To investigate the effects of LPS with IFN-γ and the flavonoid Luteolin on human stem cell derived neural cells
Cardiff Metropolitan University
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Introduction: Inflammation is a very important part of the immune reaction to pathogens such as bacteria, when the body recognises a certain stimulus, the body causes inflammation to which causes an influx of immune cells such as natural killer cells to fight the pathogen. However this inflammation is not always good and has been shown to be implicated in neurodegenerative diseases as when not properly regulated it can cause cell damage. A common stimulus which causes a large inflammatory response is lipopolysaccharide which is found on the cell surface of Gram negative bacteria such as Escherichia coli. IFN-γ also produces an inflammatory response in cells as it is a pro-inflammatory cytokine. Luteolin is a flavonoid which is found in foods such as celery and is proven to have shown anti-inflammatory as well as ant-oxidant properties. Therefore the aim of this project was to see if luteolin could reduce the amount of inflammation caused by both LPS and IFN-γ in neural stem cells (NSCs), neurons and astrocytes. Method: Neurons, astrocytes and NSCs were grown up on 12 well plates after being differentiated from NSCs, IFN-γ and LPS were added to the cells to cause inflammation and luteolin was added to try and reduce inflammation. The concentration of LPS and IFN-γ used were worked out by carrying out a PI viability assay. Inflammation was monitored by measuring MHC-I and MHC-II expression by carrying out flow cytometry on neurons only, neurons with LPS and IFN-γ, neurons with luteolin and neurons with IFN-γ, LPS and luteolin. An ELISA was carried out to measure IL-6 production of neurons under the same concentrations as the neurons used in flow cytometry. 4 Results: The results showed that for both LPS and IFN-γ concentration there was no statistical difference between the concentrations. During flow cytometry the addition of both LPS and IFN-γ caused an increase in the expression of both MHC-I and MHC-II compared to cells only in neurons. The addition of luteolin to the cells containing LPS and IFN-γ caused both MHC molecules to be expressed more than stimuli only. The results also showed that there was no significant difference between IL-6 productions between different conditions of the cells between the three cell types. Conclusion: In conclusion luteolin did not reduce inflammation in neural cells after they had been inflamed by the addition of LPS and IFN-γ instead it increased inflammation as shown by the increased MHC-II expression that was found in the flow cytometry assay. This result contradicts what has been shown in literature before about the anti-inflammatory properties of luteolin.
BSc (Hons) Biomedical Science
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