Protective Effect Of Metformin On Vascular Endothelial Cell Damage Induced By Saturated Fatty Acid

Objective: Using palmitic acid to stimulate vascular endothelial cell and change chemokines in cells,levels of adherence factor,and affect its phosphorylation-level and induce cell injury and apoptosis,to detect Intervention and protection of metformin from damage induced by palmitic acid on vascular endothelial cells and to make further research on its potential mechanism,to provide new method and target on diabetes mellitus combined with cardiovascular disease.Methods:(1)Five experimental groups were divided as control group,PA group(300?mol/L),low concentrations of metformin(1mmol/L)+PA group,moderate concentrations of metformin(2mmol/L)+PA group and high concentrations of metformin(4mmol/L)+PA group.Cultivating human umbilical vein endothelial cells for 24 hours.Using the method of CCK-8 to detect cell viability(2)Using Westenblot to detect protein expression of NF?B p65,ICAM1,I?B?,p-I?B?,AMPK and p-AMPK.(3)Using qRT-PCR to detect mRNA expression of CCL2 and ICAM1.Results:(1)Compared with blank control group,protein expression of NF?B p65 in PA group significantly increased(P<0.05).Compared with PA group,expression level is significantly lower in Met combined with PA groups(P all<0.05).Among these groups,moderate concentration group has lower amount of protein expression of NF?B p65 than the other two concentration groups(P all<0.05).(2)Protein expression of p-I?B?: Compared with blank control group,expression of p-I?B? in PA group increased significantly(P<0.05);Compared with PA group,expression level is significantly lower in Met combined with PA groups(P all<0.05).Among these groups,moderate concentration group has lower amount of protein expression of p-I?B? than the other two concentration groups(P all<0.05).(3)Compared with blank control group,protein expression of ICAM1 significantly increased(P<0.05);Compared with PA group,protein expression decreased progressively as Met gradient increased in Met combined with PA groups(P all<0.05).(4)Protein expression of p-AMPK: Compared with blank control group and PA group,protein expression significantly increased with increase of the concentration gradient of Met in Met combined with PA groups(P all <0.05).Among these groups,moderate concentration group has higher amount of protein expression of p-AMPK than the other two concentration groups(P all<0.05).(5)mRNA expression of ICAM1: Compared with blank control group,expression of ICAM1 in PA group increased a little(P<0.05);Compared with PA group,mRNA expression of ICAM1 decreased as the increase of the concentration gradient of Met in combination groups(P all<0.05).(6)mRNA expression of CCL2:Compared with blank control group,mRNA expression of CCL2 significantly increased(P<0.05);Compared with PA group,mRNA expression of CCL2 decreased as increase of the concentration gradient of Met in combination groups(P all<0.05).Conclusion: Metformin can inhibit the NF-?B pathway to reduce the inflammatory reaction of vascular endothelial cells caused by palmitic acid and it can also reduce the high expression of cell adhesion molecules and chemotactic factor to protect vascular endothelial cells.The adenosine monophosphate activated protein kinase was activated in all metformin groups,so we can suspect that the activation of AMPK may play a important role in the inhibition of NF-?B pathway.