RT info:eu-repo/semantics/doctoralThesis
T1 Análisis lipidómico de los cambios en el metabolismo lipídico que tienen lugar durante la activación de los macrófagos peritoneales de ratón por estímulos proinflamatorios
A1 Monge Bartolomé, Patricia
A2 Universidad de Valladolid. Escuela de Doctorado
K1 Bioquímica
K1 Macrophage
K1 Macrófago
K1 Phospholipase
K1 Fosfolipasa
K1 Arachidonic acid
K1 Ácido araquidónico
K1 Adrenic acid
K1 Ácido adrénico
K1 2403 Bioquímica
AB Macrophages have been studied as one of the main sources of free AA and eicosanoids. In the presence of an inflammatory stimulus, the AA contained in membrane phospholipids is hydrolyzed to be released and subsequently metabolized to its oxygenated derivatives, which perform biological functions. In this work, the mobilization of AA and AdA from phospholipids before zymosan and its opsonized homologue is studied, as well as the role played by different phospholipases in the hydrolysis and release of said n-6 fatty acid. The results indicate that AA and AdA are mainly located in PE species, mainly in plasmalogens, although a high amount is also located in PC and PI. In the presence of zymosan, macrophages experience decreases in these last two phospholipids, without large variations in PE. However, the release of AA and AdA is higher in the presence of opsonized zymosan. In addition, selective inhibitors of the phospholipases sPLA2-V, cPLA2α and iPLA2β were used in the experiments, observing the great role that cPLA2α plays in the hydrolysis and subsequent release of AA, while both cPLA2α and iPLA2β participate in the mobilization of AdA. .This study has analyzed how, in the presence of exogenous AdA, macrophages use AA in the same way as when they are not enriched with AdA, therefore, these fatty acids do not compete with each other in inflammatory conditions.On the other hand, the reason for the lack of AA mobilization from PE species has also been analyzed. Data reported in the literature have described the lack of mobilization of said fatty acid as a continuous reincorporation of this in the PE species thanks to CoA-IT. However, in this study it is observed that, although in the presence of zymosan there is no apparent release in PE due to remodeling, decreases in levels are observed in the presence of opsonized zymosan. Thanks to studies carried out using [3H]AA labeling, it has been confirmed that remodeling in the presence of opsonized zymosan is slower and the rate of remodeling is lower than that analyzed with unopsonized zymosan. These results explain the rapid recovery of PE with AA in the presence of zymosan unlike its opsonized counterpart.Due to the great involvement reported in the literature of oxidized phospholipids in inflammatory situations, a detailed analysis of their presence in murine peritoneal macrophages is performed. For its detection, techniques with great sensitivity and reproducibility have been used, such as chromatography coupled with mass spectrometry. The results indicate an increase in the levels of oxidized phospholipids under inflammatory conditions. The hydrolysis and release of these compounds is of great importance in inflammatory situations, describing the great implication of iPLA2β since, as has been verified, its inhibition prevents the release of AAox. However, it has been observed that cPLA2α also partially participates in the synthesis of oxidized phospholipids, hydrolyzing and releasing AA, which is subsequently oxidized through the LOX enzymatic pathway.Finally, a lipidomic study of the phospholipids present in peritoneal macrophages polarized to M1 and M2 was carried out in order to find differences between both phenotypes, both in their basal and stimulated forms. Although no differences were found between both polarizations and control cells in terms of AA release under inflammatory conditions, the release of fatty acids such as AdA, DHA, DPA and EPA in activated cells was different in both phenotypes, with less release observed. and therefore, greater accumulation in the M1 phenotype.
YR 2021
FD 2021
LK https://uvadoc.uva.es/handle/10324/60589
UL https://uvadoc.uva.es/handle/10324/60589
LA spa
NO Escuela de Doctorado
DS UVaDOC
RD 22-dic-2024