Dimethyl Sulfoxide Damages Mitochondrial Integrity and Membrane Potential in Cultured Astrocytes

Dimethyl sulfoxide (DMSO) is a polar organic solvent that is used to dissolve neuroprotective or neurotoxic agents in neuroscience research. However, DMSO itself also has pharmacological and pathological effects on the nervous system. Astrocytes play a central role in maintaining brain homeostasis, but the effect and mechanism of DMSO on astrocytes has not been studied. The present study showed that exposure of astrocyte cultures to 1% DMSO for 24 h did not significantly affect cell survival, but decreased cell viability and glial glutamate transporter expression, and caused mitochondrial swelling, membrane potential impairment and reactive oxygen species production, and subsequent cytochrome c release and caspase-3 activation. DMSO at concentrations of 5% significantly inhibited cell variability and promoted apoptosis of astrocytes, accompanied with more severe mitochondrial damage. These results suggest that mitochondrial impairment is a primary event in DMSO-induced astrocyte toxicity. The potential cytotoxic effects on astrocytes need to be carefully considered during investigating neuroprotective or neurotoxic effects of hydrophobic agents dissolved by DMSO.

[1]  F. Russo,et al.  DMSO-associated encephalopathy during autologous peripheral stem cell infusion: a predisposing role of preconditioning exposure to CNS-penetrating agents? , 2009, Bone Marrow Transplantation.

[2]  P. Pinton,et al.  Relation between mitochondrial membrane potential and ROS formation. , 2012, Methods in molecular biology.

[3]  J. C. de la Torre,et al.  Pharmacology of dimethyl sulfoxide in cardiac and CNS damage , 2009, Pharmacological reports : PR.

[4]  S. Goldberg,et al.  Reversible encephalopathy after cryopreserved peripheral blood stem cell infusion , 1994, American journal of hematology.

[5]  Nicola J. Allen,et al.  Neuroscience: Glia — more than just brain glue , 2009, Nature.

[6]  V. Borutaite Mitochondria as decision‐makers in cell death , 2010, Environmental and molecular mutagenesis.

[7]  S. Hébert,et al.  Dimethyl Sulfoxide Induces Both Direct and Indirect Tau Hyperphosphorylation , 2012, PloS one.

[8]  B. Zhivotovsky,et al.  Multiple pathways of cytochrome c release from mitochondria in apoptosis. , 2006, Biochimica et biophysica acta.

[9]  B. Chazotte Labeling mitochondria with TMRM or TMRE. , 2011, Cold Spring Harbor protocols.

[10]  P Windrum,et al.  Severe neurotoxicity because of dimethyl sulphoxide following peripheral blood stem cell transplantation , 2003, Bone Marrow Transplantation.

[11]  M. Mattson,et al.  Dimethyl Sulfoxide Suppresses NMDA- and AMPA-Induced Ion Currents and Calcium Influx and Protects against Excitotoxic Death in Hippocampal Neurons , 2001, Experimental Neurology.

[12]  L. Galluzzi,et al.  Mechanisms of cytochrome c release from mitochondria , 2006, Cell Death and Differentiation.

[13]  G. Cosnard,et al.  Recurrent seizure and sustained encephalopathy associated with dimethylsulfoxide-preserved stem cell infusion , 2005, Leukemia & lymphoma.

[14]  B. Lyeth,et al.  Dimethyl sulfoxide provides neuroprotection in a traumatic brain injury model. , 2008, Restorative neurology and neuroscience.

[15]  Sten Orrenius,et al.  Cytochrome c release from mitochondria proceeds by a two-step process , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[16]  Christian Steinhäuser,et al.  Astrocyte dysfunction in neurological disorders: a molecular perspective , 2006, Nature Reviews Neuroscience.

[17]  Michael P. Murphy,et al.  How mitochondria produce reactive oxygen species , 2008, The Biochemical journal.

[18]  F. H. Lopes da Silva,et al.  Glutamate transporters alterations in the reorganizing dentate gyrus are associated with progressive seizure activity in chronic epileptic rats , 2002, The Journal of comparative neurology.

[19]  R. Simon,et al.  Dimethylsulfoxide (DMSO) treatment reduces infarction volume after permanent focal cerebral ischemia in rats , 1997, Neuroscience Letters.

[20]  Jamshed Anwar,et al.  Modulating the structure and properties of cell membranes: the molecular mechanism of action of dimethyl sulfoxide. , 2007, The journal of physical chemistry. B.

[21]  G. Behre,et al.  Neurotoxicity upon infusion of dimethylsulfoxide‐cryopreserved peripheral blood stem cells in patients with and without pre‐existing cerebral disease , 2007, European journal of haematology.

[22]  S. Orrenius,et al.  Triggering and modulation of apoptosis by oxidative stress. , 2000, Free radical biology & medicine.

[23]  N. Beauchamp,et al.  Reversible leukoencephalopathy associated with re-infusion of DMSO preserved stem cells , 2000, Bone Marrow Transplantation.

[24]  G. Cavaletti,et al.  Effect on the peripheral nervous system of systemically administered dimethylsulfoxide in the rat: a neurophysiological and pathological study. , 2000, Toxicology letters.

[25]  P. Quinn,et al.  Solvation effects of dimethyl sulphoxide on the structure of phospholipid bilayers. , 1998, Biophysical chemistry.

[26]  A. Volterra,et al.  Glutamate transporters are oxidant-vulnerable: a molecular link between oxidative and excitotoxic neurodegeneration? , 1998, Trends in pharmacological sciences.

[27]  Carlota Saldanha,et al.  Multidisciplinary utilization of dimethyl sulfoxide: pharmacological, cellular, and molecular aspects. , 2003, Biochemical pharmacology.

[28]  Y. Tsujimoto,et al.  Role of the mitochondrial membrane permeability transition in cell death , 2007, Apoptosis.

[29]  M. Robinson,et al.  The role of glutamate transporters in neurodegenerative diseases and potential opportunities for intervention , 2007, Neurochemistry International.

[30]  E. Eruslanov,et al.  Identification of ROS using oxidized DCFDA and flow-cytometry. , 2010, Methods in molecular biology.

[31]  J. Phillis,et al.  Protective effects of the free radical scavengers, dimethyl sulfoxide and ethanol, in cerebral ischemia in gerbils , 1998, Neuroscience Letters.

[32]  I. Tetko,et al.  In silico approaches to prediction of aqueous and DMSO solubility of drug-like compounds: trends, problems and solutions. , 2006, Current medicinal chemistry.

[33]  G. Juhász,et al.  The effect of intraperitoneally administered dimethyl sulfoxide on absence-like epileptic activity of freely moving WAG/Rij rats , 2011, Journal of Neuroscience Methods.

[34]  J. Meldolesi,et al.  Astrocytes, from brain glue to communication elements: the revolution continues , 2005, Nature Reviews Neuroscience.

[35]  S. Sürücü,et al.  How DMSO, a widely used solvent, affects spinal cord injury. , 2008, Annals of vascular surgery.

[36]  R. Tapia Release and uptake of glutamate as related to excitotoxicity. , 1996, Revista brasileira de biologia.

[37]  R. Saboya,et al.  Neurotoxicity associated with dimethylsulfoxide-preserved hematopoietic progenitor cell infusion , 2008, Bone Marrow Transplantation.

[38]  P. Fisher,et al.  Role of Excitatory Amino Acid Transporter‐2 (EAAT2) and glutamate in neurodegeneration: Opportunities for developing novel therapeutics , 2011, Journal of cellular physiology.

[39]  Jie Ding,et al.  Phenolic Alkaloids from Menispermum dauricum Rhizome Protect against Brain Ischemia Injury via Regulation of GLT-1, EAAC1 and ROS Generation , 2012, Molecules.

[40]  J. Olney,et al.  Dimethyl sulfoxide (DMSO) produces widespread apoptosis in the developing central nervous system , 2009, Neurobiology of Disease.

[41]  M. Morgan,et al.  Microinjection of the vehicle dimethyl sulfoxide (DMSO) into the periaqueductal gray modulates morphine antinociception , 2008, Brain Research.

[42]  A. Eschalier,et al.  Behavioural assessment of dimethylsulfoxide neurotoxicity in rats. , 2002, Toxicology letters.

[43]  J. Rothstein,et al.  Traumatic Brain Injury Down‐Regulates Glial Glutamate Transporter (GLT‐1 and GLAST) Proteins in Rat Brain , 1998, Journal of Neurochemistry.

[44]  A. Colquhoun,et al.  Mitochondrial Swelling and Incipient Outer Membrane Rupture in Preapoptotic and Apoptotic Cells , 2012, Anatomical record.

[45]  G. Hu,et al.  Aquaporin-4 deficiency down-regulates glutamate uptake and GLT-1 expression in astrocytes , 2007, Molecular and Cellular Neuroscience.

[46]  S. Orrenius Mitochondrial regulation of apoptotic cell death. , 2003, Toxicology letters.

[47]  A. Beydoun,et al.  Transient global amnesia associated with the infusion of DMSO-cryopreserved autologous peripheral blood stem cells , 2008, Haematologica.

[48]  Jamshed Anwar,et al.  Molecular basis for dimethylsulfoxide (DMSO) action on lipid membranes. , 2006, Journal of the American Chemical Society.

[49]  K. Capriotti,et al.  Dimethyl sulfoxide: history, chemistry, and clinical utility in dermatology. , 2012, The Journal of clinical and aesthetic dermatology.

[50]  J. McCabe,et al.  Short and long-term motor and behavioral effects of diazoxide and dimethyl sulfoxide administration in the mouse after traumatic brain injury , 2013, Pharmacology Biochemistry and Behavior.

[51]  I. Hwang,et al.  Oxidative DNA damage and alteration of glutamate transporter expressions in the hippocampal Ca1 area immediately after ischemic insult. , 2002, Molecules and cells.