RESUMO
Adipose tissue affects metabolic-related diseases because it consists of various cell types involved in fat metabolism and adipokine release. CXC ligand 5 (CXCL5) is a member of the CXC chemokine family and is highly expressed by macrophages in white adipose tissue (WAT). In this study, we generated and investigated the function of CXCL5 in knockout (KO) mice using CRISPR/Cas9. The male KO mice did not show significant phenotype differences in normal conditions. However, proteomic analysis revealed that many proteins involved in fatty acid beta-oxidation and mitochondrial localization were enriched in the inguinal WAT (iWAT) of Cxcl5 KO mice. Cxcl5 KO mice also showed decreased protein and transcript expression of genes associated with thermogenesis, including uncoupling protein 1 (UCP1), a well-known thermogenic gene, and increased expression of genes associated with inflammation. The increase in UCP1 expression in cold conditions was significantly retarded in Cxcl5 KO mice. Finally, we found that CXCL5 treatment increased the expression of transcription factors that mediate Ucp1 expression and Ucp1 itself. Collectively, our data show that Ucp1 expression is induced in adipocytes by CXCL5, which is secreted upon ß-adrenergic stimulation by cold stimulation in M1 macrophages. Our data indicate that CXCL5 plays a crucial role in regulating energy metabolism, particularly upon cold exposure. These results strongly suggest that targeting CXCL5 could be a potential therapeutic strategy for people suffering from disorders affecting energy metabolism.
Assuntos
Tecido Adiposo Branco/metabolismo , Quimiocina CXCL5/metabolismo , Macrófagos/metabolismo , Animais , Células Cultivadas , Quimiocina CXCL5/deficiência , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos TransgênicosRESUMO
Matrix metalloproteinases (MMPs) have been implicated in the cleavage of several proinflammatory chemokines, thereby modulating their function and having an impact on the inflammatory process. However, in vivo evidence of such a role remains limited. In this study, we use IL-1ß-induced peritonitis as a model for an acute immune response, which is initiated by neutrophil influx followed by macrophage infiltration within a few hours of IL-1ß injection into the peritoneal cavity. Using single and double knockout mice for MMP-2 and MMP-9, we show that MMP-2 and MMP-9 act synergistically mainly at the initial step of neutrophil recruitment into the peritoneal cavity. The use of bone marrow chimeric mice revealed the cellular sources of MMP-2 and MMP-9 to be distinct, with resident cells being the source of the former and infiltrating leukocytes the source of the latter. We show that the omentum is the main site of neutrophil entry into the peritoneal cavity, where MMP-2 and MMP-9 act synergistically to potentiate the action of CXCL5 (ENA-78/ LIX), thereby, promoting neutrophil migration into the peritoneal cavity. To our knowledge, this is the first in vivo demonstration of MMP-2 and MMP-9 processing of a chemokine that has been directly correlated with an enhanced chemoattracting function.
Assuntos
Quimiocina CXCL5/metabolismo , Interleucina-1beta/administração & dosagem , Metaloproteinase 2 da Matriz/fisiologia , Metaloproteinase 9 da Matriz/fisiologia , Infiltração de Neutrófilos/imunologia , Peritonite/imunologia , Peritonite/patologia , Processamento de Proteína Pós-Traducional/imunologia , Doença Aguda , Animais , Quimiocina CXCL5/deficiência , Modelos Animais de Doenças , Metaloproteinase 2 da Matriz/deficiência , Metaloproteinase 9 da Matriz/deficiência , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Infiltração de Neutrófilos/genética , Neutrófilos/enzimologia , Neutrófilos/imunologia , Neutrófilos/patologia , Peritônio/imunologia , Peritônio/metabolismo , Peritônio/patologia , Peritonite/enzimologia , Processamento de Proteína Pós-Traducional/genéticaRESUMO
CXCL5, a member of the CXC family of chemokines, contributes to neutrophil recruitment during lung inflammation, but its regulation is poorly understood. Because the T cell-derived cytokine IL-17A enhances host defense by triggering production of chemokines, particularly in combination with TNF-α, we hypothesized that IL-17A would enhance TNF-α-induced expression of CXCL5. Intratracheal coadministration of IL-17A and TNF-α in mice induced production of CXCL1, CXCL2, and CXCL5, which was associated with increased neutrophil influx in the lung at 8 and 24 h. The synergistic effects of TNF-α and IL17A were greatly attenuated in Cxcl5(-/-) mice at 24 h, but not 8 h, after exposure, a time when CXCL5 expression was at its peak in wild-type mice. Bone marrow chimeras produced using Cxcl5(-/-) donors and recipients demonstrated that lung-resident cells were the source of CXCL5. Using differentiated alveolar epithelial type II (ATII) cells derived from human fetal lung, we found that IL-17A enhanced TNF-α-induced CXCL5 transcription and stabilized TNF-α-induced CXCL5 transcripts. Whereas expression of CXCL5 required activation of NF-κB, IL-17A did not increase TNF-α-induced NF-κB activation. Apical costimulation of IL-17A and TNF-α provoked apical secretion of CXCL5 by human ATII cells in a transwell system, whereas basolateral costimulation led to both apical and basolateral secretion of CXCL5. The observation that human ATII cells secrete CXCL5 in a polarized fashion may represent a mechanism to recruit neutrophils in host defense in a fashion that discriminates the site of initial injury.
Assuntos
Quimiocina CXCL5/biossíntese , Interleucina-17/fisiologia , Alvéolos Pulmonares/imunologia , Alvéolos Pulmonares/metabolismo , Fator de Necrose Tumoral alfa/fisiologia , Lesão Pulmonar Aguda/genética , Lesão Pulmonar Aguda/imunologia , Lesão Pulmonar Aguda/patologia , Animais , Inibição de Migração Celular/genética , Inibição de Migração Celular/imunologia , Células Cultivadas , Quimiocina CXCL1/biossíntese , Quimiocina CXCL2/biossíntese , Quimiocina CXCL5/deficiência , Quimiocina CXCL5/metabolismo , Quimiotaxia de Leucócito/genética , Quimiotaxia de Leucócito/imunologia , Modelos Animais de Doenças , Quimioterapia Combinada , Humanos , Mediadores da Inflamação/metabolismo , Mediadores da Inflamação/fisiologia , Interleucina-17/administração & dosagem , Interleucina-17/biossíntese , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neutrófilos/imunologia , Neutrófilos/patologia , Pneumonia Bacteriana/imunologia , Pneumonia Bacteriana/metabolismo , Pneumonia Bacteriana/patologia , Alvéolos Pulmonares/patologia , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/biossíntese , Índice de Gravidade de Doença , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Fator de Necrose Tumoral alfa/administração & dosagem , Fator de Necrose Tumoral alfa/biossínteseRESUMO
We show here high levels of expression and secretion of the chemokine CXC ligand 5 (CXCL5) in the macrophage fraction of white adipose tissue (WAT). Moreover, we find that CXCL5 is dramatically increased in serum of human obese compared to lean subjects. Conversely, CXCL5 concentration is decreased in obese subjects after a weight reduction program, or in obese non-insulin-resistant, compared to insulin-resistant, subjects. Most importantly we demonstrate that treatment with recombinant CXCL5 blocks insulin-stimulated glucose uptake in muscle in mice. CXCL5 blocks insulin signaling by activating the Jak2/STAT5/SOCS2 pathway. Finally, by treating obese, insulin-resistant mice with either anti-CXCL5 neutralizing antibodies or antagonists of CXCR2, which is the CXCL5 receptor, we demonstrate that CXCL5 mediates insulin resistance. Furthermore CXCR2-/- mice are protected against obesity-induced insulin resistance. Taken together, these results show that secretion of CXCL5 by WAT resident macrophages represents a link between obesity, inflammation, and insulin resistance.