Based On The PI3K/AKT Signaling Pathway,study The Pharmacodynamic Basis And Mechanism Of Combination Of Paeon If Lor In And Calycosin-7-glucoside Promotes Ischemic Stroke Recovery

ObjectiveBased on the PI3K/AKT signaling pathway,study the pharmacodynamic basis and mechanism of combination of paeoniflorin(PF)and calycosin7-glucoside(CG)(PF+CG)promotes ischemic stroke recovery.Methods1.Effects of PF+CG components on middle cerebral artery ischemiareperfusion(MCAO)rat model and its correlation with PI3K/AKT signaling pathwayMCAO rat model was prepared by modified suture method,and behavioral scoring,cerebral infarction area,brain tissue water content measurement,using PI3K,p-PI3K,AKT,p-AKT,Bcl-2,Bax,GSK3 β protein expression as indicators,compared the effects of low,middle and high dose PF+CG on MCAO rat models and their correlation with PI3K/AKT signaling pathway.2.Observe the effect of PI3K/AKT signaling pathway inhibitor LY294002 on the anti-ischemia-reperfusion effect of PF+CGMCAO rat model was prepared by modified suture method,and behavioral scoring,cerebral infarction area,brain tissue water content measurement,using PI3K,p-PI3K,AKT,p-AKT,Bcl-2,Bax,GSK3β protein expression as indicators,observe the effect of PI3K/AKT signaling pathway inhibitor LY294002 on the anti-ischemia-reperfusion effect of PF+CG.3.Effects of PF+CG components on oxygen-glucose deprivation and reperfusion(OGD/R)cell model and its correlation with PI3K/AKT signaling pathwayOxygen deprivation method was used to prepare the OGD/R model,CCK8 was used to determine the survival rate of HT22 cells,the contents of SOR,ROS,MDA,and LHD were determined,and apoptosis was detected by flow cytometry and mitochondrial membrane potential,using PI3K,pPI3K,AKT,p-AKT,Bcl-2,Bax,GSK-3β protein expression as indicators,compared the effects of low,middle and high dose PF+CG on OGR/R cell models and their correlation with PI3K/AKT signaling pathway.4.Observe the effect of PI3K/AKT signaling pathway inhibitor LY294002 on the anti-oxidative and glucose deprivation and reperfusion effect of PF+CGOxygen deprivation method was used to prepare the OGD/R model,CCK8 was used to determine the survival rate of HT22 cells,the contents of SOR,ROS,MDA,and LHD were determined,and apoptosis was detected by flow cytometry and mitochondrial membrane potential,using PI3K,pPI3K,AKT,p-AKT,Bcl-2,Bax,GSK-3 β protein expression as indicators,observe the effect of PI3K/AKT signaling pathway inhibitor LY294002 on the anti-oxidative and glucose deprivation effect of PF+CG.Results1.PF+CG promotes MCAO rat recovery correlates with PI3K/AKT signaling pathwayPF+CG significantly decreased neurobehavioral deficits,cerebral infarct volume,and brain edema;increased levels of PI3K,AKT,p-PI3K,p-AKT,and Bcl-2(P<0.01)and reduced BAX and GSK-3 β expression(P<0.01).PF+CG promotes MCAO rat recovery correlates with PI3K/AKT signaling pathway.2.PF+CG promotes MCAO rat recovery via the PI3K/AKT signaling pathwayLY294002 significantly increased neurobehavioral deficits,cerebral infarct volume,and brain edema;decreased levels of PI3K,AKT,p-PI3K,p-AKT,and Bcl-2(P<0.01)and enhanced BAX and GSK-3 βexpression(P<0.01).PF+CG promotes MCAO rat recovery via the PI3K/AKT signaling pathway.3.PF+CG promotes OGR/R cell recovery correlates with PI3K/AKT signaling pathwayPF+CG enhanced cell vitality;elevated levels of SOD;reduced levels of ROS,LDH,and MDA;decreased cell apoptosis rates;increased levels of PI3K,AKT,p-PI3K,p-AKT,and Bcl-2(P<0.01);and reduced BAX and GSK-3 β expression(P<0.01).PF+CG promotes OGR/R cell recovery correlates with PI3K/AKT signaling pathway.4.PF+CG promotes OGR/R cell recovery via the PI3K/AKT signaling pathwayLY294002 reduced cell vitality;reduced levels of SOD;elevated levels of ROS,LDH,and MDA;increased cell apoptosis rates;decreased levels of PI3K,AKT,p-PI3K,p-AKT,and Bcl-2(P<0.01);and enhanced BAX and GSK-3 β expression(P<0.01).PF+CG promotes OGR/R cell recovery via the PI3K/AKT signaling pathway.ConclusionThese results demonstrate that PF+CG has a positive therapeutic effect on ischemia/reperfusion and OGD/R injury,with the mechanism attributed to activation of the PI3K/AKT signaling pathway.