Androgen Receptor Regulate Experimental Autoimmune Myocarditis Mice Cardiac Fibrosis By Increasing MiRNA-125b Expression

Part 1 Attenuation of cardiac fibrosis by inhibiting androgen receptor protected the EAM mice against myocardial damageObjective:To explore the role of androgen receptor(AR)in regulating the heart fibrosis with the experimental autoimmune myocarditis(EAM).Methods: The male Balb/c mice 6-8 weeks weighted18-20 g was conducted by immunizing murine ?-cardiac myosin heavy chain sequence polypeptides.The EAM mice treated with ASC-J9 every otherday from day 8 to day 20.The heart fibrosis was detected by Masson's stain on day 21.The expression of AR in myocardium was detected immunohistochemistry.And the TGF-???-SMA was determined by western blot.Results: The AR of the EAM mice were significantly increased compared with the conrrol.the AR were singnificantly decreased used ASC-J9.The fibrosis area of EAM+ASC-J9 mice was significantly reduced compared with the EAM mice.The expression of TGF-? and ?-SMA in myocardium of EAM mice was significantly increased.The fibrosis area in the myocardium of EAM + ASC-J9 treated group was significantly reduced,TGF-? and ?-SMA was significantly reduced.Conclusion: AR was significantly increased in acute phase of EAM and AR was significantly decreased when EAM mice intraperitoneally injected with ASC-J9.EMA mice receiving ASC-J9 displayed a significant reduction in ECM deposits,The ASC-J9-mediated protection involved attenuation of cardiac fibrosis,which was confirmed Western blot-based quantifications of fibrosis-associated gene expression in the heart of EAM mice.The levels of TGF-? and ?-SMA were dramatically reduced in heart of mice treated with ASC-J9Part 2The effects of androgen receptor on fibroblasts and its mechanismObjective: To study the effects of androgen receptors on fibroblasts and their mechanisms in vitro.Methods: Fibroblasts were administered to the androgen receptor degrading agent ASC-J9,solvent control(0.0005% DMSO medium),AR overexpression plasmid(LV5-AR),AR overexpression plasmid negative control(LV5-AR-NC),AR overexpression plasmid +miR-125 b inhibitor,AR overexpression plasmid + miR-125 b inhibitor negative control,Fibroblast stimulated with TGF-? for 24 h.The expression of miRNA-125 b,collagen I and ?-SMA were detected by QRT-PCR.The expression of androgen receptor,collagen I and ?-SMA in fibroblasts stimulated by TGF-? was detected by Western Blot.Cell proliferation was detected by CCK-8.Results: Compared with the control group,the expression of collagen I and ?-SMA was significantly decreased in the TGF? + ASC-J9 group.After overexpression of AR,the expression of collagen I and ?-SMA was significantly higher than the AR overexpression negative control group.While the group of AR overexpression plasmid + miR-125 b inhibitor,compared with AR overexpression plasmid + miR-125 b inhibitor negative,Collagen I and ?-SMA expression was significantly reduced.Cell proliferation was significantly increased when AR overexpression.After AR overexpression,miR-125 b inhibitor treatment attenuated TGF-?-induced Collagen I and ?-SMA in miRNA and protein level.Moreover,the knockdown of miR-125 b could weaken CFs' proliferation.Conclusion: The use of the androgen receptor degrading agent ASC-J9 can reduce the expression of fibroblast-associated fibrotic molecules.Fibroblasts overexpress AR,increase cell proliferation,increase expression of fiber-related molecules.But overexpress AR and inhibit miRNA-125 b,fibroblast proliferation is inhibited and expression of fiber-related molecules is decreased.