The Effect Of Fingolimod On The Injury Of Vascular Endothelial Cells Induced By Advanced Glycation End Products

At present,the incidence and prevalence of diabetes is increasing year by year in the world.The complications of diabetes seriously affect the health of patients with diabetes.Among them,diabetic vascular complications has become the main cause of disability and death in patients with diabetes.The role of advanced glycation end products in the pathogenesis and development of diabetic angiopathy has attracted much attention.Advanced glycosylation end products can activate intracellular signaling pathways,induce the release of downstream inflammatory factors and damage vascular endothelial cells.However,the specific signal pathway remains to be confirmed.Some studies have shown that FTY720 may play an important role in the treatment of diabetic vascular lesions.At present,animal experiments in vivo have confirmed that FTY720 can be associated with 1-phosphoric acid on vascular endothelial cells.Sphingosine receptor 1and 3,Effects of FTY720 on the synthesis and release of downstream signaling molecules,thus improving the function of endothelial cells and protecting vascular endothelial cells.However,the effect of FTY720 on the damage induced by advanced glycation end products and its specific mechanism are not well understood.Objective:To study the protective effect and signaling pathway of FTY720 on vascular endothelial cell injury induced by advanced glycation end products.Method:EA.hy926 cell lines which were cultured in vitro were divided into groups: the control group,the AGEs injury group,and the FTY720+AGEs group.Cells in the FTY720+AGEs group were reated with 7.5 mmol/L FTY720 for 6 hours.After simultaneous treatment without any reagents for the same time in the control group and the AGEs injury group,AGEs(the final concentration was 200 mg/L)was added to AGEs injury group to observe the morphology of the cells under microscope after 12 hours.CCK-8 assay was used to detect cell viability,The content of NO in supernatant of culture medium was determined by Griess method.The concentration of IL-6 and IL-1 ?in the cell homogenate was detected by Elisa kit.Western blot assay was used to detect the level of PI3 K phosphorylation in the cell homogenate.Result: AO/EB staining showed the chromatin was green and the structure was normal,the cell morphology was normal polygonal or fusiform in the control group.Part of the cells in AGEs injury group became orange,nuclear pyknosis,swollen.FTY720+AGEs group showed green chromatin pyknosis.Cell antennae disappear,slightly rounded.The results of CCK-8 method showed that,Compared with the control group,the cell viability of the AGEs injury group and the FTY720+AGEs group was decreased(P <0.05),and that of FTY720+AGEs group was higher than that of AGEs group(P <0.05).Compared with the AGEs injury group,the concentration of NO,IL-6,IL-1 ? and the phosphorylation level of PI3 K decreased(P < 0.05)in FTY720+AGEs group.Conclusion:Our study demonstrates that AGEs can cause the release of NO,IL-6,IL-1beta and other substances by enhancing the phosphorylation of PI3 K,cause endothelial cells damage.FTY720 can inhibit the phosphorylation of PI3 K and reduce the release of NO,IL-6,IL-1? and so on,thus reduce the endothelial cell damage caused by AGEs.