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Withanone, an Active Constituent from Withania somnifera, Affords Protection Against NMDA-Induced Excitotoxicity in Neuron-Like Cells

Molecular Neurobiology volume 54, pages 5061–5073 (2017)Cite this article

Abstract

Withania somnifera has immense pharmacologic and clinical uses. Owing to its similar pharmacologic activity as that of Korean Ginseng tea, it is popularly called as Indian ginseng. In most cases, extracts of this plant have been evaluated against various diseases or models of disease. However, little efforts have been made to evaluate individual constituents of this plant for neurodegenerative disorders. Present study was carried out to evaluate Withanone, one of the active constituents of Withania somnifera against NMDA-induced excitotoxicity in retinoic acid, differentiated Neuro2a cells. Cells were pre-treated with 5, 10 and 20 μM doses of Withanone and then exposed to 3-mM NMDA for 1 h. MK801, a specific NMDA receptor antagonist, was used as positive control. The results indicated that NMDA induces significant death of cells by accumulation of intracellular Ca2+, generation of reactive oxygen species (ROS), loss of mitochondrial membrane potential, crashing of Bax/Bcl-2 ratio, release of cytochrome c, increased caspase expression, induction of lipid peroxidation as measured by malondialdehyde levels and cleavage of poly(ADP-ribose) polymerase-1 (Parp-1), which is indicative of DNA damage. All these parameters were attenuated with various doses of Withanone pre-treatment. These results suggest that Withanone may serve as potential neuroprotective agent.

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Abbreviations

NMDA:

N-methyl-D-aspartate

RA:

Retinoic acid

ROS:

Reactive oxygen species

DMSO:

Dimethyl sulfoxide

HRP:

Horse reddish peroxidase

ΔΨm:

Mitochondrial membrane potential

H2DCFDA:

2′,7′-Dichlorodihydrofluorescein diacetate

NMDARs:

N-methyl-D-aspartate receptor

DMEM:

Dulbecco’s Modified Eagle Medium

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Acknowledgment

Dr. Ahmad’s work was partly supported by the Ramalingaswamy Fellowship of Department of Biotechnology and Financial Assistance (MLP6009) as well as logistic support from the Council for Scientific and Industrial Research. Dr. Abid Hamid’s research is supported by the Department of Biotechnology (BT/PR/3140/PBD/17/656/2009). Additionally, research in both labs is supported by (BSC-0108). Mr. Dar is thankful to the University Grants Commission and Mr. Bhat is thankful to CSIR India for their Ph.D. research fellowships. Authors are thankful to director IIIM for facilitating the work. The institutional publication number of this manuscript is IIIM/1947/2016.

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Affiliations

  1. Neuropharmacology Laboratory, CSIR-Indian Institute of Integrative Medicine, Sanat Nagar, Srinagar, India

    Nawab John Dar & Muzamil Ahmad

  2. Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu, India

    Nawab John Dar, Javeed Ahmad Bhat, Parduman Raj Sharma & Abid Hamid

  3. Academy of Scientific and Innovative Research, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu, India

    Nawab John Dar, Abid Hamid & Muzamil Ahmad

  4. Natural Product Chemistry Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu, India

    Naresh Kumar Satti

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  1. Nawab John Dar
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  2. Javeed Ahmad Bhat
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  3. Naresh Kumar Satti
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  4. Parduman Raj Sharma
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  5. Abid Hamid
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  6. Muzamil Ahmad
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Correspondence to Abid Hamid or Muzamil Ahmad.

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Dar, N.J., Bhat, J.A., Satti, N.K. et al. Withanone, an Active Constituent from Withania somnifera, Affords Protection Against NMDA-Induced Excitotoxicity in Neuron-Like Cells. Mol Neurobiol 54, 5061–5073 (2017). https://doi.org/10.1007/s12035-016-0044-7

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  • DOI: https://doi.org/10.1007/s12035-016-0044-7

Keywords

  • Withanone
  • Withania somnifera
  • Glutamate
  • ROS
  • Caspase
  • Intracellular calcium
  • Cytochrome c
  • Parp-1
  • NMDA