Study on the Mechanism of Yiqi Tongmai Powder Against Cerebral Ischemia-Reperfusion Injury Based on Network Pharmacology,Molecular Docking and Experimental Verification
Objective To investigate the mechanism of Yiqi Tongmai Powder(Ginseng Radix et Rhizoma,Salviae Miltiorrhizae Radix et Rhizoma,Notoginseng Radix et Rhizoma,Hirudo,Eupolyphaga Steleophaga,Rhei Radix et Rhizoma)against cerebral ischemia-reperfusion injury(CIRI)based on network pharmacology,molecular docking and experimental verification.Methods(1)The active components of Yiqi Tongmai Powder and their action targets were screened by TCMSP,TCMID and ETCM databases,the disease related targets of CIRI were screened by GeneCards,OMIM and TTD disease databases,and the intersection targets of the above targets were obtained through Venny 2.1 online platform,that is,the potential targets of Yiqi Tongmai Powder in the treatment of CIRI.The"drugs-active components-targets"network was constructed by Cytoscape 3.7.1 software,and the potential active components of Yiqi Tongmai Powder in the treatment of CIRI were screened.Protein-protein interaction(PPI)analysis of potential targets was carried out by STRING 11.0 database to screen core targets.The GO function and KEGG pathway enrichment of potential targets were analyzed by Metascape database.AutoDockTools 1.5.7 software was used to verify the molecular docking between the key active components and the core targets.(2)The rat model of CIRI was established by middle cerebral artery occlusion and reperfusion(MCAO/R).SD rats were randomly divided into sham operation group,model group,Yiqi Tongmai Powder low-dose group(0.27 g·kg-1),Yiqi Tongmai Powder high-dose group(1.08 g·kg-1)and Nimodipine group(30 mg·kg-1),with 10 rats in each group.Pre-administration began three days before the establishment of the model,once a day for seven days.The neurological deficit of MCAO/R rats was evaluated by modified neurological deficit score(mNSS).The volume of cerebral infarction was measured by TTC staining.Nissl staining was used to observe the damage of neurons in brain tissue.ELISA method was used to detect serum inflammatory factors and oxidative stress related indexes.TUNEL staining was used to detect brain tissue apoptosis.Western Blot method was used to detect the protein expressions of Bax and Bcl-2 in brain tissue.Results(1)A total of 46 active components and 178 potential targets of Yiqi Tongmai Powder in the treatment of CIRI were obtained.The key active components such as quercetin,digitalis flavonoids,kaempferol,valine and uracil were analyzed,and the core targets such as TNF,IL-6,STAT3,VEGFA,AKT1,IL-1β,CASP3,TP53,MAPK3 and EGFR were analyzed.The potential targets are involved in inflammation,oxidative stress,cell proliferation and differentiation,apoptosis and other biological processes,including cAMP,NF-κB,PI3K-Akt and other signal pathways.The main active components quercetin,flavonoids of digitalis,kaempferol and valine have good binding activity to target proteins such as TNF,IL-6,STAT3 and VEGFA.(2)Compared with the model group,the neurological deficit score of rats in each treatment group was significantly decreased(P＜0.05,P＜0.01),the area of cerebral infarction was significantly decreased(P＜0.01),and the pathological changes of ischemic necrotic area of brain tissue were improved.The number of neurons in ischemic area of brain tissue was significantly increased(P＜0.01),and the rate of neuronal apoptosis was significantly decreased(P＜0.01).The levels of serum IL-1β,IL-6,TNF-α and MDA were significantly decreased,while the activities of SOD and GSH-Px were significantly increased(P＜0.05).The protein expression of Bax in brain tissue were significantly decreased and the protein expression of Bcl-2 significantly increased(P＜0.05).Conclusion Yiqi Tongmai Powder may play an anti-CIRI effect by reducing inflammation and oxidative stress,inhibiting cell apoptosis.
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