- Title
- Comparative study of the effect of iloprost on neuroinflammatory changes in c8-b4 microglial cells and murine model of trypanosomiasis
- Creator
- Jacobs, Ashleigh
- Subject
- Trypanosomiasis -- South Africa
- Subject
- DNA -- Methylation -- Research -- Methodology
- Subject
- Central nervous system -- Diseases
- Subject
- Nervous system -- Degeneration
- Date Issued
- 2024-04
- Date
- 2024-04
- Type
- Master's theses
- Type
- text
- Identifier
- http://hdl.handle.net/10948/64077
- Identifier
- vital:73651
- Description
- Neurodegenerative conditions significantly impact well-being and quality of life in individuals with major symptoms including mood disorders, cognitive decline, and psychiatric disturbances, often resulting from neuroinflammation triggered by immune responses to bacterial or parasitic infections such as gram-negative bacteria or Human African Trypanosomiasis. Microglia play a crucial role in both neurotoxicity and cellular processes involved in restoring the neural health. Exploring the therapeutic potential of prostacyclin and its analogues in regulating microglia responses to inflammatory insult and treating Trypanosoma brucei (T.b) infection remains an unexplored area. The aim of this study was to assess the potential neuroprotective effects of Iloprost through comparative analysis of neuroinflammatory responses in both microglial cells exposed to lipopolysaccharide (LPS) and mouse brains infected with T.b brucei. In phase I of this study both resting and LPS treated C8-B4 microglial cells were exposed to varying concentrations of Iloprost. The effects of Iloprost on LPS-induced inflammation were analysed using immunofluorescence to detect microglial activation and differentiate between pro and anti-inflammatory phenotypes. Furthermore, pro and anti-inflammatory cytokine secretion was determined using an ELISA, in addition gene expression analysis was carried out using quantitative polymerase chain reaction (qPCR). Also, DNA methylation status of C8-B4 cells exposed to LPS challenge alone or in combination with various concentrations of Iloprost were determined using bisulfite sequencing technique followed by qPCR. In phase II of the study, a total of twenty-four Albino Swiss male mice (8-10 weeks old) were divided into four treatment groups with 6 mice in each group. All treatment groups except the non-infected control were inoculated with the T.b brucei parasite. One group received a single intraperitoneal injection of Diminazene aceturate (4 mg kg-1) while the remaining group received repeated intraperitoneal injections of Iloprost (200 μg kg-1). On day ten of the study, mouse brains were removed on ice using forceps. The hippocampal tissues were dissected out and processed for quantification of gene expression changes in pro and anti-inflammatory cytokines. Overall, the findings of this study indicate that LPS-induced pro-inflammatory cytokine, TNF-α and IL-1β, secretion and gene expression is down-regulated in C8-B4 microglial cells treated with Iloprost. Furthermore, there was a significant up-regulation in the expression of anti-inflammatory genes, particularly ARG-1, CD206, BDNF and CREB in response to Iloprost treatment following LPS-induced inflammation. This study is also the first to confirm M2 microglial polarization with Iloprost treatment in both resting and LPS treated cells. However, hypermethylation at CREB and BDNF promoter regions was observed 24 hours after Iloprost treatment. Additionally, Iloprost reversed hypomethylation at the BDNF promoter region that had been induced by LPS treatment. The rodent model also indicated a downregulation in the pro-inflammatory cytokine, IL-1β, expression and upregulation of BDNF transcription in T.b brucei infected mice treated with repeated doses of Iloprost. In conclusion, determining the immunomodulatory roles of Iloprost in both in vitro and in vivo models of neuroinflammation could assist in the development of alternative therapy for neurodegenerative disease.
- Description
- Thesis (MSc) -- Faculty of Science, School of Biomolecular & Chemical Sciences, 2024
- Format
- computer
- Format
- online resource
- Format
- application/pdf
- Format
- 1 online resource (127 pages)
- Format
- Publisher
- Nelson Mandela University
- Publisher
- Faculty of Science
- Language
- English
- Rights
- Nelson Mandela University
- Rights
- All Rights Reserved
- Rights
- Open Access
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| Thumbnail | File | Description | Size | Format | |||
|---|---|---|---|---|---|---|---|
| View Details Download | SOURCE1 | Jacobs, A.pdf | 2 MB | Adobe Acrobat PDF | View Details Download |