The Role And Mechanism Of BDNF/TRK B/CREB Signaling Pathway In SK-N-SH Cells Induced By BPS

Objective:The aim is to study the toxic damage and potential toxicity mechanism of bisphenol S-induced SK-N-SH cells,and to provide a basis and theoretical basical for further study of the neurotoxicity of bisphenol S.Methods:1.According to the previous experiment,the experiment was divided into control group(DMSO)and experimental group(10,20,50,100,150,200,250,300?mol/L BPS).SK-N-SH cells were treated with different concentrations of bisphenol S for 48 hours.The cell viability was detected by MTT assay,and the median lethal concentration of SK-N-SH cells treated with bisphenol S for 48 hours was calculated.The morphological changes of SK-N-SH cells were observed under light microscope.Flow cytometry detected the apoptosis and cell cycle of SK-N-SH cells.The changes of mitochondrial membrane potential were detected by rhodamine 123 dye.The expression level of BDNF in SK-N-SH cells was detected by ELISA.The protein levels of Cleaved-caspase3,Bcl-2,Bax,TrkB,CREB and p-CREB were detected by Western Blot.2.After pretreatment with TrkB activator 7,8-DHF,SK-N-SH cells were treated with bisphenol S(250 ?mol/L BPS)and DMSO(control group)for 48 hours,the morphological changes of SK-N-SH cells were observed under light microscope;flow cytometry detected the apoptosis and of SK-N-SH cells.The changes of mitochondrial membrane potential were detected by rhodamine 123 dye.The expression level of BDNF in SK-N-SH cells was detected by ELISA.The protein levels of Cleaved-caspase3,Bcl-2,Bax,TrkB,CREB and p-CREB were detected by Western Blot.Results:1.MTT assay showed that bisphenol S could decrease the activity of SK-N-SH cells.After 48 h,bisphenol S could decrease the activity of SK-N-SH cells in a concentration-dependent manner.Bisphenol S caused morphological changes in SK-N-SH cells,cell synapses became shorter,and even cells became round;flow cytometry results showed that bisphenol S caused apoptosis in SK-N-SH cells,and occurred Cell cycle arrest,block occurs in the G1 phase.The results of rhodamine dye test showed that bisphenol S can decrease the mitochondrial membrane potential of SK-N-SH cells.The results of ELISA showed that bisphenol S decreased the expression of BDNF protein.Western Blot results showed that bisphenol S decreased the phosphorylation of TrkB and CREB,increased Bax and cleaved-caspase3,but decreased the expression of Bcl-2.2.After pretreatment with TrkB activator 7,8-DHF,the activity of SK-N-SH cells was increased compared with the bisphenol S group;flow cytometry results showed that the apoptosis rate of SK-N-SH cells was reduced;Rhodamine dye The results showed that the mitochondrial membrane potential of SK-N-SH cells increased;the ELISA results showed that the expression of BDNF protein in SK-NSH cells was increased,and the results of Western Blot showed that the phosphorylation levels of TrkB and CREB in SK-N-SH cells were increased.The expression levels of Bax and cleaved-caspase3 were decreased,and the expression level of Bcl-2 was increased.TrkB activator 7,8-DHF attenuated the toxic effects of bisphenol S on SK-NSH cells.Conclusion:1.Bisphenol S causes decreased activity of SK-N-SH cells,impairs cell morphological structure,leads to apoptosis and cell cycle arrest,and is cytotoxic.2.BDNF/TrkB/CREB signaling pathway mediates the effects of bisphenol S on SK-N-SH cell injury.