Effects And Mechanisms Of Wnt/JNK Signal Pathway Activation On Myocardial Remodeling

PART I EFFECT OF WNT/JNK SIGNAL PATHWAY ACTIVATION ON MYOCARDIAL REMODELINGObjective:To investigate the effect of Wnt/JNK signal pathway activation on myocardial remodeling.Methods:The experimental model was established by primary cultured myocardial cells of newborn SD rats.Cells were dived into four groups:1)Normal Control;2)Wnt5a;3)sp600125;4)Wnt5a+sp600125.Immunofluorescence was used to determine the cell surface;confocol was used to detect the cell cytoskeleton;JC-1 was used to detect mitochondrial membrane potential;Ca-AM was used to detect mitochondrial permeability transition;Hoechest and Flow cytometry were used to assay the level of apoptosis;Western blotting was used to detect the protein expression of c-Jun,ANF,P-MHC,Bcl-2,Bax,c-caspase-9-3,cyc,VDAC,CyPD,ANT.Results:?After incubated with Wnt5a,the activity of JNK was increased in a time-dependent manner.?Compared with the NC group,the relative cell surface in Wnt5a group was significantly increased(P<0.05);Compared with Wnt5a group,the relative cell surface in Wnt5a+sp600125 group was significantly decreased(P<0.05).?Compared with the NC group,the expression of ANF was significantly increased in Wnt5a group(P<0.05);Compared with Wnt5a group,the expression of ANF in Wnt5a+sp600125 group was significantly decreased(P<0.05).?Compared with the NC group,the percentage of cytoskeleton disrupted cells was significantly increased in Wnt5a group(P<0.05);Compared with Wnt5a group,the percentage of cytoskeleton disrupted cells was significantly decreased in Wnt5a+sp600125 group(P<0.05).?Compared with the NC group,the percentage of nucleic apoptosis,cell apoptosis,and apoptosis related protein Bax/Bcl-2,c-caspase-9-3,Cyc were significantly increased in Wnt5a group(P<0.05);Compared with Wnt5a group,the percentage of nucleic apoptosis,cell apoptosis,and apoptosis related protein Bax/Bcl-2,c-caspase-9-3,Cyc were significantly decreased in Wnt5a+sp600125 group(P<0.05).?Compared with the NC group,the MMP and expression of VDAC were significantly decreased but the opening of mPTP was increased in Wnt5a group(P<0.05);Compared with Wnt5a group,the MMP and expression of VDAC were significantly increased but the opening of mPTP was decreased in Wnt5a+sp600125 group(P<0.05).Conclusions:Wnt5a promotes myocardial remodeling via activating Wnt/JNK signal pathwey.PART II MECHANISM OF MYOCARDIAL REMODELING INDUCED BY WNT/JNK SIGNAL PATHWAY ACTIVATIONObjective:To investigate the mechanism of myocardial remodeling induced by Wnt/JNK signal pathway activation.Methods:The experimental model was established by primary cultured myocardial cells of newborn SD rats.siRNA was used to establish cells with depleted Vangl2.Cells were dived into:1)scramble-siRNA;2)scramble-siRNA+Wnt5a;3)Vangl2-siRNA;4)Vangl2-siRNA+Wnt5a.Immunofluorescence was used to determine the cell surface;confocol was used to detect the cell cytoskeleton;JC-1 was used to detect mitochondrial membrane potential;Ca-AM was used to detect mitochondrial permeability transition;Hoechest and Flow cytometry were used to assay the level of apoptosis;Western blotting was used to detect the protein expression of Vangl2,c-Jun,ANF,JNK,Bcl-2,Bax,c-caspase-9-3,cyc,VDAC.Results:? After incubated with Vangl2-siRNA,the protein expression of Vangl2 was significantly decreased(P<0.05).?Compared with the scramble-siRNA group,the relative cell surface in scramble-siRNA+Wnt5a group was significantly increased(P<0.05);Compared with scramble-siRNA+Wnt5a group,the relative cell surface in Vangl2-siRNA+Wnt5a group was significantly decreased(P<0.05).?Compared with the scramble-siRNA group,the expression of ANF and c-Jun in scramble-siRNA+Wnt5a group was significantly increased(P<0.05);Compared with scramble-siRNA+Wnt5a group,the expression of ANF and c-Jun in Vangl2-siRNA+Wnt5a group was significantly decreased(P<0.05).?Compared with scramble-siRNA group,the percentage of cytoskeleton disrupted cells was significantly increased in scramble-siRNA+Wnt5a group(P<0.05);Compared with scramble-siRNA+Wnt5a group,the percentage of cytoskeleton disrupted cells was significantly decreased in Vangl2-siRNA+Wnt5a group(P<0.05).?Compared with the scramble-siRNA group,the percentage of nucleic apoptosis,cell apoptosis,and apoptosis related protein Bax/Bcl-2,c-caspase-9-3,Cyc were significantly increased in scramble-siRNA+Wnt5a group(P<0.05);Compared with scramble-siRNA+Wnt5a group,the percentage of nucleic apoptosis,cell apoptosis,and apoptosis related protein Bax/Bcl-2,c-caspase-9-3,Cyc were significantly decreased in Vangl2-siRNA+Wnt5a group(P<0.05).?Compared with the scramble-siRNA group,the MMP and expression of VDAC were significantly decreased but the opening of mPTP was increased in scramble-siRNA+Wnt5a group(P<0.05);Compared with scramble-siRNA+Wnt5a group,the MMP and expression of VDAC were significantly increased but the opening of mPTP was decreased in Vangl2-siRNA+Wnt5a group(P<0.05).Conclusions:Vangl2 plays an essential role in myocardial remodeling activated by Wnt/JNK signal pathwey.