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Eur J Pediatr Surg 2019; 29(02): 188-195
DOI: 10.1055/s-0037-1618593
Original Article
Georg Thieme Verlag KG Stuttgart · New York

Effect of N-Acetylserotonin on TLR-4 and MyD88 Expression during Intestinal Ischemia-Reperfusion in a Rat Model

Igor Sukhotnik
1   Department of Pediatric Surgery, The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Laboratory of Intestinal Adaptation and Recovery, Bnai Zion Medical Center, Haifa, Israel
,
Yoav Ben Shahar
2   Department of Surgery, The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Carmel Medical Center, Haifa, Israel
,
Salim Halabi
3   Department of Emergency Medicine, The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Carmel Medical Center, Haifa, Israel
,
Nir Bitterman
4   The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Laboratory of Intestinal Adaptation and Recovery, Haifa, Israel
,
Tatiana Dorfman
1   Department of Pediatric Surgery, The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Laboratory of Intestinal Adaptation and Recovery, Bnai Zion Medical Center, Haifa, Israel
,
Yulia Pollak
4   The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Laboratory of Intestinal Adaptation and Recovery, Haifa, Israel
,
Arnold Coran
5   Section of Pediatric Surgery, C.S. Mott Children's Hospital, Ann Arbor, Michigan, United States
,
Arie Bitterman
2   Department of Surgery, The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Carmel Medical Center, Haifa, Israel
› Author Affiliations
Further Information

Publication History

24 May 2017

06 December 2017

Publication Date:
05 January 2018 (online)

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Abstract

Background Accumulating evidence indicates that changes in intestinal toll-like receptors (TLRs) precede histological injury in a rodent model of necrotizing enterocolitis. N-acetylserotonin (NAS) is a naturally occurring chemical intermediate in the biosynthesis of melatonin. A recent study has shown that treatment with NAS prevents gut mucosal damage and inhibits programmed cell death following intestinal ischemia-reperfusion (IR). The objective of this study was to determine the effects of NAS on TLR-4, myeloid differentiation factor 88 (Myd88), and TNF-α receptor-associated factor 6 (TRAF6) expression in intestinal mucosa following intestinal IR in a rat.

Materials and Methods Male Sprague-Dawley rats were randomly assigned to one of the four experimental groups: 1) Sham rats underwent laparotomy; 2) Sham-NAS rats underwent laparotomy and were treated with intraperitoneal (IP) NAS (20 mg/kg); 3) IR rats underwent occlusion of both superior mesenteric artery and portal vein for 20 minutes followed by 48 hours of reperfusion; and 4) IR-NAS rats underwent IR and were treated with IP NAS immediately before abdominal closure. Intestinal structural changes, mucosal TLR-4, MyD88, and TRAF6 mucosal gene, and protein expression were examined using real-time PCR, Western blot, and immunohistochemistry.

Results Significant mucosal damage in IR rats was accompanied by a significant upregulation of TLR-4, MyD88, and TRAF6 gene and protein expression in intestinal mucosa compared with control animals. The administration of NAS decreased the intestinal injury score, inhibited cell apoptosis, and significantly reduced the expression of TLR-4, MyD88, and TRAF6.

Conclusion Treatment with NAS is associated with downregulation of TLR-4, MyD88, and TRAF6 expression along with a concomitant decrease in intestinal mucosal injury caused by intestinal IR in a rat.

Keywords

N-acetylserotonin - intestine - ischemia-reperfusion - toll-like receptor 4 - MyD88
 

  • References

  • 1 Carden DL, Granger DN. Pathophysiology of ischaemia-reperfusion injury. J Pathol 2000; 190 (03) 255-266
    CrossrefPubMedGoogle Scholar
  • 2 Kurose I, Argenbright LW, Wolf R, Lianxi L, Granger DN. Ischemia/reperfusion-induced microvascular dysfunction: role of oxidants and lipid mediators. Am J Physiol 1997; 272 (6 Pt 2): H2976-H2982
    PubMedGoogle Scholar
  • 3 Wolin MS. Interactions of oxidants with vascular signaling systems. Arterioscler Thromb Vasc Biol 2000; 20 (06) 1430-1442
    CrossrefPubMedGoogle Scholar
  • 4 Mangino JE, Kotadia B, Mangino MJ. Characterization of hypothermic intestinal ischemia-reperfusion injury in dogs. Effects of glycine. Transplantation 1996; 62 (02) 173-178
    CrossrefPubMedGoogle Scholar
  • 5 Schoenberg MH, Poch B, Younes M. , et al. Involvement of neutrophils in postischaemic damage to the small intestine. Gut 1991; 32 (08) 905-912
    CrossrefPubMedGoogle Scholar
  • 6 Ikeda H, Suzuki Y, Suzuki M. , et al. Apoptosis is a major mode of cell death caused by ischaemia and ischaemia/reperfusion injury to the rat intestinal epithelium. Gut 1998; 42 (04) 530-537
    CrossrefPubMedGoogle Scholar
  • 7 Noda T, Iwakiri R, Fujimoto K, Matsuo S, Aw TY. Programmed cell death induced by ischemia-reperfusion in rat intestinal mucosa. Am J Physiol 1998; 274 (2 Pt 1): G270-G276
    PubMedGoogle Scholar
  • 8 Arends MJ, Wyllie AH. Apoptosis: mechanisms and roles in pathology. Int Rev Exp Pathol 1991; 32: 223-254
    PubMedGoogle Scholar
  • 9 Hall PA, Coates PJ, Ansari B, Hopwood D. Regulation of cell number in the mammalian gastrointestinal tract: the importance of apoptosis. J Cell Sci 1994; 107 (Pt 12): 3569-3577
    PubMedGoogle Scholar
  • 10 Medzhitov R. Toll-like receptors and innate immunity. Nat Rev Immunol 2001; 1 (02) 135-145
    CrossrefPubMedGoogle Scholar
  • 11 Akira S, Takeda K. Toll-like receptor signalling. Nat Rev Immunol 2004; 4 (07) 499-511
    CrossrefPubMedGoogle Scholar
  • 12 Poltorak A, He X, Smirnova I. , et al. Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science 1998; 282 (5396): 2085-2088
    CrossrefPubMedGoogle Scholar
  • 13 Beutler B. Endotoxin, toll-like receptor 4, and the afferent limb of innate immunity. Curr Opin Microbiol 2000; 3 (01) 23-28
    CrossrefPubMedGoogle Scholar
  • 14 Medzhitov R, Preston-Hurlburt P, Janeway Jr CA. A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. Nature 1997; 388 (6640): 394-397
    CrossrefPubMedGoogle Scholar
  • 15 Oxenkrug G. Antioxidant effects of N-acetylserotonin: possible mechanisms and clinical implications. Ann N Y Acad Sci 2005; 1053: 334-347
    CrossrefPubMedGoogle Scholar
  • 16 Yu S, Zheng J, Jiang Z. , et al. Protective effect of N-acetylserotonin against acute hepatic ischemia-reperfusion injury in mice. Int J Mol Sci 2013; 14 (09) 17680-17693
    CrossrefPubMedGoogle Scholar
  • 17 Tang XL, Takano H, Xuan YT. , et al. Hypercholesterolemia abrogates late preconditioning via a tetrahydrobiopterin-dependent mechanism in conscious rabbits. Circulation 2005; 112 (14) 2149-2156
    CrossrefPubMedGoogle Scholar
  • 18 Chomczynski P. A reagent for the single-step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. Biotechniques 1993; 15 (03) 532-534
    PubMedGoogle Scholar
  • 19 Park PO, Haglund U, Bulkley GB, Fält K. The sequence of development of intestinal tissue injury after strangulation ischemia and reperfusion. Surgery 1990; 107 (05) 574-580
    PubMedGoogle Scholar
  • 20 Costa RIDD, Rasslan R, Koike MK, Utiyama EM, Montero EFS. Bacterial translocation and mortality on rat model of intestinal ischemia and obstruction. Acta Cir Bras 2017; 32 (08) 641-647
    CrossrefPubMedGoogle Scholar
  • 21 Li Y, Xu B, Xu M. , et al. 6-Gingerol protects intestinal barrier from ischemia/reperfusion-induced damage via inhibition of p38 MAPK to NF-κB signalling. Pharmacol Res 2017; 119: 137-148
    CrossrefPubMedGoogle Scholar
  • 22 Al-Sadi R, Khatib K, Guo S, Ye D, Youssef M, Ma T. Occludin regulates macromolecule flux across the intestinal epithelial tight junction barrier. Am J Physiol Gastrointest Liver Physiol 2011; 300 (06) G1054-G1064
    CrossrefPubMedGoogle Scholar
  • 23 Basset C, Holton J, O'Mahony R, Roitt I. Innate immunity and pathogen-host interaction. Vaccine 2003; 21 (Suppl. 02) S12-S23
    CrossrefPubMedGoogle Scholar
  • 24 Tabeta K, Georgel P, Janssen E. , et al. Toll-like receptors 9 and 3 as essential components of innate immune defense against mouse cytomegalovirus infection. Proc Natl Acad Sci U S A 2004; 101 (10) 3516-3521
    CrossrefPubMedGoogle Scholar
  • 25 Stuss M, Wiktorska JA, Sewerynek E. N-acetylserotonin reduces lipopolysaccharide-induced lipid peroxidation in vitro more effectively than melatonin. Neuroendocrinol Lett 2010; 31 (04) 489-496
    PubMedGoogle Scholar
  • 26 Li Z, Nickkholgh A, Yi X. , et al. Melatonin protects kidney grafts from ischemia/reperfusion injury through inhibition of NF-kB and apoptosis after experimental kidney transplantation. J Pineal Res 2009; 46 (04) 365-372
    CrossrefPubMedGoogle Scholar
  • 27 Mathes AM. Hepatoprotective actions of melatonin: possible mediation by melatonin receptors. World J Gastroenterol 2010; 16 (48) 6087-6097
    CrossrefPubMedGoogle Scholar
  • 28 Muñoz-Casares FC, Padillo FJ, Briceño J. , et al. Melatonin reduces apoptosis and necrosis induced by ischemia/reperfusion injury of the pancreas. J Pineal Res 2006; 40 (03) 195-203
    CrossrefPubMedGoogle Scholar
  • 29 Ben Shahar Y, Sukhotnik I, Bitterman N. , et al. Effect of N-Acetylserotonin on intestinal recovery following intestinal ischemia-reperfusion injury in a rat. Eur J Pediatr Surg 2016; 26 (01) 47-53
    PubMedGoogle Scholar
  • 30 Perianayagam MC, Oxenkrug GF, Jaber BL. Immune-modulating effects of melatonin, N-acetylserotonin, and N-acetyldopamine. Ann N Y Acad Sci 2005; 1053: 386-393
    CrossrefPubMedGoogle Scholar
  • 31 Ortega-Cava CF, Ishihara S, Rumi MA. , et al. Strategic compartmentalization of Toll-like receptor 4 in the mouse gut. J Immunol 2003; 170 (08) 3977-3985
    CrossrefPubMedGoogle Scholar
  • 32 Liu Y, Zhu L, Fatheree NY. , et al. Changes in intestinal Toll-like receptors and cytokines precede histological injury in a rat model of necrotizing enterocolitis. Am J Physiol Gastrointest Liver Physiol 2009; 297 (03) G442-G450
    CrossrefPubMedGoogle Scholar
  • 33 Jilling T, Simon D, Lu J. , et al. The roles of bacteria and TLR4 in rat and murine models of necrotizing enterocolitis. J Immunol 2006; 177 (05) 3273-3282
    CrossrefPubMedGoogle Scholar
  • 34 Zhou H, Wang J, Jiang J. , et al. N-acetyl-serotonin offers neuroprotection through inhibiting mitochondrial death pathways and autophagic activation in experimental models of ischemic injury. J Neurosci 2014; 34 (08) 2967-2978
    CrossrefPubMedGoogle Scholar
  • 35 da Silveira Cruz-Machado S, Carvalho-Sousa CE, Tamura EK. , et al. TLR4 and CD14 receptors expressed in rat pineal gland trigger NFKB pathway. J Pineal Res 2010; 49 (02) 183-192
    PubMedGoogle Scholar