Neurotoxicity Research

, Volume 34, Issue 1, pp 121–136 | Cite as

2,4 Dinitrophenol Attenuates Mitochondrial Dysfunction and Improves Neurobehavioral Outcomes Postanoxia in Neonatal Rats

  • Puneet K. Samaiya
  • Gopeshwar Narayan
  • Ashok Kumar
  • Sairam Krishnamurthy


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.


Anoxia Mitochondrial function 2,4 Dinitrophenol Neurobehavior Apoptosis Animal model 


2,4 DNP

2,4 Dinitrophenol


Nitric oxide


Mitochondrial uncouplers


Mitochondrial membrane potential


Mitochondrial permeability transition


Respiratory control ratio


Electron transport chain


Lipid peroxidation


Superoxide dismutase



Cyt C

Cytochrome C


Gamma-aminobutyric acid


Central nervous system


Excitatory amino acid


Adenosine triphosphate


Adenosine diphosphate


Nicotinamide adenine dinucleotide




Reactive oxygen species


Reactive nitrogen species


B cell lymphoma 2


Bcl-2-associated X protein


Calcium ion










Tetramethylrhodamine methyl ester


(4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid)


(Ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid)


Dimethyl sulfoxide


Succinate dehydrogenase


Sodium dodecyl sulfate polyacrylamide gel electrophoresis


Polyvinylidene fluoride


Fluorescein isothiocyanate


Phosphate buffer saline




Propidium iodide


Adenosine nucleotide translocase


Endoplasmic reticulum


Mitochondria-associated ER membranes


Nitro blue tetrazolium






Analysis of variance


Funding Information

SK is thankful to Department of Biotechnology (DBT), New Delhi, India, for assistance in terms of research grant [102/IFD/SAN/4654/2011-2012].

Compliance with Ethical Standards

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.

Conflict of Interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Puneet K. Samaiya
    • 1
  • Gopeshwar Narayan
    • 2
  • Ashok Kumar
    • 3
  • Sairam Krishnamurthy
    • 1
  1. 1.Neurotherapeutics Laboratory, Department of Pharmaceutical Engineering and TechnologyIndian Institute of Technology (Banaras Hindu University)VaranasiIndia
  2. 2.Department of Molecular and Human GeneticsBanaras Hindu UniversityVaranasiIndia
  3. 3.Department of Pediatrics, Institute of Medical SciencesBanaras Hindu UniversityVaranasiIndia

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