木犀草素通过激活AMPK信号通路增强线粒体自噬，减轻LPS诱导的IPEC-J2细胞损伤。

Luteolin attenuates LPS-induced damage in IPEC-J2 cells by enhancing mitophagy via AMPK signaling pathway activation.

作者信息

Yuan Jianyun, Zhang Ke, Yang Lingling, Cheng Xinyi, Chen Jinyan, Guo Xiaoquan, Cao Huabin, Zhang Caiying, Xing Chenghong, Hu Guoliang, Zhuang Yu

机构信息

Jiangxi Provincial Key Laboratory for Animal Health, Institute of Animal Population Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, Jiangxi, China.

出版信息

Front Nutr. 2025 Mar 26;12:1552890. doi: 10.3389/fnut.2025.1552890. eCollection 2025.

DOI:10.3389/fnut.2025.1552890

PMID:40206944

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11978636/

Abstract

BACKGROUND

Luteolin (LUT), a flavonoid compound widely present in natural plants, has been extensively studied for its diverse biological properties, involving anti-inflammatory,antioxidant, anti-apoptosis and other properties.

METHODS

The aim of this study was to investigate the effect of LUT on lipopolysaccharide (LPS)-induced Intestinal Porcine Epithelial Cell line-J2 (IPEC-J2 cells) damage and its underlying mechanism.

RESULTS

The experiment showed that LPS treatment induced injury in IPEC-J2 cells, leading to tight junction disruption, ROS accumulation, and cell apoptosis. Remarkably, LUT attenuated LPS-induced IPEC-J2 cells damage by the up-regulation of Zonula Occludens-1(ZO-1), Occludin, and Claudin protein 1 (Claudin-1) protein expression levels.Besides, LUT increased the activities of CAT, and SOD and prevented LPS-induced MDA and ROS production. LUT suppressed Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation in LPS-induced IPEC-J2 cells, reducing (Interleukin-1beta) IL-1β and Interleukin-6 (IL-6) expression. Moreover, LUT attenuated LPS-induced apoptosis in IPEC-J2 cells by up-regulating expression of B-cell lymphoma 2 (Bcl-2) and down-regulating expression of Cysteine-aspartic acid protease 3 (Caspase-3), Cysteine - aspartic acid protease 9 (Caspase-9) and Bcl-2-associated X protein (Bax). Furthermore, LUT upregulated the AMP-activated protein kinase (AMPK)/Unc-51 like autophagy activating kinase (ULK) signaling pathway and Parkin-RBR E3 ubiquitin protein ligase (Parkin)/PTEN induced putative kinase 1 (PINK1)-mediated mitophagy in a dose-dependent manner. When AMPK was knocked down by short-hairpin RNA (shRNA), the protective effects of LUT against LPS-induced IPEC-J2 cell damage were weakened, as evidenced by the accumulation of excessive ROS and impaired mitophagy.

CONCLUSION

In summary, LUT exhibits the ability to protect against LPS-induced damage to intestinal tight junctions by enhancing mitophagy through AMPK activation.

摘要

背景

木犀草素（LUT）是一种广泛存在于天然植物中的黄酮类化合物，因其具有抗炎、抗氧化、抗凋亡等多种生物学特性而受到广泛研究。

方法

本研究旨在探讨LUT对脂多糖（LPS）诱导的猪肠上皮细胞系-J2（IPEC-J2细胞）损伤的影响及其潜在机制。

结果

实验表明，LPS处理可诱导IPEC-J2细胞损伤，导致紧密连接破坏、活性氧（ROS）积累和细胞凋亡。值得注意的是，LUT通过上调闭合蛋白-1（ZO-1）、闭合蛋白（Occludin）和紧密连接蛋白1（Claudin-1）的蛋白表达水平减轻了LPS诱导的IPEC-J2细胞损伤。此外，LUT提高了过氧化氢酶（CAT）和超氧化物歧化酶（SOD）的活性，并抑制了LPS诱导的丙二醛（MDA）和ROS生成。LUT抑制了LPS诱导的IPEC-J2细胞中核因子κB（NF-κB）的激活，降低了白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）的表达。此外，LUT通过上调B细胞淋巴瘤-2（Bcl-2）的表达并下调半胱天冬酶-3（Caspase-3）、半胱天冬酶-9（Caspase-9）和Bcl-2相关X蛋白（Bax）的表达减轻了LPS诱导的IPEC-J2细胞凋亡。此外，LUT以剂量依赖的方式上调了腺苷酸活化蛋白激酶（AMPK）/Unc-51样自噬激活激酶（ULK）信号通路以及帕金蛋白-RBR E3泛素蛋白连接酶（Parkin）/PTEN诱导激酶1（PINK1）介导的线粒体自噬。当用短发夹RNA（shRNA）敲低AMPK时，LUT对LPS诱导的IPEC-J2细胞损伤的保护作用减弱，过量ROS的积累和线粒体自噬受损证明了这一点。