Thymoquinone – The Natural Healer: A study into the therapeutic properties of the flavonoid, and its ability to alleviate downstream inflammatory mechanisms initiated via receptors for advanced glycated end-products
Cardiff Metropolitan University
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Introduction Receptors for advanced glycated end products play a pivotal role in the pathogenesis of inflammation when bound with their complementary ligands; advanced glycated end products. Said inflammation occurs via several downstream signalling pathways including the NF-kB pathway and can induce severe medical complications. Studies have proved a spliced product of RAGE, named soluble RAGE that has lost its membrane bound domain, circulating in the plasma acting as a decoy receptor inhibiting the detrimental pathways induced by full-length RAGE. Thymoquinone, however is a natural plant-based flavonoid widely identified for its anti-inflammatory and anti-oxidant properties and has been known to treat a broad array of diseases. This study aims to investigate the behaviour of thymoquinone on RAGE and its ability to possibly promote RAGE splicing. Methods THP-1 cells (ECACC, Salisbury) were used in order to mimic an inflammatory model as they were easy to manipulate. A viability count was carried out at varying thymoquinone concentrations in order to illustrate at which concentration the flavonoid would have an apoptotic effect. Levels of soluble RAGE were detected using the a quantitative sandwich ELISA procedure (R&D Systems), and performed on both macrophage and monocytic THP-1 cells. the protocol for which can be found in the appendix. Addition of pro-inflammatory cytokines, TNF-α and INF-γ, were used to induce RAGE expression. Flow cytometry was also performed on monocytic THP-1 cells in order to observe the behaviour of thymoquinone on full-length RAGE when administered with the same pro-inflammatory cytokines. Results Viability counts illustrated thymoquinone having detrimental effects at high concentrations and although deciding on suitable concentrations of the flavonoid, ELISA failed to show any increase in sRAGE when presented 6 alongside pro-inflammatory cytokines. However the flow cytometry successfully proved thymoquinone’s capability to downregulate full-length RAGE expression. Discussion Although there are several mechanisms by which thymoquinone could suppress RAGE expression, the exact one is yet to be identified. It is thought to be through anti-oxidant mechanisms however some evidence, shown in this study, disproves that and suggests it might also act as a decoy ligand. Nevertheless its anti-inflammatory properties seem to be apparent.
BSc (Hons) Biomedical Science
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