Paeonol Inhibits PM2.5-induced VEGF Expression In Human Bronchial Epithelial Cells

Part 1Effects of PM2.5 on Expression of VEGF in Bronchial Epithelial CellsObjectives:To investigate the effects of particulate matter(PM)2.5 on the expression of VEGF in bronchial epithelial cells.Methods:BEAS-2B cells were subcultured in DMEM medium containing 10%FBS until to 80%in fusion.After washed twice with PBS and digested by trypsin,BEAS-2B cells were inoculated on the six-well plate with the density of 6×105 cells/ml.Sterilized phosphate buffer(PBS)was used to prepare PM2.5 suspension,containing 0,25,50,100 ug/ml PM2.5,respectively.The cells were devided into control group and PM2.5 treated groups.Cells in control group were incubated in the growing medium with PM2.5 of 0 ?g/mL.Cells in other groups were stimulated with a series of concentrations(25,50 and 100 ?g/mL)of PM2.5 for 24h.Supernatants and cells were harvested at 6h,12h and 24h in each group.Enzyme linked immunosorbent assay(ElISA)was used to analyse VEGF level in supernatants,and western blot was used to detect VEGF protein level in BEAS-2B cells.Results:VEGF levels in the supernatants in PM2.5 treated groups were significant increased with a concentration-dependent manner as compared to the control group(with P value of 0.001?0.0252.0.0381,respectively).The VEGF protein levels in BEAS-2B cells induced by PM2.5 were significant higher than that in the control group,and showed concentration-dependent manner(with P value of 0.0279,0.0004 and 0.0043 respectively).Meanwhile,at the same PM2.5 density,VEGF levels in the supernatants were significant increased with a time-dependent manner.Conclusions:The expression of VEGF protein in BEAS-2B cells were induced by PM2.5 with a concentration-dependent and time-dependent manner.Part 2Effects of aeonol on the expression of VEGF in bronchial epithelial cells induced by PM2.5Objectives:To explore the effects of paeonol on the expression of VEGF in BEAS-2B cells induced by PM2.5.Methods:BEAS-2B cells were incubated in the growing medium with PM2.5 of 0,25,50 and 100 ug/ml,and treated with paeonol of 15 ?mol/L or 30 ?mol/L for 24h respectively.Supernatants were collected to detect the levels of VEGF in each group by ELISA.Results:Treatment with paeonol could not decrease the VEGF level in BEAS-2B cells at baseline(without PM2.5);however,paeonol could inhibit the expression of VEGF in BEAS-2B cells induced by PM2.5 with different density.Compared with control group,there was no significant increase of VEGF in BEAS-2B cells treated with paeonol of 15 ?mol/L or 30 ?mol/L(all P>0.05).On the contrary,when induced with PM2.5 of 25,50 and 100 ug/ml,paeonol of 15 ?mol/L could decrease the level of VEGF in the BEAS-2B cells as compared with the groups without paeonol(P value<0.001,0.008,and<0.001,respectively).The levels of VEGF in the BEAS-2B cells treated with paeonol of 30 ?mol/L were even lower than the groups with paeonol of 15 ?mol/L(P value<0.001,<0.001 and?0.015,respectively).Conclusions:Paeonol could inhibit the expression of VEGF in BEAS-2B cells induced by PM2.5.