Protective Effect of the SIRT1-Mediated NF-κB Signaling Pathway against Necrotizing Enterocolitis in Neonatal Mice

Abstract

Objective To discover the mechanism of the sirtuin 1 (SIRT1)-mediated nuclear factor-κB (NF-κB) pathway in the protection against necrotizing enterocolitis (NEC) in neonatal mice.

Materials and Methods Neonatal mice were treated with EX527 (an inhibitor of SIRT1) and/or pyrrolidine dithiocarbamate (PDTC, an inhibitor of NF-κB). The survival rate of the mice was recorded. Hematoxylin and eosin (HE) staining was performed to observe the pathological changes in the intestines. Furthermore, western blotting, enzyme-linked immunosorbent assay, and real-time quantitative polymerase chain reaction were conducted to measure the protein and gene expression, while corresponding kits were used to detect the levels of oxidative stress indicators.

Results PDTC increased the survival rate of NEC mice. When compared with the NEC+ EX527 + PDTC group, the histological NEC score was higher in the NEC + EX527 group but lower in the NEC + PDTC group. SIRT1 expression in the intestines of NEC mice was downregulated, with an increase in p65 nuclear translocation. Additionally, malondialdehyde increased and glutathione peroxidase decreased in the intestines of NEC mice, with the upregulation of interleukin (IL)-6, IL-1β, and tumor necrosis factor-α, as well as the downregulation of ZO-1, occludin, and claudin-4 in the intestines. However, the above changes could be improved by PDTC, which could be further reversed by EX527.

Conclusion SIRT1 can mitigate inflammation and the oxidative stress response and improve intestinal permeability by mediating the NF-κB pathway, playing an important role in the alleviation of NEC.

Keywords

neonatal NEC - SIRT1 - NF-κB - inflammation - oxidative stress

Availability of Data and Materials

The data that support the findings of this study are available on request form the corresponding author. The data are not publicly available due to privacy.

Publikationsverlauf

Eingereicht: 20. Juni 2022

Angenommen: 06. September 2022

Artikel online veröffentlicht:
15. November 2022

• References

• 1 Meister AL, Doheny KK, Travagli RA. Necrotizing enterocolitis: it's not all in the gut. Exp Biol Med (Maywood) 2020; 245 (02) 85-95
• 2 Alganabi M, Lee C, Bindi E, Li B, Pierro A. Recent advances in understanding necrotizing enterocolitis. F1000Res 2019; 8: F1000 Faculty Rev-107
• 3 Bellodas Sanchez J, Kadrofske M. Necrotizing enterocolitis. Neurogastroenterol Motil 2019; 31 (03) e13569
• 4 Yan X, Managlia E, Tan XD, De Plaen IG. Prenatal inflammation impairs intestinal microvascular development through a TNF-dependent mechanism and predisposes newborn mice to necrotizing enterocolitis. Am J Physiol Gastrointest Liver Physiol 2019; 317 (01) G57-G66
• 5 Rich BS, Dolgin SE. Necrotizing enterocolitis. Pediatr Rev 2017; 38 (12) 552-559
• 6 Riggle KM, Rentea RM, Welak SR, Pritchard Jr KA, Oldham KT, Gourlay DM. Intestinal alkaline phosphatase prevents the systemic inflammatory response associated with necrotizing enterocolitis. J Surg Res 2013; 180 (01) 21-26
• 7 Harris MC, D'Angio CT, Gallagher PR, Kaufman D, Evans J, Kilpatrick L. Cytokine elaboration in critically ill infants with bacterial sepsis, necrotizing enterocolitis, or sepsis syndrome: correlation with clinical parameters of inflammation and mortality. J Pediatr 2005; 147 (04) 462-468
• 8 Tang BL. Sirt1 and the mitochondria. Mol Cells 2016; 39 (02) 87-95
• 9 Chen C, Zhou M, Ge Y, Wang X. SIRT1 and aging related signaling pathways. Mech Ageing Dev 2020; 187: 111215
• 10 Alves-Fernandes DK, Jasiulionis MG. The role of SIRT1 on DNA damage response and epigenetic alterations in cancer. Int J Mol Sci 2019; 20 (13) E3153
• 11 Farghali H, Kemelo MK, Canová NK. SIRT1 modulators in experimentally induced liver injury. Oxid Med Cell Longev 2019; 2019: 8765954
• 12 Shi L, Zhang J, Wang Y, Hao Q, Chen H, Cheng X. Sirt1 regulates oxidative stress in oxygen-glucose deprived hippocampal neurons. Front Pediatr 2020; 8: 455
• 13 Tian L, Cao W, Yue R. et al. Pretreatment with Tilianin improves mitochondrial energy metabolism and oxidative stress in rats with myocardial ischemia/reperfusion injury via AMPK/SIRT1/PGC-1 alpha signaling pathway. J Pharmacol Sci 2019; 139 (04) 352-360
• 14 Kundu A, Dey P, Park JH, Kim IS, Kwack SJ, Kim HS. EX-527 prevents the progression of high-fat diet-induced hepatic steatosis and fibrosis by upregulating SIRT4 in Zucker rats. Cells 2020; 9 (05) E1101
• 15 Thirupathi A, de Souza CT. Multi-regulatory network of ROS: the interconnection of ROS, PGC-1 alpha, and AMPK-SIRT1 during exercise. J Physiol Biochem 2017; 73 (04) 487-494
• 16 Maiese K. Targeting the core of neurodegeneration: FoxO, mTOR, and SIRT1. Neural Regen Res 2021; 16 (03) 448-455
• 17 Kauppinen A, Suuronen T, Ojala J, Kaarniranta K, Salminen A. Antagonistic crosstalk between NF-κB and SIRT1 in the regulation of inflammation and metabolic disorders. Cell Signal 2013; 25 (10) 1939-1948
• 18 Zhang K, Zhang X, Lv A, Fan S, Zhang J. Saccharomyces boulardii modulates necrotizing enterocolitis in neonatal mice by regulating the sirtuin 1/NF–κB pathway and the intestinal microbiota. Mol Med Rep 2020; 22 (02) 671-680
• 19 Lawrence T. The nuclear factor NF-kappaB pathway in inflammation. Cold Spring Harb Perspect Biol 2009; 1 (06) a001651
• 20 de Gregorio E, Colell A, Morales A, Marí M. Relevance of SIRT1-NF-κB axis as therapeutic target to ameliorate inflammation in liver disease. Int J Mol Sci 2020; 21 (11) E3858
• 21 Yu Q, Dong L, Li Y, Liu G. SIRT1 and HIF1α signaling in metabolism and immune responses. Cancer Lett 2018; 418: 20-26
• 22 Gui L, Lei X, Zuo Z. Decrease of glial cell-derived neurotrophic factor contributes to anesthesia- and surgery-induced learning and memory dysfunction in neonatal rats. J Mol Med (Berl) 2017; 95 (04) 369-379
• 23 Ma Y, Kong L, Nan K. et al. Apolipoprotein-J prevents angiotensin II-induced apoptosis in neonatal rat ventricular cells. Lipids Health Dis 2015; 14: 114
• 24 Klinke M, Vincent D, Trochimiuk M. et al. Degradation of extracellular DNA significantly ameliorates necrotizing enterocolitis severity in mice. J Surg Res 2019; 235: 513-520
• 25 Council NR. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press; 2011
• 26 Le K, Chibaatar Daliv E, Wu S. et al. SIRT1-regulated HMGB1 release is partially involved in TLR4 signal transduction: a possible anti-neuroinflammatory mechanism of resveratrol in neonatal hypoxic-ischemic brain injury. Int Immunopharmacol 2019; 75: 105779
• 27 Nurmi A, Goldsteins G, Närväinen J. et al. Antioxidant pyrrolidine dithiocarbamate activates Akt-GSK signaling and is neuroprotective in neonatal hypoxia-ischemia. Free Radic Biol Med 2006; 40 (10) 1776-1784
• 28 Bein A, Eventov-Friedman S, Arbell D, Schwartz B. Intestinal tight junctions are severely altered in NEC preterm neonates. Pediatr Neonatol 2018; 59 (05) 464-473
• 29 Bai M, Lu C, An L. et al. SIRT1 relieves necrotizing enterocolitis through inactivation of hypoxia-inducible factor (HIF)-1a. Cell Cycle 2020; 19 (16) 2018-2027
• 30 Hwang JW, Yao H, Caito S, Sundar IK, Rahman I. Redox regulation of SIRT1 in inflammation and cellular senescence. Free Radic Biol Med 2013; 61: 95-110
• 31 D'Angelo G, Chimenz R, Reiter RJ, Gitto E. Use of melatonin in oxidative stress related neonatal diseases. Antioxidants 2020; 9 (06) E477
• 32 Wu NC, Wang JJ. Niacin pretreatment attenuates lung ischemia and reperfusion-induced pulmonary barrier function impairment by reducing oxidative stress and activating SIRT1 in an isolated-perfused rat lung model. Transplant Proc 2018; 50 (09) 2834-2838
• 33 Yao H, Sundar IK, Ahmad T. et al. SIRT1 protects against cigarette smoke-induced lung oxidative stress via a FOXO3-dependent mechanism. Am J Physiol Lung Cell Mol Physiol 2014; 306 (09) L816-L828
• 34 Wang XL, Li T, Li JH, Miao SY, Xiao XZ. The effects of resveratrol on inflammation and oxidative stress in a rat model of chronic obstructive pulmonary disease. Molecules 2017; 22 (09) E1529
• 35 Yang F, Yan J, Lu Y, Wang D, Liu L, Wang Z. MicroRNA-499-5p targets SIRT1 to aggravate lipopolysaccharide-induced acute lung injury. Free Radic Res 2021; 55 (01) 71-82
• 36 Buckley A, Turner JR. Cell biology of tight junction barrier regulation and mucosal disease. Cold Spring Harb Perspect Biol 2018; 10 (01) a029314
• 37 Sawada N. Tight junction-related human diseases. Pathol Int 2013; 63 (01) 1-12
• 38 Kim S, Kim GH. Roles of claudin-2, ZO-1 and occludin in leaky HK-2 cells. PLoS One 2017; 12 (12) e0189221
• 39 Reinhold AK, Rittner HL. Barrier function in the peripheral and central nervous system-a review. Pflugers Arch 2017; 469 (01) 123-134
• 40 Seo S, Miyake H, Alganabi M. et al. Vasoactive intestinal peptide decreases inflammation and tight junction disruption in experimental necrotizing enterocolitis. J Pediatr Surg 2019; 54 (12) 2520-2523
• 41 Fu C, Hao S, Xu X. et al. Activation of SIRT1 ameliorates LPS-induced lung injury in mice via decreasing endothelial tight junction permeability. Acta Pharmacol Sin 2019; 40 (05) 630-641
• 42 Qian C, Jin J, Chen J. et al. SIRT1 activation by resveratrol reduces brain edema and neuronal apoptosis in an experimental rat subarachnoid hemorrhage model. Mol Med Rep 2017; 16 (06) 9627-9635
• 43 Huang J, Liu W, Doycheva DM. et al. Ghrelin attenuates oxidative stress and neuronal apoptosis via GHSR-1α/AMPK/Sirt1/PGC-1α/UCP2 pathway in a rat model of neonatal HIE. Free Radic Biol Med 2019; 141: 322-337
• 44 Wang Z, Zhao H, Peng S, Zuo Z. Intranasal pyrrolidine dithiocarbamate decreases brain inflammatory mediators and provides neuroprotection after brain hypoxia-ischemia in neonatal rats. Exp Neurol 2013; 249: 74-82
• 45 Managlia E, Liu SXL, Yan X. et al. Blocking NF-κB activation in Ly6c⁺ monocytes attenuates necrotizing enterocolitis. Am J Pathol 2019; 189 (03) 604-618
• 46 Jing Y, Peng F, Shan Y, Jiang J. Berberine reduces the occurrence of neonatal necrotizing enterocolitis by reducing the inflammatory response. Exp Ther Med 2018; 16 (06) 5280-5285
• 47 Sun L, Sun M, Ma K, Liu J. Let-7d-5p suppresses inflammatory response in neonatal rats with necrotizing enterocolitis via LGALS3-mediated TLR4/NF-κB signaling pathway. Am J Physiol Cell Physiol 2020; 319 (06) C967-C979
• 48 Chen H, Zhang H, Zheng Y. et al. Prolyl hydroxylase 2 silencing enhances the paracrine effects of mesenchymal stem cells on necrotizing enterocolitis in an NF-κB-dependent mechanism. Cell Death Dis 2020; 11 (03) 188
• 49 Zhang L, Fan J, He J. et al. Regulation of ROS-NF-κB axis by tuna backbone derived peptide ameliorates inflammation in necrotizing enterocolitis. J Cell Physiol 2019; 234 (08) 14330-14338
• 50 Hong G, Zheng D, Zhang L. et al. Administration of nicotinamide riboside prevents oxidative stress and organ injury in sepsis. Free Radic Biol Med 2018; 123: 125-137