Investigation Of The Mechanism And Chemical Basis Of Buyang Huanwu Decoction Attenuating Cardiac Fibrosis After Myocardial Infarction By Inhibiting TGFBR1

Cardiac fibrosis,as the main feature of ventricular remolding,is caused by the excessive deposition of extracellular matrix which is produced by fibroblasts in cardiac tissue,the increases of collagen concentration and volume fraction,the change in collagen composition and the disorder in arrangement.These pathologic changes reduce the compliance of cardiac tissue and accelerate the ventricular remolding.Thus,cardiac fibrosis is the key factor of long-term cardiogenic death.Buyang Huanwu decoction,which is developed from WangQing Ren's book "Yilin Gaicuo",is a representive prescription of supplementing Qi and activating blood circulation method.However,its anti-cardiac fibrosis mechanism is still not fully understood.To further investigate the Buyang Huanwu decoction and its decomposed prescription,coronary artery ligation was performed in rats to create models of myocardial infarction and cardiac fibrosis.The possible target TGF-?1 is also studied to clarify the anti-cardiac fibrosis mechanism in regard of signalling pathway.The expression in the early differential gene analysis showed that TGFBR1 would be an anti-cardiac-fibrosis target.20 compounds in Buyang Huanwu decoction with potential binding to TGFBR1 target were identified using virtual screening technique.The virtual screening results were further optimized using machine learning.Then,isotope labelling,western blot and real time PCR were applied to detect the compounds effects on collagen synthesis,protein level and key gene expression respectiely.The results showed that the Buyang Huanwu decoction improved cardiac morphology,cardiac function and the rats' survival rate after myocardial infarction.These were achieved by the antagonism of TGF-?1/Samds signalling pathways which inhibited cardiac fibrosis and collagen deposition.The decomposed prescription of suppling Qi group showed similar effects with decreases of cardiac fibrosis after myocardial infarction like Buyang Huanwu decoction while the decomposed prescription of activating blood circulation group did not show significant change compared with control group.Astragaloside ?,was confirmed as the effective component and the antagonist of TGFBR1 signalling pathway.Compared with the control group,astragaloside ? group showed dose-dependent inhibition of rat cardiac fibroblasts collagen synthesis(P<0.0001),inhibition of TGFBR1,and phosphorylation of downstream signaling molecules Smad2 and Smad3(P<0.0001),inhibition of TGFBR1 related downstream genes CTGF and TIMP1,increasing the expression of MMP9 and the ratio of MMP9/TIMP1.To sum up,the Buyang Huanwu decoction alleviate or reverse cardiac fibrosis after myocardial fibrosis by virtual screening and machine learning,astragaloside ? was identified as an effective anti-cardiac fibrosis component in Buyang Huanwu decoction by inhibition of TGFBR1.