothelial Cells

Objects:To observe the influence of HbA_1C on endothelial cells producing eNOS, NO, t-PA, PAI-1, E-selectin and ICAM-1 and to investigate the role of HbA_1C in the development of atherosclerosis.Methods:1. Human umbilical vein endothelial cells (ECV304) cells were cultured in vitro.2. PRMI 1640 with 2% new bron calf serum(control group), haematoglobin (5.3% haematoglobin group), high HbA_1C(5.3% HbA_1C group) and lower HbA_1C(9.2% HbA_1C group) were used to incubate ECV304 cells for 24 hours(repeated in six wells).3. The activity of eNOS was measured by chemical chromatometry. The activity of NO was measured by double antibody scandwith ELISA methods. The activity of t-PA and PAI-1 were measured by nitrate reductase methods. The activity of E-selectin and ICAM-1 were measured by spectrophotometric methods.Results:1. The concentration of NO and eNOS was similar between the low HbA_1C group and the haematoglobin group with no statistic significance. The concentration of NO and eNOS in the high HbA_1C group were lower than the other groups.2. The concentration of PAI-1 was similar between the control group, the haematoglobin group and the low HbA_1C group with no statistic significance. The concentration of PAI-1 in the high HbA_1C group was significantly higher than the other groups. The concentration of t-PA in the low HbA_1C group was significantly higher than the other groups. The concentration of t-PA in the high HbA_1C group was lower than the control group and the haematoglobin group, and even lower than the low HbA_1C group.3. The concentration of sICAM-1 in the haematoglobin group and the low HbA_1C group was significantly higher than the control group. The concentration of E-selectin and ICAM-1 in the high HbA_1C group was significantly higher than the other groups.Conclusions1. HbA_1C could induce umbilical vein endothelial cells producing NO and eNOS by the means of negative concentration dependence. NO and eNOS were significantly inhibited in the high HbA_1C group.2. HbA_1C could induce umbilical vein endothelial cells producing t-PA and PAI-1 by the means of concentration dependence, with t-PA in negative dependence and PAI-1 in positive dependence.3. HbA_1C could induce umbilical vein endothelial cells producing E-selectin and sICAM-1 by the means of positive concentration dependence.4. The production of eNOS, NO, t-PA, PAI-1, E-selectin and ICAM-1 of endothelial cells elevated with increasing serum HbA_1C. It suggested that HbA_1C might be involved in the development of atherosclerosis and promote diabetic vascular disease.