Dietary transforming growth factor-beta 2 (TGF-Œ≤2) supplementation reduces methotrexate-induced intestinal mucosal injury in a rat
Authors: Arnold G Coran
Jorge Mogilner
Shani Ben-Lulu
Igor Sukhotnik
Jacob Bejar
Yulia Pollak
Journal: PLoS ONE (2012)
Discipline: Multidisciplinary
Keywords: Dietary supplementation; Transforming growth factor-beta (TGF-Œ≤); Intestinal damage; Regeneration; Methotrexate (MTX)
Affiliations: Laboratory of Intestinal Adaptation and Recovery, The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel.

Abstract:  Background/aims: Dietary supplementation with transforming growth factor-beta (TGF-Œ≤) has been proven to minimize intestinal damage and facilitate regeneration after mucosal injury. In the present study, we evaluated the effects of oral TGF-Œ≤2 supplementation on intestinal structural changes, enterocyte proliferation and apoptosis following methotrexate (MTX)-induced intestinal damage in a rat and in a cell culture model. Methods: Caco-2 cells were treated with MTX and were incubated with increasing concentrations of TGF-Œ≤2. Cell apoptosis was assessed using FACS analysis by annexin staining and cell viability was monitored using Trypan Blue assay. Male rats were divided into four experimental groups: Control rats, CONTR- TGF-Œ≤ rats were treated with diet enriched with TGF-Œ≤2, MTX rats were treated with a single dose of methotrexate, and MTX- TGF-Œ≤ rats were treated with diet enriched with TGF-Œ≤2. Intestinal mucosal damage, mucosal structural changes, enterocyte proliferation and enterocyte apoptosis were determined at sacrifice. Real Time PCR and Western blot were used to determine bax and bcl-2 mRNA, p-ERK, Œ≤-catenin, IL-1B and bax protein expression. Results: Treatment of MTX-pretreated Caco-2 cells with TGF-B2 resulted in increased cell viability and decreased cell apoptosis. Treatment of MTX-rats with TGF-Œ≤2 resulted in a significant increase in bowel and mucosal weight, DNA and protein content, villus-height (ileum), crypt-depth (jejunum), decreased intestinal-injury score, decreased level of apoptosis and increased cell proliferation in jejunum and ileum compared to the untreated MTX group. MTX-TGF-Œ≤2 rats demonstrated a lower bax mRNA and protein levels as well as increased bcl-2 mRNA levels in jejunum and ileum compared to MTX group. Treatment with TGF-Œ≤2 also led to increased pERK, IL-1B and Œ≤-catenin protein levels in intestinal mucosa. Conclusions: Treatment with TGF-Œ≤2 prevents mucosal-injury, enhances p-ERK and Œ≤-catenin induced enterocyte proliferation, inhibits enterocyte apoptosis and improves intestinal recovery following MTX-induced intestinal-mucositis in rats. 

Retracted: No

PubPeer comments:
 This paper appears to be part of a set of at least 13 papers that appear to share immunohistochemistry images. All papers list Igor Sukhotnik as an author and they span a period of 12 years. The images do not always represent the same experiments. 1. Mogilner JG et al., Journal of Pediatric Surgery (2007), DOI: 10.1016 j.jpedsurg.2007.03.035 2. Sukhotnik I et al., Cancer Biology Therapy (2009), DOI: 10.4161 cbt.8.10.8128 3. Sukhotnik I et al., Journal of Pediatric Surgery (2011), DOI: 10.1016 j.jpedsurg.2010.09.051 4. Ben-Lulu S et al., PLOS ONE (2012), DOI: 10.1371 journal.pone.0045221 5. Koppelman T et al., BMC Gastroenterology (2012), DOI: 10.1186 1471-230X-12-41 6. Koppelman T et al., British Journal of Nutrition (2013), DOI: 10.1017 S0007114512000542 7. Swaid F et al., Nutrition Metabolism (2013), DOI: 10.1186 1743-7075-10-53 8. Sukhotnik I et al., Nutrition Metabolism (2014), DOI: 10.1186 1743-7075-11-17 9. Sukhotnik I et al., PLOS ONE (2014), DOI: 10.1371 journal.pone.0110675 10. Sukhotnik I et al., European Journal of Pediatric Surgery (2016), DOI: 10.1055 s-0036-1587588 11. Ben Shahar Y et al., European Journal of Pediatric Surgery (2016), DOI: 10.1055 s-0035-1559886 12. Sukhotnik I et al., Food Nutrition Research (2018), DOI: 10.29219 fnr.v62.1327 13. Ben-Shahar Y et al., European Journal of Pediatric Surgery (2019), DOI: 10.1055 s-0039-1700550 Here are three collages of the IHC figures from these papers, with boxes of the same color highlighting panels that look more similar than expected.    , 
 Awesome, truly awesome. I wonder how much immunoblot reuse might be going on? Hereu2019s a starter  , 
 This paper is one of at least four papers by the same research group that appear to share the same ERK2 blot to represent different experiments (marked in red). Two of these papers also seem to share another blot (marked with blue boxes). The four papers are: * Ben-Lulu S et al., PLOS ONE (2012), DOI: 10.1371 journal.pone.0045221 * Sukhotnik I et al., Pediatr Surg Int (2013), DOI: 10.1007 s00383-012-3197-0 * Koppelman T et al., BMC Gastroenterology (2012), DOI: 10.1186 1471-230X-12-41 * Koppelman T et al., British Journal of Nutrition (2013), DOI: 10.1017 S0007114512000542 Here is a collage of the four figures, one from each of these papers.  ,