Aryl hydrocarbon receptor (AhR) alleviates the LPS-induced inflammatory responses in IPEC-J2 cells by activating PINK1/Parkin-mediated mitophagy

  • Inflamm Res. 2025 Jun 30;74(1):98. doi: 10.1007/s00011-025-02063-y.
Mengfei Ma  #  1  2 ,  Jianxun Guo  #  1  2 ,  Xiaoying Su  #  1  2 ,  Baocai Ma  1  2 ,  Xiufen Wang  1  2 ,  Mingyu Wangshao  1  2 ,  Kai Zhong  1  2 ,  Yueying Wang  1  2 ,  Guoyu Yang  1  2  3 ,  Yingqian Han  4  5
Affiliations
  • 1. Key Laboratory of Animal Biochemistry and Nutrition of Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, Henan, China.
  • 2. Key Laboratory of Veterinary Biotechnology of Henan Province, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, Henan, China.
  • 3. Henan University of Animal Husbandry and Economy, Zhengzhou , 450046, Henan, China.
  • 4. Key Laboratory of Animal Biochemistry and Nutrition of Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, Henan, China. [email protected].
  • 5. Key Laboratory of Veterinary Biotechnology of Henan Province, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, Henan, China. [email protected].
  • # Contributed equally.
PMID: 40583101 DOI: 10.1007/s00011-025-02063-y
Abstract

Objective: This study investigates the role of the Aryl Hydrocarbon Receptor (AhR) in lipopolysaccharide (LPS)-induced inflammatory responses in IPEC-J2 cells.

Methods: Inflammatory responses were triggered in IPEC-J2 cells using 5 μg/ml LPS. AhR was activated with tryptophan or FICZ and knocked down via RNA interference. PINK1/Parkin-mediated Mitophagy was activated using CCCP and inhibited by PINK1 knockdown. Inflammatory mediators and pathway proteins were analyzed through ELISA, RT-qPCR, western blot, and immunofluorescence. Mitochondrial function was assessed by measuring ROS, ATP, and mitochondrial membrane potential. The interaction between AhR and PINK1 was examined using dual-luciferase reporter assays.

Results: The IDO1/AhR signaling axis was activated in LPS-stimulated IPEC-J2 cells. AhR activation was found to attenuate LPS-induced inflammatory responses, whereas AhR knockdown exacerbated these responses. Mechanistic investigations demonstrated that AhR activation alleviated LPS-induced mitochondrial damage. Activating PINK1/Parkin-mediated Mitophagy successfully countered the increased inflammatory response in IPEC-J2 cells after AhR knockdown. Moreover, blocking PINK1 reversed the anti-inflammatory effects of FICZ. Dual-luciferase reporter assays revealed that AhR acts as a crucial transcription factor by directly binding to the promoter region, thereby initiating PINK1 transcription.

Conclusions: AhR reduces LPS-induced inflammatory response in IPEC-J2 cells by activating PINK1/Parkin-mediated Mitophagy, with AhR directly engaging the PINK1 promoter to enhance its transcription. Targeting AhR may present a novel strategy for the prevention and management of Escherichia coli-induced diarrhea in piglets.

Keywords
Aryl hydrocarbon receptor; IPEC-J2 cell; Inflammatory response; Lipopolysaccharide; PINK1/Parkin-mediated mitophagy.
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