Jiuzhuan Huangjing Pills alleviate fatigue by preventing energy metabolism dysfunctions in mitochondria.

Abstract

Background Fatigue exerts a profound impact on the efficiency of work and learning, as well as overall health, in a significant portion of the global population. Unfortunately, current anti-fatigue medications have fallen short in delivering satisfactory outcomes, underscoring the imperative for extensive research into the development of therapeutic interventions to effectively manage fatigue and mitigate its associated adverse effects. Purpose The aim of this study was to investigate the efficacy of dietary supplement Jiuzhuan Huangjing Pills (JHP) in improving fatigue induced by exercise and to elucidate its underlying mechanisms. Methods The weight-loaded forced swimming test was employed to establish a fatigue model in mice. C2C12 cells stimulated with H₂O₂ were employed to establish an in vitro oxidative stress model. Enzyme linked immunosorbent assays (ELISA) were conducted to measure oxidative stress, mitochondrial function, and energy metabolism-related markers in both in vivo and in vitro models. Immunofluorescence assays were performed to assess mitochondrial membrane potential and cell apoptosis. Ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) was used to identify metabolites in tissues and the JHP-derived ingredients, respectively. Network pharmacology analysis and molecular docking were applied to reveal the potential key genes and pathways targeted by the main ingredients. Results JHP significantly increased the swimming time of mice and improved abnormal changes in fatigue indicators caused by intensity exercise. Mechanistically, JHP improved fatigue by protecting against structural damage and functional disorders of mitochondria through the reduction of oxidative stress, thereby preventing cell death and enhancing energy metabolism. Consistent with JHP, the ingredients derived from JHP also displayed similar protective effects against fatigue-induced oxidative stress-mediated mitochondrial damage and cellular apoptosis. Importantly, JHP alleviated oxidative stress mainly by modulating the abundances of organic acids and lipids. The main ingredients of JHP as bioactive components exert their effects by binding to key genes involved in pathways crucial in fatigue. Conclusions Taken together, our findings demonstrated that JHP can serve as a candidate dietary supplement to improve exercise-induced fatigue without causing adverse effects, acting through the modulation at both metabolite and gene levels to ensure cellular survival and energy metabolism, ultimately enhancing overall energy production in the body.