BDMC Activates AMPK/SIRT1 Pathway Improve The Toxic Effect Of A?_1-42 On SK-N-SH Cells

[Objective]This study was to investigate the mechanism that BDMC improves the toxic effects of A?_1-42 on SK-N-SH cells.And confirm BDMC improve toxic effects of A?_1-42 on SK-N-SH cells whether through activate AMPK and up-regulates SITI1.[Methods]1.The AD cell model was constructed by inducing SK-N-SH cells by A?_1-42.The activity of SK-N-SH cells was detected by MTT method and the optimal model concentration of A?_1-42 was screened out.2.To explore the protective effect of BDMC on A?_1-42-induced SK-N-SH cells.The activity of SK-N-SH cells was detected by MTT and the optimal therapeutic concentration was screened.SOD and GSH were observed to observe the protective effect of BDMC on A?_1-42-induced SK-N-SH cells.Western blotting was used to detect the expression of p-AMPK and SIRT1 to observe the effect of BDMC on AMPK/SIRT1pathway.3.To investigate the effect of AMPK inhibitor Compound C on the protective effect of BDMC on A?_1-42-induced SK-N-SH cells.Western blotting was used to detect the expression of p-AMPK and observe the inhibitory effect of Compound C.The cell viability was detected by MTT,the SOD and GSH were detected by kit method,and the expression of SIRT1 protein was detected by western blotting.The effect of Compound C on the protective effect of BDMC on A?_1-42-induced SK-N-SH cells was observed.4.To investigate the effect of SIRT1 inhibitor EX527 on the protective effect of BDMC on A?_1-42-induced SK-N-SH cells.Western blotting was used to detect the expression of SIRT1 and observe the inhibitory effect of EX527.Cell viability was detected by MTT,SOD and GSH were detected by kit method.The effect of EX527 on the protective effect of BDMC on A?_1-42-induced SK-N-SH cells was observed.[Results]1.BDMC can protect the toxic effects of A?_1-42 on SK-N-SH cells.1.1 Treatment of SK-N-SH cells with A?_1-42?5,10,15,20?mol/L,24 h?can significantly reduce the viability of SK-N-SH cells,demonstrating that A?_1-42 has cytotoxic effects.Based on the MTT results,a subsequent study selected 10?mol/L of A?_1-42 as the drug concentration for establishing an AD cell model.1.2 Treatment of SK-N-SH cells induced by A?_1-42 by BDMC?5,10,15?mol/L,24h?can significantly improve the viability of SK-N-SH cells and screen out the optimal concentration of BDMC.We found that BDMC can increase the SOD and GSH of SK-N-SH cells induced by A?_1-42,activate AMPK phosphorylation,and up-regulate SIRT1 expression,suggesting that BDMC can antagonize A?_1-42 to SK-N-SH and may be related to the AMPK/SIRT1 pathway.2.AMPK inhibitor Compound C can reverse the protective effect of BDMC on A?_1-42-induced SK-N-SH cells.After treatment with A?_1-42-induced SK-N-SH cells by Compound C,BDMC could not improve cell viability,and could not increase SOD,GSH and SIRT1 expression,indicating that inhibition of AMPK could cause loss of BDMC.A?_1-42-induced protection of SK-N-SH cells and up-regulation of SIRT1.3.SIRT1 inhibitor EX527 can antagonize the protective effect of BDMC on A?_1-42-induced SK-N-SH cells.After treatment of SK-N-SH cells induced by A?_1-42 by EX527,BDMC lost the improvement of cell viability,and did not increase the activity of SOD and GSH,indicating that inhibition of SIRT1 could cause loss of BDMC.A?_1-42 induced protection of SK-N-SH cells.[Conclusion] BDMC may improves the cytotoxic effect of A?_1-42 on SK-N-SH cells by activating phosphorylation of AMPK and up-regulating the level of SIRT1.