Efficacy And Mechanism Of Tangxinping Capsule In Treatment Of Diabetic Cardiomyopathy Base On PI3K/AKT Signaling Pathway

Research backgroundDiabetic cardiomyopathy(DCM)is a pathological change characterized by structural and functional abnormalities of the heart in a high glucose state,and it is one of the diabetes-related cardiac complications.Tangxinping capsule,TXPC)is a Chinese medicinal preparation used for the treatment and improvement of diabetic heart disease,but its mechanism is still not very clear.Based on the project of Beijing Municipal Science and Technology Commission "Research and Development of Ten Diseases and Ten Medications-Study on the Drug Property of Tangxinping Capsule,a new drug for the treatment of diabetes combined with coronary heart disease",which integrates the research methods of network pharmacology and animal experimental verification,this study explored and verified the efficacy and mechanism of TXPC.Research purposesThe research methods of network pharmacology analysis combined with animal experimental verification were used to explore and evaluate the therapeutic effect of TXPC on DCM and the exploration of its mechanism.Research method1?Method one:In this study,the active ingredient compounds in TXPC formula were mined from the chemical information databases of TCMSP,ETCM,TCMID and BATAMN.The action targets of the components were predicted and screened out by Pubchem and Swiss Target Prediction platforms.A TXPC drug-component-target network was constructed by Cytoscape software,and the core components in the TXPC formula were analyzed and screened out by using the Network Analyze plug-in.The disease target database of OMIM,Disgenet and Genecards was used to construct the combined set of DCM targets.The intersection of that TXPC target and the DCM target is map through the R Studio software;The TXPC-DCM-target network was constructed using String online protein function analysis website,and Cytoscape software was imported for visualization,to construct PPI network,and the core targets of PPI network were screened out through Network Analyze.Module analysis is carry out on that PPI network through an MCODE plug-in;Finally,GO and KEGG analyses of the PPI network were performed using Matescape online enrichment analysis platform,to obtain the biological process(BP),molecular function(MF),cell composition(CC)and enrichment pathway analysis results of the core targets.2?Method two:In this experiment,the rat model with DCM was established by high-fat diet feeding combined with low-dose STZ intraperitoneal injection.After four weeks of high-fat diet feeding,STZ(35 mg/kg)was injected intraperitoneally at one time for 6 weeks.After STZ injection,the high-fat diet fed rats were divided into Model group(Model),TXPG,TXPZ,low dose(TXPD)group and metformin group(Met)by random number method.The normal diet group was the normal Control group(CONtrol).Rats in the normal control group and the model control group were intragastrically administered with 0.9%NaCl solution at the dose of 1 mL/100 g.The rats in the Tangxinping low,medium and high dose groups were intragastrically administered with 0.21 g/kg,0.42g/kg and 0.84g/kg TXPC powder aqueous solution lml/100g,respectively.The rats in the metformin group were intragastrically administered with 0.15g/kg metformin powder aqueous solution 1ml/100g per day.Fasting blood glucose was measured at weeks 2,4 and 6 after drug administration.After the drug administration,the levels of TC,TG,LDL-C,HDL-C,and INs in rats were measured biochemically,and the HOMA-IR value was calculated.The levels of serum CKMB,CRP,BNP,6-KPG,ET-1,TXB2,MDA in rats were detected by ELISA,and the myocardial tissue damage was observed through cardiac pathological sections to clarify the therapeutic effect of TXPC on DCM.The cardiac function was evaluated by echocardiography.The rat model of acute myocardial ischemia was established by sublingual injection of vasopressin,and the ST-T segment displacement in the ECG of rats after treatment with TXPC was observed.Further,the levels of myocardial TNF-?,IL-1?,SOD,ATP,ADP,and ATP/ADP in rats with DCM were detected histologically,and the transcription levels of myocardial NF-?Bp65,GLUT4,TGF-?1,PI3KP85,P-AKT,P-GSK3? protein,PI3KP85mRNA,P-AKT mRNA,P-GSK3?mRNA,NF-KBp65 mRNA,and GLUT4 mRNA were detected by western blot assay,in order to explore the pharmacodynamic mechanism of TXPC.Research Results1?Results of method one:There were 107 active components in Tangxinping Decoction;Core components include:quercetin,kaempferol.luteolin,etc.A total of 192 corresponding targets were obtained,including key targets such as AKT1,EGFR and SRC;Through module analysis and copolymerization,four functional modules were identified,of which,Module 1 and Module 2 contained more targets,which represented the overall therapeutic effect of Tangxinping Capsule on diabetic cardiomyopathy.A total of 961 biological processes were accumulated;75 molecular functions;60 cells are formed;218 signaling pathways,which can play a role in the treatment of DCM through PI3K/Akt signaling pathway,MAPK signaling pathway,endocrine resistance and other pathways.2?Results of method two:?TXPC could improve FPG of rats with DCM,reduce the levels of TG,TC and LDL-C in rats,and increase HDL-C level at the same time,reduce lipid deposition in myocardial tissue and play a role in regulating glucolipid metabolism;?TXPC can reduce the levels of myocardial injury indicators CRP,BNP,and CKMB,and improve myocardial fiber rupture and injury,etc.,which may play a role in myocardial protection by reducing the level of inflammation;?TXPC can reduce serum ET-I and TXB2,and increase 6-KPG level,regulate vascular endothelial cell function and prevent myocardial microangiopathy;?TXPC can reduce serum MDA level and alleviate oxidative stress injury;?TXPC can reduce the level of LVIDs in rats with DCM and increase the levels of LVIDD,SV,FS and EF,thereby maintaining the stability of the heart structure and morphology,thus playing a role in myocardial protection.?TXPC can resist ST-T elevation caused by vasopressin and improve the myocardial resistance to acute myocardial ischemia,thereby playing a role of cardiac protection.?TXPC can increase the ATP and ATP/ADP levels in rats with DCM,promote the expression of GLUT4 protein and GLUT4 mRNA transcription level,and regulate energy metabolism in rats with DCM by activating PI3K/AKT/GLUT4 signaling pathway;?TXPC reduced TNF-? level and pro-inflammatory factor IL-1? level in rats with DCM,and inhibited the expression of NF-?B protein and NF-?BmRNA transcription,which played a role in inhibiting inflammation by activating PI3K/AKT/NF-?B signaling pathway.Pet-name ruby ?TXPC can reduce the expression level of GSK3? protein in rats with DCM,and reduce the levels of large TGF-?1 and SOD in rats at the same time.It can inhibit the transcription and expression of GSK-3? protein and GSK-3?mRNA by activating PI3K/AKT/GSK-3? signaling pathway,and play a role in resisting myocardial cell injury.TXPC can promote the phosphorylation and expression of target proteins PI3K and AKT in PI3K/AKT pathway,and enhance the transcription levels of PI3KP85mRNA and p-AKT mRNA,suggesting that TXPC's therapeutic effect on DCM is related to PI3K/AKT pathway.Research Conclusions1?Network pharmacology analysis results show that TXPC can exert the therapeutic effect on DCM through PI3K/AKT signaling pathway;2?TXPC can regulate glycolipid metabolism,improve inflammation,reduce oxidative stress level and play a role in the protection of vascular endothelial cells.Protecting heart function,resisting myocardial ischemia,and protecting heart;3?The efficacy of TXPC is related to the PI3K/Akt signaling pathway.By activating the PI3K/Akt signaling pathway,it can exert the therapeutic effect on DCM through the PI3K/Akt/Glut4,PI3K/Akt/GSK-3? and PI3K/Akt/NF-?B pathways.