Network Pharmacology And Experimental Study Of Grape Polyphenols Against Diabetic Cardiomyopathy

Background:Diabetic cardiomyopathy(DCM)is an important chronic cardiovascular complication in diabetic patients.It is the metabolic disorder of myocardial tissue caused by glucose and lipid metabolism disorders in the diabetic state,resulting in changes such as myocardial hypertrophy and interstitial fibrosis.The development of congestive heart failure poses a serious threat to human health.The pathogenesis of DCM is very complex.With the deepening of research,multiple factors and links have been confirmed to be involved in the occurrence and development of DCM.There is currently no specific drug for the treatment of DCM,and the study of natural medicine may provide a new direction for its treatment.Grape polyphenol(GP)is a phenolic substance extracted from grapes,mainly including procyanidins and resveratrol,etc.,which is a natural antioxidant and free radical scavenger with anti-arteriosclerosis and cardiovascular protection effects,but its mechanism is not clear.In recent years,with the development of science and technology and the deepening of network pharmacology research,the molecular correlation between drugs and diseases has been gradually revealed.Our method on the basis of network pharmacology,by building the active ingredient-targets-disease network,system analysis at the molecular level GP main active ingredient of DCM key targets and discuss the possible mechanism of action,further validated in cell and animal level,so as to reveal the GP resistance to the molecular mechanism of diabetic cardiomyopathy,for natural medicine clinical prevention and treatment of diabetic myocardial Disease provides the theoretical basis.Objective:1.To study the target and molecular mechanism of grape polyphenols against diabetic cardiomyopathy based on network pharmacology.2.To explore the protective effect and mechanism of the main active components of GP on high glucose injury in rat cardiomyocytes(H9c2).3.To study the protective mechanism and target verification of GP on myocardial injury in diabetic mice.Method:1.Through PubMed and ten thousand party database retrieval literature collection of grape polyphenols effective active ingredients,pharmacology and application of traditional Chinese medicine system database and analysis platform(TCMSP)get grape polyphenols targets;The target genes related to diabetic cardiomyopathy were searched by GeneCards and NCBI database.Application Cytoscape3.7.1 active ingredient-targets-disease network built software analysis,in-depth analysis of protein interaction(PPI)and the Kyoto encyclopedia pathways(KEGG)gene and genome enrichment,determine the GP targets and signaling pathways of resistance to the core of diabetic cardiomyopathy.2.High sugar H9c2 cell damage of rats model was established,with seven different concentrations of grape polyphenols main active ingredients to intervene,CCK 8 method is used to detect H9c2 cell survival rate,and western blot method to detect the expression of related proteins.3.The chain of male SAMP8 mice urea with cephalosporins(50 mg/kg/d)intraperitoneal injection of diabetes model was constructed,give GP intervention in eight weeks,the determination of the type empty blood glucose,body weight and heart weight,by HE staining and Masson staining to observe each group mice myocardial tissue pathology and fibrosis,MMP9,IGF by western blot method Expression of 1R,TGF-β1,Nrf2 and other proteins.Result:1.The candidate targets of 23 main components of grape polyphenols and DCM related targets were identified,and MAPK,VEGF,nod-like receptor and other signaling pathways were regulated by key targets such as AKT,TP53,STAT3,MMP9,IGF1R,Nrf2,TGF-β1 to play the role of anti-diabetic cardiomyopathy.2.The main active components of grape polyphenols can improve the high glucose damage of rat H9c2 cells.The expression of MMP9,IGF1R and TGF-β1 was significantly increased,while the expression of Nrf2 was significantly decreased.The survival rate of MMP9,IGF1R and TGF-β1 was significantly decreased,while the expression of Nrf2 was significantly increased after the intervention of active ingredients.3.After 8 weeks of GP intervention in diabetic mice,the weight of diabetic mice was significantly increased and the myocardial fibrosis was improved.The expression of MMP9,IGF1R and TGF-β1 protein in myocardial tissue of diabetic mice was significantly increased,while the expression of Nrf2 protein was significantly decreased.GP can significantly inhibit the expression of MMP9,IGF1R and TGF-β1 protein,and increase the expression of Nrf2 protein.Conclusion:1.Based on network pharmacology research,it was found that grape polyphenols have a variety of active components,and achieve the overall effect of anti-diabetic cardiomyopathy through the organic integration of multiple targets and multiple signal transduction pathways.2.The main active components of grape polyphenols have different degrees of protection against high glucose injury in H9c2 cells,and the possible mechanism is related to the regulation of Nrf2,IGF1R,MMP9 and TGF-β1 protein expressions.3.The grape polyphenols can significantly reduce diabetic myocardial fibrosis in mice,by increasing the Nrf2 protein expression,reduces MMP9,TGF-beta 1 and IGF1R protein expression,have the effect of antioxidant,anti fibrosis,provide new treatment approach for prevention and control of diabetic cardiomyopathy,which provides the theoretical foundation for the clinical application of natural medicine.