Dataset Information

Comprehensive proteomic analysis reveals omega-3 fatty acids to counteract endotoxin-stimulated metabolic dysregulation in porcine enterocytes

ABSTRACT: Omega-3 polyunsaturated fatty acids (n-3 PUFA), such as the eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are reported to beneficially affect intestinal immunity. The intestinal epithelial layer (IEL), made up of absorptive cells as the enterocytes, is the primary site of fatty acid absorption. So far, the biological pathways modulated by n-3 PUFA in IEL during infection remain elusive. In the present study, shotgun proteomics analysis integrated with RNA sequencing (RNA-seq) technology was employed to unveil the proteomic changes induced by n-3 PUFA in enterocytes, in the presence and absence of lipopolysaccharide (LPS) stress conditions. A total of 33, 85, and 88 differential proteins were identified in the IPEC-J2 cells exposed to (i) n-3 PUFA (DHA:EPA, 1:2, 10 µM, 24 h), (ii) LPS (10 µg/mL, 24 h), or (iii) n-3 PUFA pre-treatment followed by LPS stimulation, respectively. Functional annotation and pathway analysis of the differential proteins revealed the modulation of central carbon metabolism, including glycolysis/gluconeogenesis, pentose phosphate pathway, and oxidative phosphorylation. Specifically, LPS caused metabolic dysregulation in the IPEC-J2 cells, abated upon prior treatment with n-3 PUFA. Indeed, n-3 PUFA supplementation facilitated enterocyte development and lipid homeostasis. This work provided a comprehensive picture of the biological pathways modulated by n-3 PUFA in enterocytes, both in the absence and presence of a state of endotoxin-induced metabolic dysregulation. Our findings provide important insights into the role of n-3 PUFA in the intestinal barrier, which could support better planning of nutritional strategies in managing intestinal health and immunity.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Sus Scrofa Domesticus (domestic Pig)

TISSUE(S): Cell Culture, Intestinal Epithelial Cell

SUBMITTER: Salvatore Pisanu

LAB HEAD: Salvatore Pisanu

PROVIDER: PXD036456 | Pride | 2023-12-11

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

Items per page:

1 - 5 of 56

Similar Datasets

Project description:Dietary polyunsaturated fatty acids (PUFA) act as potent natural hypolipidemics and are linked to many health benefits in humans and in animal models. Mice fed long-term a high fat diet, in which medium-chain alpha linoleic acid (ALA) was partially replaced by long-chain docosahexaenoic (DHA) and eicosapentaenoic (EPA) fatty acids, showed reduced accumulation of body fat and prevention of insulin resistance, besides increased mitochondrial beta-oxidation in white adipose tissue and decreased plasma lipids. ALA, EPA and DHA all belong to PUFA of n-3 series. The intestine is a gatekeeper organ for ingested lipids. To examine the potential contribution of the intestine in the beneficial effects of EPA and DHA, this study assessed gene expression changes using whole genome microarray analysis on small intestinal scrapings. The main biological process affected was lipid metabolism. Fatty acid uptake, peroxisomal and mitochondrial beta-oxidation, and omega-oxidation of fatty acids were all increased. Quantitative real time PCR and intestinal fatty acid oxidation measurements ([14C(U)]-palmitate) confirmed significant gene expression differences in a dose-dependent manner. Furthermore, no major changes in the expression of lipid metabolism genes were observed in colonic scrapings. In conclusion, we show that marine n-3 fatty acids regulate small intestinal gene expression patterns. Since this organ contributes significantly to whole organism energy use, this adaptation of the small intestine may contribute to the complex and observed beneficial physiological effects of these natural compounds under conditions that will normally lead to development of obesity and diabetes. Experiment Overall Design: Male 4-month-old C57BL/6J mice were maintained for 4 weeks on semisynthetic high-fat (20% wt/wt) diets differing in the composition of n-3 PUFA. Two isocaloric diets were used (n=12): control sHFf diet which contained flax-seed oil (rich in ALA) as the only lipid source, or the sHFf-F2 diet, which had the same composition except that 44 % of the lipids were replaced by a n-3 PUFA concentrate (EPA&DHA) containing 6 % EPA and 51 % DHA (EPAX 1050TG; EPAX AS, Lysaker, Norway). Quality control of RNA samples showed that four samples did not pass quality thresholds, and these samples were excluded from array analysis (two control small intestine samples and one EPA&DHA small intestine sample). The remaining RNA samples were pooled per tissue per diet group. Thus; the flax-seed (control) array was hybridized with RNA pooled from of 10 mice, the PUFA array was hybridized with RNA pooled from 11 mice.

			Action	DRS
	Control_C1.msf	Msf
	Control_C1.msfView	Msf
	Control_C1.raw	Raw
	Control_C2.msf	Msf
	Control_C2.msfView	Msf

Dataset Information

Comprehensive proteomic analysis reveals omega-3 fatty acids to counteract endotoxin-stimulated metabolic dysregulation in porcine enterocytes

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets