Adrenal cortical activation in murine colitis Denis Franchimont, Gerd Bouma, Jerome Galon, Gernot W. Wolkersdörfer, Andrea Haidan, George P. Chrousos, Stefan R. Bornstein  Gastroenterology  Volume 119, Issue 6, Pages 1560-1568 (December 2000) DOI: 10.1053/gast.2000.20235 Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 1 TNBS induces colitis in SJL/J but not in C57Bl/6 mice. Weight curves (data are expressed as mean ± SD) and colon specimens after intrarectal administration of TNBS in (A) resistant C57Bl/6 mice and (B) susceptible SJL/J mice (n = 5). □, Placebo-treated mice; ●, TNBS-injected mice. TNBS-induced colitis in SJL/J mice leads to statistically significant, ongoing weight loss (P < 0.05 compared with controls), whereas C57Bl/6 mice quickly recover from the TNBS enema. One representative of 5 experiments is shown. Gastroenterology 2000 119, 1560-1568DOI: (10.1053/gast.2000.20235) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 2 Increased corticosterone secretion during colitis. (A) Plasma corticosterone levels in TNBS colitis–susceptible SJL/J (□) and resistant C57Bl/6 (▴) mice at different time points. Data on the x-axis represent different time points (in days) after the induction of colitis. A rapid and strong increase in corticosterone levels is observed in SJL/J mice developing colitis (P < 0.0001). In the resistant strain, a small, stress-related increase in plasma corticosterone is seen (P < 0.01). The magnitude of the increase in plasma corticosterone in SJL/J mice is significantly higher than in C57Bl/6 mice (P < 0.0005). (B) Plasma ACTH levels in susceptible SJL/J (□) and resistant C57Bl/6 (▴) mice. Data are means ± SEM. The experiment is representative of 3 different independent experiments (n = 5). *P < 0.01; **P < 0.0001. Gastroenterology 2000 119, 1560-1568DOI: (10.1053/gast.2000.20235) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 3 Morphologic change of adrenals during colitis. Light microscopy of toluidine blue–stained adrenal slides from (A) placebo-treated and (B) TNBS colitis SJL/J mice. The adrenal glands from the mice with colitis showed dramatic signs of stimulation with hyperplasia of the adrenocortical cells and a marked hypervascularization (arrows) of the cortex. Gastroenterology 2000 119, 1560-1568DOI: (10.1053/gast.2000.20235) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 4 Ultrastructural changes of the adrenocortical cells. Electron microscopy of adrenocortical cells from (A) placebo-treated and (B) TNBS colitis SJL/J mice. Adrenocortical steroidogenic cells from colitis mice showed signs of marked stimulation with increased mitochondria (Mit), decreased liposomes (Lip), dilated smooth endoplasmic reticulum, and frequent formation of filopodia. Nuc, nucleus; Lym, lymphocyte; E, erythrocyte. Bar = 2 μm. Gastroenterology 2000 119, 1560-1568DOI: (10.1053/gast.2000.20235) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 5 Interaction of intra-adrenal lymphocytes (Lym) with adrenocortical cells during colitis. Lymphocytes were found in direct contact with adrenocortical cells in SJL/J mice with colitis; the adrenocortical cells extended filopodia to the cell membrane of the lymphocytes. Bar = 0.5 μm. B is a higher-magnification of the insert in A. Mit, mitochondrium; TZ, thrombocyte; E, erythrocyte; Ser, smooth endoplasmic reticulum; Rer, rough endoplasmic reticulum. Bar = 0.2 μm. Gastroenterology 2000 119, 1560-1568DOI: (10.1053/gast.2000.20235) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 6 Increased intra-adrenal and plasma IL-6 may stimulate adrenocortical cells in mice with colitis. (A) Increased intra-adrenal IL-6 and IL-1β expression in colitis. Adrenals from both control, susceptible SJL/J, and resistant C57BL/6 mice were removed and sonicated in lysis buffer. RNAs were then extracted, and cytokine expression was evaluated by RNase protection. TNF-α, IL-1β, and IL-6 expression in placebo-treated SJL/J and C57BL/6 mice (lanes 1 and 3) and in TNBS-treated SJL/J and C57BL/6 mice (lanes 2 and 4). (B) Increased plasma IL-6 during colitis. Plasma IL-6 level in control and TNBS-treated SJL/J mice (lanes 1 and 2) and in control and TNBS-treated C57BL/6 mice (lanes 3 and 4). Results are representative of 3 different independent experiments (n = 5). (C) Cytokine receptor expression in the adrenal glands. Control lane without adrenal RNAs (lane 1). IL-1RI/RII, IL-6Rα, gp130, and TNF-αRp75/p55 expression in normal adrenal glands (lane 2). (D) IL-6Rα and gp130 were equally expressed in the adrenal glands of placebo-treated (lane 1) and TNBS-treated (lane 2) SJL/J mice. (E) Direct IL-6 stimulation of corticosterone secretion by adrenocortical cells. Corticosterone production by adrenocortical Y-1 cells untreated (lane 1) or treated either with ACTH (lane 2) or with increasing doses of IL-6 (lanes 3–5). These data are from 1 experiment representative of 3 different independent experiments. Gastroenterology 2000 119, 1560-1568DOI: (10.1053/gast.2000.20235) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 7 Effect of anti–IL-6 treatment on plasma corticosterone levels and adrenal morphology. Light microscopy of toluidine blue–stained adrenal slides from (A) placebo-treated and (B) anti–IL-6–treated (same magnification) SJL/J mice with TNBS colitis. The signs of stimulation with hyperplasia of the adrenocortical cells and hypervascularization of the cortex were markedly reduced by a single injection of 1 mg of monoclonal antibody to IL-6. (C) Anti–IL-6 treatment led to a 22% reduction in plasma corticosterone levels. *P < 0.05; n = 10. Gastroenterology 2000 119, 1560-1568DOI: (10.1053/gast.2000.20235) Copyright © 2000 American Gastroenterological Association Terms and Conditions