Department of Veterinary Medicine and Animal Sciences, University of Milan, Via Dell'Università 6, 26900, Lodi, Italy.
University of Veterinary Medicine and Pharmacy in Košice, Komenského 68/73, 04181, Košice, Slovakia.
Sci Rep. 2023 Dec 7;13(1):21595. doi: 10.1038/s41598-023-48018-3.
Omega-3 polyunsaturated fatty acids (n-3 PUFA), such as the eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are reported to beneficially affect the intestinal immunity. The biological pathways modulated by n-3 PUFA during an infection, at the level of intestinal epithelial barrier remain elusive. To address this gap, we investigated the proteomic changes induced by n-3 PUFA in porcine enterocyte cell line (IPEC-J2), in the presence and absence of lipopolysaccharide (LPS) stress conditions using shotgun proteomics analysis integrated with RNA-sequencing technology. A total of 33, 85, and 88 differentially abundant proteins (DAPs) were identified in cells exposed to n-3 PUFA (DHA:EPA), LPS, and n-3 PUFA treatment followed by LPS stimulation, respectively. Functional annotation and pathway analysis of DAPs revealed the modulation of central carbon metabolism, including the glycolysis/gluconeogenesis, pentose phosphate pathway, and oxidative phosphorylation processes. Specifically, LPS caused metabolic dysregulation in enterocytes, which was abated upon prior treatment with n-3 PUFA. Besides, n-3 PUFA supplementation facilitated enterocyte development and lipid homeostasis. Altogether, this work for the first time comprehensively described the biological pathways regulated by n-3 PUFA in enterocytes, particularly during endotoxin-stimulated metabolic dysregulation. Additionally, this study may provide nutritional biomarkers in monitoring the intestinal health of human and animals on n-3 PUFA-based diets.
ω-3 多不饱和脂肪酸（n-3PUFA），如二十碳五烯酸（EPA）和二十二碳六烯酸（DHA），据报道可有益地影响肠道免疫。在感染过程中，n-3PUFA 调节的生物学途径在肠道上皮屏障水平上仍不清楚。为了解决这一差距，我们使用 shotgun 蛋白质组学分析结合 RNA 测序技术，研究了 n-3PUFA 在猪肠上皮细胞系（IPEC-J2）中诱导的蛋白质组变化，同时存在和不存在脂多糖（LPS）应激条件。在暴露于 n-3PUFA（DHA:EPA）、LPS 和 n-3PUFA 处理后再用 LPS 刺激的细胞中，分别鉴定出 33、85 和 88 个差异丰度蛋白（DAP）。DAP 的功能注释和途径分析显示，中央碳代谢的调节，包括糖酵解/糖异生、磷酸戊糖途径和氧化磷酸化过程。具体来说，LPS 导致肠细胞代谢失调，而 n-3PUFA 的预先处理减轻了这种失调。此外，n-3PUFA 的补充促进了肠细胞的发育和脂质稳态。总的来说，这项工作首次全面描述了 n-3PUFA 在肠细胞中调节的生物学途径，特别是在内毒素刺激的代谢失调期间。此外，这项研究可能为基于 n-3PUFA 的饮食中监测人类和动物肠道健康提供营养生物标志物。
