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Protective Effect of Nigella sativa Extract and Thymoquinone on Serum/Glucose Deprivation-Induced PC12 Cells Death

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Abstract

The serum/glucose deprivation (SGD)-induced cell death in cultured PC12 cells represents a useful in vitro model for the study of brain ischemia and neurodegenerative disorders. Nigella sativa L. (family Ranunculaceae) and its active component thymoquinone (TQ) has been known as a source of antioxidants. In the present study, the protective effects of N. sativa and TQ on cell viability and reactive oxygen species (ROS) production in cultured PC12 cells were investigated under SGD conditions. PC12 cells were cultured in DMEM medium containing 10% (v/v) fetal bovine serum, 100 units/ml penicillin, and 100 μg/ml streptomycin. Cells were seeded overnight and then deprived of serum/glucose for 6 and 18 h. Cells were pretreated with different concentrations of N. sativa extract (15.62–250 μg/ml) and TQ (1.17–150 μM) for 2 h. Cell viability was quantitated by MTT assay. Intracellular ROS production was measured by flow cytometry using 2′,7′-dichlorofluorescin diacetate (DCF-DA) as a probe. SGD induced significant cells toxicity after 6, 18, or 24 h (P < 0.001). Pretreatment with N. sativa (15.62–250 μg/ml) and TQ (1.17–37.5 μM) reduced SGD-induced cytotoxicity in PC12 cells after 6 and 18 h. A significant increase in intracellular ROS production was seen following SGD (P < 0.001). N. sativa (250 μg/ml, P < 0.01) and TQ (2.34, 4.68, 9.37 μM, P < 0.01) pretreatment reversed the increased ROS production following ischemic insult. The experimental results suggest that N. sativa extract and TQ protects the PC12 cells against SGD-induced cytotoxicity via antioxidant mechanisms. Our findings might raise the possibility of potential therapeutic application of N. sativa extract and TQ for managing cerebral ischemic and neurodegenerative disorders.

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Acknowledgments

The authors would like to thank Research Affairs of Mashhad University of Medical Sciences for financially supporting this work and Miss Aghaee for her assistance in preparation of N. sativa extract. We are also grateful of Dr. H. Nasirli for her assistance in flow cytometry.

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Authors and Affiliations

  1. Department of Pharmacology and Pharmacological Research Centre of Medicinal Plants, School of Medicine, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran

    S. H. Mousavi, Z. Tayarani-Najaran & H. R. Sadeghnia

  2. Medical Toxicology Research Center, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran

    S. H. Mousavi

  3. Department of Biochemistry, Payame Noor University (PNU), Mashhad, Iran

    M. Asghari

  4. Department of Modern Technologies and Sciences, Mashhad University of Medical Sciences, P.O. Box 9177948564, Mashhad, Iran

    H. R. Sadeghnia

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  1. S. H. Mousavi
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  2. Z. Tayarani-Najaran
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  4. H. R. Sadeghnia
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Correspondence to H. R. Sadeghnia.

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Mousavi, S.H., Tayarani-Najaran, Z., Asghari, M. et al. Protective Effect of Nigella sativa Extract and Thymoquinone on Serum/Glucose Deprivation-Induced PC12 Cells Death. Cell Mol Neurobiol 30, 591–598 (2010). https://doi.org/10.1007/s10571-009-9484-1

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