allopurinol and Enterocolitis--Necrotizing

An intraluminal casein model (ICM) of necrotizing enterocolitis (NEC) is able to produce small-bowel changes reminiscent of human NEC in neonatal animals. We studied bacterial translocation (BT) in NEC induced by using the ICM in neonatal piglets. We also studied whether allopurinol (AL) and N-acetylcysteine (NAC) have an effect on BT and mucosal changes in the ICM of NEC. Twenty-eight neonatal piglets were randomized into four groups. NEC was induced in 21 by injecting casein-d-gluconate into a loop of terminal ileum: group Cas (n = 7) had no premedication, in group Cas/AL (n = 7) intravenous (i.v.) Al (100 mg/kg), and in group Cas/NAC (n = 7) i.v. NAC (200 mg/kg) was given. Group Sham (n = 7) had the ileum injected with 0.9% saline with no premedication. Immediately after the injection a mesenteric lymph node (MLN) adjacent to the loop was harvested for quantitative aerobic bacterial culture; 4 h after the injection another MLN and samples of spleen, liver, kidney, and lung were harvested and cultured. Comparison of the incidence of samples with positive bacterial cultures and the number of colony-forming units (CFU) in samples was made between groups. The severity of NEC in the ileum was graded from 0 to 3 according to macroscopic and histologic findings. NEC changes in the bowel were most severe in Cas piglets, less severe in Cas/NAC piglets ( P < 0.5), and sham piglets had the least severe changes ( P < 0.05). piglets with NEC changes in the ileum had a higher incidence of BT into the MLN than piglets without NEC changes ( P < 0.05), but the difference in CFU was not significant ( P > 0.05). In Cas and Cas/NAC piglets a high incidence of BT into the MLN was noted as early at -5 min after casein injection. The incidence of BT into the MLN was significantly higher in Cas and Cas/NAC piglets than in Sham piglets ( P < 0.05), the difference in CFU being not significant ( P > 0.05). BT in Cas/Al piglets was not significantly different from that of Cas piglets ( P > 0.05), but less than in Cas/NAC piglets ( P < 0.05). Four hours after casein injection into the ileum there was significant BT into the MLN. Premedication with NAC was associated with less severe NEC changes, but neither NAC nor AL significantly affected BT.

Topics: Acetylcysteine; Allopurinol; Analysis of Variance; Animals; Animals, Newborn; Bacterial Translocation; Caseins; Disease Models, Animal; Enterocolitis, Necrotizing; Ileum; Intestinal Mucosa; Lymphatic System; Male; Swine

Necrotizing enterocolitis (NEC) is the most important gastrointestinal emergency affecting especially preterm infants and causes severe morbidities and mortality. However, there is no cure. Oxidant stress, inflammation, apoptosis, as well as prematurity are believed to responsible in the pathogenesis of the disease. Ginger and its compounds have anti-inflammatory, antimicrobial, anti-oxidant properties and immunomodulatory, cytoprotective/regenerative actions.. This study aimed to evaluate the beneficial effects of ginger on the intestinal damage in an experimental rat model of NEC.. Thirty newborn Wistar rats were divided into three groups: NEC, NEC + ginger and control in this experimental study. NEC was induced by injection of intraperitoneal lipopolysaccharide, feeding with enteral formula, hypoxia-hyperoxia and cold stress exposure. The pups in the NEC + ginger group were orally administered ginger at a dose of 1000 mg/kg/day. Proximal colon and ileum were excised. Histopathological, immunohistochemical (TUNEL for apoptosis, caspase 3 and 8) and biochemical assays including xanthine oxidase (XO), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malonaldehyde (MDA) and myeloperoxidase (MPO), tumor necrosis factor-α (TNF-α), interleukin1β (IL-1β), and interleukin 6 (IL-6) activity were evaluated.. Compared with the NEC group, the rat pups in the NEC + ginger group had better clinical disease scores and weight gain (p < 0.05). Macroscopic evaluation, Histopathologic and apoptosis assessment (TUNEL, caspase 3 and 8) releaved that severity of intestinal damage were significantly lower in the NEC + ginger group (p < 0.05). The levels of TNF-α, IL-1β and IL-6 in the ginger treated group were significantly decreased (P < 0.05). The GSH-Px and SOD levels of the ginger treated group were significantly preserved in the NEC + ginger group (p < 0.05). The tissue XO, MDA and MPO levels of the NEC + ginger group were significantly lower than those in the NEC group (P < 0.05).. Ginger therapy efficiently ameliorated the severity of intestinal damage in NEC and may be a promising treatment option.

Topics: Animals; Anti-Inflammatory Agents; Apoptosis; Caspase 3; Caspase 8; Cytokines; Enterocolitis, Necrotizing; Glutathione Peroxidase; Intestinal Mucosa; Intestines; Malondialdehyde; Peroxidase; Phytotherapy; Plant Extracts; Rats, Wistar; Superoxide Dismutase; Xanthine Oxidase; Zingiber officinale

The aim of this study was to determine the beneficial effects of Nigella sativa oil (NSO) on rats with necrotizing enterocolitis (NEC).. Thirty newborn Sprague-Dawley rats were randomly divided into three groups as NEC, NEC + NSO, and control. NEC was induced by enteral formula feeding, exposure to hypoxia-hyperoxia and cold stress. Pups in the NEC + NSO group were administered NOS at a dose of 2 ml/kg daily by intraperitoneal route from the first day until the end of the study. Proximal colon and ileum were excised for histopathologic, apoptosis (TUNEL) and biochemical evaluation, including xanthine oxidase (XO), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malonaldehyde (MDA), and myeloperoxdase (MPO) activities.. Pups in the NEC + NOS group had better clinical sickness scores and weight gain compared to the NEC group (p < 0.05). In the macroscopic assessment, histopathologic and apoptosis evaluation (TUNEL), severity of bowel damage was significantly lower in the NEC + NOS group compared to the NEC group (p < 0.05). Tissue GSH-Px and SOD levels were significantly preserved in the NEC + NSO group (p < 0.05), whereas, tissue MDA, MPO levels of the NEC + NSO group were significantly lower than those in the NEC group (p < 0.05).. NSO significantly reduced the severity of intestinal damage in NEC.

Topics: Animals; Animals, Newborn; Antioxidants; Apoptosis; Colon; Disease Models, Animal; Enterocolitis, Necrotizing; Glutathione Peroxidase; Ileum; Malondialdehyde; Medicine, Traditional; Nigella sativa; Peroxidase; Phytotherapy; Plant Oils; Rats; Rats, Sprague-Dawley; Superoxide Dismutase; Xanthine Oxidase

To determine the preventative effect of caffeic acid phenethyl ester (CAPE) in necrotizing enterocolitis (NEC) in an experimental rat model of NEC.. Thirty newborn Sprague-Dawley rats were randomly divided into three groups; as NEC, NEC + CAPE and control. NEC was induced by enteral formula feeding, subjected to hypoxia-hyperoxia and cold stress. Pups in the NEC + CAPE group were treated with CAPE at a dose of 30 mg/kg daily by intraperitoneal route from the first day to the end of the study. All pups were executed on the fourth day. Proximal colon and ileum were allocated for histopathologic and biochemical evaluation, including xanthine oxidase (XO), total antioxidant status (TAS), total oxidant status (TOS), malonaldehyde (MDA) and myeloperoxidase (MPO) activities.. The pups in the NEC + CAPE group had better histopathologic and apoptosis evaluations (TUNEL and caspase-9) and the severity of bowel damage was significantly lower in the NEC + CAPE group compared to the NEC group (P < 0.01). The clinical sickness scores and body weight in the NEC + CAPE group was significantly better compared to the NEC group (P < 0.05). Tissue MDA, MPO, XO levels and TOS were remarkably reduced in the NEC + CAPE group, however, TAS was significantly increased in the NEC + CAPE group (P < 0.05).. Treatment with CAPE reduces the intestinal damage in NEC.

Topics: Administration, Oral; Animals; Animals, Newborn; Apoptosis; Caffeic Acids; Caspase 9; Cell Count; Disease Models, Animal; Enterocolitis, Necrotizing; Ileum; Immunohistochemistry; In Situ Nick-End Labeling; NF-kappa B; Oxidative Stress; Peroxidase; Phenylethyl Alcohol; Rats; Rats, Sprague-Dawley; Treatment Outcome; Xanthine Oxidase

Evaluation of prophylactic effects of omeprazole and/or vitamin E on the formation of free oxygen radicals (FOR) and bowel histopathology in the newborn rat model of hypoxia/reoxygenation (H/R) that resembles human necrotizing enterocolitis (NEC). Eighty newborn rats were randomly divided into eight groups. H/R was done using airtight chamber. Rats were exposed to 100% CO2 for 15 min followed by a reoxygenation for the next 15 min with 100% O2. Group 1 (n = 10) was the control group. Group 2 (n = 10) rats received vitamin E. In Group 3 (n = 10) omeprazole was administrated. Group 4 (n = 10) rats received omeprazole and vitamin E. Group 5 (n = 10) rats were subjected to H/R two times for 2 days and one time for 3 days. Group 6 (n = 10) received vitamin E in addition to H/R for 5 days and in Group 7 (n = 10) omeprazole in addition to H/R for 5 days. In Group 8 (n = 10), vitamin E and omeprazole and H/R were applied for 5 days. Rats were killed at the end of the each process and bowel specimens were harvested for histopathological and biochemical investigations. We administrated vitamin E intramuscularly 300 unit/kg per day and omeprazole orally 20 mg/kg per day. Malondialdehyde (MDA), xanthine oxidase (XO), xanthine dehydogenase (XDH) and XO/(XO + XDH) were measured. Vitamin E and/or omeprazole treated rats had significantly less XO% levels than H/R only group (0.36, 0.38 and 0.57, respectively). Similarly, the MDA levels were significantly lower in vitamin E and/or omeprazole received rats than H/R only rats (88.8, 97.9 and 122.6, respectively). All rats treated with omeprazole and/or vitamin E had better biochemical and histopathological levels compared to H/R rats (p < 0.05). Histopathological results show that Group 5 (H/R only) had significantly more intestinal damage when compared with Group 6 (vitamin E + H/R), Group 7 (omeprazole + R/H) and Group 8 (vitamin E + omeprazole + H/R) (p < 0.001). Grade 2 and 3 intestinal damages were only in Group 5 and there were no statistical difference between in Groups 6, 7 and 8 (p > 0.001). Omeprazole and/or vitamin E may protect the biochemical and histopathological intestinal damage of H/R injury in rats. These drugs may be beneficial in the prophylaxis of NEC in humans as well.

Topics: Administration, Oral; Animals; Animals, Newborn; Anti-Ulcer Agents; Antioxidants; Disease Models, Animal; Drug Therapy, Combination; Enterocolitis, Necrotizing; Hypoxia; Injections, Intramuscular; Intestine, Small; Malondialdehyde; Omeprazole; Rats; Rats, Wistar; Spectrophotometry; Vitamin E; Xanthine Dehydrogenase; Xanthine Oxidase

We utilized a newborn rat model of hypoxia/reoxygenation (H/R) that resembles human necrotizing enterocolitis (NEC) to investigate the effects of omeprazole and/or gentamicin on the formation of free oxygen radicals (FOR) and bowel histopathology. For H/R, 1-day-old rats were placed into a chamber of 100% CO2 for 5 min, then they were reoxygenized for the next 5 min. The rats (n = 70) were divided into seven groups: group 1 (control), group 2 (H/R), group 3 (omeprazole), group 4 (H/R + omeprazole), group 5 (gentamicin), group 6 (H/R + gentamicin), group 7 (H/R + omeprazole + gentamicin). Gentamicin and/or omeprazole were given orally for 3 days, then all animals were killed; bowel specimens were harvested. Histopathologic injury scores (HIS) and malonyldialdehyde (MDA) and XO/(XO+XDH) rates (XO; xanthine oxidase, XDH; xanthine dehydrogenase) were measured, which reflect the FOR levels. In group 2, the HIS was significantly higher than groups 4 and 6. The mean MDA values in groups 1-7 were as follows: 54.16, 104.2, 56.85, 63.43, 62.31, 76.85, 79.13, respectively. The mean XO/(XO + XDH) levels were 0.306, 0.461, 0.286, 0.335, 0.323, 0.410, 0.375 from groups 1 -7, respectively. Group 2 rats had significantly more MDA and XO/(XO + XDH) rates versus other groups (P < 001). Histopathologic injury and biochemical results were significantly more severe in group 2 than in groups 4 and 6 (P < 001). There was no difference between groups 1 and 4 according to XO/(XO + XDH) rates. In newborn rats, H/R produces FOR, which cause serious intestinal damage. Omeprazole and/or gentamicin reduce biochemical and histopathologic bowel damage. This effect was more obvious in omeprazole treated rats. We think omeprazole may open new insights into the treatment of H/R related bowel injuries like NEC.

Topics: Animals; Animals, Newborn; Anti-Bacterial Agents; Anti-Ulcer Agents; Disease Models, Animal; Enterocolitis, Necrotizing; Free Radicals; Gentamicins; Hypoxia; Intestinal Mucosa; Intestines; Malondialdehyde; Omeprazole; Rats; Rats, Wistar; Xanthine Dehydrogenase; Xanthine Oxidase

Although bilirubin, which crosses the blood-brain barrier, can cause irreversible brain damage, it also possesses antioxidant properties that may be protective against oxidative stress. Intestinal ischemia-reperfusion (IR) injury results in cell destruction, mediated via the generation of reactive oxygen species. Although increased serum bilirubin is correlated with increased antioxidant potential in the face of hyperoxia, evidence of bilirubin-associated protective effect against IR injury remains nonspecific. We therefore sought to investigate whether hyperbilirubinemia would be protective against IR injury to the intestine.. Young adult rats were randomly assigned to one of three groups: 1) IR/control (n = 12); 2) IR/hyperbilirubinemia (n = 10), in which IR was generated while the rats were treated with a continuous infusion of bilirubin; and 3) hyperbilirubinemia controls (n = 10). Blood and intestinal tissue samples were obtained to determine serial thiobarbituric acid reducing substances (index of lipid peroxidation) and for xanthine oxidase/xanthine dehydrogenase and glutathione/glutathione disulfide ratios. Intestinal histopathology was graded from 1 (normal) to 4 (severe necrotic lesions).. Histopathologic scoring and circulating and tissue thiobarbituric acid reducing substances were highest in the IR/control animals compared with either the IR/hyperbilirubinemics or the controls. All of these are consistent with the most severe injury in this group. Xanthine oxidase/xanthine dehydrogenase ratios were not significantly different among the groups.. Hyperbilirubinemia ameliorates the extent of intestinal IR injury in our model and appears to act as an antioxidant. This study supports the concept that bilirubin possesses some beneficial properties in vivo, although no direct clinical conclusions can be drawn from these data.

Topics: Animals; Bilirubin; Disease Models, Animal; Enterocolitis, Necrotizing; Glutathione Disulfide; Intestines; Rats; Reperfusion Injury; Thiobarbituric Acid Reactive Substances; Time Factors; Xanthine Dehydrogenase; Xanthine Oxidase