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Neuroprotective Mechanisms of Methylene Blue
Author(s) -
Ryou MyoungGwi,
Choudhury Gourav Roy,
Mallet Robert T,
Yang Shaohua
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.691.9
Subject(s) - protein kinase b , pi3k/akt/mtor pathway , viability assay , chemistry , methylene blue , erythropoietin , phosphorylation , neuroprotection , apoptosis , microbiology and biotechnology , pharmacology , biochemistry , biology , endocrinology , photocatalysis , catalysis
Background The phenothiazine dye methylene blue (MB) has antioxidant properties and augments ATP production, and is selectively internalized by neurons after systemic injection into rats. Hypothesis MB protects neurons against hypoxia‐reoxygenation (H/R) injury by enhancing ATP production and activating the Akt/mTOR signaling pathway. Methods HT22 cells were subjected to 3h oxygen‐glucose deprivation (OGD; 0.5% O 2 ) followed by 24 h reoxygenation (H/R protocol). Cell viability (Calcein AM), and contents of hypoxia inducible factor‐1α, Akt/mTOR pathway components, and actin (immunoblot), erythropoietin (EPO; immunocytochemistry) and ATP content (microplate assay) were analyzed. P values <0.05 were considered statistically different. Results OGD reduced cell viability by 40% vs . normoxia, but 50 and 100 nM MB prevented H/R induced cell death. MB (10–1000 nM) concentration‐dependently increased cellular ATP content, in parallel with increased HIF‐1α content and EPO expression. MB (500 μM) increased Akt phosphorylation fourfold and mTOR phosphorylation by 45% vs. H/R control. Conclusion MB protects HT22 cells by both enhancing ATP production and activating EPO's anti‐apoptotic signaling.