The Mechanisms Of Ethanol On Plasma Enos/no Levels In Korean-chinese And Han Nationality Patients With Coronary Heart Disease In Yanbian Area

Objective:To investigate the risk factors of coronary heart disease in Korean-Chinese and Han nationality in Yanbian and the mechanisms of endothelial nitric oxide synthase(eNOS)/nitric oxide(NO)levels in patients' plasma.Methods:The patients were divided into two groups of Korean-Chinese and Han nationality,We statistical basic data and clinical results of coronary angiography.The levels of plasma NO,eNOS,neural nitric oxide synthase(nNOS)and inducible nitric oxide synthase(iNOS)were measured by enzyme linked immunosorbent assay(ELISA).According to the differences of alcohol intake and plasma eNOS/NO levels,cell experiments were carried out to explore the effect and mechanism of ethanol on eNOS/NO levels.Human umbilical vein endothelial cells(HUVECs)were cultured,and Western Blotting was used to determine the quantity-effect relationship and time-effect relationship of ethanol decreasing HUVECs eNOS protein expression.MTS assay was used to detect cell viability and flow cytometry to detect cell apoptosis under this concentration time condition.The follow-up experiments were divided into control group(Control),EtOH group(200mM concentration),Tiron(ROS scavenger)group,Tiron+EtOH group,GKT137831(NOX1/NOX4 inhibitor)group,GKT137831+EtOH group,SC-79(Akt activator)group,SC79+EtOH group.Western Blotting was used to detect the expression of Akt,eNOS and p-eNOS Ser1177 protein to explore the correlation in alcohol and oxidative stress and Akt/eNOS.Griess assay was used to determine the concentration of NO in each group of cells before and after adding eNOS inhibitor N?-Nitro-L-arginine methyl ester(L-NAME).Results:(1)The general data between the two groups in the comparison results show that alcohol consumption on Korean-Chinese nationality patients was significantly higher than that in Han patients(p<0.05);Two groups of patients with coronary angiography showed that Korean-Chinese nationality patients with multivessel disease proportion is higher than Han patients(p<0.05);Plasma nitric oxide synthase(NOS)and NO concentration test results showed that the Korean-Chinese nationality patients eNOS plasma concentration was lower than Han patients(p<0.05),the concentration of NO was lower than Han patients(p<0.05),but there were no significant differences in plasma iNOS and nNOS concentrations(p>0.05).(2)Ethanol decreased the HUVECs expression of eNOS protein,and showed time and dose dependences;200mM ethanol concentration in cultured 24h was no significant change in cell morphology compared with the control group and does not affect cell viability changes(p>0.05)and no cell toxicity(p>0.05);Preconditioning Tiron,GKT137831 did not impact ethanol reduced the expression of eNOS protein;SC-79 pretreated the cells to inhibit the effect of ethanol on the expression of eNOS protein(p<0.01)and p-eNOS Ser1177 protein(p<0.01).In the absence of L-NAME,the formation of NO in group EtOH was significantly lower than that in group Control(p<0.01),but there was no significant difference in NO formation between SC-79 and SC-79+EtOH groups(p>0.05),and higher than that in EtOH group.In the case of adding L-NAME,the yield of Control,SC-79 and SC-79+EtOH groups decreased significantly(p<0.05,p<0.01 and p<0.05),while NO yield of EtOH group was not significantly different from that of absence of L-NAME(p>0.05).Conclusion:1.The degree of CHD in Korean-Chinese nationality is heavier than that of Han patients,and it may be closely related to the high consumption of drinking in Korean-Chinese nationality.2.Ethanol can reduce the eNOS derived NO release of HUVECs by down-regulating the Akt/eNOS pathway,which may provide a new theoretical basis for the harmful effects of drinking on CHD.