| Objective:To investigate effects of EMs on synthesis and secretion of vasoactive substances by human umbilical vein endothelial cells (HUVECs) in the condition of advanced glycation end products (AGEs).Methods:HUVECs were isolated from freshly collected umbilical cords, stimulated with AGEs-bovine serum albumin (AGEs-BSA), bovine serum albumin (BSA) or both AGEs-BSA and EMs, respectively. Then, HUVEC survival rate was calculated by MTT assay, the levels of NO, endothelial nitric oxide synthase and inducible nitric oxide synthase (eNOS, iNOS) were detected by colorimetric analysis, contents of endothelin-1 (ET-1) were detected by ELISA. Furthermore, expression of eNOS, ET-1 mRNA was measured by reverse transcription polymerase chain reaction (RT-PCR). In addition, expression of p38 mitogen-activated protein kinase (p38MAPK) was detected by immunofluorescence assay.Results:Cell viability was time dependently decreased by exposed to AGEs-BSA (P＜0.01), AGEs-BSA can time dependently increase NO, iNOS, ET-1 concentration and up regulate ET-1 mRNA expression, decrease eNOS secretion and down regulate eNOS mRNA expression (P<0.05), and elevate the fluorescence intensity of p38MAPK in nucelus. While cell viability, secretion of NO,iNOS,ET-1, mRNA expression of ET-1 was significantly decreased after incubation with EMs compared to that with AGEs-BSA (P<0.05), secretion and mRNA expression of eNOS had opposite changes (P<0.05), which in a time and concentration dependent manner. EMs also decreaed the fluorescence intensity of p38MAPK in nucelus compared to AGEs-BSA group.Conclusion:It was demonstrated that AGEs-BSA decreased the cell viability of HUVECs, and impaired the function of secreting endothelial active factors such as NO,eNOS,iNOS,ET-1. EMs has a certain protection effect on AGEs-BSA-induced injury in HUVECs as time and concentration dependent manner, and a inhibitory effect on the expression of p38MAPK in the nucelus activated by AGEs-BSA.
|
PDF Full Text Request