Skip to main content
SpringerLink
Log in

Psychological stress exacerbates NSAID-induced small bowel injury by inducing changes in intestinal microbiota and permeability via glucocorticoid receptor signaling

  • Original Article—Alimentary Tract
  • Published:
Journal of Gastroenterology Aims and scope Submit manuscript

Abstract

Background

Nonsteroidal anti-inflammatory drugs (NSAIDs) are popular painkillers, but they have serious side effects, not only in the upper gastrointestinal tract but also in the small intestine. It is well known that psychological stress may exacerbate various gastrointestinal diseases. The aim of this study was to determine whether psychological stress exacerbates NSAID enteropathy and to determine the possible underlying mechanisms for this.

Methods

Experiment 1: mice were exposed to water avoidance stress (WAS) or sham stress for 1 h per day for 8 consecutive days, and then enteropathy was induced by indomethacin. Experiment 2: cecal contents from stress (−) or (+) mice were transplanted into mice that had received antibiotics and in which NSAID enteropathy had been induced without WAS. Experiment 3: mifepristone, a glucocorticoid receptor antagonist, was injected before WAS for 8 days. Small intestinal injury, mRNA expression of TNFα, intestinal permeability, and the microbial community were assessed.

Results

Psychological stress exacerbated NSAID enteropathy and increased intestinal permeability. Psychological stress induced changes in the ileal microbiota that were characterized by increases in the total number of bacteria and the proportion of Gram-negative bacteria. The increased susceptibility to NSAIDs and intestinal permeability due to WAS was transferable via cecal microbiota transplantation. The increased permeability and aggravation of NSAID enteropathy caused by WAS were blocked by the administration of mifepristone.

Conclusions

This study demonstrated a relationship between NSAID enteropathy and psychological stress, and showed the utility of studying the intestinal microbiota in order to elucidate the pathophysiology of NSAID enteropathy. It also showed the impact of stress on the intestinal microbiota and the mucosal barrier in gastrointestinal diseases.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1a–e
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

NSAIDs:

Nonsteroidal anti-inflammatory drugs

HPA-axis:

Hypothalamic–pituitary–adrenal axis

MGB-axis:

Microbiota–gut–brain axis

WAS:

Water avoidance stress

DMSO:

Dimethyl sulfoxide

RT-PCR:

Reverse transcription polymerase chain reaction

FITC-dextran:

Fluorescein isothiocyanate dextran

T-RFLP:

Terminal restriction fragment length polymorphism

PBS:

Phosphate-buffered saline

FMT:

Fecal microbiota transplantation

TLR:

Toll-like receptor

GPB:

Gram-positive bacteria

GNB:

Gram-negative bacteria

NLRP6:

NOD-like receptor family pyrin domain containing 6

CRH:

Corticotropin-releasing hormone

References

  1. Turk DC. Clinical effectiveness and cost-effectiveness of treatments for patients with chronic pain. Clin J Pain. 2002;18:355–65.

    Article  PubMed  Google Scholar 

  2. Gibson GR, Whitacre EB, Ricotti CA. Colitis induced by nonsteroidal anti-inflammatory drugs: report of four cases and review of the literature. Arch Intern Med. 1992;152:625–32.

    Article  CAS  PubMed  Google Scholar 

  3. Matsumoto T, Kudo T, Esaki M, et al. Prevalence of non-steroidal anti-inflammatory drug-induced enteropathy determined by double-balloon endoscopy: a Japanese multicenter study. Scand J Gastroenterol. 2008;43:490–6.

  4. Dorais J, Gregoire G, LeLorier J. Gastrointestinal damage associated with nonsteroidal antiinflammatory drugs. N Engl J Med. 1992;327:1882–3.

    Article  CAS  PubMed  Google Scholar 

  5. Boelsterli UA, Redinbo MR, Saitta KS. Multiple NSAID-induced hits injure the small intestine: underlying mechanisms and novel strategies. Toxicol Sci. 2013;131:654–67.

    Article  CAS  PubMed  Google Scholar 

  6. Syer SD, Blackler RW, Martin R, et al. NSAID enteropathy and bacteria: a complicated relationship. J Gastroenterol. 2015;50:387–93.

    Article  CAS  PubMed  Google Scholar 

  7. Collins SM, Bercik P. The relationship between intestinal microbiota and the central nervous system in normal gastrointestinal function and disease. Gastroenterology. 2009;136:2003–14.

    Article  PubMed  Google Scholar 

  8. Mayer EA, Knight R, Mazmanian SK, et al. Gut microbes and the brain: paradigm shift in neuroscience. J Neurosci. 2014;34:15490–6.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Aguilera M, Vergara P, Martinez V. Stress and antibiotics alter luminal and wall-adhered microbiota and enhance the local expression of visceral sensory-related systems in mice. Neurogastroenterol Motil. 2013;25:e515–29.

    Article  CAS  PubMed  Google Scholar 

  10. Higashiyama M, Hokari R, Kurihara C, et al. Indomethacin-induced small intestinal injury is ameliorated by cilostazol, a specific PDE-3 inhibitor. Scand J Gastroenterol. 2012;47:993–1002.

    Article  CAS  PubMed  Google Scholar 

  11. Yamada S, Naito Y, Takagi T, et al. Reduced small-intestinal injury induced by indomethacin in interleukin-17A-deficient mice. J Gastroenterol Hepatol. 2011;26:398–404.

    Article  CAS  PubMed  Google Scholar 

  12. Ueda T, Hokari R, Higashiyama M, et al. Beneficial effect of an omega-6 PUFA-rich diet in non-steroidal anti-inflammatory drug-induced mucosal damage in the murine small intestine. World J Gastroenterol. 2015;21:177–86.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Chiu CJ, McArdle AH, Brown R, et al. Intestinal mucosal lesion in low-flow states. I. A morphological, hemodynamic, and metabolic reappraisal. Arch Surg. 1970;101:478–83.

    Article  CAS  PubMed  Google Scholar 

  14. Ishida T, Miki I, Tanahashi T, et al. Effect of 18beta-glycyrrhetinic acid and hydroxypropyl gammacyclodextrin complex on indomethacin-induced small intestinal injury in mice. Eur J Pharmacol. 2013;714:125–31.

    Article  CAS  PubMed  Google Scholar 

  15. Sun Y, Zhang M, Chen CC, et al. Stress-induced corticotropin-releasing hormone-mediated NLRP6 inflammasome inhibition and transmissible enteritis in mice. Gastroenterology. 2013;144:1478–87.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Matsuki T, Watanabe K, Fujimoto J, et al. Quantitative PCR with 16S rRNA-gene-targeted species-specific primers for analysis of human intestinal bifidobacteria. Appl Environ Microbiol. 2004;70:167–73.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Hayashi H, Sakamoto M, Benno Y. Fecal microbial diversity in a strict vegetarian as determined by molecular analysis and cultivation. Microbiol Immunol. 2002;46:819–31.

    Article  CAS  PubMed  Google Scholar 

  18. Sakamoto M, Hayashi H, Benno Y. Terminal restriction fragment length polymorphism analysis for human fecal microbiota and its application for analysis of complex bifidobacterial communities. Microbiol Immunol. 2003;47:133–42.

    Article  CAS  PubMed  Google Scholar 

  19. Guo X, Xia X, Tang R, et al. Development of a real-time PCR method for firmicutes and bacteroidetes in faeces and its application to quantify intestinal population of obese and lean pigs. Lett Appl Microbiol. 2008;47:367–73.

    Article  CAS  PubMed  Google Scholar 

  20. Bacchetti De Gregoris T, Aldred N, Clare AS, et al. Improvement of phylum- and class-specific primers for real-time PCR quantification of bacterial taxa. J Microbiol Methods. 2011;86:351–6.

    Article  CAS  PubMed  Google Scholar 

  21. Gregory JC, Buffa JA, Org E, et al. Transmission of atherosclerosis susceptibility with gut microbial transplantation. J Biol Chem. 2015;290:5647–60.

    Article  CAS  PubMed  Google Scholar 

  22. Wang Z, Klipfell E, Bennett BJ, et al. Gut flora metabolism of phosphatidylcholine promotes cardiovascular disease. Nature. 2011;472:57–63.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Collins SM. Stress and the gastrointestinal tract IV. Modulation of intestinal inflammation by stress: basic mechanisms and clinical relevance. Am J Physiol Gastrointest Liver Physiol. 2001;280:G315–8.

    CAS  PubMed  Google Scholar 

  24. Zheng G, Wu SP, Hu Y, et al. Corticosterone mediates stress-related increased intestinal permeability in a region-specific manner. Neurogastroenterol Motil. 2013;25:e127–39.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Rakoff-Nahoum S, Paglino J, Eslami-Varzaneh F, et al. Recognition of commensal microflora by toll-like receptors is required for intestinal homeostasis. Cell. 2004;118:229–41.

    Article  CAS  PubMed  Google Scholar 

  26. Zoppi S, Madrigal JL, Perez-Nievas BG, et al. Endogenous cannabinoid system regulates intestinal barrier function in vivo through cannabinoid type 1 receptor activation. Am J Physiol Gastrointest Liver Physiol. 2012;302:G565–71.

    Article  CAS  PubMed  Google Scholar 

  27. Muccioli GG, Naslain D, Backhed F, et al. The endocannabinoid system links gut microbiota to adipogenesis. Mol Syst Biol. 2010;6:392.

    Article  PubMed  PubMed Central  Google Scholar 

  28. Shen L. Tight junctions on the move: Molecular mechanisms for epithelial barrier regulation. Ann NY Acad Sci. 2012;1258:9–18.

    Article  PubMed  PubMed Central  Google Scholar 

  29. Zareie M, Johnson-Henry K, Jury J, et al. Probiotics prevent bacterial translocation and improve intestinal barrier function in rats following chronic psychological stress. Gut. 2006;55:1553–60.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Sheehan D, Moran C, Shanahan F. The microbiota in inflammatory bowel disease. J Gastroenterol. 2015;50:495–507.

    Article  CAS  PubMed  Google Scholar 

  31. Goldsmith JR, Sartor RB. The role of diet on intestinal microbiota metabolism: downstream impacts on host immune function and health, and therapeutic implications. J Gastroenterol. 2014;49:785–98.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Lewis K, Lutgendorff F, Phan V, Soderholm JD, et al. Enhanced translocation of bacteria across metabolically stressed epithelia is reduced by butyrate. Inflamm Bowel Dis. 2010;16:1138–48.

    Article  PubMed  Google Scholar 

  33. Wu Z, Nybom P, Magnusson KE. Distinct effects of vibrio cholerae haemagglutinin/protease on the structure and localization of the tight junction-associated proteins occludin and ZO-1. Cell Microbiol. 2000;2:11–7.

    Article  CAS  PubMed  Google Scholar 

  34. Simovitch M, Sason H, Cohen S, et al. EspM inhibits pedestal formation by enterohaemorrhagic Escherichia coli and enteropathogenic E. coli and disrupts the architecture of a polarized epithelial monolayer. Cell Microbiol. 2010;12:489–505.

  35. Ng KM, Ferreyra JA, Higginbottom SK, et al. Microbiota-liberated host sugars facilitate post-antibiotic expansion of enteric pathogens. Nature. 2013;502:96–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Zyrek AA, Cichon C, Helms S, et al. Molecular mechanisms underlying the probiotic effects of Escherichia coli nissle 1917 involve ZO-2 and PKCzeta redistribution resulting in tight junction and epithelial barrier repair. Cell Microbiol. 2007;9:804–16.

    Article  CAS  PubMed  Google Scholar 

  37. Ewaschuk JB, Diaz H, Meddings L, et al. Secreted bioactive factors from Bifidobacterium infantis enhance epithelial cell barrier function. Am J Physiol Gastrointest Liver Physiol. 2008;295:G1025–34.

  38. Karczewski J, Troost FJ, Konings I, et al. Regulation of human epithelial tight junction proteins by Lactobacillus plantarum in vivo and protective effects on the epithelial barrier. Am J Physiol Gastrointest Liver Physiol. 2010;298:G851–9.

  39. Anderson RC, Cookson AL, McNabb WC, et al. Lactobacillus plantarum MB452 enhances the function of the intestinal barrier by increasing the expression levels of genes involved in tight junction formation. BMC Microbiol. 2010;10:316.

  40. Cario E, Gerken G, Podolsky DK. Toll-like receptor 2 enhances ZO-1-associated intestinal epithelial barrier integrity via protein kinase C. Gastroenterology. 2004;127:224–38.

    Article  CAS  PubMed  Google Scholar 

  41. Moussaoui N, Braniste V, Ait-Belgnaoui A, et al. Changes in intestinal glucocorticoid sensitivity in early life shape the risk of epithelial barrier defect in maternal-deprived rats. PLoS One. 2014;9:e88382.

    Article  PubMed  PubMed Central  Google Scholar 

  42. Kelly SP, Chasiotis H. Glucocorticoid and mineralocorticoid receptors regulate paracellular permeability in a primary cultured gill epithelium. J Exp Biol. 2011;214:2308–18.

    Article  CAS  PubMed  Google Scholar 

  43. Hagiwara M, Kataoka K, Arimochi H, et al. Role of unbalanced growth of gram-negative bacteria in ileal ulcer formation in rats treated with a nonsteroidal anti-inflammatory drug. J Med Invest. 2004;51:43–51.

    Article  PubMed  Google Scholar 

  44. Kent TH, Cardelli RM, Stamler FW. Small intestinal ulcers and intestinal flora in rats given indomethacin. Am J Pathol. 1969;54:237–49.

    CAS  PubMed  PubMed Central  Google Scholar 

  45. Dalby AB, Frank DN, St Amand AL, et al. Culture-independent analysis of indomethacin-induced alterations in the rat gastrointestinal microbiota. Appl Environ Microbiol. 2006;72:6707–15.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Watanabe T, Higuchi K, Kobata A, et al. Non-steroidal anti-inflammatory drug-induced small intestinal damage is Toll-like receptor 4 dependent. Gut. 2008;57:181–7.

  47. Xu D, Gao J, Gillilland M 3rd, et al. Rifaximin alters intestinal bacteria and prevents stress-induced gut inflammation and visceral hyperalgesia in rats. Gastroenterology. 2014;146:484–96.

    Article  CAS  PubMed  Google Scholar 

  48. Anand PK, Malireddi RK, Lukens JR, et al. NLRP6 negatively regulates innate immunity and host defence against bacterial pathogens. Nature. 2012;488:389–93.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This study was supported by a Health and Labor Sciences research grant into research on intractable diseases, from the Ministry of Health, Labor and Welfare, Japan (http://www.mhlw.go.jp/stf/seisakunitsuite/bunya/hokabunya/kenkyujigyou/).

Author information

Authors and Affiliations

  1. Division of Gastroenterology, Department of Internal Medicine, National Defense Medical College, 3-2 Namiki, Tokorozawa-shi, Saitama, 359-8513, Japan

    Kenichi Yoshikawa, Chie Kurihara, Hirotaka Furuhashi, Takeshi Takajo, Koji Maruta, Yuichi Yasutake, Hirokazu Sato, Kazuyuki Narimatsu, Yoshikiyo Okada, Masaaki Higashiyama, Chikako Watanabe, Kengo Tomita & Ryota Hokari

  2. Division of Gastroenterology and Hepatology, Internal Medicine, The Jikei University School of Medicine, 3-25-8 Nishi-Shimbashi, Minato-ku, Tokyo, 105-8461, Japan

    Kenichi Yoshikawa & Hisao Tajiri

  3. Department of Endoscopic and Photodynamic Medicine, National Defense Medical College, Saitama, Japan

    Shunsuke Komoto & Shigeaki Nagao

  4. National Defense Medical College, Saitama, Japan

    Soichiro Miura

Authors
  1. Kenichi Yoshikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Chie Kurihara
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hirotaka Furuhashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Takeshi Takajo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Koji Maruta
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yuichi Yasutake
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hirokazu Sato
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Kazuyuki Narimatsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Yoshikiyo Okada
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Masaaki Higashiyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Chikako Watanabe
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Shunsuke Komoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Kengo Tomita
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Shigeaki Nagao
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Soichiro Miura
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Hisao Tajiri
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Ryota Hokari
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kenichi Yoshikawa.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yoshikawa, K., Kurihara, C., Furuhashi, H. et al. Psychological stress exacerbates NSAID-induced small bowel injury by inducing changes in intestinal microbiota and permeability via glucocorticoid receptor signaling. J Gastroenterol 52, 61–71 (2017). https://doi.org/10.1007/s00535-016-1205-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00535-016-1205-1

Keywords

Search

Navigation