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Membrane peroxidation by lipopolysaccharide and iron-ascorbate adversely affects Caco-2 cell function: beneficial role of butyric acid^1,2,3

¹ From the Division of Gastroenterology, Hepatology, and Nutrition, Centre de Recherche, Sainte Justine Hospital, and the Departments of Nutrition (FC, SB, MLM, and EL), Pediatrics (EGS), and Biochemistry (ED), Université de Montréal, Montreal, Canada, and the Department of Anatomy and Cell Biology, Université de Sherbrooke, Sherbrooke, Canada (CA).

Background: Membrane lipid peroxidation may play a role in immune-mediated bowel diseases.

Objective: We examined the effects of lipopolysaccharide (LPS), a ubiquitous endotoxin mediator of gram-negative bacteria, alone and in combination with iron-ascorbate, on enterocyte function. Furthermore, we assessed the antioxidant capacity of butylated hydroxytoluene (BHT) and butyric acid, which are known to play a significant role in the welfare of intestinal mucosa.

Design: Differentiated intestinal Caco-2 cells were used to study the induction of membrane peroxidation by LPS (100 µg/mL) and iron-ascorbate (0.2 and 2 mmol/L, respectively) and to examine the beneficial effects of BHT and butyric acid.

Results: A significant dose-dependent increase in malondialdehyde, accompanied by lower apical membrane fluidity and significantly decreased sucrase activity, was observed when Caco-2 cells were incubated with LPS. LPS also augmented paracellular permeability ([¹⁴C]polyethylene glycol flux), prostaglandin E₂ production, and cyclooxygenase-2 (EC 1.14.99.1) expression. These abnormalities were exacerbated by the coadministration of iron-ascorbate, but most of them were suppressed by butyric acid and BHT.

Conclusion: Bacterial endotoxin and prooxidants may overwhelm antioxidant defenses and become deleterious to enterocyte function, whereas butyric acid and BHT may provide antioxidant protection.

Key Words: Iron-ascorbate • membrane fluidity • permeability • cyclooxygenase-2 • prostaglandin E₂ • PGE₂ • lipopolysaccharide • butylated hydroxytoluene • BHT • butyric acid • Caco-2 cells

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