Human Cell

, Volume 32, Issue 1, pp 12–21 | Cite as

Subcutaneously administered adrenomedullin exerts a potent therapeutic effect in a murine model of ulcerative colitis

  • Yuta Kinoshita
  • Seiya Arita
  • Haruka Murazoe
  • Kazuo Kitamura
  • Shinya Ashizuka
  • Kyoko Inagaki-OharaEmail author
Research Article


Adrenomedullin (AM) exerts a potent anti-inflammatory effect. Intrarectal or consecutive intravenous administrations of AM reduce pathological manifestations in rodent colitis models. However, in clinical applications, a safer administration route that provides stronger alleviation of patient burden is preferred. We investigated whether subcutaneously administered AM is effective against dextran sulfate sodium (DSS)-induced colitis. C57BL/6J mice were administered 1% DSS in drinking water and received AM at 8, 40 or 80 nmol/kg subcutaneously once a day for 7 consecutive days. Subcutaneously administered AM significantly and dose-dependently ameliorated body weight loss, diarrhea, and histological severity of colonic inflammation in DSS-treated mice. The AM therapeutic effect was associated with the upregulation of the production of autocrine AM, and expression of cAMP, c-fos, KLF4, and downregulation of STAT3 and NF-κB p65 phosphorylation, as well as a decrease in proinflammatory cytokine expression in the colon. Subcutaneous AM treatment potently attenuated DSS-induced colitis, which suggests that AM administered subcutaneously in ulcerative colitis (UC) patients may decrease diseases burden and improve quality of life.


Adrenomedullin Anti-inflammation Colitis Subcutaneous injection Goblet cells 



This work was supported by Grants-in-Aid for Scientific Research (C) from the Ministry of Education, Culture, Sports, Science and Technology of Japan (16K09316). We would like to thank all of members of the Division of Host Defense of the Prefectural University of Hiroshima for animal care support.

Compliance with ethical standards

Conflict of interest

Kazuo Kitamura holds the stock of Himuka AM Pharma Corp. Yuta Kinoshita, Seiya Arita, Haruka Murazoe, Shinya Ashizuka, and Kyoko Inagaki-Ohara have no conflicts of interest to declare.

Ethical approval

All animal experiments were conducted in accordance with the guidelines of the Animal Research Committee of the Prefectural University of Hiroshima Animal Care and Use Committee.

Supplementary material

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Supplementary material 5 (DOCX 63 KB)


  1. 1.
    El Mourabet M, El-Hachem S, Harrison JR, et al. Anti-TNF antibody therapy for inflammatory bowel disease during pregnancy: a clinical review. Curr Drug Targets. 2010;11:234–41.CrossRefGoogle Scholar
  2. 2.
    Park SC, Jeen YT. Anti-integrin therapy for inflammatory bowel disease. World J Gastroenterol. 2018;24:1868–80.CrossRefGoogle Scholar
  3. 3.
    Terzic J, Grivennikov S, Karin E, et al. Inflammation and colon cancer. Gastroenterology. 2010;138:2101-14 e5.CrossRefGoogle Scholar
  4. 4.
    Eaden JA, Abrams KR, Mayberry JF. The risk of colorectal cancer in ulcerative colitis: a meta-analysis. Gut. 2001;48:526–35.CrossRefGoogle Scholar
  5. 5.
    Gillen CD, Walmsley RS, Prior P, et al. Ulcerative colitis and Crohn’s disease: a comparison of the colorectal cancer risk in extensive colitis. Gut. 1994;35:1590–2.CrossRefGoogle Scholar
  6. 6.
    Chouraki V, Savoye G, Dauchet L, et al. The changing pattern of Crohn’s disease incidence in northern France: a continuing increase in the 10- to 19-year-old age bracket (1988–2007). Aliment Pharmacol Ther. 2011;33:1133–42.CrossRefGoogle Scholar
  7. 7.
    Lopez J, Martinez A. Cell and molecular biology of the multifunctional peptide, adrenomedullin. Int Rev Cytol. 2002;221:1–92.CrossRefGoogle Scholar
  8. 8.
    Poyner DR, Sexton PM, Marshall I, et al. International Union of Pharmacology. XXXII. The mammalian calcitonin gene-related peptides, adrenomedullin, amylin, and calcitonin receptors. Pharmacol Rev. 2002;54:233–46.CrossRefGoogle Scholar
  9. 9.
    Kitamura K, Kangawa K, Kawamoto M, et al. Adrenomedullin: a novel hypotensive peptide isolated from human pheochromocytoma. Biochem Biophys Res Commun. 1993;192:553–60.CrossRefGoogle Scholar
  10. 10.
    Martinez-Herrero S, Martinez A. Adrenomedullin regulates intestinal physiology and pathophysiology. Domest Anim Endocrinol. 2016;56(Suppl):66–83.CrossRefGoogle Scholar
  11. 11.
    Gonzalez-Rey E, Fernandez-Martin A, Chorny A, et al. Therapeutic effect of urocortin and adrenomedullin in a murine model of Crohn’s disease. Gut. 2006;55:824–32.CrossRefGoogle Scholar
  12. 12.
    Ashizuka S, Inagaki-Ohara K, Kuwasako K, et al. Adrenomedullin treatment reduces intestinal inflammation and maintains epithelial barrier function in mice administered dextran sulphate sodium. Microbiol Immunol. 2009;53:573–81.CrossRefGoogle Scholar
  13. 13.
    Martinez-Herrero S, Larrayoz IM, Narro-Iniguez J, et al. Lack of adrenomedullin aggravates acute TNBS-induced colitis symptoms in mice, especially in females. Front Physiol. 2017;8:1058.CrossRefGoogle Scholar
  14. 14.
    Martinez-Herrero S, Larrayoz IM, Narro-Iniguez J, et al. Lack of adrenomedullin results in microbiota changes and aggravates azoxymethane and dextran sulfate sodium-induced colitis in mice. Front Physiol. 2016;7:595.Google Scholar
  15. 15.
    Ashizuka S, Inatsu H, Kita T, et al. Adrenomedullin therapy in patients with refractory ulcerative colitis: a case series. Dig Dis Sci. 2016;61:872–80.CrossRefGoogle Scholar
  16. 16.
    Ashizuka S, Kita T, Inatsu H, et al. Adrenomedullin: a novel therapy for intractable ulcerative colitis. Inflamm Bowel Dis. 2013;19:E26-7.CrossRefGoogle Scholar
  17. 17.
    Nagata S, Yamasaki M, Kitamura K. Anti-inflammatory effects of PEGylated human adrenomedullin in a mouse DSS-induced colitis model. Drug Dev Res. 2017;78:129–34.CrossRefGoogle Scholar
  18. 18.
    Inagaki-Ohara K, Okamoto S, Takagi K, et al. Leptin receptor signaling is required for high-fat diet-induced atrophic gastritis in mice. Nutr Metab (Lond). 2016;13:7.CrossRefGoogle Scholar
  19. 19.
    Inagaki-Ohara K, Sasaki A, Matsuzaki G, et al. Suppressor of cytokine signalling 1 in lymphocytes regulates the development of intestinal inflammation in mice. Gut. 2006;55:212–9.CrossRefGoogle Scholar
  20. 20.
    Najjar YG, Rayman P, Jia X, et al. Myeloid-derived suppressor cell subset accumulation in renal cell carcinoma parenchyma is associated with intratumoral expression of IL1beta, IL8, CXCL5, and Mip-1alpha. Clin Cancer Res. 2017;23:2346–55.CrossRefGoogle Scholar
  21. 21.
    Inagaki-Ohara K, Chinen T, Matsuzaki G, et al. Mucosal T cells bearing TCRgammadelta play a protective role in intestinal inflammation. J Immunol. 2004;173:1390–8.CrossRefGoogle Scholar
  22. 22.
    DeRoche TC, Xiao SY, Liu X. Histological evaluation in ulcerative colitis. Gastroenterol Rep (Oxf). 2014;2:178–92.CrossRefGoogle Scholar
  23. 23.
    Koga K, Takaesu G, Yoshida R, et al. Cyclic adenosine monophosphate suppresses the transcription of proinflammatory cytokines via the phosphorylated c-Fos protein. Immunity. 2009;30:372–83.CrossRefGoogle Scholar
  24. 24.
    Thomson LM, Kapas S, Carroll M, et al. Autocrine role of adrenomedullin in the human adrenal cortex. J Endocrinol. 2001;170:259–65.CrossRefGoogle Scholar
  25. 25.
    Maki T, Ihara M, Fujita Y, et al. Angiogenic and vasoprotective effects of adrenomedullin on prevention of cognitive decline after chronic cerebral hypoperfusion in mice. Stroke. 2011;42:1122–8.CrossRefGoogle Scholar
  26. 26.
    Grivennikov S, Karin E, Terzic J, et al. IL-6 and Stat3 are required for survival of intestinal epithelial cells and development of colitis-associated cancer. Cancer Cell. 2009;15:103–13.CrossRefGoogle Scholar
  27. 27.
    Fyderek K, Strus M, Kowalska-Duplaga K, et al. Mucosal bacterial microflora and mucus layer thickness in adolescents with inflammatory bowel disease. World J Gastroenterol. 2009;15:5287–94.CrossRefGoogle Scholar
  28. 28.
    Wenzel UA, Magnusson MK, Rydstrom A, et al. Spontaneous colitis in Muc2-deficient mice reflects clinical and cellular features of active ulcerative colitis. PLoS One. 2014;9:e100217.CrossRefGoogle Scholar
  29. 29.
    Neurath MF. New targets for mucosal healing and therapy in inflammatory bowel diseases. Mucosal Immunol. 2014;7:6–19.CrossRefGoogle Scholar
  30. 30.
    Hasnain SZ, Tauro S, Das I, et al. IL-10 promotes production of intestinal mucus by suppressing protein misfolding and endoplasmic reticulum stress in goblet cells. Gastroenterology. 2013;144:357–68 e9.CrossRefGoogle Scholar
  31. 31.
    Katz JP, Perreault N, Goldstein BG, et al. The zinc-finger transcription factor Klf4 is required for terminal differentiation of goblet cells in the colon. Development. 2002;129:2619–28.Google Scholar
  32. 32.
    Ghaleb AM, Laroui H, Merlin D, et al. Genetic deletion of Klf4 in the mouse intestinal epithelium ameliorates dextran sodium sulfate-induced colitis by modulating the NF-kappaB pathway inflammatory response. Inflamm Bowel Dis. 2014;20:811–20.CrossRefGoogle Scholar
  33. 33.
    Gersemann M, Becker S, Kubler I, et al. Differences in goblet cell differentiation between Crohn’s disease and ulcerative colitis. Differentiation. 2009;77:84–94.CrossRefGoogle Scholar
  34. 34.
    Prossomariti A, Scaioli E, Piazzi G, et al. Short-term treatment with eicosapentaenoic acid improves inflammation and affects colonic differentiation markers and microbiota in patients with ulcerative colitis. Sci Rep. 2017;7:7458.CrossRefGoogle Scholar
  35. 35.
    Rowe J, Finlay-Jones JJ, Nicholas TE, et al. Inability of histamine to regulate TNF-alpha production by human alveolar macrophages. Am J Respir Cell Mol Biol. 1997;17:218–26.CrossRefGoogle Scholar
  36. 36.
    Spehlmann ME, Eckmann L. Nuclear factor-kappa B in intestinal protection and destruction. Curr Opin Gastroenterol. 2009;25:92–9.CrossRefGoogle Scholar
  37. 37.
    Suzuki A, Hanada T, Mitsuyama K, et al. CIS3/SOCS3/SSI3 plays a negative regulatory role in STAT3 activation and intestinal inflammation. J Exp Med. 2001;193:471–81.CrossRefGoogle Scholar
  38. 38.
    Bollrath J, Phesse TJ, von Burstin VA, et al. gp130-mediated Stat3 activation in enterocytes regulates cell survival and cell-cycle progression during colitis-associated tumorigenesis. Cancer Cell. 2009;15:91–102.CrossRefGoogle Scholar
  39. 39.
    Takeda K, Clausen BE, Kaisho T, et al. Enhanced Th1 activity and development of chronic enterocolitis in mice devoid of Stat3 in macrophages and neutrophils. Immunity. 1999;10:39–49.CrossRefGoogle Scholar
  40. 40.
    Hsieh FC, Cheng G, Lin J. Evaluation of potential Stat3-regulated genes in human breast cancer. Biochem Biophys Res Commun. 2005;335:292–9.CrossRefGoogle Scholar
  41. 41.
    Lim SY, Ahn SH, Park H, et al. Transcriptional regulation of adrenomedullin by oncostatin M in human astroglioma cells: implications for tumor invasion and migration. Sci Rep. 2014;4:6444.CrossRefGoogle Scholar
  42. 42.
    Hall J, Guo G, Wray J, et al. Oct4 and LIF/Stat3 additively induce Kruppel factors to sustain embryonic stem cell self-renewal. Cell Stem Cell. 2009;5:597–609.CrossRefGoogle Scholar
  43. 43.
    Ou L, Shi Y, Dong W, et al. Kruppel-like factor KLF4 facilitates cutaneous wound healing by promoting fibrocyte generation from myeloid-derived suppressor cells. J Invest Dermatol. 2015;135:1425–34.CrossRefGoogle Scholar

Copyright information

© Japan Human Cell Society and Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  • Yuta Kinoshita
    • 1
  • Seiya Arita
    • 1
  • Haruka Murazoe
    • 1
  • Kazuo Kitamura
    • 2
  • Shinya Ashizuka
    • 2
  • Kyoko Inagaki-Ohara
    • 1
    Email author
  1. 1.Division of Host Defense, Department of Life Sciences, Faculty of Life and Environmental SciencesPrefectural University of HiroshimaShobaraJapan
  2. 2.First Department of Internal MedicineUniversity of MiyazakiKiyotakeJapan

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