Intestine and Brain TLR-4 Modulation Following N-acetyl-cysteine Treatment in NEC Rodent Model

Overview

Journal Sci Rep

Specialty Science

Date 2023 May 22

PMID 37217588

Authors

Ron Beloosesky

Ola Gutzeit

Yuval Ginsberg

Nizar Khatib

Michael G Ross

Zeev Weiner

Osnat Zmora

Affiliations

Soon will be listed here.

Abstract

Necrotizing enterocolitis (NEC) brain injury is mediated through Toll-like receptor 4 (TLR4) on the intestinal epithelium and brain microglia. Our aim was to determine whether postnatal and/or prenatal NAC can modify NEC associated intestinal and brain TLR4 expression and brain glutathione levels in a rat model of NEC. Newborn Sprague-Dawley rats were randomized into three groups: Control (n = 33); NEC (n = 32)-hypoxia and formula feeding; and NEC-NAC (n = 34)-received NAC (300 mg/kg IP) in addition to NEC conditions. Two additional groups included pups of dams treated once daily with NAC (300 mg/kg IV) for the last 3 days of pregnancy: NAC-NEC (n = 33) or NAC-NEC-NAC (n = 36) with additional postnatal NAC. Pups were sacrificed on the fifth day, and ileum and brains harvested for TLR-4 and glutathione protein levels. Brain and ileum TLR-4 protein levels were significantly increased in NEC offspring as compared to control (brain 2.5 ± 0.6 vs. 0.88 ± 0.12 U and ileum 0.24 ± 0.04 vs. 0.09 ± 0.01, p < 0.05). When NAC was administered only to dams (NAC-NEC) a significant decrease in TLR-4 levels was demonstrated in both offspring brain (1.53 ± 0.41 vs. 2.5 ± 0.6 U, p < 0.05) and ileum (0.12 ± 0.03 vs. 0.24 ± 0.04 U, p < 0.05) as compared to NEC. The same pattern was demonstrated when NAC was administered only or postnatally. The decrease in brain and ileum glutathione levels observed in NEC offspring was reversed with all NAC treatment groups. NAC reverses the increase in ileum and brain TLR-4 levels and the decrease in brain and ileum glutathione levels associated with NEC in a rat model, and thus may protect from NEC associated brain injury.

Citing Articles

Use of N-Acetylcysteine in Preterm Neonates with Enteral Feeding Intolerance and Intestinal Obstruction: A Case Series and Review of the Literature.

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PMID: 39062322 PMC: 11276329. DOI: 10.3390/children11070873.

Modification of Experimental Model of Necrotizing Enterocolitis (NEC) in Rat Pups by Single Exposure to Hypothermia and Hypoxia and Impact of Mother's Milk on Incidence of Disease.

Wolski M Med Sci Monit. 2024; 30:e943443.

PMID: 38678319 PMC: 11064733. DOI: 10.12659/MSM.943443.

References

Gutziet O, Iluz R, Ben Asher H, Segal L, Ben Zvi D, Ginsberg Y . Maternal N-Acetyl-Cysteine Prevents Neonatal Hypoxia-Induced Brain Injury in a Rat Model. Int J Mol Sci. 2021; 22(24). PMC: 8709193. DOI: 10.3390/ijms222413629. View

Wiest D, Chang E, Fanning D, Garner S, Cox T, Jenkins D . Antenatal pharmacokinetics and placental transfer of N-acetylcysteine in chorioamnionitis for fetal neuroprotection. J Pediatr. 2014; 165(4):672-7.e2. PMC: 4177316. DOI: 10.1016/j.jpeds.2014.06.044. View

Mihi B, Good M . Impact of Toll-Like Receptor 4 Signaling in Necrotizing Enterocolitis: The State of the Science. Clin Perinatol. 2019; 46(1):145-157. PMC: 6383801. DOI: 10.1016/j.clp.2018.09.007. View

Anderson K . Toll signaling pathways in the innate immune response. Curr Opin Immunol. 2000; 12(1):13-9. DOI: 10.1016/s0952-7915(99)00045-x. View

Riggs B, Bronstein A, Kulig K, Archer P, Rumack B . Acute acetaminophen overdose during pregnancy. Obstet Gynecol. 1989; 74(2):247-53. View

Beloosesky R, Weiner Z, Ginsberg Y, Ross M . Maternal N-acetyl-cysteine (NAC) protects the rat fetal brain from inflammatory cytokine responses to lipopolysaccharide (LPS). J Matern Fetal Neonatal Med. 2011; 25(8):1324-8. DOI: 10.3109/14767058.2011.632793. View

Beloosesky R, Weiner Z, Khativ N, Maravi N, Mandel R, Boles J . Prophylactic maternal n-acetylcysteine before lipopolysaccharide suppresses fetal inflammatory cytokine responses. Am J Obstet Gynecol. 2009; 200(6):665.e1-5. DOI: 10.1016/j.ajog.2009.01.032. View

Beloosesky R, Gayle D, Amidi F, Nunez S, Babu J, Desai M . N-acetyl-cysteine suppresses amniotic fluid and placenta inflammatory cytokine responses to lipopolysaccharide in rats. Am J Obstet Gynecol. 2006; 194(1):268-73. DOI: 10.1016/j.ajog.2005.06.082. View

M S Durieux A, Fernandes C, Murphy D, Labouesse M, Giovanoli S, Meyer U . Targeting Glia with N-Acetylcysteine Modulates Brain Glutamate and Behaviors Relevant to Neurodevelopmental Disorders in C57BL/6J Mice. Front Behav Neurosci. 2015; 9:343. PMC: 4677305. DOI: 10.3389/fnbeh.2015.00343. View

10.

Neu J . Necrotizing enterocolitis: the search for a unifying pathogenic theory leading to prevention. Pediatr Clin North Am. 1996; 43(2):409-32. PMC: 7127724. DOI: 10.1016/s0031-3955(05)70413-2. View

11.

Blakely M, Lally K, McDonald S, Brown R, Barnhart D, Ricketts R . Postoperative outcomes of extremely low birth-weight infants with necrotizing enterocolitis or isolated intestinal perforation: a prospective cohort study by the NICHD Neonatal Research Network. Ann Surg. 2005; 241(6):984-9. PMC: 1359076. DOI: 10.1097/01.sla.0000164181.67862.7f. View

12.

Kelly N, Friend K, Boyle P, Zhang X, Wong C, Hackam D . The role of the glutathione antioxidant system in gut barrier failure in a rodent model of experimental necrotizing enterocolitis. Surgery. 2004; 136(3):557-66. DOI: 10.1016/j.surg.2004.05.034. View

13.

Yazji I, Sodhi C, Lee E, Good M, Egan C, Afrazi A . Endothelial TLR4 activation impairs intestinal microcirculatory perfusion in necrotizing enterocolitis via eNOS-NO-nitrite signaling. Proc Natl Acad Sci U S A. 2013; 110(23):9451-6. PMC: 3677476. DOI: 10.1073/pnas.1219997110. View

14.

Liu T, Zhang L, Joo D, Sun S . NF-κB signaling in inflammation. Signal Transduct Target Ther. 2017; 2. PMC: 5661633. DOI: 10.1038/sigtrans.2017.23. View

15.

Hendrickson R . What is the most appropriate dose of -acetylcysteine after massive acetaminophen overdose?. Clin Toxicol (Phila). 2019; 57(8):686-691. DOI: 10.1080/15563650.2019.1579914. View

16.

Soliman A, Michelsen K, Karahashi H, Lu J, Meng F, Qu X . Platelet-activating factor induces TLR4 expression in intestinal epithelial cells: implication for the pathogenesis of necrotizing enterocolitis. PLoS One. 2010; 5(10):e15044. PMC: 2955554. DOI: 10.1371/journal.pone.0015044. View

17.

Baek O, Cilieborg M, Nguyen D, Bering S, Thymann T, Sangild P . Sex-Specific Survival, Growth, Immunity and Organ Development in Preterm Pigs as Models for Immature Newborns. Front Pediatr. 2021; 9:626101. PMC: 7905020. DOI: 10.3389/fped.2021.626101. View

18.

Park E, Lee G, Choi S, Choi S, Chae S, Kang S . Correlation between extracellular glutamate release and neuronal cell death in an eleven vessel occlusion model in rat. Brain Res. 2010; 1342:160-6. DOI: 10.1016/j.brainres.2010.04.054. View

19.

Hunter C, Upperman J, Ford H, Camerini V . Understanding the susceptibility of the premature infant to necrotizing enterocolitis (NEC). Pediatr Res. 2007; 63(2):117-23. DOI: 10.1203/PDR.0b013e31815ed64c. View

20.

Zmora O, Gutzeit O, Segal L, Boulos S, Millo Z, Ginsberg Y . Maternal N-acetyl-cysteine prevents neonatal brain injury associated with necrotizing enterocolitis in a rat model. Acta Obstet Gynecol Scand. 2020; 100(5):979-987. DOI: 10.1111/aogs.14054. View