Rol de PAR2 en macrófagos similares a M1 asociados con AR

Identificador:  TFG:2194

Handle: https://hdl.handle.net/20.500.11797/TFG2194

Autores:  Fortuny Martí, Lídia

Resumen:
Rheumatoid Arthritis (RA) is an autoimmune disease characterised by chronic inflammation in the joints. M1-like macrophages have been associated with the synovial inflammation observed in the pathology due to the release of pro-inflammatory cytokines. Nowadays, the main treatments for RA target the inflammatory pathways. While several therapies with different mechanisms of action exist, they do not work in all patients. Also, RA patients sometimes develop antibodies against these drugs. Protease-activated receptor 2 (PAR2) has been proposed as a possible therapeutic target since the receptor has been linked to the severity of RA. However, the mechanism by which PAR2 drives RA pathology is unclear and until recently, a suitable inhibitor has not been available. The aim of this study was to characterise the function of PAR2 in M1-like macrophages and investigate its possible inhibition with the recently reported PAR2 inhibitor, AZ8838. THP-1 cells were differentiated with phorbol-12-myristate-13-acetate (PMA) to an M1-like macrophage phenotype. The cells were stimulated with several molecules (lipopolysaccharide (LPS) and the PAR2 activating peptides FLIGRLO-NH2 and SLIGKV-NH2) in the presence or absence of AZ8838. Culture supernatants were collected after 24 hours. ELISA was used to measure the cytokine profile, with changes in cell morphology also analysed. Cell viability was investigated using an MTT assay. PMA differentiated macrophages released high amounts of pro-inflammatory cytokines related to RA (IL-6, IL-8 and TNFα). Moreover, the activation of PAR2 enhanced the inflammatory response as evidence by IL-6 secretion, suggesting that the inhibition of PAR2 in M1-like macrophages is a potential treatment for this disease. However, AZ8838 was not able to inhibit PAR2 activity at the concentrations studied. Therefore, different concentrations of AZ8838 and macrophage PAR2 activators should be tried in future investigations. Finally, neither AZ8388 or any of the stimulators used affect the viability of the cells.