Study On Molecular Mechanism Of Injury Induced By PM2.5 In HUVEC And Beas-2B Cells

Objective:To explore the molecular mechanism of ANGII and its receptors' activation in mediating cell damage effects in human umbilical vein endothelial cells(HUVEC)induced by arsenite exposure;To investigate the contribution of p53-dependent autophagy to the induction of inflammatory factors expression in the human bronchial epithelial cells(Beas-2B)induced by particulate matter 2.5(PM2.5).Methods:1.HUVEC cells were cultured in vitro and treated with arsenite.ang? atl,at2 and hif1 ?mRNA expression levels were tested by reverse transcriotion-polymerase chain reaction(RT-PCR)and quantificational real-time polymerase chain reaction(Real-time PCR);the transactivation of AP-1 was detected by dual-luciferase reporter analysis system;expression levels of ANGII,sICAM-1 and IL-8 were tested by enzyme-linked immuno sorbent assay(ELISA);flow cytometric analysis was used to detect expression levels of VCAM-1,E-selectin,P-selectin and the adhesion of U937 cells and HUVEC cells;Western Blotting was used to test induction of c-Jun and ATF2 activaion and expression of ANGII,AT1,AT2,HIF-1?,IRE 1? and XBP1.2.PM2.5 samples were collected in Wuhan;the induction of autophagic flux was detected by flow cytometric analysis;autophagy was detected by Laser scanning confocal microscope;p53 transactivity and the transcription of vegf gene were tested by dual-luciferase reporter analysis system;Western Blotting was used to detect activation of p53 and STAT3,induction of LC3B?/II,Beclin-1,Tigar,Sestrin-2,DRAM and VEGF expressions.Results:1.ANGII and its receptors mediated inflammatory responses in human umbilical vein endothelial cells(HUVEC)induced by arsenite exposure.(1)Arsenite exposure upregulated ANGII and its receptors expression in HUVEC cells.(2)HIF1? was responsible for arsenite-induced ANGII and its receptors expression in HUVEC cells.(3)ER stress mediated by IRE-1/XBP-1 was responsible for HIF1? accumulation in HUVEC cells under arsenite exposure.(4)ER stress mediated by IRE-1/XBP-1 was responsible for ANGII and its receptors expression in HUVEC cells under arsenite exposure.(5)Arsenite upregulated IL-8 and E-Selectin,VCAM-1,P-Selectin,sICAM-1 expression in HUVEC cells.(6)Arsenite promoted the adhesion of U937 cells and HUVEC cells(7)ANGII and its receptors were responsible for the adhesive molecules and IL-8 expression in HUVEC cells under arsenite exposure.(8)ANGII and its receptors were responsible for the adhesion of U937 cells and HUVEC cells in HUVEC cells under arsenite exposure.2.p53-dependent autophagy mediated induction of the inflammatory factor VEGF expression in bronchial epithelial cells(Beas-2B)induced by PM2.5.(1)PM2.5 induced p53 activation in Beas-2B cells.(2)PM2.5 induced autophagy in Beas-2B cells.(3)PM2.5 induced autophagy depended on p53 activation in Beas-2B cells.(4)p53-dependent autophagy was responsible for PM2.5-induced the inflammatory factor VEGF expression.Conclusions:1.(1)ANGII and its receptors are responsible for IL-8,E-Selectin,VCAM-1,P-Selectin and sICAM-1 expression in HUVEC cells under arsenite exposure.ANGII and its receptors are responsible for the adhesion of U937 cells and HUVEC cells in HUVEC cells under arsenite exposure.(2)HIF1? and ER stress responses signal molecules IRE-1/XBP-1 are upstream signaling molecules of ANGII and its receptors,IRE-1/XBP-1 is upstream signaling molecule of HIF1?.2.PM2.5 induces autophagy depended on p53 activation in Beas-2B cells;PM2.5-induced autophagy response is responsible for VEGF expression.VEGF involves in the process of injury induced by PM2.5 in the bronchial epithelial cells.