Accelerated Intestinal Epithelial Cell Turnover Correlates with Stimulated BMP Signaling Cascade following Intestinal Ischemia–Reperfusion in a Rat

Abstract

Introduction Bone morphogenetic proteins (BMPs) are a family of proteins that regulate proliferation and differentiation of intestinal epithelial cells. The purpose of this study was to evaluate the role of BMP signaling following intestinal ischemia–reperfusion (IR) in a rat model.

Materials and Methods Male Sprague-Dawley rats were divided into four experimental groups: Sham-24 and Sham-48 rats underwent laparotomy and were sacrificed 24 or 48 hours later, respectively; IR-24 and IR-48 rats underwent occlusion of superior mesenteric artery and portal vein for 30 minutes followed by 24 or 48 hours of reperfusion, respectively. Enterocyte proliferation and apoptosis were determined at sacrifice. BMP-related genes and protein expression were determined using real-time polymerase chain reaction, Western blot, and immunohistochemistry for 48 hours followed by IR.

Results IR rats demonstrated a significant increase in BMP2 (twofold increase, p < 0.05), BMP4 (sevenfold increase), STAT3 (70% increase), BMPR1 (70% increase) messenger ribonucleic acid levels in jejunum and was accompanied by a significant increase in BMP2 and BMP4 protein levels in jejunum (sixfold increase) (Western blot) and upward increase in the number of BMP-positive cells (by immunohistochemistry) in jejunal (48% increase) and ileal (56% increase) villi compared with Sham-48 animals. Elevation in BMP2 and BMP4 levels was associated with increased rates of cell proliferation and increased cell apoptosis.

Conclusion Forty-eight hours following intestinal IR in rats, BMP signaling pathway was stimulated. The increase in BMP signaling pathway activity correlates with accelerated cell turnover.

Keywords

intestinal ischemia–reperfusion - bone morphogenetic protein - stem cells - proliferation - apoptosis

• References

• 1 Carden DL, Granger DN. Pathophysiology of ischaemia-reperfusion injury. J Pathol 2000; 190 (03) 255-266
• 2 Wolin MS. Interactions of oxidants with vascular signaling systems. Arterioscler Thromb Vasc Biol 2000; 20 (06) 1430-1442
• 3 Mangino JE, Kotadia B, Mangino MJ. Characterization of hypothermic intestinal ischemia-reperfusion injury in dogs. Effects of glycine. Transplantation 1996; 62 (02) 173-178
• 4 Schoenberg MH, Poch B, Younes M. , et al. Involvement of neutrophils in postischaemic damage to the small intestine. Gut 1991; 32 (08) 905-912
• 5 Yamamoto S, Tanabe M, Wakabayashi G, Shimazu M, Matsumoto K, Kitajima M. The role of tumor necrosis factor-alpha and interleukin-1beta in ischemia-reperfusion injury of the rat small intestine. J Surg Res 2001; 99 (01) 134-141
• 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
• 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 (02) G270-G276
• 8 Ben-Shahar Y, Pollak Y, Bitterman A, Coran AG, Bejar IN, Sukhotnik I. Sonic hedgehog signaling controls gut epithelium homeostasis following intestinal ischemia-reperfusion in a rat. Pediatr Surg Int 2019; 35 (02) 255-261
• 9 de Santa Barbara P, van den Brink GR, Roberts DJ. Development and differentiation of the intestinal epithelium. Cell Mol Life Sci 2003; 60 (07) 1322-1332
• 10 Roberts DJ, Smith DM, Goff DJ, Tabin CJ. Epithelial-mesenchymal signaling during the regionalization of the chick gut. Development 1998; 125 (15) 2791-2801
• 11 Bitgood MJ, McMahon AP. Hedgehog and Bmp genes are coexpressed at many diverse sites of cell-cell interaction in the mouse embryo. Dev Biol 1995; 172 (01) 126-138
• 12 Hogan BL. Bone morphogenetic proteins: multifunctional regulators of vertebrate development. Genes Dev 1996; 10 (13) 1580-1594
• 13 Bandyopadhyay A, Yadav PS, Prashar P. BMP signaling in development and diseases: a pharmacological perspective. Biochem Pharmacol 2013; 85 (07) 857-864
• 14 Shroyer NF, Wong MH. BMP signaling in the intestine: cross-talk is key. Gastroenterology 2007; 133 (03) 1035-1038
• 15 Hardwick JCH, Van Den Brink GR, Bleuming SA. , et al. Bone morphogenetic protein 2 is expressed by, and acts upon, mature epithelial cells in the colon. Gastroenterology 2004; 126 (01) 111-121
• 16 Sukhotnik I, Berkowitz D, Dorfman T. , et al. The role of the BMP signaling cascade in regulation of stem cell activity following massive small bowel resection in a rat. Pediatr Surg Int 2016; 32 (02) 169-174
• 17 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
• 18 Sukhotnik I, Brod V, Lurie M. , et al. The effect of 100% oxygen on intestinal preservation and recovery following ischemia-reperfusion injury in rats. Crit Care Med 2009; 37 (03) 1054-1061
• 19 Sampath TK, Maliakal JC, Hauschka PV. , et al. Recombinant human osteogenic protein-1 (hOP-1) induces new bone formation in vivo with a specific activity comparable with natural bovine osteogenic protein and stimulates osteoblast proliferation and differentiation in vitro. J Biol Chem 1992; 267 (28) 20352-20362
• 20 Radhakrishnan RS, Radhakrishnan GL, Radhakrishnan HR. , et al. Pretreatment with bone morphogenetic protein-7 (BMP-7) mimics ischemia preconditioning following intestinal ischemia/reperfusion injury in the intestine and liver. Shock 2008; 30 (05) 532-536
• 21 Ge C, Yang Q, Zhao G, Yu H, Kirkwood KL, Franceschi RT. Interactions between extracellular signal-regulated kinase 1/2 and p38 MAP kinase pathways in the control of RUNX2 phosphorylation and transcriptional activity. J Bone Miner Res 2012; 27 (03) 538-551
• 22 Kosinski C, Li VS, Chan AS. , et al. Gene expression patterns of human colon tops and basal crypts and BMP antagonists as intestinal stem cell niche factors. Proc Natl Acad Sci U S A 2007; 104 (39) 15418-15423