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Effects of Cannabidiol and Hypothermia on Short-Term Brain Damage in New-Born Piglets after Acute Hypoxia-Ischemia.

Lafuente H, Pazos MR, Alvarez A, Mohammed N, Santos M, Arizti M, Alvarez FJ, Martinez-Orgado JA - Front Neurosci (2016)

Bottom Line: 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 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.

View Article: PubMed Central - PubMed

Affiliation: Neonatology Research Group, Biocruces Health Research Institute Bizkaia, Spain.

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.

No MeSH data available.


Related in: MedlinePlus

Effect of the treatments on the characteristics of oxidative stress. (A) A representative image of immunoblotting indicating protein carbonyl formation (Oxyblot) using anti-DNPH antibody. Samples were obtained using brain tissue from 1- to 2-day-old piglets after hypoxic-ischemic insult, treated with normothermia or hypothermia and administered with either vehicle or cannabidiol. (B) Relative protein carbonyl levels were quantified by densitometric analyses of the blots. The content of oxidized proteins was normalized by total Red Ponceau-stained protein loading and expressed as OxyBlot/Red Ponceau ratio (see Materials and Methods for details). Bars represent mean ± SEM of 6–8 experiments. (a.u.) arbitrary units; (*) P < 0.05 vs. NV group by Kruskall-Wallis one-factorial analysis of variance; (***) P < 0.05 vs. all groups by Kruskall-Wallis one-factorial analysis of variance.
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Figure 3: Effect of the treatments on the characteristics of oxidative stress. (A) A representative image of immunoblotting indicating protein carbonyl formation (Oxyblot) using anti-DNPH antibody. Samples were obtained using brain tissue from 1- to 2-day-old piglets after hypoxic-ischemic insult, treated with normothermia or hypothermia and administered with either vehicle or cannabidiol. (B) Relative protein carbonyl levels were quantified by densitometric analyses of the blots. The content of oxidized proteins was normalized by total Red Ponceau-stained protein loading and expressed as OxyBlot/Red Ponceau ratio (see Materials and Methods for details). Bars represent mean ± SEM of 6–8 experiments. (a.u.) arbitrary units; (*) P < 0.05 vs. NV group by Kruskall-Wallis one-factorial analysis of variance; (***) P < 0.05 vs. all groups by Kruskall-Wallis one-factorial analysis of variance.

Mentions: A lower mean value of OxyBlot/Red Ponceau ratio in the analyzed proteins from the parietal cortex was observed in the HV group than in NV group (Figure 3), indicating a reduction of protein carbonylation. A single dose of CBD after HI (NC group) induced a similar effect on this ratio as in the HV group (Figure 3). The mean levels of OxyBlot/Red Ponceau ratio were lower in the HC group than in the HV or NC groups, suggesting that administration of hypothermia together with CBD led to an additive effect.


Effects of Cannabidiol and Hypothermia on Short-Term Brain Damage in New-Born Piglets after Acute Hypoxia-Ischemia.

Lafuente H, Pazos MR, Alvarez A, Mohammed N, Santos M, Arizti M, Alvarez FJ, Martinez-Orgado JA - Front Neurosci (2016)

Effect of the treatments on the characteristics of oxidative stress. (A) A representative image of immunoblotting indicating protein carbonyl formation (Oxyblot) using anti-DNPH antibody. Samples were obtained using brain tissue from 1- to 2-day-old piglets after hypoxic-ischemic insult, treated with normothermia or hypothermia and administered with either vehicle or cannabidiol. (B) Relative protein carbonyl levels were quantified by densitometric analyses of the blots. The content of oxidized proteins was normalized by total Red Ponceau-stained protein loading and expressed as OxyBlot/Red Ponceau ratio (see Materials and Methods for details). Bars represent mean ± SEM of 6–8 experiments. (a.u.) arbitrary units; (*) P < 0.05 vs. NV group by Kruskall-Wallis one-factorial analysis of variance; (***) P < 0.05 vs. all groups by Kruskall-Wallis one-factorial analysis of variance.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4940392&req=5

Figure 3: Effect of the treatments on the characteristics of oxidative stress. (A) A representative image of immunoblotting indicating protein carbonyl formation (Oxyblot) using anti-DNPH antibody. Samples were obtained using brain tissue from 1- to 2-day-old piglets after hypoxic-ischemic insult, treated with normothermia or hypothermia and administered with either vehicle or cannabidiol. (B) Relative protein carbonyl levels were quantified by densitometric analyses of the blots. The content of oxidized proteins was normalized by total Red Ponceau-stained protein loading and expressed as OxyBlot/Red Ponceau ratio (see Materials and Methods for details). Bars represent mean ± SEM of 6–8 experiments. (a.u.) arbitrary units; (*) P < 0.05 vs. NV group by Kruskall-Wallis one-factorial analysis of variance; (***) P < 0.05 vs. all groups by Kruskall-Wallis one-factorial analysis of variance.
Mentions: A lower mean value of OxyBlot/Red Ponceau ratio in the analyzed proteins from the parietal cortex was observed in the HV group than in NV group (Figure 3), indicating a reduction of protein carbonylation. A single dose of CBD after HI (NC group) induced a similar effect on this ratio as in the HV group (Figure 3). The mean levels of OxyBlot/Red Ponceau ratio were lower in the HC group than in the HV or NC groups, suggesting that administration of hypothermia together with CBD led to an additive effect.

Bottom Line: 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 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.

View Article: PubMed Central - PubMed

Affiliation: Neonatology Research Group, Biocruces Health Research Institute Bizkaia, Spain.

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.

No MeSH data available.


Related in: MedlinePlus