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Thymoquinone Protects Neurons and Astrocytes in Hippocampal Primary Cell Culture from Lead Toxicity
Author(s) -
Rao Muddanna,
Shivanandan Smitha
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.708.5
Subject(s) - thymoquinone , neuroprotection , hippocampal formation , doublecortin , chemistry , toxicity , neurotoxicity , neun , pharmacology , andrology , medicine , antioxidant , biochemistry , immunohistochemistry , dentate gyrus , organic chemistry
Thymoquinone (TQ) is the active constituent of Nigella sativa (NS) seeds. Quinines, of which TQ is the most abundant, accounts for most of the pharmacological properties of NS. TQ possesses anti‐inflammatory, antioxidant and neuroprotective properties. Present study was aimed to investigate the role of TQ in protection of newly born neurons and astrocytes against lead induced toxicity in hippocampal cell culture. Fetal hippocampal primary cultures were divided into four groups: i) Normal Control (NC, n=6) ‐ no treatment, ii) Lead only (LO, n=6)‐ media was replaced with media containing 20µM lead nitrate, iii) Thymoquinone TQ, n=6)‐ media containing 10µM TQ, iv) Lead + TQ, (L+TQ, n=6) ‐ media containing 20µM lead nitrate and 10µM TQ. Cell survival was analyzed by counting number of cells in randomly selected fields. Culture dishes were stained for doublecortin and GFAP immunostaining and the number of neurons and astrocytes were quantified. Data were analyzed with one way ANOVA. The LO cultures showed significantly decreased number of surviving cells compared to NC, TQ, and L+ TQ groups (20‐30% decrease, P<0.001). Cells in the LO group had fewer and shorter processes. The cell death count was significantly decreased (40%, P<0.001) in cultures treated with lead and TQ, compared to LO group. The cultures treated with L+TQ showed significantly increased number of doublecortin positive neurons (40%, P<0.001) and GFAP immunopositive astrocytes (50%, P<0.001) compared to LO group. These data suggest that TQ can counteract lead toxicity and enhance cell survival and differentiation into neurons and astrocytes in culture. (We acknowledge RCF, Faculty of medicine, Kuwait University (Grant No. SRUL02/13 technical support for confocal and phase contrast microscopy).