title: Inducible and neuronal nitric oxide synthases exert contrasting effects during rat intestinal recovery following fasting.
abstract: We investigated the effects of endogenous inducible (iNOS) and neuronal nitric oxide synthase on recovery from intestinal mucosal atrophy caused by fasting-induced apoptosis and decreased cell proliferation during refeeding in rats. Rats were divided into five groups, one of which was fed ad libitum, and four of which underwent 72 h of fasting, followed by refeeding for 0, 6, 24, and 48 h, respectively. iNOS and neuronal nitric oxide synthase mRNA and protein levels in jejunal tissues were measured, and mucosal height was histologically evaluated. Apoptotic indices, interferon-γ (IFN-γ) transcription levels, nitrite levels (as a measure of nitric oxide [NO] production),8-hydroxydeoxyguanosine formation (indicating reactive oxygen species [ROS] levels), crypt cell proliferation, and the motility indices (MI) were also estimated. Associations between mucosal height and NOS protein levels were determined using Spearman's rank correlation test. Notably, we observed significant increases in mucosal height and in neuronal nitric oxide synthase mRNA and protein expression as refeeding time increased. Indeed, there was a significant positive correlation between neuronal nitric oxide synthase protein level and mucosal height during the 48-h refeeding period ( r = 0.725, P < 0.01). Conversely, iNOS mRNA and protein expression decreased according to refeeding time, with a significant negative correlation between iNOS protein level and mucosal height being recorded during the 48-h refeeding period ( r = -0.898, P < 0.01). We also noted a significant negative correlation between jejunal neuronal nitric oxide synthase and iNOS protein concentrations over this same period ( r = -0.734, P < 0.01). Refeeding also restored the decreased jejunal MI caused by fasting. Our finding suggests that refeeding likely repairs fasting-induced jejunal atrophy by suppressing iNOS expression and subsequently inhibiting NO, ROS, and IFN-γ as apoptosis mediators, and by promoting neuronal nitric oxide synthase production and inducing crypt cell proliferation via mechanical stimulation. Impact statement Besides providing new data confirming the involvement of iNOS and nNOS in intestinal mucosal atrophy caused by fasting, this study details their expression and function during recovery from this condition following refeeding. We demonstrate a significant negative correlation between iNOS and nNOS levels during refeeding, and associate this with cell proliferation and apoptosis in crypts and villi. These novel findings elucidate the relationship between these NOS isoforms and its impact on recovery from intestinal injury. A mechanism is proposed comprising the up-regulation of nNOS activity by mechanical stimulation due to the presence of food in the intestine, restricting iNOS-associated apoptosis and promoting cell proliferation and gut motility. Our investigation sheds light on the molecular basis behind the repercussions of total parenteral nutrition on intestinal mucosal integrity, and more importantly, the beneficial effects of early enteral feeding.
authors.affiliation: Division of Pathophysiology, Faculty of Pharmaceutical Science, Department of Clinical Dietetics and Human Nutrition, Josai University, Saitama 350-0295, Japan.
authors.forename: Junta
authors.initials: J
authors.lastname: Ito
authors.affiliation: Division of Pathophysiology, Faculty of Pharmaceutical Science, Department of Clinical Dietetics and Human Nutrition, Josai University, Saitama 350-0295, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Uchida
authors.affiliation: Division of Pathophysiology, Faculty of Pharmaceutical Science, Department of Clinical Dietetics and Human Nutrition, Josai University, Saitama 350-0295, Japan.
authors.forename: Naomi
authors.initials: N
authors.lastname: Machida
authors.affiliation: Division of Pathophysiology, Faculty of Pharmaceutical Science, Department of Clinical Dietetics and Human Nutrition, Josai University, Saitama 350-0295, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Ohtake
authors.affiliation: Division of Pathophysiology, Faculty of Pharmaceutical Science, Department of Clinical Dietetics and Human Nutrition, Josai University, Saitama 350-0295, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Saito
authors.affiliation: Division of Pathophysiology, Faculty of Pharmaceutical Science, Department of Clinical Dietetics and Human Nutrition, Josai University, Saitama 350-0295, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Kobayashi
mesh_terms: D000818:Animals; D017209:Apoptosis; D001284:Atrophy; D049109:Cell Proliferation / Q000502:physiology; D004435:Eating / Q000502:physiology; D005215:Fasting / Q000502:physiology*; D007413:Intestinal Mucosa / Q000737:chemistry / Q000201:enzymology / Q000473:pathology* / Q000503:physiopathology; D007422:Intestines; D008297:Male; D009569:Nitric Oxide / Q000032:analysis; D052248:Nitric Oxide Synthase Type I / Q000378:metabolism / Q000502:physiology*; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar
chemical_list: D009569:Nitric Oxide; D052248:Nitric Oxide Synthase Type I; D052247:Nitric Oxide Synthase Type II
keywords: Intestinal atrophy; apoptosis; inducible nitric oxide synthase; neuronal nitric oxide synthase; refeeding