The role of reperfusion injury in occlusive intestinal ischemia of the neonate: Malonaldehyde-derived fluorescent products and correlation of histology

doi:10.1016/0022-4804(91)90061-P

Journal of Surgical Research

Volume 51, Issue 1, July 1991, Pages 1-4

https://doi.org/10.1016/0022-4804(91)90061-P Get rights and content

Abstract

Oxygen free radical reperfusion products may play a critical role in neonatal occlusive intestinal ischemia. We report a comparative analysis of light microscopy- and malonaldehyde (MDA)-derived fluorescent products as a measure of lipid peroxidation in occlusive intestinal ischemia in the rat. Weanling rats (n = 25) underwent cross clamping of the common mesenteric artery followed by various intervals of reperfusion; blood was sampled from the common mesenteric vein and the ileum was simultaneously biopsied. Blind-light microscopic scoring of the ischemic intestine was used. Fluorescent products were extracted using a chloroform/methanol/acidic water solvent extraction and measured with a spectrophotofluorometer using excitation/emission wavelengths of 360 and 430 nm, respectively. A trend was observed with prolonged reperfusion. Accumulation of fluorescent products correlated directly with the interval of reperfusion. Graded intervals of vascular occlusion produced progressive intestinal injury, but light microscopic analysis was not a sensitive index to distinguish the influence of graded reperfusion intervals. These data confirm a role for both ischemia and reperfusion in occlusive intestinal injury in the neonate and suggest that MDA accumulation may be a sensitive index of the reperfusion component of such injury.

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Cited by (36)

Therapeutic and preventative effects of ankaferd blood stopper in an experimental necrotizing enterocolitis model
2019, Biomedicine and Pharmacotherapy
Citation Excerpt :
Reoxygenation after hypoxia also leads to an increase in ROS formation [30]. Through the oxidation of membrane lipids, DNA, and proteins, ROS have a major role in cellular injury and subsequent tissue damage and development of NEC [2,5,17,30,31]. TAS, TOS, and OSI are important indicators of global changes in oxidation status [27].
Necrotizing enterocolitis (NEC) is a major neonatal health problem that especially affects preterm infants and causes severe morbidity and mortality. Although its pathogenesis is not fully understood, important risk factors include prematurity, oxidative stress, inflammation, and apoptosis. Ankaferd Blood Stopper® (ABS) has antioxidant, antiinflammatory, antimicrobial, antiapoptotic, and wound healing accelerant properties. In this study, we aimed to investigate whether treatment with ABS reduced the severity of NEC in rat pups in an experimental NEC model. Thirty-six newborn Wistar albino rat pups were randomly assigned to the control, NEC + saline, or NEC + ABS groups. NEC was induced by intraperitoneal injection of lipopolysaccharide, feeding with hyperosmolar enteral formula, and exposure to hypoxia/hyperoxia and cold stress. ABS was administered intraperitoneally to the pups in the NEC + ABS group daily starting on day 1 of the study at a dose of 2 ml/kg by diluting 2 ml with saline at a ratio of 1:3. All pups were sacrificed on day 4. The terminal ileum including the proximal colon was removed for histopathological and immunohistochemical examination and biochemical analysis. Macroscopic assessment and intestinal injury scores were lower in NEC + ABS group compared to the NEC + saline group (p < 0.05). Immunohistochemical evaluations of caspase-3, -8, and -9 revealed significantly reduced apoptosis in the NEC + ABS group compared to the NEC + saline group (p = 0.001). Total oxidant status, oxidative stress index, tumor necrosis factor α and interleukin-1β levels, and lipid, protein, and deoxyribonucleic acid oxidation products were significantly lower in the NEC + ABS group compared to NEC + saline group (p < 0.001 for all), while total antioxidant status, glutathione, and superoxide dismutase levels were higher in the NEC + ABS group (p < 0.001, p < 0.001, p = 0.01, respectively). ABS treatment has the potential to effectively reduce the severity of intestinal damage in NEC due to its antioxidant, antiinflammatory, and antiapoptotic properties. Therefore, NEC may be an alternative option for treatment.
Protective effects of colchicine in an experimental model of necrotizing enterocolitis in neonatal rats
2013, Journal of Surgical Research
Citation Excerpt :
In this regard, our findings are consistent with previous reports because we also demonstrated significantly higher levels of intestinal IL-1β in the saline-treated NEC group compared with rats in the control and colchicine-treated NEC groups, which suggests that colchicine may be responsible for suppressing expression of this cytokine in tissues. Oxygen-derived free radicals may be involved in the pathogenesis of hypoxia/reoxygenation injury in the intestine [33,34]. After antigenic stimulation or triggering of the immune pathway, migration of leukocytes commences with the subsequent release of cytokines, leading to excess generation of reactive nitrogen and oxygen species [35].
The pathophysiology of necrotizing enterocolitis (NEC) includes the massive production of endogenous cytokines with exaggerated activation of inflammatory pathways. Colchicine has been used as an anti-inflammatory agent. The aim of this study was to investigate the possible beneficial effects of colchicine in a neonatal rat model of NEC.
We randomly divided rat pups into three groups: a control group, a saline-treated NEC group, and a colchicine-treated NEC group. We induced NEC by hyperosmolar enteral formula feeding and exposure to hypoxia/reoxygenation after cold stress. Intestinal samples were harvested for biochemical and histopathologic analyses.
The grade of intestinal injury of pups in the saline-treated NEC group was significantly higher than in the control and colchicine-treated groups (P < 0.001 and 0.003, respectively). The median level of intestinal malondialdehyde was significantly higher in the saline-treated NEC group compared with the control group (P = 0.006) or the colchicine-treated NEC group (P = 0.015). We observed significantly higher activity levels of intestinal superoxide dismutase and glutathione peroxidase in the colchicine-treated NEC group compared with the saline-treated NEC group (P = 0.033 and 0.030, respectively). The tissue levels of tumor necrosis factor-α and interleukin-1β were significantly higher in the saline-treated NEC group compared with the colchicine-treated NEC group (P < 0.001 and 0.003, respectively).
We observed that in this model of NEC, colchicine had favorable effects on intestinal histologic and biochemical changes.
Antioxidant effects of N-acetylcysteine in a neonatal rat model of necrotizing enterocolitis
2012, Journal of Pediatric Surgery
Hypoxia and ischemia appear to play an important role in the pathogenesis of necrotizing enterocolitis (NEC), which may be related to oxygen-derived free radical formation. This study was designed to evaluate the role of oxidative stress and potentially beneficial effects of N-acetylcysteine (NAC) in a neonatal rat model of NEC.
Thirty Wistar albino rat pups were randomly divided into 3 groups: group 1, control; group 2, NEC and saline; and group 3, NEC and NAC treatment. Necrotizing enterocolitis was induced by hyperosmolar enteral formula feeding and exposure to hypoxia after cold stress at 4°C and oxygen. The pups were killed on the fourth day, and their intestinal tissues were harvested for biochemical and histopathologic analysis.
Mucosal injury scores and intestinal malondialdehyde levels in group 2 were found to be significantly higher than that in other groups (P ≤ .05). Intestinal superoxide dismutase activities in group 3 were significantly higher than that in group 2 (P = .018). Intestinal tissue tumor necrosis factor α levels were significantly reduced with NAC treatment in group 3 compared with group 2 (P < .003).
It is likely that oxidative stress and inflammatory mediators contribute to the pathogenesis of NEC and that NAC has a protective effect on intestinal injury through its antiinflammatory and antioxidant properties.
Comparison of the Effects of Sodium Nitroprusside and L-Carnitine in Experimental Ischemia-Reperfusion Injury in Rats
2007, Transplantation Proceedings
Citation Excerpt :
The spectrophotometric analysis of MDA is an important marker of in vivo lipid peroxidation. Estimation of MDA reacting with amine groups on phospholipids by photometric analysis of the fluorescence effect is frequently used as we did in our study.14,15 The MDA level is considered to be a reliable indicator of an I/R event, increasing with the severity of the injury.16
The aim of this study was to evaluate the protective effect of sodium nitroprusside (SNP) as a nitric oxide (NO) donor and L-carnitine intraperitoneal administration to treat experimental ischemia-reperfusion (I/R) in rats.
Rats were divided into four groups, each one consisting of 10 animals. Group 1 was subjected to a sham operation. In group 2, an I/R process was applied to the rats. In group 3, SNP (5 mg/kg) and in group 4, L-carnitine (500 mg/kg) was administered in addition to the I/R process. Ileal tissue samples were obtained for analysis of tissue malonyl dialdehyde (MDA) and for histopathologic examination.
By histopathologic examination, the I/R group showed a significant difference from the SNP and L-carnitine groups (P < .05). There was no difference between the sham, the SNP, and the L-carnitine groups (P > .05). SNP used as an NO donor produced a significant decrease in MDA levels. There was a significant difference between the MDA levels of the SNP and the I/R groups (P < .05). Also, the difference between this group and the I/R group was significant (P < .05).
SNP helped to both prevent and reduce mucosal damage in terms of histological and tissue MDA levels. Since the results of the L-carnitine group and the SNP group were similar, L-carnitine was as effective as exogenous NO.
Protective effects of recombinant human interleukin-10 on intestines of hypoxia-induced necrotizing enterocolitis in immature rats
2002, Journal of Pediatric Surgery
Citation Excerpt :
Clinically, the association of perinatal or neonatal hypoxia with NEC has been well established.15,26 Several studies define a role for oxygen-derived free radicals in the pathogenesis of ischemia-reperfusion or hypoxia-reoxygenation injury in the bowel.27,28 These oxygen-derived free radicals damage tissue by peroxidation of unsaturated lipids within the cellular and mitochondrial membranes.
Background/Purpose: The role of cytokines in the pathogenesis of hemodynamic instability or tissue destruction in patients with necrotizing enterocolitis (NEC) remains undefined. The aim of this study was to determine the effects of recombinant human interleukin-102 (rhIL-10) on intestines of hypoxia-induced necrotizing enterocolitis in immature rats. Methods: The study was performed on 1-day-old Sprague Dawley rat pups. Group 1 (n = 8) served as nonhypoxic controls. Group 2 (untreated, n = 11) rats were subjected to hypoxia-reoxygenation (H/O) and then were returned to their mothers. Group 3 (rhIL-10 treated, n = 10) rats were subjected to H/O, were returned to their mothers, and were treated with rhIL-10 (75 μg/kg subcutaneously) for the next 3 days. All animals were killed on day 4, and intestine specimens were obtained to determine the tissue level of malondialdehyde (MDA) and histologic changes. Results: The microscopic lesions in the untreated rats were virtually the same as those seen in neonatal NEC, with destruction of villi and crypts, and in some cases extension to the muscularis. In contrast, in the rats treated with rhIL-10, lesions were limited essentially to the very tips of the villi. Intestinal injury score was significantly less in the rhIL-10–treated rats than in the untreated rats (P <.05). In the rhIL-10–treated group, Malondialdehyde (MDA) levels were not significant in comparison to the control group. In the untreated group, MDA levels were significantly increased when compared with the control and the rhIL-10–treated groups (P <.001 and P <.05, respectively). Conclusion: RhIL-10 has a protective effect on intestinal injury in NEC in an experimental model. J Pediatr Surg 37:1330-1333. Copyright 2002, Elsevier Science (USA). All rights reserved.
Effect of bilirubin in ischemia/reperfusion injury on rat small intestine
2001, Journal of Pediatric Surgery
Citation Excerpt :
Lipid peroxidation, in turn, results in production of conjugated dienes and MDA.1-3 Thus, MDA level is considered to be a reliable indicator of an ischemia-reperfusion event, increasing with the severity of the injury.14 In the current study, the significant low tissue MDA levels in groups B1-IR and B2-IR compared with S-IR (U = 36, P < .05), show that bilirubin prevents lipid peroxidation in intestinal I/R.
Purpose: The aim of this study was to determine the effects of bilirubin in experimental small intestinal ischemia/reperfusion (I/R) injury in rats. Methods: Thirty rats were divided into 5 groups (n = 6). In group S, saline and in group B, bilirubin, 20 mg/kg were infused via the jugular vein without an additional procedure. In groups S-IR, saline, B₁-IR and B₂-IR, 10 and 20 mg/kg/h of bilirubin were infused for 2 hours, respectively. In these groups, an I/R procedure was done after infusions by occluding the superior mesenteric artery for 45 minutes followed by 1 hour of reperfusion. After reperfusion, the small intestines were resected for histopathologic and malondialdehyde (MDA) assessments. Mucosal lesions were scored between 0 and 5. Malondialdehyde levels and histopathologic grades were analyzed statistically. Results: Mucosal injury was severe in S-IR (grade 4 to 5), mild in B₁-IR (grade 0 to 3) and none in B₂-IR group (grade 0). Grades of group S-IR were higher than those of B₁-IR and B₂-IR statistically (P <.05). Tissue MDA levels of the S-IR group were significantly higher than those of B₁-IR and B₂-IR groups (U = 36, P < .05). Bilirubin levels correlated inversely with MDA levels (r = −0.94). Conclusions: Bilirubin effectively prevents intestinal I/R injury in rat. This observation is consistent with the hypotheses regarding bilirubin as an antioxidant, having a role in the body defense. J Pediatr Surg 36:1764-1767. Copyright © 2001 by W.B. Saunders Company.

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^☆: Presented at the Annual Meeting of the Association for Academic Surgery, Louisville, KY, November 15–18, 1989.

²: Surgeon-in-Chief, Children's Hospital Medical Center, University of Cincinnati College of Medicine, Cincinnati, OH 45221.

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Journal of Surgical Research

The role of reperfusion injury in occlusive intestinal ischemia of the neonate: Malonaldehyde-derived fluorescent products and correlation of histology☆

Abstract

Role of oxygen in the cellular damage induced by reoxygenation of hypoxic heart

J. Mol. Cell. Cardiol.

Oxygen free radical-mediated myocardial and vascular dysfunction

Am. J. Physiol.

Contributions of ischemia and reperfusion to mucosal lesion formation

Am. J. Physiol.

On the pathophysiology of intestinal ischemic injury

Acta Chir. Scand.

Xanthine oxidase inhibitors attenuate ischemia-induced vascular permeability changes in the cat intestine

Gastroenterology

Xanthine oxidase Type D in the intestine and other organs of the rat

Biochem. J.

Conversion of xanthine dehydrogenase to oxidase in ischemic rat intestine: A reevaluation

Am. J. Physiol.

Therapeutic and preventative effects of ankaferd blood stopper in an experimental necrotizing enterocolitis model

Protective effects of colchicine in an experimental model of necrotizing enterocolitis in neonatal rats

Antioxidant effects of N-acetylcysteine in a neonatal rat model of necrotizing enterocolitis

Comparison of the Effects of Sodium Nitroprusside and L-Carnitine in Experimental Ischemia-Reperfusion Injury in Rats

Protective effects of recombinant human interleukin-10 on intestines of hypoxia-induced necrotizing enterocolitis in immature rats

Effect of bilirubin in ischemia/reperfusion injury on rat small intestine