Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
JCR Impact Factor (IF) – 2.1
5-Year Impact Factor – 2.2
Scopus CiteScore – 3.4 (CiteScore Tracker 3.4)
Index Copernicus  – 161.11; MEiN – 140 pts

ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
Periodicity – monthly

Download original text (EN)

Advances in Clinical and Experimental Medicine

2020, vol. 29, nr 5, May, p. 535–545

doi: 10.17219/acem/121008

Publication type: original article

Language: English

License: Creative Commons Attribution 3.0 Unported (CC BY 3.0)

Download citation:

  • BIBTEX (JabRef, Mendeley)
  • RIS (Papers, Reference Manager, RefWorks, Zotero)

Antifungal itraconazole ameliorates experimental autoimmune encephalomyelitis through a novel mechanism of action

Huifen Huang1,B,C,D,F, Xiaolin Tian2,B,C,F, Xiao Peng1,C,F, Liangtong Huang1,D,E,F, Lerong Mei1,C,F, Yanli Zhan1,D,F, Siying Chen1,B,F, Huihua Wu1,B,F, Guofang Wei1,D,E,F, Xueli Cai1,A,F

1 Department of Neurology, Lishui Hospital, Zhejiang University School of Medicine, China

2 Department of Rehabitation Medicine, Second Hospital of Tianjin Medical University, China


Background. Multiple sclerosis (MS) is an autoimmune disease characterized by a loss of myelin, limb disabilities and dysregulation of gene expression. Unfortunately, there still is no treatment to cure MS.
Objectives. To explore a novel way to treat MS using currently available antifungal drugs.
Material and Methods. We built an experimental autoimmune encephalomyelitis (EAE) model to mimic MS and tested the effect of an antifungal drug – itraconazole – on EAE by comparing it with a phosphate-buffered saline (PBS) control group. We assessed the animal limb deficits with Weaver’s scoring and used histology staining (including luxol fast blue (LFB) and hematoxylin & eosin (H&E) methods) to determine the demyelination in the spinal tissues. We also performed western blotting to quantify the expression changes of proteins related to endoplasmic reticulum (ER) stress response and apoptosis.
Results. The limb disabilities were greatly diminished and the demyelination in the spinal tissues of the EAE mice was mostly reduced following itraconazole treatment. The hyperactivation of the ER stress response and apoptosis pathway in EAE was also significantly diminished by the itraconazole treatment. In addition, the AMPK pathway was downregulated in EAE, its expression level bi-directionally affected the activity of the ER stress response, and its downregulation removed the beneficial effect of itraconazole.
Conclusion. Our study revealed a new method for treating MS using currently approved antifungal drugs.

Key words

apoptosis, EAE, itraconazole, ER stress, AMPK

References (41)

  1. Browne P, Chandraratna D, Angood C, et al. Atlas of Multiple Sclerosis 2013: A growing global problem with widespread inequity. Neurology. 2014;83(11):1022–1024.
  2. Goverman J. Autoimmune T cell responses in the central nervous system. Nat Rev Immunol. 2009;9(6):393–407.
  3. Constantinescu CS, Farooqi N, O’Brien K, Gran B. Experimental autoimmune encephalomyelitis (EAE) as a model for multiple sclerosis (MS). Br J Pharmacol. 2011;164(4):1079–1106.
  4. Rivers TM, Sprunt DH, Berry GP. Observations on attempts to produce acute disseminated encephalomyelitis in monkeys. J Exp Med. 1933;58(1):39–53.
  5. Farooqi N, Gran B, Constantinescu CS. Are current disease-modifying therapeutics in multiple sclerosis justified on the basis of studies in experimental autoimmune encephalomyelitis? J Neurochem. 2010;115(4):829–844.
  6. Han MH, Hwang SI, Roy DB, et al. Proteomic analysis of active multiple sclerosis lesions reveals therapeutic targets. Nature. 2008;451(7182):1076–1081.
  7. Steinman L. A molecular trio in relapse and remission in multiple sclerosis. Nat Rev Immunol. 2009;9(6):440–447.
  8. English C, Aloi JJ. New FDA-approved disease-modifying therapies for multiple sclerosis. Clin Ther. 2015;37(4):691–715.
  9. Kamarehei M, Kabudanian Ardestani S, Firouzi M, et al. Increased expression of endoplasmic reticulum stress-related caspase-12 and CHOP in the hippocampus of EAE mice. Brain Res Bull. 2019;147:174–182.
  10. Schroder M. Endoplasmic reticulum stress responses. Cell Mol Life Sci. 2008;65(6):862–894.
  11. Walter P, Ron D. The unfolded protein response: From stress pathway to homeostatic regulation. Science. 2011;334(6059):1081–1086.
  12. Haze K, Yoshida H, Yanagi H, Yura T, Mori K. Mammalian transcription factor ATF6 is synthesized as a transmembrane protein and activated by proteolysis in response to endoplasmic reticulum stress. Mol Biol Cell. 1999;10(11):3787–3799.
  13. Marciniak SJ, Yun CY, Oyadomari S, et al. CHOP induces death by promoting protein synthesis and oxidation in the stressed endoplasmic reticulum. Genes Dev. 2004;18(24):3066–3077.
  14. Reimold AM, Iwakoshi NN, Manis J, et al. Plasma cell differentiation requires the transcription factor XBP-1. Nature. 2001;412(6844):300–307.
  15. Benito-Leon J, Laurence M. The role of fungi in the etiology of multiple sclerosis. Front Neurol. 2017;8:535.
  16. Faergemann J. Treatment of sebopsoriasis with itraconazole. Mykosen. 1985;28(12):612–618.
  17. Boehncke WH, Schon MP. Psoriasis. Lancet. 2015;386(9997):983–994.
  18. Ahmad N, Mukhtar H. Cytochrome p450: A target for drug development for skin diseases. J Invest Dermatol. 2004;123(3):417–425.
  19. Denmeade SR, Isaacs JT. The SERCA pump as a therapeutic target: Making a “smart bomb” for prostate cancer. Cancer Biol Ther. 2005;4(1):14–22.
  20. Berard JL, Wolak K, Fournier S, David S. Characterization of relapsing-remitting and chronic forms of experimental autoimmune encephalomyelitis in C57BL/6 mice. Glia. 2010;58(4):434–445.
  21. Grames MS, Jackson KL, Dayton RD, Stanford JA, Klein RL. Methods and tips for intravenous administration of adeno-associated virus to rats and evaluation of central nervous system transduction. J Vis Exp. 2017(126):55994. doi:10.3791/55994
  22. Weaver A, Goncalves da Silva A, Nuttall RK, et al. An elevated matrix metalloproteinase (MMP) in an animal model of multiple sclerosis is protective by affecting Th1/Th2 polarization. FASEB J. 2005;19(12):1668–1670.
  23. Dasilva AG, Yong VW. Expression and regulation of matrix metalloproteinase-12 in experimental autoimmune encephalomyelitis and by bone marrow derived macrophages in vitro. J Neuroimmunol. 2008;199(1–2):24–34.
  24. Groves AK, Barnett SC, Franklin RJ, et al. Repair of demyelinated lesions by transplantation of purified O-2A progenitor cells. Nature. 1993;362(6419):453–455.
  25. Richter-Landsberg C, Heinrich M. OLN-93: A new permanent oligodendroglia cell line derived from primary rat brain glial cultures. J Neurosci Res. 1996;45(2):161–173.
  26. Prasad R, Giri S, Nath N, Singh I, Singh AK. 5-aminoimidazole-4-carboxamide-1-beta-4-ribofuranoside attenuates experimental autoimmune encephalomyelitis via modulation of endothelial-monocyte interaction. J Neurosci Res. 2006;84(3):614–625.
  27. Meares GP, Qin H, Liu Y, Holdbrooks AT, Benveniste EN. AMP-activated protein kinase restricts IFN-gamma signaling. J Immunol. 2013;190(1):372–380.
  28. Mangalam AK, Rattan R, Suhail H, et al. AMP-activated protein kinase suppresses autoimmune central nervous system disease by regulating M1-type macrophage-Th17 axis. J Immunol. 2016;197(3):747–760.
  29. Macchi B, Marino-Merlo F, Nocentini U, et al. Role of inflammation and apoptosis in multiple sclerosis: Comparative analysis between the periphery and the central nervous system. J Neuroimmunol. 2015;287:80–87.
  30. Mangano K, Nicoletti A, Patti F, et al. Variable effects of cyclophosphamide in rodent models of experimental allergic encephalomyelitis. Clin Exp Immunol. 2010;159(2):159–168.
  31. Bechtold DA, Kapoor R, Smith KJ. Axonal protection using flecainide in experimental autoimmune encephalomyelitis. Ann Neurol. 2004;55(5):607–616.
  32. Tabas I, Ron D. Integrating the mechanisms of apoptosis induced by endoplasmic reticulum stress. Nat Cell Biol. 2011;13(3):184–190.
  33. Carrasco DR, Sukhdeo K, Protopopova M, et al. The differentiation and stress response factor XBP-1 drives multiple myeloma pathogenesis. Cancer Cell. 2007;11(4):349–360.
  34. Wang M, Kaufman RJ. The impact of the endoplasmic reticulum protein-folding environment on cancer development. Nat Rev Cancer. 2014;14(9):581–597.
  35. Hetz C, Mollereau B. Disturbance of endoplasmic reticulum proteostasis in neurodegenerative diseases. Nat Rev Neurosci. 2014;15(4):233–249.
  36. Lin W, Lin Y, Li J, et al. Oligodendrocyte-specific activation of PERK signaling protects mice against experimental autoimmune encephalo­myelitis. J Neurosci. 2013;33(14):5980–5991.
  37. Paintlia AS, Paintlia MK, Mohan S, Singh AK, Singh I. AMP-activated protein kinase signaling protects oligodendrocytes that restore central nervous system functions in an experimental autoimmune encephalomyelitis model. Am J Pathol. 2013;183(2):526–541.
  38. Wang P, Xu TY, Guan YF, et al. Nicotinamide phosphoribosyltransferase protects against ischemic stroke through SIRT1-dependent adenosine monophosphate-activated kinase pathway. Ann Neurol. 2011;69(2):360–374.
  39. Mihaylova MM, Shaw RJ. The AMPK signalling pathway coordinates cell growth, autophagy and metabolism. Nat Cell Biol. 2011;13(9):1016–1023.
  40. Nath N, Khan M, Paintlia MK, Singh I, Hoda MN, Giri S. Metformin attenuated the autoimmune disease of the central nervous system in animal models of multiple sclerosis. J Immunol. 2009;182(12):8005–8014.
  41. Singh I, Samuvel DJ, Choi S, Saxena N, Singh AK, Won J. Combination therapy of lovastatin and AMP-activated protein kinase activator improves mitochondrial and peroxisomal functions and clinical disease in experimental autoimmune encephalomyelitis model. ­Immunology. 2018;154(3):434–451.