P11 Signaling pathways involved in exogenous hydrogen sulfide and donepezil regulation of APP alpha metabolism

Abstract

Objective To observe the effects of PI3-K, MAPKs and JNK on APP α metabolism treated with exogenous hydrogen sulfide (H 2 S) and Donepezil in PC12 cell. Methods PC12 cell were cultured in vitro and NaHS were exogenous H 2 S donor. Cells were pretreated with SP600125 (SP, JNK inhibitor 20 μM), LY294002 (LY, PI3-K inhibitor, 20 μM) or SB203580 (SB, MAPK inhibitor, 30 μM), before NaHS (100 μM) or Donepezil (20 μM) was added in the medium.24 h later, A β 40, A β 42, sAPPa, sAPPs protein levels were measured by ELISA. The APP, ADAM10, ADAM17 protein were measured by Western blotting. Results (1) The APP protein level is not inhibited by SP, LY and SB ( p > 0.05). However, SP and LY inhibit ADAM10 and ADAM17 protein expression. (2) SB has no obvious effect on A β 40, A β 42, sAPP β , sAPP α protein expression ( p > 0.05), whereas LY and SP enhance A β 40, A β 42, sAPP β protein level and reduce sAPP α protein expression ( p 2 S enhancement of ADAM10; SB, SP and LY reduce Donepezil modulation of ADAM10 and ADAM17. (4) Compared with control, SP and LY inhibit H 2 S reduction of A β 40, A β 42, sAPP β protein expression ( p β 40, A β 42, sAPP β protein expression ( p 2 S enhancement of sAPP α protein expression ( p α protein expression ( p Conclusion Exogenous H 2 S up-regulates the expression of ADAM10 and ADAM17 and sAPP α , and effectively reduce the production of A β 40, A β 42 and sAPP β in vitro in PC12. The PI3-K or JNK signal pathway may be involved in the regulation. Donepezil has similar effects as H 2 S, which involves PI3-K, JNK or p38 MAPK signaling.