Study On Signal Pathways Involved In Vascular Endothelial Cell Migration Induced By Endosulfan

Environmental pollution is closely related to human diseases.Persistent organic pollutants(POPs)typified by endosulfan are important risk factors for cardiovascular disease.Atherosclerosis(AS)is the common pathological basis of many cardiovascular diseases.Vascular endothelial cell migration plays an important role in the early formation of AS,angiogenesis and vascular remodeling within the AS plaque.Previous studies found that low-dose endosulfan(0.1-20 ?M)could reduce the production of nitric oxide(NO)that has an inhibitory effect on the migration of vascular endothelial cells.Therefore,in this study the effects and signaling' pathways of cell migration as well as the post-transtriptional regulatory mechanism were explored in vascular endothelial cells exposed to low-dose endosulfan.In this study,the effect of low-dose endosulfan on vascular endothelial cell migration was first investigated.Wound healing and Transwell assay were carried out to determine cell migration ability.The results showed that endosulfan enhanced cell migration abilities in vascular endothelial cells.Then the molecular mechanism of cell migration in endosulfan-exposed vascular endothelial cells was explored.To examine mRNA and protein expression alterations in signaling pathway of cell migration,qRT-PCR and Western blot were performed in 20 ?M endosulfan-exposed cells in the presence or absence of inhibitors.The results showed that.endosulfan increased the expression of prote in tyrosine phosphatase 4 A3(PTP4A3)in a dose-dependent manner.Specific inhibitor for PTP4A3 could reduce the expression of PTP4A3 and inhibit endosulfan-induced cell migration and activation of multiple signaling pathways related to endothelial cell migration including MAPK/ERK,PI3K/AKT and NF-?B signaling pathway.Endosulfan also caused oxidative stress response,but antioxidant(NAC)treatment had no effect on cell migration induced by endosulfan.Finally,the post-transcriptional regulatory mechanism in endosulfan-induced upregulation of PTP4A3 was analyzed.qRT-PCR results showed that endosulfan exposure resulted in the decrease of hsa-miR-140-5p and the increase of KCNQ1OT1.Bio informatics analysis was utilized to screen out IncRNA KCNQ10T1 that can share hsa-miR-140-5p with PTP4A3 to form target corresponding relationship.Double luciferase reporter assay confirmed that KCNQ10T1 acted as a target to bind with hsa-miR-140-5p and PTP4A3 might be the target gene of hsa-miR-140-5p.In summary,exposure to low-dose endosulfan can promote vascular endothelial cellmigration and increase the expression of PTP4A3 through lncRNA-miRNA regulatory mechanism.Endosulfan activated MAPK/ERK,PI3K/AKTand NF-?B signaling pathways mediated by PTP4A3 in endothelial cell migration This study is of great scientific significance for better understanding the health risk of long-term exposure to POPs and revealing potential impacts on cardiovascular disease.