Mechanism by which hydrogen-rich water mitigates exercise-induced fatigue: activation of the immunoresponsive gene 1-itaconate/nuclear factor erythroid 2-related factor 2/heme oxygenase-1 pathway

Zhang, Yinyin1,#; Ying, Yajing1,#; Zu, Xianpeng2,#; Ding, Lingling3,#; Shi, Xuan4; Wang, Jing1; Li, Xiangtong1; Li, Chujian1,5; Zhou, Qicheng1; Shen, Hui1; Li, Hongxia1,*; Lu, Hongtao1,*; Cheng, Jin3,*

Author Information

1Department of Naval Medicine, Naval Medical University, Shanghai, China

2School of Pharmacy, Naval Medical University, Shanghai, China

3Internal Medicine III (Nephrology and Endocrinology), Naval Medical Center, Naval Medical University, Shanghai, China

4Department of Anesthesiology, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China

5College of Food Science and Engineering, Qingdao Agricultural University, Qingdao, Shandong Province, China

*Correspondence to: Jin Cheng, MD, chengjin455@126.com; Hongtao Lu, PhD, hongtao_lu1990@163.com; Hongxia Li, PhD, hongxialove617@126.com.

#Both authors contributed equally to this work.Funding:This study was supported by the Medical Science and Technology Youth Cultivation Program (No. 21QNPY035), Foundation of Naval Medical University (Nos. 2022MS003, 2022QN019), National Natural Science Foundation of China (No. 82004215), Shanghai Municipal Health Commission Project (No. 20204Y0326), and Foundation of Naval Medical Center of PLA (No. 22M3201).

Abstract

Exercise-induced fatigue limits athletic performance. Molecular hydrogen is an effective treatment for relieving fatigue, but the exact mechanism is not clear. In our study, a mouse model of fatigue was established to explore the molecular mechanism by which hydrogen-rich water reduces exercise-induced fatigue. The results showed that hydrogen-rich water improved the motor function of fatigue mice, reduced the levels of fatigue-related biomarkers (blood urea nitrogen, lactate, and creatine kinase), and alleviated gastrocnemius muscle injury. Furthermore, ultrahigh-performance liquid chromatography–mass spectrometry revealed that hydrogen-rich water upregulated the expression of immune response gene 1 (IRG1), increased the abnormally reduced levels of itaconic acid due to fatigue, and subsequently activated the downstream nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) pathway. Finally, C2C12 cells exposed to an IRG1 inhibitor (IRG1-IN) or 4-octyl itaconic acid (4-OI) were treated with hydrogen-rich water, indicating that hydrogen-rich water effectively upregulated the expression of Nrf2 and HO-1 in cells. In summary, hydrogen-rich water alleviates exercise-induced fatigue by activating the IRG1-itaconic acid/Nrf2/HO-1 pathway and inhibiting oxidative stress.

摘要

运动性疲劳会限制运动表现。分子氢是缓解疲劳的有效方法，但其确切机制尚不明确。本研究通过建立小鼠疲劳模型，探讨富氢水减轻运动性疲劳的分子机制。结果表明，富氢水改善了疲劳小鼠的运动功能，降低了疲劳相关生物标志物（血尿素氮、乳酸和肌酸激酶）的水平，并减轻了腓肠肌损伤。此外，超高效液相色谱-质谱分析显示，富氢水上调了免疫反应基因1（IRG1）的表达，增加了因疲劳而异常降低的衣康酸水平，进而激活下游核因子E2相关因子2（Nrf2）/血红素加氧酶1（HO-1）通路。最后，对暴露于IRG1抑制剂（IRG1-IN）或4-辛基衣康酸（4-OI）的C2C12细胞进行富氢水处理，结果表明富氢水能有效上调细胞中Nrf2和HO-1的表达。综上所述，富氢水通过激活IRG1-衣康酸/Nrf2/HO-1通路并抑制氧化应激来减轻运动性疲劳。