Ma Mengfei, Guo Jianxun, Su Xiaoying, Ma Baocai, Wang Xiufen, Wangshao Mingyu, Zhong Kai, Wang Yueying, Yang Guoyu, Han Yingqian
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.
Key Laboratory of Veterinary Biotechnology of Henan Province, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, Henan, China.
Inflamm Res. 2025 Jun 30;74(1):98. doi: 10.1007/s00011-025-02063-y.
This study investigates the role of the aryl hydrocarbon receptor (AhR) in lipopolysaccharide (LPS)-induced inflammatory responses in IPEC-J2 cells.
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.
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.
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.
本研究探讨芳烃受体（AhR）在脂多糖（LPS）诱导的IPEC-J2细胞炎症反应中的作用。
使用5μg/ml LPS在IPEC-J2细胞中引发炎症反应。用色氨酸或FICZ激活AhR，并通过RNA干扰将其敲低。使用CCCP激活PINK1/Parkin介导的线粒体自噬，并通过敲低PINK1来抑制。通过酶联免疫吸附测定（ELISA）、逆转录-定量聚合酶链反应（RT-qPCR）、蛋白质免疫印迹法和免疫荧光分析炎症介质和信号通路蛋白。通过测量活性氧（ROS）、三磷酸腺苷（ATP）和线粒体膜电位来评估线粒体功能。使用双荧光素酶报告基因检测法检测AhR与PINK1之间的相互作用。
在LPS刺激的IPEC-J2细胞中，吲哚胺2,3-双加氧酶1（IDO1）/AhR信号轴被激活。发现AhR激活可减轻LPS诱导的炎症反应，而AhR敲低则加剧了这些反应。机制研究表明，AhR激活减轻了LPS诱导的线粒体损伤。激活PINK1/Parkin介导的线粒体自噬成功对抗了AhR敲低后IPEC-J2细胞中炎症反应的增加。此外，阻断PINK1可逆转FICZ的抗炎作用。双荧光素酶报告基因检测显示，AhR通过直接结合启动子区域作为关键转录因子，从而启动PINK1转录。
AhR通过激活PINK1/Parkin介导的线粒体自噬降低LPS诱导的IPEC-J2细胞炎症反应，AhR直接作用于PINK1启动子以增强其转录。靶向AhR可能为预防和治疗仔猪大肠杆菌性腹泻提供一种新策略。
