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


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.



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

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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

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