Study Of Therapeutic Effect And Underlying Mechanisms Of PI3K Inhibitor ZSTK474 In Mice With Cerebral Ischemia/Reperfusion Injury

Background: In cerebral ischemia/reperfusion injury,inflammation and immune response play an important role,and more and more attention in recent years.The studies have shown that adjustment after cerebral ischemia/reperfusion injury of inflammation,immune response,can improve ischemia after a series of reactions,thus plays the brain protective effect.Among them,the microglia/macrophages as the inherent immune cells of the central nervous system,is considered to be the most potential regulation and control targets.The damaged central nervous system(CNS),such as cerebral ischemia/reperfusion injury,microglia/macrophages can reversibly change their phenotype to "detrimental" state or" restorative" status,so as to control inflammation,play a role of brain protection.So,to seek a kind of compound,through regulation of microglia/macrophages phenotype,thereby inhibiting ischemia/reperfusion injury caused by a series of inflammation,immune response,improve the prognosis of cerebral ischemia/reperfusion injury,has become a hot spot of research.Methods: Our research using 6-8 weeks C57 / BL6 male mice as the research object.The mice were randomly divided into shame group?control group and ZSTK474 treatment group,a total of three groups.To Longa line optimization method to block the mice model of middle cerebral artery,1 h ischemia and reperfusion for 24 h.The shame group and the control group given equivalent of PBS lavage,ZSTK474 group gavaged according to the 200 mg/kg,6 h after ischemia/reperfusion,1 day 1,for three consecutive days.We investigated the effect of ZSTK474 by assessing nerve function score groups?TTC staining to evaluate cerebral infarction volume?HE dyed the detection of the number of inflammatory cells infiltrating central injury?immunofluorescence observations Iba1,GFAP,CD16/32,CD206 expression of ischemic penumbra?Rt-PCR detection of IL-?,IL-6,IL-1,TNF-?,IL-10 and TGF-?gene expression;and ELISA test IL-?,IL-6,IL-1,TNF-?,IL-10 and TGF � ?expression in protein levels.At last,by Western blot test AKT,P-AKT,p70S6 K and P-p70S6 K expression.Finally using Tukey's test and the Mann-Whitney U test statistical analysis was carried out on the components,compare their differences.Results: Our study shows that,compared with control group,ZSTK474 treatment group obviously improve nerve function score,reduce the area of cerebral infarction,reduce inflammatory cell infiltration of the ischemia/reperfusion injury in mice model.The ZSTK474 treatment group,meanwhile,inhibite microglia and astrocyte cell proliferation,regulating microglial/macrophage differentiation at the same time,the regulation of microglial/macrophages in a"restorative" status,thereby inhibiting the secretion of proinflammatory factor,promote the generation of anti-inflammatory factors,play a key action of brain protection;Our study found that ZSTK474 treatment group p-AKT and their downstream products p-p70S6 K expression was reduced,ZSTK474 as PI3 K inhibitors,the mechanism about the brain protection may be related to regulate PI3K/AKT/mTOR pathway.Conclusion: Cerebral ischemia/reperfusion injury in mice model,ZSTK474 can inhibit the proliferation of microglia/macrophages,regulating microglial/macrophage differentiation,and inhibiting proinflammatory factor secretion,promote the anti-inflammatory factor,by adjusting the cerebral ischemia/reperfusion injury induced inflammation,immune response,give play to the role of brain protection.ZSTK474 regulate PI3K/AKT/mTORC1 signalling pathways to acted the brain protection.Therefore,ZSTK474 may represent a therapeutic intervention with potential for circumventing the catastrophic aftermath of ischemic stroke.