NCS1 Mediated Golgi Apparatus Stress To Promote Vascular Smooth Muscle Cell Migration Induced By Apelin-13

Aim: Apelin as an endogenous ligand for APJ,and apelin / APJ system plays a vital role in cardiovascular diseases.Our previous report that apelin-13 promotes the proliferation of vascular smooth muscle cells through PI3K-Akt,ERK1 / 2,14-3-3,NOX4 \ ROS and Notch3 signal pathways.It has been reported that apelin can induce the migration of rat vascular smooth muscle cells,but the effects and the mechanisms in human vascular smooth muscle cells are not clear.Studies have shown that oxidative stress play an important role in cell migration.Golgi apparatus stresss is a type of oxidative stress and wherther it also mediates smooth muscle cell migration.Therefore,the purpose of this study is to investigate the role of apelin-13 in human aortic vascular smooth muscle cells.To explore the new molecular mechanism of NCS1-induced Golgi apparatus stress-mediated apelin-13 to promote the migration of aortic vascular smooth muscle cells.And to find new and feasible mechanism and target for the treatment of vascular proliferative diseases such as hypertension and atherosclerosis.Methods:1.Western Blot: The expression of NCS1,GOLPH3 and ?-tubulin in human aortic smooth muscle cells were detected;2.Wound healing detects cell migration;3.Transwell assay of the effect of apelin-13 on the migration of HA-VSMCs;4.The Golgi morphology of HA-VSMCs was observed by transmission electron microscopy;5.Laser confocal observation of GOLPH3 in subcellular localization;6.RNA interference: design RNA interference chain,respectively,interfere with HA-VSMCs in the expression of NCS1,GOLPH3;7.Immunohistochemistry(IHC)method were used to detect the expression of apelin,APJ,NCS1 and GOLPH3 proteins in the vascular smooth muscle tissue of the arterial arteries and double kidney-double clip rat hypertensive coronary artery vascular smooth muscle tissue.Results:1.Wound healing and transwell experiments show that apelin-13 promote migration of vascular smooth muscle cells;2.Apelin-13 concentration and time-dependent promote the expression of Golgi stress marker GOLPH3;3.APJ inhibitor F13 A inhibites the expression of apelin-13-induced GOLPH3;4.Apelin-13 alters GOLPH3-labeled Golgi subcellular localization;5.Apelin-13 changes the Golgi structure;6.Apelin-13 concentration and time-dependent enhance the expression of NCS1;7.APJ inhibitor F13 A inhibits the expression of NCS1 in human vascular smooth muscle cells induced by apelin-13;8.NCS1-siRNA down-regulates the expression of GOLPH3 in human vascular smooth muscle cells induced by apelin-13;9.NCS1-siRNA inhibits apelin-13-induced changes in Golgi structure;10.NCS1-siRNA or GOLPH3-siRNA inhibits the migration of VSMCs induced by apelin-13;11.Apelin,APJ,NCS1 and GOLPH3 were positive in the vascular smooth muscle tissue of atherosclerotic coronary artery in autopsy,which was significantly higher than that in normal coronary artery smooth muscle tissue;The expression of apelin,APJ and GOLPH3 in kidney-double clip rats hypertensive coronary artery vascular smooth muscle tissue was significantly increased,and the proliferation and thickening of the vessel wall were obvious.Conclusion: Apelin-13 through NCS1 regulated Golgi apparatus stress to promote migration of human aortic vascular smooth muscle cells.