Downregulation of intestinal multidrug resistance transporter 1 in obese mice: Effect on its barrier function and role of TNF-α receptor 1 signaling

• Published in: Nutrition (Burbank, Los Angeles County, Calif.) Vol. 111; p. 112050
• Main Authors: Barranco, María Manuela, Perdomo, Virginia Gabriela, Zecchinati, Felipe, Manarin, Romina, Massuh, Greta, Sigal, Nicolás, Vignaduzzo, Silvana, Mottino, Aldo Domingo, Villanueva, Silvina Stella Maris, García, Fabiana
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.2023; Elsevier Limited
• Subjects: Animals; Bioavailability; Cytokines; Diet; Diet, High-Fat; Down-Regulation; Drug Resistance, Multiple; Enzyme-linked immunosorbent assay; Enzymes; Gene expression; Glucose; High fat diet; Immunohistochemistry; Inflammation; Inflammatory response; Insulin resistance; Intestine; Laboratory animals; Male; Mdr-1; Metabolic disorders; Metabolic syndrome; Mice; Mice, Inbred C57BL; Mice, Obese; Multidrug resistance; Multidrug resistant organisms; Obesity; Oral administration; P-Glycoprotein; Polymerase chain reaction; Proteins; Receptors; Rhodamine; Rhodamine 123; RNA, Messenger; Signal transduction; Signaling; Small intestine; Substrates; TNFR1; Tumor Necrosis Factor-alpha - metabolism; Tumor necrosis factor-TNF; Tumor necrosis factor-α; Variance analysis; Western blotting; United States > US; Argentina; ABC; C; P-gp; R1KO; R123; HFD; ITT; Inflammation; IPGTT; BBM; TNFR1; AUC; CYP3A; Mdr-1; MetS; TNF-α; High-fat diet; ELISA; w/v
• ISSN: 0899-9007; 1873-1244
• DOI: 10.1016/j.nut.2023.112050

•A high-fat diet induced downregulation of intestinal multidrug resistance transporter 1 (Mdr-1).•High-fat diet–induced Mdr-1downregulation had negative outcomes for its barrier function.•Tumor necrosis factor α signaling played a role in Mdr-1 downregulation induced by a high-fat diet.•Impairment of the Mdr-1 barrier function may have consequences for drug bioavailability. Multidrug resistance transporter 1 (Mdr-1) is a relevant component of the intestinal transcellular barrier that decreases absorption of oral drugs, thus modulating their bioavailability. Obese patients with metabolic disorders take medications that are subjected to intestinal metabolism and the Mdr-1–dependent barrier. This study evaluated the effect of a high-fat diet (HFD; 40% fat for 16 wk) on Mdr-1 expression and transport activity in C57BL/6 (C57) male mice. Comparable studies were performed in tumor necrosis factor α (TNF-α) receptor 1 knockout mice (R1KO) to delineate a possible role of TNF-α signaling. mRNA expression was evaluated by real-time polymerase chain reaction and protein levels by western blotting and immunohistochemistry. Mdr-1 activity was assessed using the everted intestinal sac model, with rhodamine 123 as the substrate. Statistical comparisons were made using the Student t test or one-way analysis of variance followed by the post hoc Tukey test. Mdr-1 protein, as well as its corresponding Mdr1a and Mdr1b mRNA, was decreased in C57-HFD mice compared with controls. Immunohistochemical studies confirmed downregulation of Mdr-1 in situ. These results correlated with a 48% decrease in the basolateral to apical transport of rhodamine 123. In contrast, R1KO-HFD modified neither intestinal Mdr-1 mRNA nor its protein expression or activity. In addition, C57-HFD showed elevated intestinal TNF-α mRNA and protein (enzyme-linked immunosorbent assay) levels, whereas R1KO-HFD was undetectable or had a lower increase, respectively. This study demonstrated an impairment of the Mdr-1 intestinal barrier function induced by HFD as a consequence of downregulation of both Mdr-1 gene homologues, resulting in impaired Mdr-1 protein expression. Inflammatory response mediated by TNF-α receptor 1 signaling was likely involved.