Adenosine monophosphate-activated protein kinase/mammalian target of rapamycin-dependent autophagy protects human dental pulp cells against hypoxia.

摘要: Abstract Introduction Human dental pulp cells (HDPCs) are recalcitrant to hypoxic stress. We investigated whether hypoxia-induced autophagy of HDPCs offered these a survival advantage and the underlying mechanism this resistance. Methods The viability apoptosis were examined after exposure hypoxia by Vi-CELL cell analyzer flow cytometry. Autophagy was assessed using immunofluorescence, acridine orange staining, real-time polymerase chain reaction, Western blotting. Either 3-methyladenine or expression vectors encoding dominant negative ATG5 used inhibit autophagy. Rapamycin as an autophagic inducer. To explore mechanisms autophagy, adenosine monophosphate–activated protein kinase (AMPK)/mammalian target rapamycin (mTOR) pathway hypoxia-inducible transcription factor-1 suppressed chemical inhibitors Compound C YC-1, respectively. Results had no effect on resulted in increasing acidic vesicular organelle-positive cells, autophagosome formation, up-regulation genes. Inhibition with 3- methyladenine abrogated protective effects HDPCs. phosphorylation AMPK up-regulated, whereas mTOR down-regulated hypoxia-treated HDPCs, which both attenuated C. Furthermore, treatment rather than YC-1 reduced Conclusions Our results suggested that might be cytoprotective against stress via AMPK/mTOR signaling pathway.