Effect Of Tongqiao Huoxue Decoction On Autophagy Of Ischemic Stroke Neurons Based On Network Pharmacologye

Objective:With the aging of the population,the incidence of ischemic stroke in China is on the rise,and the mechanism of action of ischemic stroke has not been fully clarified.At present,a number of studies have found that autophagy is involved in the disease process of ischemic stroke,and a large number of studies have confirmed that Tongqiao Huoxue Decoction has neuroprotective effects in the treatment of ischemic stroke,but its specific mechanism of action cannot be determined.In this project,the role targets and molecular mechanisms of Tongqiao Huoxue Decoction in the treatment of ischemic stroke were analyzed by network pharmacology prediction,and the specific mechanism of Tongqiao Huoxuo decoction in the treatment of ischemic stroke was explored based on animal experimental verification.Methods:1.Network pharmacology:use network pharmacology methods to predict the possible targets and molecular mechanisms of Tongqiao Huoxue decoction in the treatment of ischemic stroke,and obtain the intersection target of Tongqiao Huoxue decoction and ischemic stroke;The"drug-target-disease" protein interaction network was constructed,the core targets of Tongqiao Huoxue decoction for the treatment of ischemic stroke were screened,GO and KEGG enrichment analysis was carried out,and the core pathway was verified by animal experiments.2.Animal experiments:male mice with SPF-level SD were randomly divided into control group(Control),model group(MCAO),low-dose group of Tongqiao Huoxue Decoction(TQHXD-L),medium-dose group(TQHXD-M),and high-dose group(TQHXD-H)of Tongqiao Huoxue Soup,and the Middle cerebral Artery Occlusion(MCAO)model was prepared by wire embolism,and then perfused for 24 h.The degree of neurological deficit in rats was assessed on days 1,4 and 7,respectively.2,3,5-triphenyltetrazolium chloride(TTC)staining to observe cerebral infarction in rats;Real-time PCR(RT-qPCR)detected the relative expression of autophagic factors Beclin 1 and LC3.Western Blot detects the expression of the corresponding pathway protein.Results:1.Through SwissTargetPrediction target prediction,233 targets of Tongqiao Huoxue Decoction were obtained;Through GeneCards and OMIM database screening,4287 disease targets of ischemic stroke were obtained;By mapping each other,183 potential targets for the treatment of ischemic stroke were obtained.The core targets mainly include AKT 1、JUN、TP53、HSP90AA1、MAKK 1,etc.;Through GO and KEGG enrichment analysis,it was shown that GO biological functions were mainly based on lipopolysaccharide reactions,responses to bacteria-derived seeds,and oxidative stress reactions,and the signaling pathway was mainly PI3K/AKT signaling pathway.2.Compared with the control group,the neural function of rats in the model group was significantly weakened.Compared with the model group,the damaged nerve function of rats in the TQHXD gavage group was significantly improved.3.TTC staining results:Compared with the control group,rats in the model group had severe infarction foci,and the volume of cerebral infarction in the TQHXD gavage group was significantly smaller than that in the model group.Among them,the TQHXD-M group rat infarction area was the smallest.4.RT-qPCR results:Compared with the control group,the relative expression of Beclin 1 mRNA and LC3 mRNA in the model group and TQHXD-L group was significantly reduced,and the difference was statistically significant.Compared with the model group,the relative expression of Beclin 1 mRNA in TQHXD-M group was significantly increased,and the relative expression of LC3 mRNA in TQHXD-M group was significantly increased,and the difference was statistically significant.5.Western Blot results:The relative expression of AKT and mTOR proteins did not change significantly,compared with the control group,the relative expression of p-AKT and p-mTOR proteins in the model group was increased,and the relative expression of p-AKT and p-mTOR proteins in the TQHXD gavage group decreased.Compared with the control group,the relative relative expression of Beclin 1 protein in the model group,TQHXD-L and TQHXD-H groups decreased,and the relative expression of Beclin 1 in the model group and TQHXD-L rats decreased significantly,and the relative expression of LC3 protein in the model group was statistically significant.Compared with the rats in the model group,the relative expression of Beclin 1 and LC3 proteins in the TQHXD-M group was significantly increased,and the difference was statistically significant.Conclusions:1.PI3K/AKT signaling pathway is one of the core pathways for the treatment of ischemic stroke.2.Tongqiao Huoxue Decoction may inhibit the activation of the PI3K/AKT/mTOR signaling pathway in ischemic stroke by downregulating the expression of p-AKT and p-mTOR and upregulating the expression of Beclin 1 and LC3,promoting the activation of neuronal autophagy and exerting neuroprotective effects.