The Effect And Mechanism Of Dapagliflozin On Pressure Overload-induced Cardiac Remodeling

Part ?: Analysis of the Effect of Dapagliflozin on Ang ? Stimulated Cardic Fibroblasts by Proteomics and Secretomics?Objective? Quantitative proteomics technique was used to analyze the direct effects of Dapagliflozin(DAPA)intervention on cardiac fibroblasts(CFs)proteins and secreted proteins stimulated by Ang ?,so as to explore the relationship between these differential expressed proteins or secreted proteins and ventricular remodeling to analyze the possible mechanisms.?Method? After Isobaric tags for relative and absolute quantification,i TRAQ identification,Proteome Discoverer 2.2 was used for database retrieval;GO classification tool was used to classify the biological processes,cell components and molecular functions of these identified proteins;the Cluster Profiler software was used to perform GO enrichment analysis on the identified differential proteins.The main biological functions of differential proteins were determined by significance analysis;String software was used to analyze the protein-protein interactions.?Result? 1.There were 210 differential expressed proteins between the control group and the Ang ? group,of which 62 differential expressed proteins were up-regulated and 148 were downregulated.After DAPA intervention,12 of the up-regulated proteins were back,and 31 of the down-regulated proteins were raised.374 of differential expressed secreted proteins were identified between the control group and the Ang ? group,of which 198 differential expressed proteins were up-regulated and 176 were down-regulated.After DAPA intervention,57 of the up-regulated proteins were back,and 37 of the down-regulated proteins were raised.The result suggested that DAPA has a wide range of interventions for Ang ? stimulated fibroblasts;2.The differential proteins and differential secreted proteins after DAPA intervention are closely related with cadiac fibrosis,cardiomyocyte hypertrophy and apoptosis,suggesting that DAPA may may regulate these processes under Ang ? stimulated and provide us a basis for the further research;3.The differentially expressed proteins and secreted proteins are associated with the MAPKs signaling pathway,suggesting that DAPA may play a role in cardiomyocytes and cardiac fibroblasts through the above signaling pathways.?Conclusion? Proteomics and secretomics provide us an overall and comprehensive understanding of the direct effects of DAPA on CFs and the effect of paracrine on cardiomyocytes.Part ? The Protective Effect of Dapagliflozin on Ventricular Remodeling Induced by Pressure Overload in Mice?Objective? To verify the analysis results of proteomics and secretomics,a model of pressure overload induced ventricular remodeling was constructed by transverse aortic constriction(TAC),and early administration of DAPA was conducted to investigate the role of DAPA in non-diabetic ventricular remodeling and possible signal transduction mechanisms.?Method? After the pressure overload-induced ventricular remodeling model was successfully constructed,DAPA was administrated for 4 weeks,and then small animal cardiac ultrasound was used to assess left ventricular systolic function and ventricular remodeling.The changes of myocardial structure were observed with HE staining.Masson staining,Sirius red staining and immunohistochemistry were used to detect the distribution and expression of collagen I and collagen ?I in myocardial interstitial and perivascular;CCK8 colorimetric assay and EDU proliferation assay were used to evaluate the proliferation of CFs after Ang ? stimulation and DAPA intervention.Real-time quantitative PCR was used to evaluate the expression of procollagen I and procollagen ?I after DAPA intervention.The migration of CFs was evaluated by scratch test and Transwell method.the apoptosis of cardiomyocytes was detected by Tunnel assay.The expression of MAPKs signaling pathways and Fox O1 were detected by Western blot.?Result? 1.Pressure overload induced ventricular remodeling model has the descent of left ventricular systolic function at the fourth week after construction.After DAPA intervention,the degree of ventricular enlargement was reduced and the descent of left ventricular systolic in TAC mice were improved;2.DAPA intervention reduced myocardial hypertrophy caused by pressure overload;3.Myocardial and perivascular fibrosis were reduced in TAC+ DAPA group.In addition,DAPA may interfere with the proliferation of CFs,collagen expression and migration to attenuated fibrosis;4.DAPA treatment attenuated the apoptosis of cardiomyocytes in TAC mice;5.Fibrosis and cardiac hypertrophy-associated P38 MAPKs and JNK signaling pathway may be involved in the process of DAPA intervention in pressure overload-induced ventricular remodeling.DAPA may reduce cardiomyocyte apoptosis through the inhibition of phosphorylation of Fox O1.?Conclusion? This part of the results confirmed that DAPA improved ventricular systolic dysfunction caused by non-diabetic myocardial remodeling,mainly by reducing cardiomyocyte hypertrophy,myocardial interstitial fibrosis and cardiomyocyte apoptosis.Therefore,the application of DAPA may be a good choice for preventing ventricular remodeling.