Exploring Effect of Zhenwu Decoction(真武汤)on Structural Remodeling-Electrical Remodeling of Cardiomyocytes in Heart Failure Based on p38-MAPK Pathway
Objective Zhenwu Decoction(真武汤)can improve arrhythmia in the treatment of chronic heart failure(HF).The purpose of this study is to explore the mechanism of Zhenwu Decoction in the treatment of HF-related arrhythmias from the point of view of p38-MAPK pathway regulating hypertrophic myocardial structural remodeling and electrical remodeling.Methods H9c2 cardiomyocytes were induced by Angiotensin Ⅱ(Ang Ⅱ)to establish the cardiac hypertrophy model.The experi-ment was divided into six groups:normal group(N),model group(M),Zhenwu Decoction low dose group(D),middle dose group(Z),high dose group(G)and inhibitor group(S).The normal group(N)was cultured in high glucose medium containing fetal bovine serum,and the model group(M)was treated with Ang Ⅱ medium containing 10-6 mol/L concentration.The low(D),middle(Z)and high(G)dose groups were treated with 2％,4％and 8％volume fraction of Zhenwu Decoction containing serum and Ang Ⅱ culture medium,respectively,and the inhibitor group(S)was treated with medium containing p38MAPK inhibitor SB203580.After 48 hours of treatment,the growth of cardiomyocytes in each group was observed under microscope,the content of cardiac hypertrophy marker Atrial Natriuretic Peptide(ANP)was measured by Enzyme Linked Immunosorbent Assay(ELISA)method,the senescence of cardiomyocytes was detected by β-galactosidase staining,the expressions of Connexin 43(Cx43),p38 Mitogen-ctivated Protein Kinase(p38-MAPK),Matrix Metalloproteinase 2(MMP2),Matrix Metalloproteinase 9(MMP9),Tissue Inhibitor of Metalloproteinase 1(TIMP1)and p53 protein in cardiomyocytes were detected by Westernblot method,and the mRNA transcription levels of ANP,Cx43,p-Cx43,p38-MAPK,MMP2,MMP9 and TIMP1 were quantitatively analyzed by Quantitative Real-time PCR(qPCR).The expression and distribution of Cx43 protein were detected by immunofluorescence.Results High dose of Zhenwu Decoction could reduce the surface area of hypertrophic cardiomyocytes,but the middle and low do-ses had no obvious effect.Zhenwu Decoction could reduce the levels of ANP and mRNA,which was a marker of cardiac hypertro-phy,and its effect was positively correlated with the dose.Zhenwu Decoction can improve the senescence of mast cells,and the effect was similar to that of inhibitors.Zhenwu Decoction can reduce the expression levels of Cx43,p38-MAPK,MMP2,MMP9,p53,p-Cx43 protein and mRNA transcription levels of Cx43,p38-MAPK,MMP2 and MMP9 in hypertrophic cardiomyocytes,and increase the expression levels of TIMP1 and mRNA transcription level in a dose-dependent manner.Zhenwu Decoction can alleviate the disorder of Cx43 distribution on the surface of mast cells and reduce the lateral-lateral translocation distribution of this protein.Conclusion Zhenwu Decoction can regulate the senescence and fibrosis of myocardial mast cells through p38 signal pathway,regulate the metabolic balance of extracellular matrix,maintain the quantity,phosphorylation level and distribution loca-tion of Cx43 in cardiomyocytes,improve the gap junction remodeling,realize the unified regulation of HF myocardial structural re-modeling and electrical remodeling,and achieve the purpose of treating heart failure and related arrhythmias.
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