J Appl Biomed 17:107-114, 2019 | DOI: 10.32725/jab.2019.008

Methylene blue elicits non-genotoxic H2O2 production and protects brain mitochondria from rotenone toxicity

Artem P. Gureev1,*, Ekaterina A. Shaforostova1, Denis A. Laver1, Victoria G. Khorolskaya1, Mikhail Yu. Syromyatnikov1, Vasily N. Popov1,2
1 Voronezh State University, Faculty of Medicine and Biology, Department of Genetics, Cytology and Bioengineering, Voronezh, Russia
2 Voronezh State University of Engineering Technologies, Voronezh, Russia

Methylene blue (MB) is a promising compound with a broad range of neuroprotective activity. One of therapeutic effects is the activation of mitochondrial biogenesis via Nrf2/ARE signaling cascade. Probably, mild oxidative stress caused by MB-depended H2O2 production is a trigger for activation of this signaling cascade. So mechanistically, MB can be regarded as prooxidant. We investigated the dose-dependent H2O2 production in intact brain mitochondria and showed the increase in the H2O2 production after adding as little as 50 nM MB. We have not found genotoxic effect of therapeutic concentration of MB to mitochondrial genome. 100 μM MB selectively damaged fragments of mitochondrial DNA, which correlated with the number of purine-T-G-purine (RTGR)-sequences in studied fragments. Furthermore, 20 μM MB combined with the red light caused the formation of singlet oxygen, which strongly damaged mitochondrial DNA in all studied fragments. We did not observe mitochondrial DNA lesions in brain after single intraperitoneal injection of MB in the concentration of 50 mg/kg. Furthermore, we showed the neuroprotective properties of MB pretreatments after rotenone injection. Therefore, we suggest that MB-induced mild oxidative stress does not have genotoxic effect on mitochondrial DNA.

Keywords: DNA damage; Methylene blue; mtDNA; Neuroprotection; Rotenone; Toxicity
Grants and funding:

This work was supported in part by the grant of the President of the Russian Federation for young scientists (Project MK3173.2019.11); Ministry of Education and Science of the Russian Federation (State Assessment N 6.4656.2017/8.9); Russian Fund for Basic Research (grant 19-44-360011 r_a).

Conflicts of interest:

The authors confirm that the study causes no conflict of interests as related to funding, commercial activity, intellectual property rights, or other conceivable issues.

Received: January 10, 2018; Accepted: May 14, 2019; Prepublished online: June 17, 2019; Published: June 28, 2019  Show citation

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Gureev AP, Shaforostova EA, Laver DA, Khorolskaya VG, Syromyatnikov MY, Popov VN. Methylene blue elicits non-genotoxic H2O2 production and protects brain mitochondria from rotenone toxicity. J Appl Biomed. 2019;17(2):107-114. doi: 10.32725/jab.2019.008. PubMed PMID: 34907732.
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