Regulation Of Fibroblast Growth Factor On Fibroblast Proliferation And Transdifferentiation

BackgroundThe cardiac repair after myocardial infarction is mainly performed by cardiac fibroblasts(CFs)increment,transdifferentiation,and extracellular matrix synthesis.However,due to the over-activation of myocardial fibroblasts,a large amount of collagen will be secreted and deposited in the myocardial stroma,resulting in the imbalance of collagen in the myocardium,thus exacerbating myocardial fibrosis and seriously affecting the physiological function of the heart.Fibroblast growth factor(FGF)plays a role in cell metabolism,proliferation,differentiation and migration through complex cell signaling pathways,and is a potential class of cardiac repair and treatment factors that can protect cardiac cells and promote angiogenesis.However,the regulatory mechanism of FGF on CFs remains unclear.ObjectiveTo investigate the regulatory function of fibroblast growth factor FGF on the proliferation and transdifferentiation of CFs into myofibroblast(MFs).Methods1.Preparation of FGFs protein and bioactivity assay.2.Primary rat myocardial fibroblasts were isolated and cultured,and the morphology of the primary cells was observed under the microscope to determine whether the cells were CFs.3.Normal CFs was cultured in vitro,and immunofluorescence method was used to detect the expression differences of 1-sma and type ? collagen in CFs of different generations of rats.4.CCK8 assay was used to detect the effect of different FGFs proteins on CFs proliferation in rats.5.CFs was cultured in vitro after adding different FGFs proteins,and the expression of the protein ?-SMA was detected by western blotting,which was confirmed by immunofluorescence.Results1.3d and 6m old rat heart CFs were separated,and the cell morphology of the primary cells was typical CFs morphology after 24h adherent culture.The P1 cells were identified by DDR2 and the isolated cells were confirmed to be CFs.2.The CFs of both newborn and adult rats were activated to increase the number of MFs with the increase of culture algebra.3.FGF1,2,10,20 had no proliferative effect on CFs.After adding different FGFs to the culture,the CFs expression of position-sma decreased,the number of CFs activated myofibroblasts decreased,and the inhibition effect of FGF2 was the most obvious,followed by FGF1,FGF10 and 20 were not obvious.Conclusion1.In vitro cultured rats,CFs expression increased with the increase of algebra,indicating that the number of CFs activated myofibroblasts increased.2.FGF has no pro-proliferation effect on CFs.FGF1 and FGF2 inhibited the activation of CFs into myofibroblasts,but FGF10 and FGF20 did not significantly inhibit the activation of CFs.