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Methylene Blue Mitigates Ethanol Exacerbation of Ischemia‐Reperfusion Injury of Cardiomyocytes and Brain Cells
Author(s) -
Ryou Myounggwi,
Winters Ali,
Robert Mallet,
Yang Shaohua
Publication year - 2015
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.29.1_supplement.635.4
Subject(s) - viability assay , ischemia , chemistry , reactive oxygen species , reperfusion injury , pharmacology , biochemistry , cell , biology , medicine
Ethanol (EtOH) intoxication is a major risk factor for myocardial and brain ischemia. Methylene blue (MB) could mitigate EtOH exacerbation of ischemia‐reperfusion injury. Methods HT22 cells, primary microvascular endothelial cells (MVEC), astrocytes and cardiomyocytes (CM) were treated with 100 mM EtOH ± 100 nM MB for 5 days prior to 4 h oxygen and glucose deprivation (OGD, 0.1% O 2 ) and 24 h reoxygenation. Cell viability was assayed with Calcein AM. FITC‐Dextran leakage and transendothelial electrical resistance evaluated integrity of an in vitro blood brain barrier (BBB) model composed of MVEC and astrocytes. Generation of reactive oxygen species (ROS) was monitored with mitosox stain and DCF‐DA assay. ATP concentration was measured by spectrophotometry. Aquaporin 4 and hypoxia inducible factor‐1α (HIF‐1α) contents were assessed by immunoblot. Results EtOH was severely detrimental to both cell viability and BBB integrity vs. OGD control. EtOH significantly increased ROS in a manner blunted by MB. ATP depletion during OGD in HT22 cells and CM was appreciably worsened by EtOH, but MB cotreatment restored ATP content to that of EtOH‐free OGD. EtOH decreased HIF‐1α and increased aquaporin 4. Conclusion EtOH increased the vulnerability of all four cell types to OGD‐reoxygenation insult. MB mitigated this EtOH toxicity by minimizing ROS generation, increasing ATP availability, and maintaining HIF‐1α, a critical factor for cellular adaptation to the ischemic environment.