Cancer Chemotherapy and Pharmacology

, Volume 84, Issue 1, pp 105–116 | Cite as

Orally administered salecan ameliorates methotrexate-induced intestinal mucositis in mice

  • Yan Gao
  • Qi Sun
  • Xiao Yang
  • Weiling Lu
  • Yang Zhao
  • Wenhao Ge
  • Yunxia Yang
  • Xi Xu
  • Jianfa ZhangEmail author
Original Article



Methotrexate (MTX) is a widely used cancer chemotherapy agent. The efficacy of MTX is often limited by serious side effects, such as intestinal mucositis. The aim of this study was to evaluate the protective effect of water-soluble β-glucan salecan on MTX-induced intestinal toxicity in mice.


Intestinal mucositis was induced in C57BL/6 mice by intraperitoneal injection of MTX for two consecutive days. Mice were orally administrated with saline or salecan for 6 days before MTX injection and continued to the end of the study. Several histological and biochemical parameters were measured in the jejunum.


Orally administration of salecan improved the severity of intestinal mucositis in a dose-dependent manner, as evidenced by the well-maintained mucosal architecture and body weight in salecan-treated groups. Salecan treatment inhibited MTX-induced oxidative stress and effectively scavenged free radicals both in vitro and in vivo. Metabolomics analysis revealed that salecan treatment reversed the intestinal metabolic profiling changes in mice with MTX-induced mucositis. Salecan treatment modulated the innate immunity through the regulation of TLR and Dectin1 expression in the jejunum, thus protecting mice from MTX-induced intestinal damage.


Salecan has potential advantages in the treatment of MTX-induced intestinal mucositis, and its protective effect is mainly attributed to its antioxidant and immunomodulatory properties.


Methotrexate Intestine mucositis Salecan Antioxidant Metabonomics 



This work was supported by the grant from National Nature Science Foundation of China (Numbers 31671220, 31471111).

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

280_2019_3854_MOESM1_ESM.pdf (91 kb)
Supplementary material 1 (PDF 90 kb)
280_2019_3854_MOESM2_ESM.tif (160 kb)
Supplementary Fig. 1 The relative viability of B16F10 cells after being exposed to MTX and salecan at different concentrations for 24 h by a MTT assay. Data are shown as the mean ± SEM, n = 6, #p < 0.05, ##p < 0.01, compared to control group; *p < 0.05, **p < 0.01, compared to MTX (10 μg/ml) group. (TIFF 159 kb)
280_2019_3854_MOESM3_ESM.tif (598 kb)
Supplementary Fig. 2 Typical 500 MHz 1H NMR spectra of the small intestine of mice from the control group, MTX group, and high-dose salecan group were in black, green and red, respectively. Labeled metabolites: 1. Isoleucine, 2. Leucine, 3. Valine, 4. Lactate, 5. Threonine, 6. Alanine, 7. Lysine, 8. Acetate, 9. Glutamate, 10. Glutathione, 11. Succinate, 12. Methionine, 13. Creatine phosphate, 14. O-Phosphocholine, 15. Taurine, 16. Glycine, 17. Glucose, 18. Uridine, 19. Inosine, 20. Fumarate, 21. Tyrosine, 22. Phenylalanine, 23. Histidine, 24. 3-Methylxanthine, 25. Adenosine, 26. AMP. (TIFF 597 kb)


  1. 1.
    Jolivet J, Cowan KH, Curt GA, Clendeninn NJ, Chabner BA (1983) The pharmacology and clinical use of methotrexate. N Engl J Med 309(18):1094–1104. CrossRefGoogle Scholar
  2. 2.
    Edwin SL, Chan MD, Bruce N, Cronstein MD (2013) Mechanisms of action of methotrexate. Bull Hosp Jt Dis 71(Suppl 1):S5–S8Google Scholar
  3. 3.
    Grosflam JW, Weinblatt ME (1991) Methotrexate: mechanism of action, pharmacokinetics, clinical indications, and toxicity. Curr Opin Rheumatol 3(3):363–368CrossRefGoogle Scholar
  4. 4.
    Pico J-L, Avila-Garavito A, Naccache P (1998) Mucositis: its occurrence, consequences, and treatment in the oncology setting. Oncologist 3(6):446–451Google Scholar
  5. 5.
    Miyazono YG, Horie FT (2004) Oxidative stress contributes to methotrexate-induced small intestinal toxicity in rats. Scand J Gastroenterol 39(11):1119–1127. CrossRefGoogle Scholar
  6. 6.
    Gao F, Horie T (2002) A synthetic analog of prostaglandin E1 prevents the production of reactive oxygen species in the intestinal mucosa of methotrexate-treated rats. Life Sci 71(9):1091–1099. CrossRefGoogle Scholar
  7. 7.
    Yuncu ME, Koruk A, Sari M, Bagci I, Inaloz CS (2004) Effect of vitamin A against methotrexate-induced damage to the small intestine in rats. Med Princ Pract 13(6):346–352. CrossRefGoogle Scholar
  8. 8.
    Maeda T, Miyazono Y, Ito K, Hamada K, Sekine S, Horie T (2010) Oxidative stress and enhanced paracellular permeability in the small intestine of methotrexate-treated rats. Cancer Chemother Pharmacol 65(6):1117–1123. CrossRefGoogle Scholar
  9. 9.
    van Vliet MJ, Harmsen HJM, de Bont ESJM, Tissing WJE (2010) The role of intestinal microbiota in the development and severity of chemotherapy-induced mucositis. PLoS Pathog 6(5):e1000879. CrossRefGoogle Scholar
  10. 10.
    Kayali H, Ozdag MF, Kahraman S, Aydin A, Gonul E, Sayal A, Odabasi Z, Timurkaynak E (2005) The antioxidant effect of β-Glucan on oxidative stress status in experimental spinal cord injury in rats. Neurosurg Rev 28(4):298–302. CrossRefGoogle Scholar
  11. 11.
    Chen X, Xu X, Zhang L, Zeng F (2009) Chain conformation and anti-tumor activities of phosphorylated (1 → 3)-β-d-glucan from Poria cocos. Carbohyd Polym 78(3):581–587. CrossRefGoogle Scholar
  12. 12.
    Zhou M, Wang Z, Chen J, Zhan Y, Wang T, Xia L, Wang S, Hua Z, Zhang J (2014) Supplementation of the diet with Salecan attenuates the symptoms of colitis induced by dextran sulphate sodium in mice. Br J Nutr 111(10):1822–1829. CrossRefGoogle Scholar
  13. 13.
    Zhou M, Jia P, Chen J, Xiu A, Zhao Y, Zhan Y, Chen P, Zhang J (2013) Laxative effects of Salecan on normal and two models of experimental constipated mice. BMC Gastroenterol 13(1):52. CrossRefGoogle Scholar
  14. 14.
    Xiu A, Zhou M, Zhu B, Wang S, Zhang J (2011) Rheological properties of Salecan as a new source of thickening agent. Food Hydrocolloids 25(7):1719–1725. CrossRefGoogle Scholar
  15. 15.
    Chang CJ, Lin CS, Lu CC, Martel J, Ko YF, Ojcius DM, Tseng SF, Wu TR, Chen YY, Young JD, Lai HC (2015) Ganoderma lucidum reduces obesity in mice by modulating the composition of the gut microbiota. Nat Commun 6:7489. CrossRefGoogle Scholar
  16. 16.
    Beckonert O, Keun HC, Ebbels TM, Bundy J, Holmes E, Lindon JC, Nicholson JK (2007) Metabolic profiling, metabolomic and metabonomic procedures for NMR spectroscopy of urine, plasma, serum and tissue extracts. Nat Protoc 2(11):2692–2703. CrossRefGoogle Scholar
  17. 17.
    Craig A, Cloarec O, Holmes E, Nicholson JK, Lindon JC (2006) Scaling and normalization effects in NMR spectroscopic metabonomic data sets. Anal Chem 78(7):2262–2267. CrossRefGoogle Scholar
  18. 18.
    Pears MR, Cooper JD, Mitchison HM, Mortishire-Smith RJ, Pearce DA, Griffin JL (2005) High resolution 1H NMR-based metabolomics indicates a neurotransmitter cycling deficit in cerebral tissue from a mouse model of Batten disease. J Biol Chem 280(52):42508–42514. CrossRefGoogle Scholar
  19. 19.
    Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J Roy Stat Soc Ser B (Methodol) 57(1):289–300Google Scholar
  20. 20.
    Dok-Go H, Lee KH, Kim HJ, Lee EH, Lee J, Song YS, Lee Y-H, Jin C, Lee YS, Cho J (2003) Neuroprotective effects of antioxidative flavonoids, quercetin, (+)-dihydroquercetin and quercetin 3-methyl ether, isolated from Opuntia ficus-indica var. saboten. Brain Res 965(1):130–136. CrossRefGoogle Scholar
  21. 21.
    Marklund S, Marklund G (1974) Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47(3):469–474. CrossRefGoogle Scholar
  22. 22.
    Smirnoff N, Cumbes QJ (1989) Hydroxyl radical scavenging activity of compatible solutes. Phytochemistry 28(4):1057–1060. CrossRefGoogle Scholar
  23. 23.
    Shinomol GK, Muralidhara (2007) Differential induction of oxidative impairments in brain regions of male mice following subchronic consumption of Khesari dhal (Lathyrus sativus) and detoxified Khesari dhal. Neurotoxicology 28(4):798–806. CrossRefGoogle Scholar
  24. 24.
    Brown GD (2006) Dectin-1: a signalling non-TLR pattern-recognition receptor. Nat Rev Immunol 6(1):33–43. CrossRefGoogle Scholar
  25. 25.
    Sonis ST, Elting LS, Keefe D, Peterson DE, Schubert M, Hauer-Jensen M, Bekele BN, Raber-Durlacher J, Donnelly JP, Rubenstein EB, Mucositis Study Section of the Multinational Association for Supportive Care in C, International Society for Oral O (2004) Perspectives on cancer therapy-induced mucosal injury: pathogenesis, measurement, epidemiology, and consequences for patients. Cancer 100(9 Suppl):1995–2025. CrossRefGoogle Scholar
  26. 26.
    Chang CJ, Lin JF, Chang HH, Lee GA, Hung CF (2013) Lutein protects against methotrexate-induced and reactive oxygen species-mediated apoptotic cell injury of IEC-6 cells. PLoS One 8(9):e72553. CrossRefGoogle Scholar
  27. 27.
    Huang CC, Hsu PC, Hung YC, Liao YF, Liu CC, Hour CT, Kao MC, Tsay GJ, Hung HC, Liu GY (2005) Ornithine decarboxylase prevents methotrexate-induced apoptosis by reducing intracellular reactive oxygen species production. Apoptosis 10(4):895–907. CrossRefGoogle Scholar
  28. 28.
    Jahovic N, Çevik H, Şehirli AÖ, Yeğen BÇ, Şener G (2003) Melatonin prevents methotrexate-induced hepatorenal oxidative injury in rats. J Pineal Res 34(4):282–287. CrossRefGoogle Scholar
  29. 29.
    Sheehan D, Meade G, Foley VM, Dowd CA (2001) Structure, function and evolution of glutathione transferases: implications for classification of non-mammalian members of an ancient enzyme superfamily. Biochem J 360(Pt 1):1–16CrossRefGoogle Scholar
  30. 30.
    Fijlstra M, Schierbeek H, Voortman G, Dorst KY, van Goudoever JB, Rings EHHM, Tissing WJE (2012) Continuous enteral administration can enable normal amino acid absorption in rats with methotrexate-induced gastrointestinal mucositis. J Nutr 142(11):1983–1990. CrossRefGoogle Scholar
  31. 31.
    McHardy IH, Goudarzi M, Tong M, Ruegger PM, Schwager E, Weger JR, Graeber TG, Sonnenburg JL, Horvath S, Huttenhower C, McGovern DPB, Fornace AJ, Borneman J, Braun J (2013) Integrative analysis of the microbiome and metabolome of the human intestinal mucosal surface reveals exquisite inter-relationships. Microbiome 1(1):17. CrossRefGoogle Scholar
  32. 32.
    Stringer AM, Gibson RJ, Bowen JM, Logan RM, Ashton K, Yeoh ASJ, Al-Dasooqi N, Keefe DMK (2009) Irinotecan-induced mucositis manifesting as diarrhoea corresponds with an amended intestinal flora and mucin profile. Int J Exp Pathol 90(5):489–499. CrossRefGoogle Scholar
  33. 33.
    Stringer AM, Gibson RJ, Logan RM, Bowen JM, Yeoh ASJ, Hamilton J, Keefe DMK (2009) Gastrointestinal microflora and mucins may play a critical role in the development of 5-fluorouracil-induced gastrointestinal mucositis. Exp Biol Med 234(4):430–441. CrossRefGoogle Scholar
  34. 34.
    Zhou B, Xia X, Wang P, Chen S, Yu C, Huang R, Zhang R, Wang Y, Lu L, Yuan F, Tian Y, Fan Y, Zhang X, Shu Y, Zhang S, Bai D, Wu L, Xu H, Yang L (2018) Induction and amelioration of methotrexate-induced gastrointestinal toxicity are related to immune response and gut microbiota. EBioMed 33:122–133. CrossRefGoogle Scholar
  35. 35.
    Tilg H, Kaser A (2011) Gut microbiome, obesity, and metabolic dysfunction. J Clin Investig 121(6):2126–2132. CrossRefGoogle Scholar
  36. 36.
    Thorpe DW, Stringer AM, Gibson RJ (2013) Chemotherapy-induced mucositis: the role of the gastrointestinal microbiome and toll-like receptors. Exp Biol Med 238(1):1–6. CrossRefGoogle Scholar
  37. 37.
    Santaolalla R, Abreu MT (2012) Innate immunity in the small intestine. Curr Opin Gastroenterol 28(2):124–129. CrossRefGoogle Scholar
  38. 38.
    Rakoff-Nahoum S, Paglino J, Eslami-Varzaneh F, Edberg S, Medzhitov R (2004) Recognition of commensal microflora by toll-like receptors is required for intestinal homeostasis. Cell 118(2):229–241. CrossRefGoogle Scholar
  39. 39.
    Wong DV, Lima-Junior RC, Carvalho CB, Borges VF, Wanderley CW, Bem AX, Leite CA, Teixeira MA, Batista GL, Silva RL, Cunha TM, Brito GA, Almeida PR, Cunha FQ, Ribeiro RA (2015) The adaptor protein Myd88 is a key signaling molecule in the pathogenesis of irinotecan-induced intestinal mucositis. PLoS One 10(10):e0139985. CrossRefGoogle Scholar
  40. 40.
    Kaczmarek A, Brinkman BM, Heyndrickx L, Vandenabeele P, Krysko DV (2012) Severity of doxorubicin-induced small intestinal mucositis is regulated by the TLR-2 and TLR-9 pathways. J Pathol 226(4):598–608. CrossRefGoogle Scholar
  41. 41.
    Brown GD, Herre J, Williams DL, Willment JA, Marshall ASJ, Gordon S (2003) Dectin-1 mediates the biological effects of β-Glucans. J Exp Med 197(9):1119–1124. CrossRefGoogle Scholar
  42. 42.
    Taylor PR, Tsoni SV, Willment JA, Dennehy KM, Rosas M, Findon H, Haynes K, Steele C, Botto M, Gordon S, Brown GD (2007) Dectin-1 is required for β-glucan recognition and control of fungal infection. Nat Immunol 8:31–38. CrossRefGoogle Scholar
  43. 43.
    Brown GD, Taylor PR, Reid DM, Willment JA, Williams DL, Martinez-Pomares L, Wong SYC, Gordon S (2002) Dectin-1 is a major β-Glucan receptor on macrophages. J Exp Med 196(3):407–412. CrossRefGoogle Scholar
  44. 44.
    Underhill DM (2007) Collaboration between the innate immune receptors dectin-1, TLRs, and Nods. Immunol Rev 219(1):75–87. CrossRefGoogle Scholar
  45. 45.
    Sener G, Eksioglu-Demiralp E, Cetiner M, Ercan F, Yegen BC (2006) Beta-glucan ameliorates methotrexate-induced oxidative organ injury via its antioxidant and immunomodulatory effects. Eur J Pharmacol 542(1–3):170–178. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Yan Gao
    • 1
  • Qi Sun
    • 1
  • Xiao Yang
    • 1
  • Weiling Lu
    • 1
  • Yang Zhao
    • 1
  • Wenhao Ge
    • 1
  • Yunxia Yang
    • 1
  • Xi Xu
    • 1
  • Jianfa Zhang
    • 1
    Email author
  1. 1.Center for Molecular Metabolism, Nanjing University of Science and TechnologyNanjingPeople’s Republic of China

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