title: Effects of Cannabidiol and Hypothermia on Short-Term Brain Damage in New-Born Piglets after Acute Hypoxia-Ischemia.
abstract: Hypothermia is a standard treatment for neonatal encephalopathy, but nearly 50% of treated infants have adverse outcomes. Pharmacological therapies can act through complementary mechanisms with hypothermia improving neuroprotection. Cannabidiol could be a good candidate. Our aim was to test whether immediate treatment with cannabidiol and hypothermia act through complementary brain pathways in hypoxic-ischemic newborn piglets. Hypoxic-ischemic animals were randomly divided into four groups receiving 30 min after the insult: (1) normothermia and vehicle administration; (2) normothermia and cannabidiol administration; (3) hypothermia and vehicle administration; and (4) hypothermia and cannabidiol administration. Six hours after treatment, brains were processed to quantify the number of damaged neurons by Nissl staining. Proton nuclear magnetic resonance spectra were obtained and analyzed for lactate, N-acetyl-aspartate and glutamate. Metabolite ratios were calculated to assess neuronal damage (lactate/N-acetyl-aspartate) and excitotoxicity (glutamate/Nacetyl-aspartate). Western blot studies were performed to quantify protein nitrosylation (oxidative stress), content of caspase-3 (apoptosis) and TNFα (inflammation). Individually, the hypothermia and the cannabidiol treatments reduced the glutamate/Nacetyl-aspartate ratio, as well as TNFα and oxidized protein levels in newborn piglets subjected to hypoxic-ischemic insult. Also, both therapies reduced the number of necrotic neurons and prevented an increase in lactate/N-acetyl-aspartate ratio. The combined effect of hypothermia and cannabidiol on excitotoxicity, inflammation and oxidative stress, and on cell damage, was greater than either hypothermia or cannabidiol alone. The present study demonstrated that cannabidiol and hypothermia act complementarily and show additive effects on the main factors leading to hypoxic-ischemic brain damage if applied shortly after the insult.
authors.affiliation: Neonatology Research Group, Biocruces Health Research Institute Bizkaia, Spain.
authors.forename: Hector
authors.initials: H
authors.lastname: Lafuente
authors.affiliation: Group of Cannabinoids Research on Neonatal Pathologies, Research Institute Puerta de Hierro Majadahonda Madrid, Spain.
authors.forename: Maria R
authors.initials: MR
authors.lastname: Pazos
authors.affiliation: Department of Cell Biology, University of the Basque Country Leioa, Spain.
authors.forename: Antonia
authors.initials: A
authors.lastname: Alvarez
authors.affiliation: Group of Cannabinoids Research on Neonatal Pathologies, Research Institute Puerta de Hierro Majadahonda Madrid, Spain.
authors.forename: Nagat
authors.initials: N
authors.lastname: Mohammed
authors.affiliation: Group of Cannabinoids Research on Neonatal Pathologies, Research Institute Puerta de Hierro Majadahonda Madrid, Spain.
authors.forename: Martín
authors.initials: M
authors.lastname: Santos
authors.affiliation: Neonatology Research Group, Biocruces Health Research Institute Bizkaia, Spain.
authors.forename: Maialen
authors.initials: M
authors.lastname: Arizti
authors.affiliation: Neonatology Research Group, Biocruces Health Research Institute Bizkaia, Spain.
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Alvarez
authors.affiliation: Group of Cannabinoids Research on Neonatal Pathologies, Research Institute Puerta de Hierro MajadahondaMadrid, Spain; Department of Neonatology, Hospital Clínico San Carlos-Instituto de Investigación Sanitaria San Carlos (IdISSC)Madrid, Spain.
authors.forename: Jose A
authors.initials: JA
authors.lastname: Martinez-Orgado
keywords: cannabidiol; hypothermia; hypoxic-ischemic encephalopathy; neonatal brain; neurodevelopment; neuroprotection; newborn animal