Gomisin N attenuated cerebral ischemia-reperfusion injury through inhibition of autophagy by activating the PI3K/AKT/mTOR pathway

Ischemic stroke is a major global cause of mortality and permanent disability. Studies have shown that autophagy is essential to maintain cell homeostasis and inevitably lead to neuronal damage after cerebral ischemia. Gomisin N (GN), lignin isolated from Schisandra chinensis, possesses multiple pharmacological activities. However, there is no research on the potential of GN for neuroprotection in ischemic stroke.The current work aimed to explore the potential therapeutic possibilities of GN on ischemic stroke and investigate the underlying molecular mechanisms.The neuroprotective effects of GN on PC12 cells induced by oxygen glucose deprivation/reoxygenation (OGD/R) and mice with middle cerebral artery occlusion/reperfusion (MCAO/R) injury were investigated.On day 3 after ischemia, the infarct volume and neurological function were assessed. The level of autophagy was measured in vivo and in vitro using Transmission electron microscopy (TEM) and Monodansylcadaverine (MDC) staining. The interaction between GN and PI3K/AKT/mTOR pathway was investigated by molecular docking. Additionally, the expressions of critical proteins in the PI3K/AKT/mTOR signaling pathway and autophagy markers were determined by western blotting.In compared to the Model group, GN might considerably improve the neurological and locomotor function following a stroke, as well as lower the volume of the cerebral infarct volume and the number of autophagosomes. GN therapy may suppress autophagy by activating the PI3K/Akt/mTOR signaling pathway in the penumbra. In vitro, MDC and TEM results showed that GN treatment obviously suppressed autophagy. Meanwhile, GN downregulated LC3II/LC3I expression ratio while upregulated the p62 expression level. In further studies, GN dramatically boosted the expression ratios of p-PI3K/PI3K, p-AKT/AKT, and p-mTOR/mTOR proteins in PC12 cells following OGD/R damage. However, the PI3K inhibitor (LY294002) reversed the increase of p-PI3K/PI3K, p-AKT/AKT, and p-mTOR/mTOR expression ratio induced by GN administration. Also, LY294002 significantly partially attenuated GN induced reduction of autophagy and increase of cell viability compared with GN treatment alone.Here, we first demonstrate the neuroprotective effects of GN on MCAO mice and OGD/R induced PC12 cells injury. A possible mechanism by which GN prevents ischemic stroke is proposed: GN could restrain autophagy by stimulating the PI3K/AKT/mTOR signaling pathways. More effects and mechanisms of GN on the rehabilitation of ischemic stroke are worthy to be explored in the future.