Abstract: Objective To investigate the effects of Xinshuai Heji (XSHJ) on energy metabolism and autophagic flux disruption in hypoxia-exposed HL-1 cardiomyocytes, and to observe the concomitant changes in the expression of glycogen phosphorylase, muscle-associated(PYGM) and thrombospondin-1(Thbs1). Methods A hypoxia- induced injury model was established in HL-1 cells with interventions of XSHJ at various concentrations and empagliflozin as a positive control. Cell viability and apoptosis were assessed. Mitochondrial network morphology and membrane potential were observed by fluorescence staining. The oxygen consumption rate (OCR) was measured using a Seahorse analyzer, along with biochemical markers of energy and redox metabolism. Autophagic flux was evaluated utilizing a tandem fluorescent reporter assay. Furthermore, the protein expression levels of PYGM, Thbs1, and autophagy-related markers were determined via Western blot analysis. Results Compared with the model group, XSHJ dose-dependently increased cell viability, reduced apoptosis, and ameliorated the damaged mitochondrial network structure. XSHJ also elevated basal OCR and intracellular levels of ATP, NADPH, and GSH, while decreasing reactive oxygen species (ROS) accumulation. Concurrently, XSHJ reversed the hypoxia-induced reduction in the LC3-Ⅱ/Ⅰ ratio and the abnormal accumulation of p62 protein, accompanied by increased numbers of autophagosomes and autolysosomes. Protein expression profiling revealed that XSHJ upregulated PYGM and downregulated Thbs1 expression. These protective effects were comparable to those of empagliflozin. Conclusions XSHJ alleviated hypoxia-induced cardiomyocyte injury, improved energy metabolism, and promoted the restoration of autophagic flux. These protective effects may be associated with the involvement of PYGM and Thbs1, though the underlying mechanisms require further investigation.

Key words: Xinshuai Heji, ischemic heart failure, PYGM, autophagic flux, energy metabolism