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The Antioxidant and Anti-inflammatory Basis of the Neuroprotective Effect of Thymoquinone in Activated BV-2 Murine Microglia Cells
Başlık:
The Antioxidant and Anti-inflammatory Basis of the Neuroprotective Effect of Thymoquinone in Activated BV-2 Murine Microglia Cells
Yazar:
Cobourne-Duval, Makini K., author.
ISBN:
9780438009264
Yazar Ek Girişi:
Fiziksel Tanımlama:
1 electronic resource (152 pages)
Genel Not:
Source: Dissertation Abstracts International, Volume: 79-10(E), Section: B.
Advisors: Karam FA Soliman Committee members: Tanise Jackson; Ebenezer Oriaku; R.Renee Reams; Ngozi H. Ugochukwu.
Özet:
Neuroinflammation and excessive microglial activation are pathological markers of a number of central nervous system (CNS) diseases. Dysregulated, chronic activation of microglia, the brain's resident macrophages, induces the release of excessive amounts of reactive oxygen species (ROS) and pro-inflammatory cytokines and furthermore has been implicated in several neurodegenerative diseases such as Alzheimer's, currently the most common cause of neurodegeneration and dementia in the elderly. Thymoquinone (TQ), one of the major bioactive components identified in the natural product Nigella sativa seed oil, has been shown to exhibit antioxidant, anti-inflammatory, and neuroprotective effects. In this study, we investigated the antioxidant effects of TQ on LPS/IFN gamma or hydrogen peroxide-activated microglia by assessing the levels of specific oxidative stress markers, the activities of selected antioxidant enzymes, as well as profiling 84 key genes related to oxidative stress via real-time reverse transcription (RT2) PCR array. We also investigated TQ's ability to suppress the pro-inflammatory response using multi-analyte ELISArray for inflammatory cytokines and TQ's effect on the gene expression of NF kappa B signaling targets in BV-2 microglial cells activated with lipopolysaccharide (LPS) and interferon gamma (IFN gamma) by profiling 84 key genes via real-time reverse transcription (RT2 ) PCR array. Furthermore, we performed quantitative proteomic analysis using Orbitrap/Q-Exactive Proteomic LC-MS/MS (Liquid chromatography-mass spectrometry) to detect any additional proteins that were differentially expressed amongst the treatment groups. Our results showed that TQ treatment (12.5 uM for 24 hours) of LPS/IFNgamma-activated BV-2 microglial cells significantly decreased the cellular production of both superoxide and nitric oxide 4 fold (p < 0.0001) and 6 fold (p < 0.0001), respectively. In the hydrogen peroxide-activated microglia, TQ treatment also significantly decreased the cellular production of superoxide 3 fold (p < 0.0001) and hydrogen peroxide levels ~20% (p < 0.05). Moreover, TQ treatment significantly decreased oxidative stress in the LPS/IFN gamma-activated microglia as evidenced by the 2-fold decrease of lipid hydroperoxides levels (p < 0.0001) and 2.5 fold increase antioxidant glutathione levels (p < 0.0001). Likewise in the hydrogen peroxide-activated microglia, TQ significantly decreased lipid hydroperoxides 8-fold (p < 0.0001) and significantly increased glutathione 15% (p < 0.05). Antioxidant enzyme activities of superoxide dismutase (SOD) and catalase (Cat), in the TQ-treated microglial cells also reflected a reduced oxidative stress status in the cellular environment. SOD and Cat activities were 6 fold (p < 0.0001) and 5 fold (p < 0.0001) lower, respectfully, for the TQ-treated, LPS/INF gamma-activated microglia compared to the untreated. Similarly, SOD and Cat activities were 5 fold (p < 0.0001) and 3 fold (p < 0.01) lower, respectively, for the TQ-treated, hydrogen peroxide-activated microglia compared to the untreated. Profiling of the selected 84 genes related to oxidative stress via RT2 PCR array confirmed TQ's antioxidant effects. Results showed that TQ treatment of the LPS/IFN gamma-activated microglia significantly downregulated specific pro-oxidant genes, most markedly inducible nitric oxide synthase (NOS2) and chemokine (C-C) motif ligand 5 (CCL5/RANTES) > 100 fold and > 6.5 fold, respectively, as well as enhanced the up- or downregulation of specific genes related to the cells' natural antioxidant defense against LPS/IFN gamma activation. Upon assessing the anti-inflammatory effects, TQ treatment significantly (P < 0.0001) reduced the expression of inflammatory cytokines, IL-2 = 38%, IL-4 = 19%, IL-6 = 83%, IL-10 = 23%, and IL-17a = 29%, in the activated microglia compared to the untreated, activated which expression levels were significantly elevated than the control microglia: IL-2 = 127%, IL-4 = 151%, IL-6 = 670%, IL-10 = 133%, IL-17a = 127%. Upon assessing the gene expression of NF kappa B signaling targets, this study also demonstrated that TQ treatment activated microglia resulted in > 7 fold down-regulation of several NF kappa B signaling targets genes, including complement factor B (CFB), interleukin 6 (IL6), chemokine (C-C motif) ligand 3 (CXCL3), chemokine (C-C) motif ligand 5 (CCL5/RANTES) compared to the untreated, activated microglia. This modulation in gene expression counteracts the > 10 fold upregulation of these same genes observed in the activated microglia compared to the controls. Furthermore, our results showed that TQ treatment of LPS/IFN gamma-activated BV-2 microglial cells resulted in an increased expression of 4 neuroprotective proteins: glutaredoxin-3 (21 fold), biliverdin reductase A (15 fold), 3-mercaptopyruvate sulfurtransferase (11 fold), and mitochondrial lon protease (> 8 fold) compared to the untreated, activated cells. These studies show that TQ treatment reduced the levels of oxidative stress in the microglia as indicated with a reduction in the levels of ROS, lipid hydroperoxides, and increased levels of glutathione, despite lower antioxidant enzyme activity. Moreover, TQ treatment modulated the expression of specific pro- and antioxidant genes to support a reduction in oxidative stress in the cellular microenvironment. (Abstract shortened by ProQuest.).
Notlar:
School code: 0872
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Yer Numarası | Demirbaş Numarası | Shelf Location | Lokasyon / Statüsü / İade Tarihi |
---|---|---|---|
XX(678397.1) | 678397-1001 | Proquest E-Tez Koleksiyonu | Arıyor... |
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