Paeonol inhibits apoptosis of vascular smooth muscle cells via up-regulation of autophagy by activating class III PI3K/Beclin-1 signaling pathway

Aims: The cross talk between autophagy and apoptosis of vascular smooth muscle cells (VSMCs) plays a vital role in the development of atherosclerosis (AS). Paeonol is isolated from the radix of Cortex Moutan with anti-atherosclerotic and anti-apoptosis effects. However, the mechanisms of paeonol on VSMCs apoptosis are still not fully understood. In this study, we aimed to explore whether paeonol could inhibit VSMCs apoptosis though modulating VSMCs autophagy. Materials and methods: The proteins expressions were detected by western blotting. Autophagosomes and apoptotic body formation in VSMCs was observed by transmission electron microscopy (TEM). VSMCs autophagy was detected by monodansylcadaverine (MDC) staining using fluorescence microscopy, while VSMCs apoptosis was determined by 4',6-diamidino-2-phenylindole (DAPI) and flow cytometry. Key findings: We found that paeonol could significantly increase LC3II protein level, decrease p62 and cleaved caspase-3 proteins levels in aorta of AS mice and ox-LDL-injured VSMCs. Paeonol could augment the number of autophagosomes and reduce the amount of apoptotic bodies in ox-LDL-injured VSMCs. Moreover, paeonol obviously induced VSMCs autophagy compared to ox-LDL group and remarkably suppressed VSMCs apoptosis. However, the effects of paeonol on VSMCs apoptosis could be reversed obviously by 3-MA, the autophagy inhibitor. Furthermore, paeonol could activate class III PI3K-Beclin-1 pathway significantly. Gene silencing of either class III PI3K or Beclin-1 could reverse the effects of paeonol on VSMCs autophagy and apoptosis. Significance: Based on our results, paeonol could induce VSMCs autophagy by activating class III PI3K/Beclin-1 signaling pathway, thus ultimately inhibiting VSMCs apoptosis.