The Effect And Mechanism Of Poly(ADP-Ribose) Polymerase-1on Apoptosis Of Endothelial Cell Inducing By Aldosterone

Background:Cardiovascular disease is the major cause of disability and death throughout the world. The underlying pathology is atherosclerosis, which develops over many years and is usually advanced by the time symptoms occur, generally in middle age. Atherosclerosis is a multifactorial disease involving the interplay of genetic and environmental factors, with a complex pathogenesis. The formation of atherosclerosis is currently considered as the result of a series of inflammatory response caused by vascular endothelial injury from a number of risk factors. Endothelial cell dysfunction is not just an early manifestation before the formation of atherosclerosis, also plays a very important role in the development process of the formation of atherosclerosis.In recent years, the role of aldosterone in the formation of atherosclerosis, which is an important component of the renin-angiotensin-aldosterone system (RAAS), is receiving more and more attention from scholars.It is reported that the plasma concentrations of aldosterone in patients with heart failure, congenital heart disease or cardiac arrhythmia are elevated. Aldosterone can increase the lesion area of atherosclerosis, which is confirmed by vitro and animal experiments. The increased plasma concentration of aldosterone is correlated with endothelial cell dysfunction. In the animal model of primary aldosteronism, aldosterone can lead to aortic endothelial cell apoptosis, and this effect is independent of blood pressure. These results all indicated that aldosterone plays a very important role in the development process of the formation of atherosclerosis, but the mechanism is still not established yet.Poly [ADP-ribose] polymerase1(PARP-1) also known as NAD+ADP-ribosyltransferase1or poly [ADP-ribose] synthase1is an enzyme that widely presents in eukaryotic cells. PARP1works by modifying nuclear proteins by poly ADP-ribosylation, which is one of the important ways of protein post-translational modification. And PARP1is involved in differentiation, proliferation, and tumor transformation. The hypothesis is that endothelial cell dysfunction caused by aldosterone is mediated through apoptosis, which results from high PARP1activity which is increased by aldosterone.Objective:Observe the effect of aldosterone on apoptosis of endothelial cell and activity of PARP1. Explore the effect of increased PARP1activity on the apoptosis of endothelial cell caused by aldosterone treatment. Study the mechanism of endothelial cell dysfunction resulted from aldosterone.Methods:1. Endothelial cells are cultured with or without aldosterone of different concentrations or for different times. Cell apoptosis is detected by TUNEL assay and Flow-cytometry. Cell proliferation is detected by cell counting. The activity of PARP1is detected.2.Endothelial cells are cultured in normal medium, in medium with aldosterone, or in medium with aldosterone and aldosterone receptor antagonist. Cell apoptosis is detected by TUNEL assay and Flow-cytometry. Cell proliferation is detected by cell counting. The activity of PARP1is detected.3.Endothelial cells are cultured in normal medium, in medium with aldosterone, or in medium with aldosterone and PARP-1inhibitor. Cell apoptosis is detected by TUNEL assay and Flow-cytometry. Cell proliferation is detected by cell counting. The activity of PARP1is detected.Results:1.Aldosterone treatment decreases cell proliferation and increases apoptosis in endothelial cells. Aldosterone treatment decreases PARP1activity.2.Endothelial cells treated with aldosterone and aldosterone receptor antagonist grow faster and have lower PARP1activity than cells treated with aldosterone.3.Endothelial cells treated with aldosterone and PARP1inhibitor grow faster and have lower PARP1activity than cells treated with aldosterone.Conclusion:1. Aldosterone decreases endothelial cell proliferation and increases apoptosis and PARP1activity in dose and time dependent manners.2.Aldosterone receptor antagonist blocks the effect of aldosterone treatment in endothelial cells in cell proliferation, apoptosis and PARP1activity.3.PARP1inhibitor blocks the effect of aldosterone treatment in endothelial cells in cell proliferation, apoptosis and PARP1activity.Endothelial cell dysfunction caused by aldosterone is mediated through apoptosis, which results from high PARP1activity which is increased by aldosterone.