Vitamin D regulates claudin-2 and claudin-4 expression in active ulcerative colitis by p-Stat-6 and Smad-7 signaling

• Published in: International journal of colorectal disease 2020-07, Vol.35 (7), p.1231-1242
• Main Authors: Domazetovic, Vladana, Iantomasi, Teresa, Bonanomi, Andrea Giovanni, Stio, Maria
• Format: Article
• Language: English
• Subjects: Alfacalcidol; Analysis; Biopsy; Calcifediol; Calcitriol; Colon; Diet therapy; Ethical aspects; Gastroenterology; Hepatology; Immunosuppressive agents; Immunotherapy; Inflammatory bowel diseases; Internal Medicine; Intestine; Medicine; Medicine & Public Health; Mucosa; Original Article; Phosphorylation; Proctology; Proteins; siRNA; Smad protein; Surgery; Tight junctions; Transfection; Ulcerative colitis; Vitamin D; Vitamin D3; Vitamin deficiency
• ISSN: 0179-1958; 1432-1262
• DOI: 10.1007/s00384-020-03576-0

Purpose The tight junctions (TJ) responsible for the integrity of the intestinal barrier are altered in patients with inflammatory bowel disease (IBD), but the physiopathological mechanisms that lead to this alteration are not yet clear. The aim of this study was to determine whether vitamin D, which regulates the integrity of the epithelial barrier by expressing TJ proteins, reduces claudin-2 (Cl-2) levels by inhibiting Stat-6 phosphorylation and whether it increases claudin-4 (Cl-4) levels by blocking Smad-7 activity. Methods Biopsies were obtained from inflamed and non-inflamed tracts of the right side colon (caecum or ascending colon) from the same patient with active UC. All the patients were affected by a recent flare-up of ulcerative rectocolitis (RCU), with no previous biologic or immunosuppressive therapy, and all the biopsies were obtained before any treatments. The biopsies were cultured in the presence or not of 1,25-dihydroxyvitamin D 3 (1,25(OH) 2 D 3 ). We also used T84 cells as an in vitro model to perform transfection experiments with Stat-6 and Smad-7. Results Our results indicate that 1,25(OH) 2 D 3 is able to regulate CL-2 and CL-4 protein levels, which are increased and reduced in the intestinal mucosa of UC patients, respectively. In the biopsies obtained from UC patients 1,25(OH) 2 D 3 reduces Cl-2 levels by blocking Stat-6 phosphorylation and increases Cl-4 levels by blocking Smad-7 activity. T84 cells, transfected with siRNA of Stat-6 and Smad-7, showed reduced Cl-2 levels and increased Cl-4 levels, confirming that 1,25(OH) 2 D 3 regulates Cl-2 and Cl-4 by decreasing p-Stat-6 and Smad-7 levels. Conclusions Our results indicate that the effects of vitamin D on Cl-2 and Cl-4 are mediated by p-Stat-6 and Smad-7 signal, respectively. The study suggests that vitamin D administration to UC patients could be a useful therapeutic intervention, given that vitamin D deficiency is found in these patients.