益气通脉方调控PI3K/AKT信号通路改善脑缺血再灌注损伤大鼠神经元凋亡OACSTPCD

Objective:Based on the signal pathway of phosphatidylinositol 3-kinases/protein kinase B(PI3K/AKT),to explore the mechanism of Yiqi Tongmai Formula on neural function damage and apoptosis in middle cerebral artery occlusion and reperfusion(MCAO/R)model rats.Method:Twelve healthy SPF grade male SD rats were selected as the sham surgery group from 68 individuals,while the remaining rats were used to prepare MCAO/R models using an improved suture method.36 successful MACO/R model rats were divided into model group,Yiqi Tongmai Formula group(1.08 g·kg-1)and Nimodipine tablet group(10.8 mg·kg-1),with12 rats in each group.The rats were given corresponding drug intervention,with a gastric lavage volume of 10 mL·kg-1 for 14 consecutive days,once a day.Rats in the sham surgery group and model group were given equal volumes of physiological saline by gavage.Use the Modified Neurological Severity Score(mNSS)method to evaluate the neural function of rats;HE staining was used to observe the mor-phological changes of brain tissue in each group of rats;TUNEL staining was used to detect cell apoptosis;Western blot was used to de-tect the protein expression levels of PI3K,p-PI3K,AKT,p-AKT,B cell lymphoma-2(Bcl-2),and Bcl-2 associated X genes(Bcl-2 associated X,Bax)in ischemic penumbra brain tissue;RT qPCR was used to detect the expression levels of Bax mRNA and Bcl-2 mRNA in rat brain tissue.Results:Compared with the sham surgery group,the mNSS score,neuronal apoptosis rate,relative ex-pression levels of Bax protein and genes in the model group rats were significantly increased(P＜0.01),while the ratios of p-PI3K/PI3K and p-AKT/AKT were significantly down regulated(P＜0.01).The relative expression levels of Bcl-2 protein and genes were significantly down regulated(P＜0.01);Compared with the model group,the mNSS score of rats in the Yiqi Tongmai Formula group was significantly reduced(P＜0.05)on the 3rd,7th,and 14th day,while the apoptosis rate of neurons,relative expression of Bax pro-tein and genes were significantly reduced(P＜0.05).The ratio of p-PI3K/PI3K,p-AKT/AKT was significantly increased(P＜0.01),and the relative expression levels of Bcl-2 protein and genes were significantly increased(P＜0.01).HE staining showed that the number of nerve cells in the brain tissue of the model group rats was significantly reduced,the nuclei were pyknotic or disappeared,the cells were significantly ruptured,cavities and inflammatory cell infiltration were observed,some neurons were necrotic and degenera-ted,arranged irregularly,interstitial edema was observed,and the hierarchy was unclear.The number of brain tissue cells in the Yiqi Tongmai Formula group of rats significantly increased,the morphology of nuclear shrinkage improved,the number of cavities and inflam-matory factors decreased,interstitial edema decreased,and the number of degenerated and necrotic nerve cells significantly decreased.The morphology and structure of brain tissue cells were significantly restored.Conclusion:Yiqi Tongmai Formula can activate the PI3K/AKT signaling pathway,regulate the expression of downstream apoptosis related factors Bax and Bcl-2,and thus exert anti apoptotic and protective effects on damaged brain tissue after MCAO/R.