Role Of MiRNA199a-5p In Glycolysis And Autophagy Of Endothelial Cells Under Chronic Oxidative Stress

Background Arteriosclerosis is an important pathological basis of cardiovascular and cerebrovascular diseases and a chronic inflammatory disease characterized by the accumulation of lipids and inflammatory cells in the middle and large artery walls.Oxidative damage of vascular endothelium is considered as the first and key step of atherosclerosis.In the physiological state,vascular endothelial cells have important physiological functions,such as the exchange of substances inside and outside blood vessels,synthesis and release of vasoactive mediators.In recent years,more and more studies at home and abroad have shown that oxidative stress,lox-1 and autophagy are intermingled in the formation and development of atherosclerosis.In the oxidative stress environment,lox-1 participates in the processes of ox-ldl binding,phagocytosis and protein decomposition,so as to play the role of scavenging receptor.Autophagy is a biological process in which cells degrade abnormal proteins and pathogens in damaged organelles under the influence of various stimuli in the external environment.MicroRNA(miRNA)is a class of endogenous non-coding RNA with a length of about 18-25 nucleotides,which can regulate the degradation of mRNA or inhibit translation and regulate the expression of target genes at the post-transcriptional level.MiRNA perform many biological functions in vivo,such as participating in cell proliferation,differentiation and apoptosis.Studies have found that many miRNA expression profiles in endothelial cells are changed under oxidative stress,among which mirna199a-5p is the most significant.However,the mechanism by which miRNA affects the autophagy and glycolysis levels of endothelial cells in the context of chronic oxidative stress remains unclear.Therefore,we investigated the effect of miRNA199a-5p on endothelial micro glycolysis and autophagy under chronic oxidative stress.In this study,human umbilical vein endothelial cells were taken as the research object to study the role of miRNA199a-5p in glycolysis and autophagy of endothelial cells under chronic oxidative stress.Objective To investigate the role of miRNA199a-5p in glycolysis and autophagy of endothelial cells under chronic oxidative stress.Methods Endothelial cells under chronic oxidative stress were established in vitro,and the effects of mirna199a-5p agonists and inhibitors were transfected into endothelial cells.PCR and Western Blot were used to detect the effects of miRNA199a-5p on glycolysis and autophagy of endothelial cells.Western Blot analysis showed that the content of lox-1 in the agonist group decreased and that in the inhibitor group increased when compared with that in the control group under normal conditions.Compared with the control group,HK2 content in the agonist group increased.HK2 protein content decreased in inhibitor group.Normal circumstances,the autophagy protein LC3 Ⅱ/Ⅰ content is higher in agonist group,lower in inhibitor group.We also found that beclin-1,P62 and ATG4 levels were decreased in the agonist group and increased in the inhibitor group.In the context of chronic oxidative stress,compared with the control group,the content of lox-1 was decreased in the agonist group and increased in the inhibitor group.However,we found that compared with the control group,the content of HK2 in the agonist group decreased,while that in the inhibitor group increased.Chronic oxidative stress environment,the autophagy protein LC3 Ⅱ/Ⅰ content is higher in agonist group,lower in inhibitor group.We also found that beclin-1,P62 and ATG4 levels were decreased in the agonist group and increased in the inhibitor group.Compared with the normal environment,HK2 content reduced in chronic oxidative stress environment,LC3 Ⅱ/Ⅰ content increases,Beclin-1 levels,reduce the content of P62,ATG4 content increased.At the same time,we obtained the same experimental results as Western Blot by determining the activity of hexokinase.Conclusion MiRNA199a-5p is overexpressed in endothelial cells under chronic oxidative stress.In the context of chronic oxidative stress,overexpression of miRNA199a-5p can reduce the intracellular glycolysis level,while inhibition of miRNA199a-5p can increase the intracellular glycolysis level.In the context of chronic oxidative stress,overexpression of miRNA199a-5p can enhance intracellular autophagy to maintain cell energy demand and homeostasis of the internal environment,while inhibition of miRNA199a-5p can reduce intracellular autophagy.