Protective Effect Of FUNDC1 On Human Vascular Endothelial Cells Damaged By Lipotoxicity

Objective: This study used adenovirus to overexpress FUNDC1 and treat EA.hy926 human vascular endothelial cells with palmitic acid to establish a model of high-fat injury endothelial cells.To explore the effect of FUNDC1 on high-fat injury EA.hy926 human vascular endothelial cells and its role possible mechanism.Methods:(1)Establish EA.hy926 human vascular endothelial cell model with high-fat damage: select concentrations of 0.1,0.15,0.2,0.25,0.3,0.5,0.7 mmol/L palmitic acid solution to stimulate EA.hy926 cells,palmitic acid stimulated the cells for 24 h.(2)Protective effects of FUNDC1 on high-fat injury EA.hy926 cells: In vitro culture of EA.hy926 human vascular endothelial cells grew to about 70%-80%.There are four groups of cells: control group,high-fat Group(0.2 mmo/L palmitic acid),high-fat+empty group(transfection with empty adenovirus pretreatment for 24 h+0.2mmol/L palmitic acid),high-fat+FUNDC1 group(transfection with FUNDC1 adenovirus pre-treatment for 24 h+0.2 mmol/L palmitic acid)performed CCK8 test to detect cell proliferation activity in each group;DCFH-DA reactive oxygen kit to detect changes in reactive oxygen content of cells in each group;Changes of mitochondrial membrane potential measured by JC-1 method;The kit was used to detect the ATP content of each group of cells;Detection of P62,LC3,Bcl-2,Bax in each group by western blot.Results:(1)Cells with a concentration of 0.2 mmol/L of palmitic acid stimulated the cells for 24 h,and the cell viability was significantly reduced compared with the control group(P<0.01).(2)The cell viability of the high-fat group and the high-fat+empty group decreased significantly compared with the control group(P<0.01),between the high-fat group and the high-fat+empty group,there was no significant difference(P>0.05);The high-fat+FUNDC1 group survival rate was significantly higher than that of the high-fat group(P<0.01).(3)The content of active oxygen in the high-fat group and the high-fat+empty group increased significantlycompared with the control group(P<0.01),and between the high-fat group and the high-fat+empty group,there was no significant difference(P>0.05);The content of active oxygen in the high-fat+FUNDC1 group was lower than that in the high-fat group significantly(P<0.01).(4)The membrane potential depolarization ratio of the high-fat group and the high-fat+empty group was significantly increased compared with the control group(P<0.01),between the high-fat group and the high-fat+empty group,there was no significant difference(P>0.05);the ratio of membrane potential depolarization in the high-fat+FUNDC1 group was lower than the high-fat group significantly(P<0.01).(5)The ATP content of the high-fat group and the high-fat+empty group was decreased significantly compared with the control group(P<0.01),between the high-fat group and the high-fat+empty group,there was no significant difference(P>0.05);The content of ATP in the high-fat+FUNDC1 group was higher than the high-fat group significantly(P<0.05).(6)The amount of LC3,P62,and Bax in the high-fat group and the high-fat+empty group increased significantly compared with the control group(P<0.05),and the amount of Bcl-2 decreased significantly(P<0.05),between the high-fat+empty groups,there was no significant difference(P>0.05);the amount of LC3,P62,and Bax in the high-fat+FUNDC1 group was lower than the high-fat group significantly(P<0.05),and the amount of Bcl-2increased significantly(P<0.05).Conclusions:(1)Under high-fat conditions,EA.hy926 cell growth is inhibited,ROS content is increased,depolarization ratio of mitochondrial membrane potential is decreased,and ATP content is reduced.Overexpression of FUNDC1 can improve high-fat on EA.hy926 cells Damage.(2)Overexpression of FUNDC1 can promote autophagy of EA.hy926 vascular endothelial cells and reduce the expression of apoptotic proteins.