Guo Xudong, Yuan Xiaoyue, Xu Zhiyong, Liu Jianhua, Lv Rongrong, Gao Yiqin, Xu Wenjing, Ji Dejun, Guo Yuting
Jiangsu Vocational Institute of Commerce, Nanjing, 211168, Jiangsu, China.
Laboratory of Animal Physiology and Molecular Nutrition, Jiangsu Key Laboratory of Animal Genetic Breeding and Molecular Design, College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, China.
Stress Biol. 2025 Jul 14;5(1):47. doi: 10.1007/s44154-025-00242-9.
Given that lactoferrin (LF) exerts an excellent protection of intestinal homeostasis, the underlying mechanisms, especially epigenetic regulations, are still unknown. This study aimed to investigate the effects of dietary LF epigenetically modulates the oxidative genes by histone modifications to ameliorate ileum inflammation of mice exposed to DON contaminated diet. As expected, we found in the morphology analysis that DON exposure increased ileum crypt depth (CD) and villus width (VW) but reduced villus height (VH) and VH: CD ratio compared to those of the vehicle group. Consistently, the elevated ROS and MDA, along with the decreased ATP, SOD, CAT, GSH, and complex I, III, V were observed in the DON-exposed mice ileum. In contrast, LF markedly ameliorated the impairments of morphological and biochemical indexes. Next, we conducted transcriptome analysis to explore the changed signaling pathways using the ileum RNA of the mice treated with DON or LF. Firstly, the cell cycle pathway genes were significantly downregulated in the DON-exposed mice, and LF improved the cell cycle profile. Again, gene ontology analysis showed that inflammation and oxidative stress were significantly activated by DON exposure, and these were recovered when the DON-exposed mice were supplemented with an LF diet. Consistent with these findings, the signaling pathways of the reduced oxidative phosphorylation and elevated TNFα were also observed to be ameliorated by LF treatment. Importantly, histone modifications, including acetylation, methylation, and lactylation were suggested to be the vital players involved in the DON or LF treatment, in which LF significantly increased the loss of histone modifications on these genes. With a bioinformatics analysis and validation by qRT-PCR, the nuclear receptor NR5A2 was selected as a key master in the ileum of mice stimulated by DON. LF performed the benefit function on the NR5A2-mediated oxidative stress genes Ncoa4 and Prdx3 in the DON-exposed mice. Moreover, a ChIP-qPCR was used to verify that histone marks involving H3K9ac, H3K18ac, H3k27ac, H3K4me1, H3K9la, and H3K18la facilitated the epigenetic regulation of NR5A2-modulated actions. We conclude that dietary LF effectively ameliorated ileum lesions induced by DON in mice by modulating oxidative genes Ncoa4 and Prdx3 through histone modifications.
鉴于乳铁蛋白（LF）对肠道稳态具有出色的保护作用，但其潜在机制，尤其是表观遗传调控机制仍不清楚。本研究旨在探讨日粮LF通过组蛋白修饰对氧化基因进行表观遗传调控，以改善暴露于脱氧雪腐镰刀菌烯醇（DON）污染日粮的小鼠回肠炎症。正如预期的那样，我们在形态学分析中发现，与对照组相比，DON暴露增加了回肠隐窝深度（CD）和绒毛宽度（VW），但降低了绒毛高度（VH）和VH:CD比值。同样，在DON暴露的小鼠回肠中观察到活性氧（ROS）和丙二醛（MDA）升高，同时三磷酸腺苷（ATP）、超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽（GSH）以及复合物I、III、V降低。相比之下，LF显著改善了形态学和生化指标的损伤。接下来，我们使用DON或LF处理的小鼠回肠RNA进行转录组分析，以探索变化的信号通路。首先，DON暴露的小鼠中细胞周期通路基因显著下调，而LF改善了细胞周期谱。此外，基因本体分析表明，DON暴露显著激活了炎症和氧化应激，当给DON暴露的小鼠补充LF日粮时，这些情况得到了恢复。与这些发现一致，LF处理也改善了氧化磷酸化降低和肿瘤坏死因子α（TNFα）升高的信号通路。重要的是，包括乙酰化、甲基化和乳酰化在内的组蛋白修饰被认为是参与DON或LF处理的关键因素，其中LF显著增加了这些基因上组蛋白修饰的缺失。通过生物信息学分析和实时定量聚合酶链反应（qRT-PCR）验证，核受体NR5A2被选为DON刺激的小鼠回肠中的关键主控因子。LF对DON暴露小鼠中NR5A2介导的氧化应激基因Ncoa4和Prdx3发挥有益作用。此外，染色质免疫沉淀-定量聚合酶链反应（ChIP-qPCR）用于验证涉及H3K9ac、H3K18ac、H3k27ac、H3K4me1、H3K9la和H3K18la的组蛋白标记促进了NR5A2调节作用的表观遗传调控。我们得出结论，日粮LF通过组蛋白修饰调节氧化基因Ncoa4和Prdx3，有效改善了DON诱导的小鼠回肠损伤。
