Inhibition Of GSK-3β Alleviates Cerebral Ischemia/reperfusion Injury By Suppressing NLRP3 Inflammasome Activation Through Autophagy

Background:Ischemic cerebrovascular disease is a common disease that seriously affects human health and has the characteristics of high morbidity,mortality and disability.The pathogenesis of cerebral ischemia-reperfusion injury involves a variety of energy metabolism disorders in the brain tissue,and the formation of a cascade cascade aggravates the damage,leading to more neuronal cell death.Recent studies have shown that activation of NLRP3 inflammatory complex is a key link in neuronal cell injury.Its activation can promote the inflammatory cascade after cerebral ischemia-reperfusion and aggravate cerebral ischemia-reperfusion injury.Therefore,it is of great significance to find out the regulation of inflammatory complex and elucidate its regulatory mechanism.Objective:In this study,we examined whether NLRP3 inflammasome-derivedinflammation could be ameliorated by GSK-3β inhibition in a cerebral ischemia reperfusion injury model and assessed whether autophagy is involved in this process.Methods:(1)The Longa line plug method was used to establish the model of middle cerebral artery embolization in SD rats.The neurological function scores in each group were observed.The infarct volume of rats was detected by TTC staining.(2)Using the specific GSK-3β siRNA and intracerebroventricular injection of siRNA 24 h before the establishment of MCAO model,the lateral ventricle injection was performed 6 hours before the establishment of MCAO model using GSK-3β inhibitor SB216763,using autophagy inhibitor 3-MA was injected into the lateral ventricle 30 minutes before the establishment of the MCAO model.(3)Western blot analyze the expression of GSK-3β,GSK-3β(p-Tyr216),NLRP3 inflammasome and its downstream inflammatory factors cleaved-caspase-1,IL-1β,IL-18,autophagy-associated proteins LC3B-I,LC3B-II and p62.(4)The concentration of IL-β and IL-18 protein was detected by ELISA.(5)Transmission electron microscopy showed the inflammation of neurons and the number of autophagosomes in each experimental group.Results:(1)Inhibition of GSK-3β activity can alleviate cerebral ischemia-reperfusion injury in rats.(2)After inhibiting the activity of GSK-3β,the activity of NLRP3 inflammatory complex decreased,and the expression of downstream inflammatory factors cleaved-caspase-1,IL-1β,IL-18 decreased,the volume of cerebral infarction decreased significantly,and behavioral defects were also observed.Improved.(3)After cerebral ischemia-reperfusion injury in rats,the activity of GSK-3β was inhibited and autophagy was enhanced.(4)After the autophagy was up-regulated on the basis of inhibition of GSK-3β activity,the activity of NLRP3 inflammatory complex and the expression levels of downstream inflammatory factors cleaved-caspase-1,IL-1β and IL-18 were increased.Conclusion:(1)Inhibition of GSK-3β activity can alleviate cerebral ischemia-reperfusion injury in rats by down-regulating the activity of NLRP3 inflammatory complex and its downstream inflammatory factors cleaved-caspase-1,IL-1β,IL-18.(2)Inhibition of GSK-3β activity is to reduce the activity of NLRP3 inflammatory complex by enhancing autophagy activity.