Effect Of Coreopsis Basalis Nutt On High Glucose-Induced HUVEC Cells

Objective:1.To investigate the effects of chlorogenic acid and isochlorogenic acid on the oxidative stress of HUVEC cells damaged by high glucose.2.To investigate the effects of the coreopsis tinctoria nutt flavonoids,marein,flavanomarein on the oxidative stress and the expression of VEGF,ICAM-1 and MCP-1 proteins of HUVEC cells damaged by high glucose.3.To investigate the effect of marein and flavanomarein on the AMPK/mTOR pathway of human umbilical vein endothelial cells(HUVEC)damaged by high glucose;Methods:MTT and CCK-8 were used to detect cell viability;For oxidative stress,MDA(malondialdehyde),SOD(superoxide dismutase).CAT(catalase),GSH-PX(glutathione-peroxidase)activity were assessed by Enzyme-linked immunosorbent assay(ELISA)method,detection of the protein expression levels AMPK(Adenosine monophosphate activated protein kinase),p-AMPK(Phosphorylated adenosine activating protein kinase.),mTOR(mammalian target of rapamycin),VEGF(Vascular endothelial growth factor),ICAM-1(Intercellular cell adhesion molecule-1)and MCP-1(Monocyte chemoattractant protein-1)were detected by Western Blotting.Results:1.Compared with the normal control,the cell viability of HUVEC was significantly decreased in different concentrations of high glucose groups(P<0.05);but increased in the groups with the treatment of chlorogenic acid and isochlorogenic acid A,when compared with high glucose group(P<0.05).Meanwhile,MDA content increased sharply in HG groups(P<0.05),but the activities of SOD,CAT and GSH-PX decreased dramatically.With the administration of chlorogenic acid and isochlorogenic acid A,all antioxidative marker will increase significantly(P<0.05);Results from Western blotting showed that the protein expression of VEGF,ICAM-1 and MCP-1 increased 24 h after high glucose treatment(P<0.05).2.Compared with the normal control,the cell viability of HUVEC was significantly decreased in high glucose model groups(P<0.05),but increased in the groups with the treatment of marein and flavanomarein,when compared with high glucose group(P<0.05).Meanwhile,MDA content increased sharply in HG groups(P<0.05),but the activities of SOD,CAT and GSH-PX decreased dramatically.With the administration of marein and flavanomarein,all antioxidative marker will increase significantly(P<0.05);Interestingly,marein and flavanomarein treatment will inhibit the effect of HG on these proteins expression(P<0.05).For inflammatory marker,the protein expression of VEGF,ICAM-1,MCP-1 were increased sharply in HG groups when compared with NG,while marein and flavanomarein will decrease their expression.3.while the ratio of p-AMPK/AMPK protein in HG groups was decreased(P<0.05),and the mTOR protein expression was increased(P<0.05).Conclusion:1.Chlorogenic acid and isochlorogenic acid A can protect HUVEC cells by increasing the viability of HUVEC cells and improving the oxidative stress.2.Marein and flavanomarein can reduce the oxidative stress in high glucose-injured cells,down-regulate the expression of inflammatory factors VEGF,ICAM-1 and MCP-1 to exert protective effect on HUVEC cells.3.Marein and flavanomarein can up-regulate the activity of AMPK,and increase the ratio of p-AMPK/AMPK to exert protective effect on HUVEC cells.