title: Effects of Ranolazine on Astrocytes and Neurons in Primary Culture.
abstract: Ranolazine (Rn) is an antianginal agent used for the treatment of chronic angina pectoris when angina is not adequately controlled by other drugs. Rn also acts in the central nervous system and it has been proposed for the treatment of pain and epileptic disorders. Under the hypothesis that ranolazine could act as a neuroprotective drug, we studied its effects on astrocytes and neurons in primary culture. We incubated rat astrocytes and neurons in primary cultures for 24 hours with Rn (10-7, 10-6 and 10-5 M). Cell viability and proliferation were measured using trypan blue exclusion assay, MTT conversion assay and LDH release assay. Apoptosis was determined by Caspase 3 activity assay. The effects of Rn on pro-inflammatory mediators IL-β and TNF-α was determined by ELISA technique, and protein expression levels of Smac/Diablo, PPAR-γ, Mn-SOD and Cu/Zn-SOD by western blot technique. In cultured astrocytes, Rn significantly increased cell viability and proliferation at any concentration tested, and decreased LDH leakage, Smac/Diablo expression and Caspase 3 activity indicating less cell death. Rn also increased anti-inflammatory PPAR-γ protein expression and reduced pro-inflammatory proteins IL-1 β and TNFα levels. Furthermore, antioxidant proteins Cu/Zn-SOD and Mn-SOD significantly increased after Rn addition in cultured astrocytes. Conversely, Rn did not exert any effect on cultured neurons. In conclusion, Rn could act as a neuroprotective drug in the central nervous system by promoting astrocyte viability, preventing necrosis and apoptosis, inhibiting inflammatory phenomena and inducing anti-inflammatory and antioxidant agents.
authors.affiliation: Department of Physiology, School of Medicine, University of Valencia, Spain.
authors.forename: Martin
authors.initials: M
authors.lastname: Aldasoro
authors.affiliation: Department of Physiology, School of Medicine, University of Valencia, Spain.
authors.forename: Sol
authors.initials: S
authors.lastname: Guerra-Ojeda
authors.affiliation: Department of Physiology, School of Medicine, University of Valencia, Spain.
authors.forename: Diana
authors.initials: D
authors.lastname: Aguirre-Rueda
authors.affiliation: Department of Physiology, School of Medicine, University of Valencia, Spain.
authors.forename: M Dolores
authors.initials: MD
authors.lastname: Mauricio
authors.affiliation: Department of Physiology, School of Medicine, University of Valencia, Spain.
authors.forename: Jose M
authors.initials: JM
authors.lastname: Vila
authors.affiliation: Department of Physiology, School of Medicine, University of Valencia, Spain.
authors.forename: Patricia
authors.initials: P
authors.lastname: Marchio
authors.affiliation: Department of Physiology, School of Medicine, University of Valencia, Spain.
authors.forename: Antonio
authors.initials: A
authors.lastname: Iradi
authors.affiliation: Department of Physiology, School of Medicine, University of Valencia, Spain.
authors.forename: Constanza
authors.initials: C
authors.lastname: Aldasoro
authors.affiliation: Department of Physiology, School of Medicine, University of Valencia, Spain.
authors.forename: Adrian
authors.initials: A
authors.lastname: Jorda
authors.affiliation: Department of Physiology, School of Medicine, University of Valencia, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Obrador
authors.affiliation: Department of Physiology, School of Medicine, University of Valencia, Spain.
authors.forename: Soraya L
authors.initials: SL
authors.lastname: Valles
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D051017:Apoptosis Regulatory Proteins; D001253:Astrocytes / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D053583:Interleukin-1beta / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D047495:PPAR gamma / Q000378:metabolism; D000069458:Ranolazine / Q000494:pharmacology*; D051381:Rats; D013482:Superoxide Dismutase / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; D002352:Carrier Proteins; C489013:DIABLO protein, rat; D053583:Interleukin-1beta; D024101:Mitochondrial Proteins; D047495:PPAR gamma; D014409:Tumor Necrosis Factor-alpha; D000069458:Ranolazine; D013482:Superoxide Dismutase; D053148:Caspase 3