title: Neuroprotection by JM-20 against oxygen-glucose deprivation in rat hippocampal slices: Involvement of the Akt/GSK-3β pathway.
abstract: Cerebral ischemia is the third most common cause of death and a major cause of disability worldwide. Beyond a shortage of essential metabolites, ischemia triggers many interconnected pathophysiological events, including excitotoxicity, oxidative stress, inflammation and apoptosis. Here, we investigated the neuroprotective mechanisms of JM-20, a novel synthetic molecule, focusing on the phosphoinositide-3-kinase (PI3K)/Akt survival pathway and glial cell response as potential targets of JM-20. For this purpose, we used organotypic hippocampal slice cultures exposed to oxygen-glucose deprivation (OGD) to achieve ischemic/reperfusion damage in vitro. Treatment with JM-20 at 0.1 and 10 μM reduced PI incorporation (indicative of cell death) after OGD. OGD decreased the phosphorylation of Akt (pro-survival) and GSK 3β (pro-apoptotic), resulting in respective inhibition and activation of these proteins. Treatment with JM20 prevented the reduced phosphorylation of these proteins after OGD, representing a shift from pro-apoptotic to pro-survival signaling. The OGD-induced activation of caspase-3 was also attenuated by JM-20 treatment at 10 μM. Moreover, in cultures treated with JM-20 and exposed to OGD conditioning, we observed a decrease in activated microglia, as well as a decrease in interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α release into the culture medium, while the level of the anti-inflammatory IL-10 increased. GFAP immunostaining and IB4 labeling showed that JM-20 treatment significantly augmented GFAP immunoreactivity after OGD, when compared with cultures exposed to OGD only, suggesting the activation of astroglial cells. Our results confirm that JM-20 has a strong neuroprotective effect against ischemic injury and suggest that the mechanisms involved in this effect may include the modulation of reactive astrogliosis, as well as neuroinflammation and the anti-apoptotic cell signaling pathway.
authors.affiliation: Centro de Investigación y Desarrollo de Medicamentos, Ave 26, No. 1605 Boyeros y Puentes Grandes, CP 10600 La Habana, Cuba.
authors.forename: Jeney
authors.initials: J
authors.lastname: Ramírez-Sánchez
authors.affiliation: Programa de Pós-graduação em Bioquímica, Departamento de Bioquímica, ICBS, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2600-Anexo I, Porto Alegre, RS 90035-003, Brazil.
authors.forename: Elisa Nicoloso
authors.initials: EN
authors.lastname: Simões Pires
authors.affiliation: Centro de Investigación y Desarrollo de Medicamentos, Ave 26, No. 1605 Boyeros y Puentes Grandes, CP 10600 La Habana, Cuba.
authors.forename: Yanier
authors.initials: Y
authors.lastname: Nuñez-Figueredo
authors.affiliation: Centro de Estudio para las Investigaciones y Evaluaciones Biológicas, Instituto de Farmacia y Alimentos, Universidad de La Habana, ave. 23 # 21425 e/214 y 222, La Coronela, La Lisa CP 13600, La Habana, Cuba.
authors.forename: Gilberto L
authors.initials: GL
authors.lastname: Pardo-Andreu
authors.affiliation: Centro de Investigación y Desarrollo de Medicamentos, Ave 26, No. 1605 Boyeros y Puentes Grandes, CP 10600 La Habana, Cuba.
authors.forename: Luis Arturo
authors.initials: LA
authors.lastname: Fonseca-Fonseca
authors.affiliation: Laboratorio de Síntesis Orgánica de La Facultad de Química de La Universidad de La Habana, Zapata s/n entre G y Carlitos Aguirre, Vedado Plaza de la Revolución, CP 10400, La Habana, Cuba.
authors.forename: Alberto
authors.initials: A
authors.lastname: Ruiz-Reyes
authors.affiliation: Laboratorio de Síntesis Orgánica de La Facultad de Química de La Universidad de La Habana, Zapata s/n entre G y Carlitos Aguirre, Vedado Plaza de la Revolución, CP 10400, La Habana, Cuba.
authors.forename: Estael
authors.initials: E
authors.lastname: Ochoa-Rodríguez
authors.affiliation: Laboratorio de Síntesis Orgánica de La Facultad de Química de La Universidad de La Habana, Zapata s/n entre G y Carlitos Aguirre, Vedado Plaza de la Revolución, CP 10400, La Habana, Cuba.
authors.forename: Yamila
authors.initials: Y
authors.lastname: Verdecia-Reyes
authors.affiliation: Centro de Investigación y Desarrollo de Medicamentos, Ave 26, No. 1605 Boyeros y Puentes Grandes, CP 10600 La Habana, Cuba.
authors.forename: René
authors.initials: R
authors.lastname: Delgado-Hernández
authors.affiliation: Programa de Pós-graduação em Bioquímica, Departamento de Bioquímica, ICBS, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2600-Anexo I, Porto Alegre, RS 90035-003, Brazil; Departamento de Bioquímica, PPG em Bioquímica, PPG em Educação em Ciência, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2600 anexo, Porto Alegre, RS 90035-003, Brazil.
authors.forename: Diogo O
authors.initials: DO
authors.lastname: Souza
authors.affiliation: Programa de Pós-graduação em Bioquímica, Departamento de Bioquímica, ICBS, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2600-Anexo I, Porto Alegre, RS 90035-003, Brazil; Departamento de Bioquímica, PPG em Bioquímica, PPG em Educação em Ciência, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2600 anexo, Porto Alegre, RS 90035-003, Brazil. Electronic address: 00007090@ufrgs.br.
authors.forename: Christianne
authors.initials: C
authors.lastname: Salbego
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001569:Benzodiazepines / Q000494:pharmacology*; D016923:Cell Death / Q000187:drug effects*; D005947:Glucose / Q000378:metabolism; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism*; D000071679:Glycogen Synthase Kinase 3 beta; D006624:Hippocampus / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D009474:Neurons / Q000187:drug effects; D000066829:Neuroprotection / Q000187:drug effects; D018696:Neuroprotective Agents / Q000494:pharmacology; D009525:Niacin / Q000031:analogs & derivatives* / Q000494:pharmacology; D010100:Oxygen / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D017208:Rats, Wistar
chemical_list: C000591456:3-ethoxycarbonyl-2-methyl-4-(2-nitrophenyl)-4,11-dihydro-1H-pyrido(2,3-b)(1,5)benzodiazepine; D018696:Neuroprotective Agents; D001569:Benzodiazepines; D009525:Niacin; D019869:Phosphatidylinositol 3-Kinases; D000071679:Glycogen Synthase Kinase 3 beta; C000605508:Gsk3b protein, rat; D051057:Proto-Oncogene Proteins c-akt; D038362:Glycogen Synthase Kinase 3; D005947:Glucose; D010100:Oxygen
keywords: Akt; GSK-3β; JM-20; Neuroinflammation; Neuroprotection; Oxygen–glucose deprivation