护心康片通过AMPK/mTOR/p70S6K通路调控自噬改善大鼠慢性心力衰竭的作用机制OA

Objective To investigate the mechanism of Huxinkang Tablets in improving chronic heart failure(CHF)in rats via regulating autophagy.Methods Nineteen major chemical constitu-ents were identified from Huxinkang Tablets using ultra-performance liquid chromatography-quadru-pole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS).A"component-target-pathway"interaction network was constructed using network pharmacology methods,and 30 core targets associated with heart failure were identified.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses revealed significant enrichment of these targets in the AMP-activa-ted protein kinase/mammalian target of rapamycin(AMPK/mTOR)autophagy regulatory signaling axis.A CHF model was established by ligating the left anterior descending coronary artery in rats.Forty-eight rats were randomly divided into six groups:blank,model,captopril(0.013 5 g/kg),and low-,medium-,and high-dose Huxinkang Tablet groups(0.41,0.82,and 1.64 g/kg,respectively),with eightrats in each group.After four weeks of continuous administration,serum levels of atrial na-triuretic peptide(ANP),B-type natriuretic peptide(BNP),superoxide dismutase(SOD),malon-dialdehyde(MDA),tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1β were meas-ured.Masson staining was used to observe myocardial pathological changes,and Western blotting was performed to assess protein expression in the AMPK/mTOR/70 kDa ribosomal protein S6 kinase(AMPK/mTOR/p70S6K)signaling pathway,thereby systematically validating the network pharma-cology predictions.Results Network pharmacology predictions indicated a strong correlation be-tween core targets and the AMPK/mTOR autophagy pathway.Animal experiments revealed that the model group exhibited myocardial fiber rupture and disorganization,left ventricular dilation,and a significant reduction in left ventricular ejection fraction(LVEF)compared with the blank group(P＜0.05).Serum levels of BNP,ANP,TNF-α,IL-6,IL-1β,and MDA were significantly ele-vated,while SOD activity was significantly decreased(P＜0.05).Western blot results showed that p-AMPK protein expression in myocardial tissue was significantly reduced,whereas p-mTOR,p-p70S6K,the autophagy marker LC3-Ⅱ,and p62 protein expression were significantly increased in the model group(P＜0.01).The high-dose Huxinkang Tablet group significantly improved myo-cardial pathological damage,alleviated ventricular remodeling,increased LVEF,and reduced serum levels of inflammatory and oxidative stress markers.Additionally,it significantly upregulated p-AMPK protein expression,and downregulated p-mTOR,p-p70S6K,LC3-Ⅱ,and p62 protein levels(P＜0.01).Conclusion The multiple active constituents in Huxinkang Tablets may act on key targets such as AMPK and mTOR,regulate the AMPK/mTOR/p70S6K signaling pathway,re-store myocardial energy metabolism homeostasis,promote normal autophagy pathway function,and alleviate autophagic flux blockade,thereby reducing oxidative stress and inflammatory responses and exerting cardioprotective effects.