Study On The Effect Of MiRNA-199a-5p On The Human Vascular Endothelial Cells Function And The Mechanism Of Astragalus Polysaccharides Intervention

ObjectiveOur work is to investigate the mechanism of vascular endothelial injury induced by miRNA-199a-5p through regulation the ATF6 pathway of endoplasmic reticulum stress.Then we can provide experimental basis for further elucidating the pathophysiological mechanism of vascular endothelial dysfunction in cardiovascular diseases.At the same time,we intend to reveal the mechanism of APS in treating vascular endothelial injury,which is a compound of replenishing qi and activating blood in traditional Chinese medicine,and to provide new treatment strategies and therapeutic targets for the prevention and treatment of cardiovascular diseases related to endothelial injury of traditional Chinese medicine.MethodsBy transient transfection technique,we construct the miRNA-199a-5p differential expression vascular endothelial cell model and use the Astragalus Polysaccharide to intervent the constructed model.Then we detect the vascular endothelial function related indicators and the expression of ATF6 signaling pathway-related genes in endoplasmic reticulum stress in the model.Among them,the changes of Endothelin-1(ET-1),Von Willebrand Factor(vWF)and Thrombomodulin(TM)are determined by Enzyme-linked immunosorbent assay(ELISA),while Nitric Oxide(NO)are determined by nitrate reductase method.The expression of GRP78,ATF6 and CHOP are detected by Reverse Transcription Polymerase Chain Reaction(RT-PCR)and Western Blot.ResultsThe detection results of the vascular endothelial function related indicators certify that in the miRNA-199a-5p low expression group,the secretion of ET,vWF and TM are increased,while decreased of NO,compared with the NC and APS group.And the differences are statistically significant(P<0.05),indicating that there is endothelium damage in miRNA-199a-5p low expression group.Similarly,the expression of GRP78,ATF6 and CHOP are significantly increased in miRNA-199a-5p low expression groupcompared with the NC and APS group(P<0.05).Conclusion1.miRNA-199a-5p has protective effect on vascular endothelial cell;2.The low expression of miRNA-199a-5p can activate the ATF6 pathway of endoplasmic reticulum stress,and then lead to vascular endothelial injury,which may be an important pathogenesis of cardiovascular diseases related to endothelial injury.3.APS can inhibit the activation of ATF6 pathway,thereby alleviating endothelial damage,modulating the endoplasmic reticulum stress and have a good therapeutic effect on blood stasis syndrome.Thus,APS may be a potential drug for the treatment of cardiovascular disease related to endothelial injury.