The investigation of monocyte/macrophage polarisation and its effects on inflammation
Cardiff Metropolitan University
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Macrophage activation can be defined across two polarisatíon states, M1 and M2. It has been suggested influencing monocyte polarisation to the M2 phenotype could have a beneficial impact on inflammatory conditions such as atherosclerosis and diabetes type 2 . Evidence has indicated that PPAR-y has a positive correlation with M2 expression. Recently, studies have shown conjugated lineolic acid (CLA) to have anti-atherosclerotic and antidiabetic effects . The goal of our study was to examine whether CLA exerts anti-inflammatory properties by mediating polarisation to an M2 phenotype, similar to the reported PPAR-y activators, thiazolidinediones (TZDs) . From the human monocytic line, THP-I, macrophage-like phenotype cells were induced with the addition of PMA. This was followed by treatment of either CLA 100uM, rosiglitazone 1uM or IL-13. The effect on the expression of the M2 marker CD206 (MR) was then analysed through flow cytometry. Investigations in line with this involved the effect on MR expression with CLA 20uM and before the differentiation of THP-I cells (pre-treatment). Additionally, the effect of CLA on the pro-inflammatory mediators, IL-8 and TNF-α was also observed. CLA was shown to upregulate the expression of the M2 marker CD206, suggesting promotion towards an M2 phenotype. Interestingly, results demonstrated a synergistic effect between CLA and IL-13. It was also shown that MR expression and CLA have a dose-responsive relationship. Furthermore, post-treament of THP-I cells appears more effective in enhancing MR expression. CLA's anti-inflammatory property correlated with the decrease in TNF-α. Unexpectedly, PPAR-y agonist Rosiglitazone did not have an effect on MR expression but did decrease TNF-α. Neither CLA nor Rosiglitazone decreased IL-8. These results highlight the need to further analyse the effect of CLA on other M2 markers, pro-inflammatory and anti-inflammatory mediators. Further analysis is also required to determine whether the effect of CLA is through PPAR-y. In conclusion, this study supports the potential role of CLA in mediating macrophage polarisation towards a beneficial M2 phenotype.
BSc (Hons) Applied Biomedical Science
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