The Effective Components Of Yixintai Regulate The Proliferation And Apoptosis Of Cardiac Fibroblasts In Rabbits With Heart Failure

Objective:To take myocardial fibroblasts as the research object in vitro,observe the effects of different doses of Yixintai's effective components on the proliferation and apoptosis of myocardial fibroblasts,and explore the effects of Yixintai's effective components on the regulation of myocardial fibroblasts proliferation and apoptosis Study of possible mechanisms.Methods:Primary and subcultured CFs of ordinary healthy New Zealand rabbits were identified by vimentin combined with immunofluorescence staining;CCK-8 method was used to detect the proliferation activity of CFs at each time point in each group,expressed as absorbance?OD value?.The experiment is divided into?1?control group:myocardial fibroblasts;?2?model group?Ang II group?:myocardial fibroblasts+serum-free culture for 48h+Ang II 10^-7mol/L treatment for 12h,?3?low dosage group:myocardial fibroblasts+Yixin Thai effective component 48h lower,?4?medium dosage group:myocardial fibroblasts+Yixintai effective component 48h,?5?high dosage group:myocardial fibroblasts+Yixintai effective component 48h higher,?6?Losartan Potassium group:myocardial fibroblasts Cell+Losartan potassium group 48h,?7?Ang II+low dosage:myocardial fibroblasts+Yixintai effective component 48h+Ang II 10^-7mol/L treatment for 12h,?8?Ang II+medium dosage group:myocardial fibroblasts+Yixin Among the effective components of Thai,48h+Ang II 10^-7mol/L treatment for 12h,?9?Ang II+high dosage group:myocardial fibroblast+Yixintai effective component 48h+Ang II 10^-7mol/L treatment for 12h,?10?Ang II+Losartan Potassium group:Myocardial fibroblasts+losartan potassium group 48h+Ang II10^-7mol/L treatment for 12h.Use inverted phase contrast microscope,cell count to observe the cell morphology and growth status,and then use flow cytometry to measure the apoptosis rate and cell cycle;use Western blot to detect the expression of Bax and Bcl-2 protein in each group;use RT-PCR to detect each Group collagen1a,collagen3a m RNA expression.Results:1.The cells are fusiform under the microscope,without spontaneous pulsation Vimentin staining was positive,and the purity of CFs was higher.CCK-8 method was used to detect the proliferation of CFs in each group:compared with the control group,the OD value of the model group was significantly increased?P<0.05?;compared with the model group,the Ang II+medium dosage group,Ang II+high dosage group,Ang II The OD value of the+Losartan Potassium group decreased?P<0.05?,and the OD value of the Ang II+high dosage group decreased significantly?P<0.05?.2.The results of flow cytometry detection of apoptosis rate and cell cycle:when the effective components of Yixintai act on CFs cells,with the increase of the concentration of effective components of Yixintai,the proportion of apoptotic cells is obvious Rise.Compared with the control group,the percentage of cells in the G0/G1 phase of the model group decreased,and the percentage of cells in the S phase and G2/M phase increased.Compared with the model group,the percentage of G0/G1 phase cells increased in the Ang II+low dosage group,Ang II+medium dosage group,Ang II+high dosage group,and Losartan Potassium group,while the percentage of cells in S phase and G2/M phase decreased.The percentage of cells in the G0/G1 phase of the Ang II+high dosage group increased significantly,while the percentage of cells in the S phase and G2/M phase decreased significantly.3.Western blot method to detect the expression of Bax and Bcl-2 protein in each group:Compared with the control group,the expression of Bax in the model group decreased?P<0.05?,and the expression of Bcl-2 increased?P<0.05?.Compared with the model group,Bax expression was increased in the Ang II+medium dosage group,Ang II+high dosage group,and Losartan Potassium group?P<0.05?,and Bcl-2 expression was decreased?P<0.05?,of which Ang II+high dosage group Bax expression Significantly increased?P<0.05?,Bcl-2 expression decreased significantly?P<0.05?.4.RT-PCR detection of collagen1a and collagen3a m RNA expression in each group:Compared with the control group,the expression levels of collagen1a,collagen3a m RNA in the model group increased?P<0.05?.Compared with the model group,the m RNA expression of collagen1a and collagen3a in Ang II+medium dosage group,Ang II+high dosage group,Losartan Potassium group decreased?P<0.05?,and the expression of collagen1a and collagen3a m RNA in Ang II+high dosage group decreased significantly?P<0.05?.Conclusion:1.The effective components of Yixintai can block the proliferation cycle of CFs cells induced by Ang II.2.The effective components of Yixintai can induce apoptosis of CFs.3.The effective components of Yixintai can inhibit the expression of collagen1a and collagen3a m RNA at the transcription and translation levels,thereby exerting its anti-MF effect.4.The effective component of Yixintai can increase the expression of Bax,reduce the expression of Bcl-2,induce the apoptosis of CFs,and thus exert its anti-MF effect.