Abstract
Following anoxia, a rapid and marked mitochondrial-linked cell death occurs in the cerebral cortex of newborn rats which leads to insult advancement within a couple of days and causes lifelong neurobehavioral abnormalities. The present study investigated the role of 2,4 dinitrophenol (2,4 DNP) in three doses, i.e.,1, 2.5, and 5 mg/kg on anoxia-induced time-dependent mitochondrial dysfunction and associated neurobehavioral outcome using a well-established global model of anoxia. Briefly, rat pups of 30-h age (P2) were subjected to two episodes of anoxia (10 min each) at 24 h of the time interval in an enclosed chamber supplied with 100% N2 and immersed in a water bath (35–37 °C) to avoid hypothermia. Results demonstrated that the uncoupler 2,4 DNP, in the dose 2.5 and 5 mg/kg injected i.p. within 5 min after second anoxic episode significantly (P < 0.05) preserved mitochondrial function on day 7 preferentially by maintaining mitochondrial membrane potential (MMP) and inhibiting mitochondrial permeability transition (MPT) pore. Further, 2,4 DNP preserved mitochondrial function by improving different states of mitochondrial respiration (s2, s3, s4, s5), respiratory control ratio (RCR), antioxidant enzyme system like superoxide dismutase (SOD) and catalase (CAT), and mitochondrial complex enzymes (I, II, IV, V) after anoxia. Furthermore, a marked decrease in the levels of expression of cytochrome C (cyt C) and pro-apoptotic (Bcl-2 family) and apoptotic (caspase-9/3) proteins was observed on day 7 indicating that the treatment with 2,4 DNP prevented mitochondrial dysfunction and further insult progression (day 1 to day 7). Moreover, 2,4 DNP decreased the apoptotic cell death on day 7 and overall improved the neurobehavioral outcomes like reflex latency and hanging latency which suggests its role in treating neonatal anoxia.
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Abbreviations
- 2,4 DNP:
-
2,4 Dinitrophenol
- NO:
-
Nitric oxide
- UPCs:
-
Mitochondrial uncouplers
- MMP`:
-
Mitochondrial membrane potential
- MPT:
-
Mitochondrial permeability transition
- RCR:
-
Respiratory control ratio
- ETC:
-
Electron transport chain
- LPO:
-
Lipid peroxidation
- SOD:
-
Superoxide dismutase
- CAT:
-
Catalase
- Cyt C:
-
Cytochrome C
- GABA:
-
Gamma-aminobutyric acid
- CNS:
-
Central nervous system
- EAA:
-
Excitatory amino acid
- ATP:
-
Adenosine triphosphate
- ADP:
-
Adenosine diphosphate
- NADH:
-
Nicotinamide adenine dinucleotide
- N2 :
-
Nitrogen
- ROS:
-
Reactive oxygen species
- RNS:
-
Reactive nitrogen species
- Bcl-2:
-
B cell lymphoma 2
- Bax:
-
Bcl-2-associated X protein
- Ca2+ :
-
Calcium ion
- s:
-
State
- P:
-
Pyruvate
- M:
-
Malate
- i.p.:
-
Intraperitoneal
- TMRM:
-
Tetramethylrhodamine methyl ester
- HEPES:
-
(4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid)
- EGTA:
-
(Ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid)
- DMSO:
-
Dimethyl sulfoxide
- SDH:
-
Succinate dehydrogenase
- SDS-PAGE:
-
Sodium dodecyl sulfate polyacrylamide gel electrophoresis
- PVDF:
-
Polyvinylidene fluoride
- FITC:
-
Fluorescein isothiocyanate
- PBS:
-
Phosphate buffer saline
- ECL:
-
Chemiluminescence
- PI:
-
Propidium iodide
- ANT:
-
Adenosine nucleotide translocase
- ER:
-
Endoplasmic reticulum
- MAMs:
-
Mitochondria-associated ER membranes
- NBT:
-
Nitro blue tetrazolium
- dfz:
-
Diformazan
- MDA:
-
Malondialdehyde
- ANOVA:
-
Analysis of variance
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SK is thankful to Department of Biotechnology (DBT), New Delhi, India, for assistance in terms of research grant [102/IFD/SAN/4654/2011-2012].
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The experimental procedures were approved by the Institutional Animal Ethical Committee of BHU (Protocol No. Dean/11-12/CAEC/328). All experiments were performed as per guidelines of laboratory animal care (National Research Council US Committee for the Update of the Guide for the Care and Use of Laboratory Animals 2011) guidelines.
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Samaiya, P.K., Narayan, G., Kumar, A. et al. 2,4 Dinitrophenol Attenuates Mitochondrial Dysfunction and Improves Neurobehavioral Outcomes Postanoxia in Neonatal Rats. Neurotox Res 34, 121–136 (2018). https://doi.org/10.1007/s12640-018-9873-7
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DOI: https://doi.org/10.1007/s12640-018-9873-7