Journal of Exercise & Organ Cross Talk
Journal of Eexercise & Organ Cross Talk

High-intensity interval training and ketone ester supplementation attenuate hepatic oxidative stress and inflammation through modulation of the Keap1/Nrf2/NF-κB axis in Western diet-fed mice

Document Type : Original Article

Authors

Maryam Sarsangi Aliabad 1
Mousa Khalafi 1
Aref Habibi Maleki 2

1 Department of Sport Sciences, Faculty of Humanities, University of Kashan, Kashan, Iran

2 Physiology Research Center, Iran University of Medical Sciences, Tehran, Iran

https://doi.org/10.22122/jeoct.2026.584193.1207
Abstract
Western diet (WD) consumption promotes oxidative stress and inflammation and plays a central role in the development of fatty liver disease. This study investigated the effects of high-intensity interval training (HIIT), ketone ester supplementation, and their combination on hepatic Keap1, Nrf2, and NF-κB protein levels in WD-fed mice. Thirty male C57BL/6J mice were randomly assigned to five groups (n=6): normal diet (ND), WD, WD plus ketone ester (WD+KE), WD plus HIIT (WD+HIIT), and WD plus HIIT combined with ketone ester supplementation (WD+HIIT+KE). Except for the ND group, all animals consumed a WD for eight weeks. HIIT was performed for four weeks (three sessions/week), and ketone ester was administered daily by oral gavage. Hepatic protein levels of Keap1, Nrf2, and NF-κB were determined using Western blotting. WD feeding significantly increased hepatic Keap1 and NF-κB protein levels and reduced Nrf2 levels compared with the ND group. HIIT and ketone ester, both independently and in combination, significantly decreased NF-κB and increased Nrf2 protein levels compared with the WD group. In addition, HIIT alone and in combination with ketone ester supplementation significantly reduced hepatic Keap1 levels (p<0.05). Overall, both interventions alleviated WD-induced alterations in hepatic oxidative stress and inflammatory signaling. The combined intervention elicited the most pronounced molecular responses, suggesting potential synergistic effects of HIIT and ketone ester supplementation in attenuating Western diet-induced oxidative stress and inflammation.

What is already known on this subject?

• Western diet consumption is strongly associated with metabolic disorders, including the non-alcoholic fatty liver disease (NAFLD).

primarily through the induction of oxidative stress and chronic low-grade inflammation.

• Disruption of hepatic redox homeostasis and activation of inflammatory signaling pathways, particularly NF-κB, are key mechanisms involved in NAFLD progression.

• The Keap1/Nrf2 signaling pathway is a central regulator of cellular antioxidant defense, and its dysregulation contributes to increased susceptibility to oxidative damage in metabolic diseases.

• High-intensity interval training (HIIT) improves metabolic health, enhances mitochondrial function, and reduces oxidative stress and inflammation in both clinical and preclinical models.

• Ketone ester supplementation has emerged as a metabolic intervention that improves mitochondrial efficiency and modulates oxidative and inflammatory signaling pathways.

• Although exercise-based and nutritional strategies have independently shown protective effects against diet-induced metabolic disturbances, their combined impact on hepatic redox regulation remains insufficiently understood. 

 

What this study adds?

• This study provides new evidence that high-intensity interval training (HIIT) and ketone ester supplementation modulate hepatic redox and inflammatory signaling pathways in a Western diet-induced mouse model. 

• HIIT, ketone ester supplementation, and their combination significantly improved the Keap1/Nrf2/NF-κB signaling axis, with HIIT showing particularly strong effects on Keap1 regulation. 

• Ketone ester supplementation enhanced Nrf2 activation and reduced NF-κB protein levels, supporting its antioxidant and anti-inflammatory potential. 

• The combined intervention did not show clear additive effects across all markers, but resulted in more consistent improvements in hepatic redox homeostasis.

Keywords

Western diet
Non-alcoholic fatty liver disease
High-intensity interval training
Ketone ester
Oxidative stress
Inflammation
Subjects

Acknowledgements

This study is derived from a Master’s thesis in Exercise Physiology at the University of Kashan. We sincerely thank the faculty members of the Department of Sports Sciences at the University of Kashan and the Physiology Research Center of the Iran University of Medical Sciences for their invaluable support.

Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Compliance with ethical standards

Conflict of interest The authors declare that there is no conflict of interest in the present research.

Ethical approval All experimental procedures involving animals were conducted in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and were approved by the Ethics Committee of the Faculty of Sports Sciences, University of Tehran (Approval No. IR.UT.SPORT. REC.1403.112; approved on 04.01.2025).

Informed consent Animal study.

Author contributions 

Conceptualization: Conceptualization: M.Kh.; Methodology: M.Kh., A.H.M.; Software: M.kh., A.H.M; Validation: M.Kh.; Formal analysis: M.Kh., A.H.M.; Investigation: M.kh., M.S.A.; Resources: M.K.; Data curation: M.S.A.; Writing-original draft: M.kh., A.H.M.; Writing - review & editing: M.K., A.H.M.; Visualization: M.kh.; Supervision: M.Kh.; Project administration: M.Kh.; Funding acquisition: M.Kh., M.S.A.   

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Journal of Exercise & Organ Cross Talk
Volume 6, Issue 3
Summer 2026
Pages 167-176

  • Receive Date 01 May 2026
  • Revise Date 07 June 2026
  • Accept Date 16 June 2026

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