Advertisement

Molecular Medicine

, Volume 21, Issue 1, pp 420–429 | Cite as

Methylene Blue Reduces Acute Cerebral Ischemic Injury via the Induction of Mitophagy

  • Yao Di
  • Yun-Ling He
  • Tong Zhao
  • Xin Huang
  • Kui-Wu Wu
  • Shu-Hong Liu
  • Yong-Qi Zhao
  • Ming Fan
  • Li-Ying Wu
  • Ling-Ling Zhu
Research Article

Abstract

The treatment of stroke is limited by a short therapeutic window and a lack of effective clinical drugs. Methylene blue (MB) has been used in laboratories and clinics since the 1890s. Few studies have reported the neuroprotective role of MB in cerebral ischemia-reperfusion injury. However, whether and how MB protects against acute cerebral ischemia (ACI) injury was unclear. In this study, we investigated the effect of MB on this injury and revealed that MB protected against ACI injury by augmenting mitophagy. Using a rat middle cerebral artery occlusion (MCAO) model, we demonstrated that MB improved neurological function and reduced the infarct volume and necrosis after ACI injury. These improvements depended on the effect of MB on mitochondrial structure and function. ACI caused the disorder and disintegration of mitochondrial structure, while MB ameliorated the destruction of mitochondria. In addition, mitophagy was inhibited at 24 h after stroke and MB augmented mitophagy. In an oxygen-glucose deprivation (OGD) model in vitro, we further revealed that the elevation of mitochondrial membrane potential (MMP) by MB under OGD conditions mediated the augmented mitophagy. In contrast, exacerbating the decline of MMP during OGD abolished the MB-induced activation of mitophagy. Taken together, MB promotes mitophagy by maintaining the MMP at a relatively high level, which contributes to a decrease in necrosis and an improvement in neurological function, thereby protecting against ACI injury.

Notes

Acknowledgments

We are grateful to Chang-Hong Ren and Zhi-Feng Gao, Department of Hypoxia/Ischemia, Xuanwu Hospital, Capital Medical University, for their excellent technical assistance in preparing the rat MCAO model and analyzing the scores of neurological dysfunction. This work was supported by the National Basic Research Programs of China (2012CB518200, 2011CB910800), the National Natural Science Foundation of China (81071066),81000856 and 31271211), and the Integrated Drug Discovery Technology Platform of National Science and Technology Major Projects for “Major New Drugs Innovation and Development” (2012ZX09J12201-005).

Supplementary material

10020_2015_2101420_MOESM1_ESM.pdf (2.4 mb)
Supplementary material, approximately 2457 KB.

References

  1. 1.
    Wainwright M, Crossley KB. (2002) Methylene Blue — a therapeutic dye for all seasons? J. Chemother. 14:431–43.CrossRefGoogle Scholar
  2. 2.
    Tretter L, Horvath G, Holgyesi A, Essek F, Adam-Vizi V. (2014) Enhanced hydrogen peroxide generation accompanies the beneficial bioenergetic effects of methylene blue in isolated brain mitochondria. Free Radic. Biol. Med. 77:317–30.CrossRefGoogle Scholar
  3. 3.
    Oz M, Lorke DE, Hasan M, Petroianu GA. (2011) Cellular and molecular actions of Methylene Blue in the nervous system. Med. Res. Rev. 31:93–117.CrossRefGoogle Scholar
  4. 4.
    Callaway NL, Riha PD, Bruchey AK, Munshi Z, Gonzalez-Lima F. (2004) Methylene blue improves brain oxidative metabolism and memory retention in rats. Pharmacol. Biochem. Behav. 77:175–81.CrossRefGoogle Scholar
  5. 5.
    Zhang X, Rojas JC, Gonzalez-Lima F. (2006) Methylene blue prevents neurodegeneration caused by rotenone in the retina. Neurotox. Res. 9:47–57.CrossRefGoogle Scholar
  6. 6.
    Riha PD, Bruchey AK, Echevarria DJ, Gonzalez-Lima F. (2005) Memory facilitation by methylene blue: dose-dependent effect on behavior and brain oxygen consumption. Eur. J. Pharmacol. 511:151–8.CrossRefGoogle Scholar
  7. 7.
    Rojas JC, Bruchey AK, Gonzalez-Lima F. (2012) Neurometabolic mechanisms for memory enhancement and neuroprotection of methylene blue. Prog. Neurobiol 96:32–45.CrossRefGoogle Scholar
  8. 8.
    Kelner MJ, Bagnell R, Hale B, Alexander NM. (1988) Potential of methylene blue to block oxygen radical generation in reperfusion injury. Basic Life Sci. 49:895–8.PubMedGoogle Scholar
  9. 9.
    Cawein M, Behlen CH 2nd, Lappat EJ, Cohn JE. (1964) Hereditary diaphorase deficiency and methemoglobinemia. Arch Intern Med 113:578–85.CrossRefGoogle Scholar
  10. 10.
    Draize JH. (1933) Sodium tetrathionate and methylene blue in cyanide and carbon monoxide poisoning. Science. 78:145.CrossRefGoogle Scholar
  11. 11.
    Wen Y, et al. (2011) Alternative mitochondrial electron transfer as a novel strategy for neuroprotection. J. Biol. Chem. 286:16504–15.CrossRefGoogle Scholar
  12. 12.
    Miclescu A, Sharma HS, Martijn C, Wiklund L. (2010) Methylene blue protects the cortical blood-brain barrier against ischemia/reperfusion-induced disruptions. Crit. Care Med. 38:2199–206.CrossRefGoogle Scholar
  13. 13.
    Shen Q, et al. (2013) Neuroprotective efficacy of methylene blue in ischemic stroke: an MRI study. PLoS One. 8:e79833.CrossRefGoogle Scholar
  14. 14.
    Gropen TI, et al. (2006) Quality improvement in acute stroke: the New York State Stroke Center Designation Project. Neurology. 67:88–93.CrossRefGoogle Scholar
  15. 15.
    Ertracht O, Malka A, Atar S, Binah O. (2014) The mitochondria as a target for cardioprotection in acute myocardial ischemia. Pharmacol. Ther. 142:33–40.CrossRefGoogle Scholar
  16. 16.
    Kim SY, et al. (2014) Inhibition of cyclophilin D by cyclosporin A promotes retinal ganglion cell survival by preventing mitochondrial alteration in ischemic injury. Cell Death Dis. 5:e1105.CrossRefGoogle Scholar
  17. 17.
    Ikeda Y, et al. (2015) Endogenous drp1 mediates mitochondrial autophagy and protects the heart against energy stress. Circ. Res. 116:264–78.CrossRefGoogle Scholar
  18. 18.
    Lemasters JJ. (2005) Selective mitochondrial autophagy, or mitophagy, as a targeted defense against oxidative stress, mitochondrial dysfunction, and aging. Rejuvenation Res. 8:3–5.CrossRefGoogle Scholar
  19. 19.
    Youle RJ, Narendra DP. (2011) Mechanisms of mitophagy. Nat. Rev. Mol. Cell Biol. 12:9–14.CrossRefGoogle Scholar
  20. 20.
    Ashrafi G, Schlehe JS, LaVoie MJ, Schwarz TL. (2014) Mitophagy of damaged mitochondria occurs locally in distal neuronal axons and requires PINK1 and Parkin. J. Cell Biol. 206:655–70.CrossRefGoogle Scholar
  21. 21.
    Liu L, et al. (2012) Mitochondrial outer-membrane protein FUNDC1 mediates hypoxia-induced mitophagy in mammalian cells. Nat. Cell Biol. 14:177–85.CrossRefGoogle Scholar
  22. 22.
    Kim I, Lemasters JJ. (2011) Mitophagy selectively degrades individual damaged mitochondria after photoirradiation. Antioxid. Redox. Signal. 14:1919–28.CrossRefGoogle Scholar
  23. 23.
    Wu H, et al. (2014) The BCL2L1 and PGAM5 axis defines hypoxia-induced receptor-mediated mitophagy. Autophagy. 10:1712–25.CrossRefGoogle Scholar
  24. 24.
    Kurihara Y, et al. (2012) Mitophagy plays an essential role in reducing mitochondrial production of reactive oxygen species and mutation of mitochondrial DNA by maintaining mitochondrial quantity and quality in yeast. J. Biol. Chem. 287:3265–72.CrossRefGoogle Scholar
  25. 25.
    Bin-Umer MA, McLaughlin JE, Butterly MS, McCormick S, Tumer NE. (2014) Elimination of damaged mitochondria through mitophagy reduces mitochondrial oxidative stress and increases tolerance to trichothecenes. Proc. Natl. Acad. Sci. U. S. A. 111:11798–803.CrossRefGoogle Scholar
  26. 26.
    Schapira AH, Olanow CW, Greenamyre JT, Bezard E. (2014) Slowing of neurodegeneration in Parkinson’s disease and Huntington’s disease: future therapeutic perspectives. Lancet 384:545–55.CrossRefGoogle Scholar
  27. 27.
    Shiba-Fukushima K, et al. (2014) Phosphorylation of mitochondrial polyubiquitin by PINK1 promotes parkin mitochondrial tethering. PLoS. Genet. 10:e1004861.CrossRefGoogle Scholar
  28. 28.
    Santos RX, et al. (2011) Mitophagy in neurodegeneration: an opportunity for therapy? Curr. Drug Targets. 12:790–9.CrossRefGoogle Scholar
  29. 29.
    Rubinsztein DC, Marino G, Kroemer G. (2011) Autophagy and aging. Cell. 146:682–95.CrossRefGoogle Scholar
  30. 30.
    Martinez-Vicente M, et al. (2010) Cargo recognition failure is responsible for inefficient autophagy in Huntington’s disease. Nat. Neurosci. 13:567–76.CrossRefGoogle Scholar
  31. 31.
    Lemasters JJ, et al. (1998) The mitochondrial permeability transition in cell death: a common mechanism in necrosis, apoptosis and autophagy. Biochim. Biophys. Acta. 1366:177–96.CrossRefGoogle Scholar
  32. 32.
    Hollville E, Carroll RG, Cullen SP, Martin SJ. (2014) Bcl-2 family proteins participate in mitochondrial quality control by regulating Parkin/PINK1-dependent mitophagy. Mol. Cell. 55:451–66.CrossRefGoogle Scholar
  33. 33.
    Strappazzon F, et al. (2015) AMBRA1 is able to induce mitophagy via LC3 binding, regardless of PARKIN and p62/SQSTM1. Cell Death Differ. 22:419–32.CrossRefGoogle Scholar
  34. 34.
    Zhang JH, et al. (2015) 5-HMF prevents against oxidative injury via APE/Ref-1. Free Radic. Res. 49:86–94.CrossRefGoogle Scholar
  35. 35.
    Belayev L, Alonso OF, Busto R, Zhao W, Ginsberg MD. (1996) Middle cerebral artery occlusion in the rat by intraluminal suture. Neurological and pathological evaluation of an improved model. Stroke. 27:1616–22; discussion 1623.CrossRefGoogle Scholar
  36. 36.
    Baskin YK, Dietrich WD, Green EJ. (2003) Two effective behavioral tasks for evaluating sensorimotor dysfunction following traumatic brain injury in mice. J. Neurosci. Methods. 129:87–93.CrossRefGoogle Scholar
  37. 37.
    Lin TN, He YY, Wu G, Khan M, Hsu CY. (1993) Effect of brain edema on infarct volume in a focal cerebral ischemia model in rats. Stroke 24:117–21.CrossRefGoogle Scholar
  38. 38.
    Liu H, et al. (2015) Targeting heat-shock protein 90 with ganetespib for molecularly targeted therapy of gastric cancer. Cell Death Dis. 6:e1595.CrossRefGoogle Scholar
  39. 39.
    Back SA, et al. (2007) Hypoxia-ischemia preferentially triggers glutamate depletion from oligodendroglia and axons in perinatal cerebral white matter. J. Cereb. Blood Flow Metab. 27:334–47.CrossRefGoogle Scholar
  40. 40.
    Flameng W, Borgers M, Daenen W, Stalpaert G. (1980) Ultrastructural and cytochemical correlates of myocardial protection by cardiac hypothermia in man. J. Thorac. Cardiovasc. Surg. 79:413–24.PubMedGoogle Scholar
  41. 41.
    Li Y, et al. (2011) Spatiotemporal pattern of neuronal injury induced by DFP in rats: a model for delayed neuronal cell death following acute OP intoxication. Toxicol. Appl. Pharmacol. 253:261–9.CrossRefGoogle Scholar
  42. 42.
    Cheng X, et al. (2014) A TrxR inhibiting gold(I) NHC complex induces apoptosis through ASK1-p38-MAPK signaling in pancreatic cancer cells. Mol. Cancer. 13:221.CrossRefGoogle Scholar
  43. 43.
    Moskowitz MA, Lo EH, Iadecola C. (2010) The science of stroke: mechanisms in search of treatments. Neuron. 67:181–98.CrossRefGoogle Scholar
  44. 44.
    Narayan N, et al. (2012) The NAD-dependent deacetylase SIRT2 is required for programmed necrosis. Nature. 492:199–204.CrossRefGoogle Scholar
  45. 45.
    Wang Z, Jiang H, Chen S, Du F, Wang X. (2012) The mitochondrial phosphatase PGAM5 functions at the convergence point of multiple necrotic death pathways. Cell. 148:228–43.CrossRefGoogle Scholar
  46. 46.
    Singhal A, Morris VB, Labhasetwar V, Ghorpade A. (2013) Nanoparticle-mediated catalase delivery protects human neurons from oxidative stress. Cell Death Dis. 4:e903.CrossRefGoogle Scholar
  47. 47.
    Poteet E, et al. (2012) Neuroprotective actions of methylene blue and its derivatives. PLoS One. 7:e48279.CrossRefGoogle Scholar
  48. 48.
    Lopez-Valdes HE, et al. (2014) Memantine enhances recovery from stroke. Stroke. 45:2093–100.CrossRefGoogle Scholar
  49. 49.
    Coulibaly B, et al. (2015) Efficacy and safety of triple combination therapy with artesunate-amodiaquine-methylene blue for falciparum malaria in children: a randomized controlled trial in burkina faso. J. Infect. Dis. 211:689–697.CrossRefGoogle Scholar
  50. 50.
    Congdon EE, et al. (2012) Methylthioninium chloride (methylene blue) induces autophagy and attenuates tauopathy in vitro and in vivo. Autophagy. 8:609–22.CrossRefGoogle Scholar
  51. 51.
    Kubli DA, Gustafsson AB. (2012) Mitochondria and mitophagy: the yin and yang of cell death control. Circ. Res. 111:1208–21.CrossRefGoogle Scholar
  52. 52.
    Bagkos G, Koufopoulos K, Piperi C. (2014) A new model for mitochondrial membrane potential production and storage. Med. Hypotheses. 83:175–81.CrossRefGoogle Scholar
  53. 53.
    Gabrielli D, Belisle E, Severino D, Kowaltowski AJ, Baptista MS. (2004) Binding, aggregation and photochemical properties of methylene blue in mitochondrial suspensions. Photochem. Photobiol. 79:227–32.CrossRefGoogle Scholar

Copyright information

© The Author(s) 2015

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, and provide a link to the Creative Commons license. You do not have permission under this license to share adapted material derived from this article or parts of it.

The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this license, visit (https://doi.org/creativecommons.org/licenses/by-nc-nd/4.0/)

Authors and Affiliations

  • Yao Di
    • 1
  • Yun-Ling He
    • 1
  • Tong Zhao
    • 1
  • Xin Huang
    • 1
  • Kui-Wu Wu
    • 1
  • Shu-Hong Liu
    • 1
  • Yong-Qi Zhao
    • 1
  • Ming Fan
    • 1
    • 2
    • 3
  • Li-Ying Wu
    • 1
  • Ling-Ling Zhu
    • 1
    • 2
  1. 1.Department of Cognitive ScienceBeijing Institute of Basic Medical SciencesBeijingPeople’s Republic of China
  2. 2.Co-innovation Center of NeuroregenerationNantong UniversityNantongPeople’s Republic of China
  3. 3.Beijing Institute for Brain DisordersBeijingPeople’s Republic of China

Personalised recommendations