British Journal of Nutrition

Full Papers

Human and Clinical Nutrition

Reversal of severe methotrexate-induced intestinal damage using enteral n-3 fatty acids

Tal Koppelmanna1a2, Yulia Pollaka1, Jorge Mogilnera3, Jacob Bejara4, Arnold G. Corana5 and Igor Sukhotnika1a3 c1

a1 Laboratory of Intestinal Adaptation and Recovery, The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Bnai Zion Medical Center, Haifa, Israel

a2 Department of Surgery, Bnai Zion Medical Center, Haifa, Israel

a3 Department of Pediatric Surgery, Bnai Zion Medical Center, 47 Golomb Street, PO Box 4940, Haifa 31048, Israel

a4 Department of Pathology, Bnai Zion Medical Center, Haifa, Israel

a5 Section of Pediatric Surgery, C.S. Mott Children's Hospital, University of Michigan Medical School, Ann Arbor, MI, USA


Growing evidence suggests that n-3 PUFA and their specific lipid mediators can reduce the activity of inflammatory processes. In the present study, we evaluated the effects of oral n-3 PUFA supplementation on intestinal structural changes, enterocyte proliferation and apoptosis during methotrexate (MTX)-induced intestinal damage in the rat. A total of thirty-two male rats were divided into four experimental groups: control (CONTR) rats; CONTR-n-3 PUFA rats treated with oral administration of n-3 PUFA at a dose of 300 μg/kg once per d 72 h before and 72 h following vehicle injection; MTX rats treated with a single dose of MTX; MTX-n-3 PUFA rats treated with oral n-3 PUFA following the injection of MTX. Intestinal mucosal damage, mucosal structural changes, enterocyte proliferation and enterocyte apoptosis determined 72 h following MTX injection. Real-time PCR was used to determine B-cell lymphoma 2 (Bcl2)-associated X protein (Bax) and Bcl2 mRNA expression. Western blotting was used to determine phosphorylated extracellular signal-related kinase, β-catenin, Bax and Bcl2 protein levels. MTX-n-3 PUFA rats demonstrated a greater jejunal and ileal bowel weight, greater ileal mucosal weight, greater ileal mucosal DNA and protein levels, greater villus height in the jejunum and ileum and crypt depth in the ileum, compared with MTX animals. A significant decrease in enterocyte apoptosis in the ileum of MTX-n-3 PUFA rats (v. MTX) was accompanied by decreased Bax mRNA and protein expression and increased Bcl2 mRNA levels. Thus, the treatment with oral n-3 PUFA prevented mucosal injury and improved intestinal recovery following MTX-injury in rats.

(Received October 10 2011)

(Revised January 16 2012)

(Accepted January 26 2012)

(Online publication March 28 2012)

Key Words:

  • Methotrexate;
  • Mucositis;
  • Intestine;
  • n-3 PUFA;
  • Rats


c1 Corresponding author: Dr I. Sukhotnik, fax +972 4 8359620, email


  Abbreviations: BrdU, bromodeoxyuridine; ERK, extracellular signal-related kinase; MAPK, mitogen-activated protein kinase