Peroxidized Linoleic Acid, 13-HPODE, Alters Gene Expression Profile in Intestinal Epithelial Cells

Abstract

Lipid peroxides (LOOHs) abound in processed food and have been implicated in the pathology of diverse diseases including gut, cardiovascular, and cancer diseases. Recently, RNA Sequencing (RNA-seq) has been widely used to profile gene expression. To characterize gene expression and pathway dysregulation upon exposure to peroxidized linoleic acid, we incubated intestinal epithelial cells (Caco-2) with 100 µM of 13-hydroperoxyoctadecadienoic acid (13-HPODE) or linoleic acid (LA) for 24 h. Total RNA was extracted for library preparation and Illumina HiSeq sequencing. We identified 3094 differentially expressed genes (DEGs) in 13-HPODE-treated cells and 2862 DEGs in LA-treated cells relative to untreated cells. We show that 13-HPODE enhanced lipid metabolic pathways, including steroid hormone biosynthesis, PPAR signaling, and bile secretion, which alter lipid uptake and transport. 13-HPODE and LA treatments promoted detoxification mechanisms including cytochrome-P450. Conversely, both treatments suppressed oxidative phosphorylation. We also show that both treatments may promote absorptive cell differentiation and reduce proliferation by suppressing pathways involved in the cell cycle, DNA synthesis/repair and ribosomes, and enhancing focal adhesion. A qRT-PCR analysis of representative DEGs validated the RNA-seq analysis. This study provides insights into mechanisms by which 13-HPODE alters cellular processes and its possible involvement in mitochondrial dysfunction-related disorders and proposes potential therapeutic strategies to treat LOOH-related pathologies.

Dietary triacylglycerol hydroperoxide is not absorbed, yet it induces the formation of other triacylglycerol hydroperoxides in the gastrointestinal tract

“…This reaction may be triggered by the intake of TGOOH. Previously, dietary lipid hydroperoxides have been reported to cause oxidative stress that induces pro-inflammatory changes in the gastrointestinal tract [ 32 , 33 , 38 , 39 ]. Separately, other studies reported that an inflamed intestinal mucosa is highly infiltrated with neutrophils ( e.g., in ulcerative colitis patients [ 40 , 41 ]), which generate 1 O 2 via the myeloperoxidase system of activated phagocytosis as the initial defense mechanism against tissue damage [ [42] , [43] , [44] ].…”

Serum Untargeted Metabolomics Reveal Potential Biomarkers of Progression of Diabetic Retinopathy in Asians

“…Although the mechanism of 13-PHODE in the pathogenesis of diabetic retinopathy has not been clarified so far, 13-PHODE has been found to induce inflammatory responses in animal models, which could significantly induce pro-inflammatory gene expression of TNF-α and MCP-1 in vitro , most notably in differentiated intestinal epithelial cells ( Keewan et al, 2020 ). The possible involvement of 13-PHODE in mitochondrial dysfunction-related disorders has also been reported ( Faizo et al, 2021 ). Further studies are needed to confirm our findings.…”

DataSheet1.PDF

“…Although the mechanism of 13-PHODE in the pathogenesis of diabetic retinopathy has not been clarified so far, 13-PHODE has been found to induce inflammatory responses in animal models, which could significantly induce pro-inflammatory gene expression of TNF-α and MCP-1 in vitro, most notably in differentiated intestinal epithelial cells (Keewan et al, 2020). The possible involvement of 13-PHODE in mitochondrial dysfunctionrelated disorders has also been reported (Faizo et al, 2021). Further studies are needed to confirm our findings.…”