The Study Of The Damage Effect And Mechanism Of SCD40L On Human Coronary Artery Endothelial Cells Under Chronic Intermittent Hypoxia

Objective:Obstructive sleep apnea hypopnea syndrome?OSAHS?is the main manifestation of chronic intermittent hypoxia caused by recurrent nocturnal apnea and hypoventilation.It is one of independent risk factors forcardiovascular and cerebrovascular diseases,such as,coronary atherosclerotic heart disease,hypertension,pulmonary heart disease and stroke.In order to explore the effectof chronic intermittent hypoxiaon human coronary artery endothelial cellsdysfunction,we have established a chronic intermittent hypoxia model of human coronary artery endothelial cells?HCAEC?and check the role of recombinant human CD40L?sCD40L?in the HCAEC dysfunction.Methods:The human coronary endothelial cells were cultured.According to the requirements of the experiment,it was divided into four groups:group Control and group sCD40L,group CIH and group CIH+sCD40L.group Control and group sCD40L?80ng/m L sCD40L incubated in HCAEC for 30h?,group CIH?cultured in the chronic intermittent hypoxia stimulated for 6h:37?,94%N₂+5%CO₂+1%O₂culture for 15min;reperfusion in 5%CO2+air culture for 15min.The cycle operation?and group CIH+sCD40L?80ng/m L sCD40L incubated in HCAEC for 24h,and then cultured in the chronic intermittent hypoxia stimulated for 6h?.The application of Western blot detect the protein content of human coronary artery endothelial cells,for example:CD40,hypoxia inducible factor-1 alpha?HIF-1 alpha?,cyclooxygenase-2?COX-2?,endothelial nuclear factor kappa B?NF-K B?and intercellular adhesion molecule-1?ICAM-1?.The application of flow cytometry detect coronary arterial endothelial cell of apoptosis.Results:there was no statistically significant difference in the expression of CD40protein between the groups?P>0.05?.The expression of HIF-1 and COX-2 protein in human coronary artery endothelial cells showed no significant difference between group control and group sCD40L?P>0.05?.And group CIH+sCD40L was higher than group CIH.And group CIH+sCD40L was higher than group sCD40L?P<0.05?.The expression of NF-kappa B and ICAM-1 protein showed that the group sCD40L was higher than that in the group control,and the group CIH+sCD40L was higher than that in the group CIH?P<0.05?,group CIH+sCD40L was higher than group sCD40L?P<0.05?.The results of cell apoptosis measured by flow cytometry showed that group sCD40L was higher than that of group control,and that of group CIH+s CD40L was higher than that in group CIH,and that in group CIH+sCD40L was higher than that in group sCD40L?P<0.01?.Conclusion:sCD40L stimulated HCAEC toup-regulatethe expression of NF-kappa B,ICAM-1 protein,and has no obvious effect on the expression of COX-2 protein,HIF-1 alpha;In the chronic intermittent hypoxia condition,sCD40L stimulated HCAEC up-regulated the expression of HIF-1 alpha,COX-2,NF-kappa B,ICAM-1protein,apoptosis increased,and promote probably coronary artery endothelial cell dysfunction.