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      Histological analysis of the intestinal wall of newborn rats submitted to hypoxia and reoxygenation to evaluate the protective effect of N-Acetylcysteine 1

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          Abstract

          Purpose

          To evaluate the effect of N-Acetylcysteine (NAC) in newborn rats submitted to hypoxia and reoxygenation (H/R) conditions in an experimental model of necrotizing enterocolitis.

          Methods

          Eight pregnant rats and their 70 cubs were used (5 groups) and exposed to H/R conditions and received NAC at different times. The animals in the H/R groups were placed in a gas chamber (100% CO 2) for 10 minutes and then reoxygenated for 10 minutes (100% O 2), twice a day for the first three days of life, with a six-hour span between events. On the third day of life, the animals were anesthetized, laparotomized and the intestines were resected.

          Results

          The H/R and NAC groups showed changes in the intestinal wall in relation to the number, height and width of the villi when compared to the control group (p<0.0001), but with better preservation of structures in the NAC group. There were no differences between groups regarding the number (%) of mitoses.

          Conclusion

          The administration of NAC decreased the lesions in the intestinal wall of rats submitted to H/R, therefore suggesting that this drug can be used to prevent the development of necrotizing enterocolitis in newborns.

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          Most cited references 38

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          Necrotising enterocolitis.

           B Stoll,  W. Lin (2006)
          Necrotising enterocolitis is one of the most common gastrointestinal emergencies in newborn infants. Here we review the epidemiology, clinical presentation, and pathophysiology of the disease, as well as strategies for diagnosis, management, and prevention. Necrotising enterocolitis is one of the most devastating and unpredictable diseases affecting premature infants. Despite decades of research, its pathogenesis remains unclear; diagnosis can be difficult; and treatment is challenging. We will need to improve our understanding of intestinal defences in premature infants, dietary and bacterial factors, and genetic effects that could predispose infants to necrotising enterocolitis before we can develop new strategies for prevention and treatment.
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            The sequence of development of intestinal tissue injury after strangulation ischemia and reperfusion.

            Tissue injury at reperfusion has been reported after partial ischemia. However, previous attempts to demonstrate a component of injury caused by reperfusion after total ischemia have failed. This study was performed to evaluate the hypothesis that in such situations the extent of the tissue injury caused by ischemia itself prevented detection of a reperfusion component. Rats were subjected to near-total intestinal ischemia by means of a hydrostatic pressure clamp that produced preferential venous occlusion (strangulation) for periods from 1 to 90 minutes. Tissue injury was evaluated microscopically by a blinded examiner. Ischemic periods of 20 minutes or less did not induce detectable tissue injury. Longer durations of ischemia caused villous injury: the longer the period of ischemia, the more extensive the tissue injury. However, there was no exacerbation of injury seen after reperfusion, regardless of the duration of ischemia. In a separate series of rats, total arterial occlusion was employed without concomitant venous congestion. Such isolation arterial occlusion of 40 to 60 minutes' duration was followed by a statistically significant exacerbation of tissue injury at reperfusion. Thus total intestinal ischemia may be followed by reperfusion injury if there is no concomitant congestion and if ischemic injury is not too extensive.
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              Epidermal growth factor reduces the development of necrotizing enterocolitis in a neonatal rat model.

              Necrotizing enterocolitis (NEC) is the most common gastrointestinal disease of prematurely born infants. Maternal milk plays an important protective role against NEC development and is the major source of epidermal growth factor (EGF) for neonates. The aim of this study was to examine the effect of orally administered EGF on the incidence of NEC in a neonatal rat model. Newborn rats were artificially fed either with growth factor-free rat milk substitute (RMS) or RMS supplemented with 500 ng/ml of EGF (RMS+EGF). Experimental NEC was induced by exposure to asphyxia and cold stress. Development of NEC was evaluated by gross and histological scoring of damage in the ileum. Ileal EGF receptor (EGF-R), EGF, and transforming growth factor-alpha mRNA expression was assessed by RT competitive-PCR, and the EGF-R was localized by immunohistochemistry. EGF supplementation of formula reduced the incidence and severity of NEC in rats (13/16 RMS vs. 4/13 RMS+EGF). Ileal EGF-R mRNA expression was markedly increased in the RMS group compared with RMS+EGF. Enhanced EGF-R expression in the RMS group was localized predominantly in the epithelial cells of injured ileum. These data suggest a new potential therapeutic approach for the prevention and treatment of NEC.
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                Author and article information

                Journal
                Acta Cir Bras
                Acta Cir Bras
                acb
                Acta Cirúrgica Brasileira
                Sociedade Brasileira para o Desenvolvimento da Pesquisa em Cirurgia
                0102-8650
                1678-2674
                12 June 2020
                2020
                : 35
                : 4
                Affiliations
                [I ]Fellow Master degree, Postgraduate Program in Obstetrics, Universidade Federal de São Paulo (UNIFESP), Brazil. Acquisition of data, manuscript writing, final approval.
                [II ]PhD, Department of Obstetrics, UNIFESP, Sao Paulo-SP, Brazil. Supervision, final approval.
                [III ]PhD, Department of Morphology and Genetics, UNIFESP, Sao Paulo-SP, Brazil. Analysis and interpretation of data, final approval.
                [IV ]PhD, Division of Pediatric Surgery, UNIFESP, Sao Paulo-SP, Brazil. Analysis and interpretation of data, final approval.
                [V ]PhD, Division of General and Trauma Surgery, Universidade de São Paulo (USP), Brazil. Critical revision final approval.
                [VI ]PhD, Department of Obstetrics, UNIFESP, Sao Paulo-SP, Brazil. Manuscript writing, final approval.
                [VII ]PhD, Division of Pediatric Surgery, UNIFESP, Sao Paulo-SP, Brazil. Conception and design of the study, final approval.
                Author notes
                Correspondence: Prof. Edward Araujo Júnior Rua Belchior de Azevedo, 156/111 Torre Vitoria 05089-030 São Paulo – SP Brasil Tel.: (55 11)3796-5944 araujojred@ 123456terra.com.br

                Conflict of interest: none

                Article
                00201
                10.1590/s0102-865020200040000001
                7292620
                32555935

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                Page count
                Figures: 2, Tables: 5, Equations: 0, References: 30
                Categories
                Original Article
                Experimental Surgery

                enterocolitis, necrotizing, hypoxia, acetylcysteine, rats

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