2025_The Potential Neuroinflammatory Attenuation Effects Of Thymoquinone Against Mdma-Induced Microglial Activation In Bv2 Mouse Cell Line
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| collectionurl | https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection3 |
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| country | Malaysia |
| date | 2025-04-20 09:11 |
| format | General Document |
| id | 17233 |
| institution | UniSZA |
| originalfilename | 17233_c9711aae4b39de1.pdf |
| person | Nor Suliana Binti Mustafa |
| recordtype | oai_dc |
| resourceurl | https://intelek.unisza.edu.my/intelek/pages/view.php?ref=17233 |
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| spelling | 17233 https://intelek.unisza.edu.my/intelek/pages/view.php?ref=17233 https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection3 General Document Malaysia Library Staff (Top Management) Library Staff (Management) Library Staff (Support) Terengganu Faculty of Medicine English application/pdf 1.7 Microsoft® Word 2019 Server storage Scanned document UniSZA Private Access UniSZA Thymoquinone Copyright©PWB2025 Neuroprotection 308 Dissertations, Academic Oxidative Stress UniSZA Nor Suliana Binti Mustafa Neuroinflammation Microglial Activation MDMA (3,4-Methylenedioxymethamphetamine) BV2 Cell Line Anti-Inflammatory Agents Neurotoxicity Cell Culture Studies Neuroinflammation – Prevention and Control Microglia – Physiology Thymoquinone – Therapeutic use Nervous System – Degeneration – Pathophysiology 2025_The Potential Neuroinflammatory Attenuation Effects Of Thymoquinone Against Mdma-Induced Microglial Activation In Bv2 Mouse Cell Line Ecstasy (3,4-methylenedioxymethamphetamine; MDMA) is known to induce neurotoxicity. The abuse of MDMA causes excessive activation of the microglia, which is deleterious to the neurons. In their response to disturbances around neurons, microglia can differentiate into the M1 phenotype. As an activated M1 microglia, the microglia produce and release pro-inflammatory mediators. In contrast, microglia can take on an M2 phenotype, in which the microglial cell releases mediators that can serve as an anti-inflammatory signal to initiate recovery from an insult. Hence, a modulation between pro-inflammatory states and anti-inflammatory states of microglia makes up an important strategy to reduce the impairments made by MDMA on neuronal cells. Furthermore, the molecular mechanisms underlying the treatments and modulations of MDMA-induced microglial activation are scarce and limited. This study aims to investigate the effects of TQ on the transcriptome profile of MDMA-induced microglial activation and its mechanism in modulating the activation of microglial cells by using mouse BV2 microglial cells. The MTT assay was used to determine the cytotoxic dose of MDMA and TQ. The integrity of the cells after being challenged by TQ was evaluated using the Lactate Dehydrogenase (LDH) assay. Next, microglial activation was induced with the optimum concentrations of MDMA. The experimental group was divided into: i) Control (untreated), ii) MDMA, iii) MDMA+TQ, and iv) TQ control (TQ only). TQ was pre-treated one hour before MDMA stimulation. The evaluation of the morphological changes of microglia was done using visualization under a microscope. TNF-α protein concentrations were determined using an Enzyme-Linked Immunosorbent Assay (ELISA), and the functional genes expressed were identified via transcriptome profiling using RNA Sequencing (RNA-Seq) techniques. The data were analyzed by a one-way ANOVA test followed by Tukey's post-hoc test. The data were expressed as the mean ± SEM. Treatment differences were considered statistically significant at P ≤ 0.05. Differential gene expression analysis was performed using the DESeq2R package (1.20.0). The resulting P values were adjusted using Benjamini and Hochberg's approach for controlling the false discovery rate. Genes with an adjusted P-value ≤ 0.05 were assigned as differentially expressed. The results showed that exposure to 500 μg/mL of MDMA at 24 hours induced BV2 microglial cell activation, as observed by the morphological changes, and a significant increase in TNF-α level (P ≤ 0.05). Subsequently, a 2.0 μg/mL concentration of TQ was selected for the experimental groups as it was less toxic and showed no significant effects on LDH activity. TQ pre-treatment in the MDMA+TQ group appeared to have attenuated the morphological changes of microglia without a significant effect on TNF-α levels as compared to the MDMA group. RNA-Seq analysis revealed that TQ reduced inflammatory responses in MDMA-induced microglial activation by inhibiting the production of several pro-inflammatory genes such as Cxcl2, Ptgs2, C3ar1, Nfkbia, Il1a, Cxcl10, and Serpinf2 (P ≤ 0.05). This study concluded that TQ possesses neuroprotective effects against MDMA-induced microglial activation by stabilizing the effects of microglia. The use of TQ could attenuate the expression of certain inflammatory genes that appear to play pivotal roles in the initiation and/or progression of MDMA neurotoxicity. 2025-04-20 09:11 uuid:8CA35512-B637-40A2-978A-413976973B21 17233_c9711aae4b39de1.pdf Thesis |
| spellingShingle | 2025_The Potential Neuroinflammatory Attenuation Effects Of Thymoquinone Against Mdma-Induced Microglial Activation In Bv2 Mouse Cell Line |
| state | Terengganu |
| subject | Dissertations, Academic Neuroinflammation – Prevention and Control Microglia – Physiology Thymoquinone – Therapeutic use Nervous System – Degeneration – Pathophysiology |
| summary | Ecstasy (3,4-methylenedioxymethamphetamine; MDMA) is known to induce neurotoxicity. The abuse of MDMA causes excessive activation of the microglia, which is deleterious to the neurons. In their response to disturbances around neurons, microglia can differentiate into the M1 phenotype. As an activated M1 microglia, the microglia produce and release pro-inflammatory mediators. In contrast, microglia can take on an M2 phenotype, in which the microglial cell releases mediators that can serve as an anti-inflammatory signal to initiate recovery from an insult. Hence, a modulation between pro-inflammatory states and anti-inflammatory states of microglia makes up an important strategy to reduce the impairments made by MDMA on neuronal cells. Furthermore, the molecular mechanisms underlying the treatments and modulations of MDMA-induced microglial activation are scarce and limited. This study aims to investigate the effects of TQ on the transcriptome profile of MDMA-induced microglial activation and its mechanism in modulating the activation of microglial cells by using mouse BV2 microglial cells. The MTT assay was used to determine the cytotoxic dose of MDMA and TQ. The integrity of the cells after being challenged by TQ was evaluated using the Lactate Dehydrogenase (LDH) assay. Next, microglial activation was induced with the optimum concentrations of MDMA. The experimental group was divided into: i) Control (untreated), ii) MDMA, iii) MDMA+TQ, and iv) TQ control (TQ only). TQ was pre-treated one hour before MDMA stimulation. The evaluation of the morphological changes of microglia was done using visualization under a microscope. TNF-α protein concentrations were determined using an Enzyme-Linked Immunosorbent Assay (ELISA), and the functional genes expressed were identified via transcriptome profiling using RNA Sequencing (RNA-Seq) techniques. The data were analyzed by a one-way ANOVA test followed by Tukey's post-hoc test. The data were expressed as the mean ± SEM. Treatment differences were considered statistically significant at P ≤ 0.05. Differential gene expression analysis was performed using the DESeq2R package (1.20.0). The resulting P values were adjusted using Benjamini and Hochberg's approach for controlling the false discovery rate. Genes with an adjusted P-value ≤ 0.05 were assigned as differentially expressed. The results showed that exposure to 500 μg/mL of MDMA at 24 hours induced BV2 microglial cell activation, as observed by the morphological changes, and a significant increase in TNF-α level (P ≤ 0.05). Subsequently, a 2.0 μg/mL concentration of TQ was selected for the experimental groups as it was less toxic and showed no significant effects on LDH activity. TQ pre-treatment in the MDMA+TQ group appeared to have attenuated the morphological changes of microglia without a significant effect on TNF-α levels as compared to the MDMA group. RNA-Seq analysis revealed that TQ reduced inflammatory responses in MDMA-induced microglial activation by inhibiting the production of several pro-inflammatory genes such as Cxcl2, Ptgs2, C3ar1, Nfkbia, Il1a, Cxcl10, and Serpinf2 (P ≤ 0.05). This study concluded that TQ possesses neuroprotective effects against MDMA-induced microglial activation by stabilizing the effects of microglia. The use of TQ could attenuate the expression of certain inflammatory genes that appear to play pivotal roles in the initiation and/or progression of MDMA neurotoxicity. |
| title | 2025_The Potential Neuroinflammatory Attenuation Effects Of Thymoquinone Against Mdma-Induced Microglial Activation In Bv2 Mouse Cell Line |
| title_full | 2025_The Potential Neuroinflammatory Attenuation Effects Of Thymoquinone Against Mdma-Induced Microglial Activation In Bv2 Mouse Cell Line |
| title_fullStr | 2025_The Potential Neuroinflammatory Attenuation Effects Of Thymoquinone Against Mdma-Induced Microglial Activation In Bv2 Mouse Cell Line |
| title_full_unstemmed | 2025_The Potential Neuroinflammatory Attenuation Effects Of Thymoquinone Against Mdma-Induced Microglial Activation In Bv2 Mouse Cell Line |
| title_short | 2025_The Potential Neuroinflammatory Attenuation Effects Of Thymoquinone Against Mdma-Induced Microglial Activation In Bv2 Mouse Cell Line |
| title_sort | 2025_the potential neuroinflammatory attenuation effects of thymoquinone against mdma-induced microglial activation in bv2 mouse cell line |