RESUMEN
Chronic inflammation is the underlying cause of many diseases, including type 1 diabetes, obesity, and non-alcoholic fatty liver disease. Macrophages are continuously recruited to tissues during chronic inflammation where they exacerbate or resolve the pro-inflammatory environment. Although leukotriene B4 receptor 2 (BLT2) has been characterized as a low affinity receptor to several key eicosanoids and chemoattractants, its precise roles in the setting of inflammation and macrophage function remain incompletely understood. Here we used zebrafish and mouse models to probe the role of BLT2 in macrophage function during inflammation. We detected BLT2 expression in bone marrow derived and peritoneal macrophages of mouse models. Transcriptomic analysis of Ltb4r2-/- and WT macrophages suggested a role for BLT2 in macrophage migration, and studies in vitro confirmed that whereas BLT2 does not mediate macrophage polarization, it is required for chemotactic function, possibly mediated by downstream genes Ccl5 and Lgals3. Using a zebrafish model of tailfin injury, we demonstrated that antisense morpholino-mediated knockdown of blt2a or chemical inhibition of BLT2 signaling impairs macrophage migration. We further replicated these findings in zebrafish models of islet injury and liver inflammation. Moreover, we established the applicability of our zebrafish findings to mammals by showing that macrophages of Ltb4r2-/- mice have defective migration during lipopolysaccharide stimulation in vivo. Collectively, our results demonstrate that BLT2 mediates macrophage migration during inflammation, which implicates it as a potential therapeutic target for inflammatory pathologies.
Asunto(s)
Movimiento Celular , Macrófagos , Receptores de Leucotrieno B4 , Animales , Ratones , Inflamación/genética , Inflamación/metabolismo , Leucotrieno B4/genética , Leucotrieno B4/metabolismo , Macrófagos/citología , Macrófagos/metabolismo , Receptores de Leucotrieno B4/genética , Receptores de Leucotrieno B4/metabolismo , Pez Cebra/genética , Pez Cebra/metabolismoRESUMEN
Leukotriene B4 (LTB4) is a lipid mediator rapidly generated from arachidonic acid in response to various stimuli. This lipid mediator exerts its biological activities by binding to cognate receptors. Two LTB4 receptors have been cloned; BLT1 and BLT2 as a high- and a low-affinity receptors, respectively. In numerous analyses, physiological and pathophysiological importance of LTB4 and cognate receptors in various diseases has been clarified. For example, disruption of the BLT1 gene or treatment with blockers for this receptor reduced various diseases such as rheumatoid arthritis and bronchial asthma in mice, in contrast BLT2 deficiency facilitated several diseases in the small intestine and the skin. These data support the idea that BLT1 blockers and BLT2 agonists could be useful for the cure of these diseases. Thus, various drugs targeting each receptor are being developed by many pharmaceutical companies. In this review, we focus on our current knowledge of the biosynthesis and physiological roles of LTB4 through cognate receptors. We further describe the effects of these receptor deficiencies on several pathophysiological conditions, including the potential of LTB4 receptors as therapeutic targets for the cure of the diseases. Moreover, current information on the structure and post-translational modification of BLT1 and BLT2 is discussed.
Asunto(s)
Artritis Reumatoide , Leucotrieno B4 , Ratones , Animales , Leucotrieno B4/genética , Leucotrieno B4/metabolismo , Leucotrieno B4/farmacología , Piel/metabolismo , Receptores de Leucotrieno B4/genética , Receptores de Leucotrieno B4/metabolismoRESUMEN
Neutrophils form cellular clusters or swarms in response to injury or pathogen intrusion. Yet, intracellular signaling events favoring this coordinated response remain to be fully characterized. Here, we show that calcium signals play a critical role during mouse neutrophil clustering around particles of zymosan, a structural fungal component. Pioneer neutrophils recognizing zymosan or live Candida albicans displayed elevated calcium levels. Subsequently, a transient wave of calcium signals in neighboring cells was observed followed by the attraction of neutrophils that exhibited more persistent calcium signals as they reached zymosan particles. Calcium signals promoted LTB4 production while the blocking of extracellular calcium entry or LTB4 signaling abrogated cluster formation. Finally, using optogenetics to manipulate calcium influx in primary neutrophils, we show that calcium signals could initiate recruitment of neighboring neutrophils in an LTB4-dependent manner. Thus, sustained calcium responses at the center of the cluster are necessary and sufficient for the generation of chemoattractive gradients that attract neutrophils in a self-reinforcing process.
Asunto(s)
Señalización del Calcio , Calcio , Leucotrieno B4 , Neutrófilos , Animales , Calcio/metabolismo , Candida albicans/inmunología , Leucotrieno B4/genética , Leucotrieno B4/fisiología , Ratones , Neutrófilos/inmunología , Zimosan/inmunologíaRESUMEN
Leukotriene B4 (LTB4) is generated by the enzymatic oxidation of arachidonic acid, which is then released from the cell membrane and acts as a potent activator of leukocytes and other inflammatory cells. Numerous studies have demonstrated the physiological and pathophysiological significance of this lipid in various diseases. LTB4 exerts its activities by binding to its specific G protein-coupled receptors (GPCRs): BLT1 and BLT2. In mouse disease models, treatment with BLT1 antagonists or BLT1 gene ablation attenuated various diseases, including bronchial asthma, arthritis, and psoriasis, whereas BLT2 deficiency exacerbated several diseases in the skin, cornea, and small intestine. Therefore, BLT1 inhibitors and BLT2 activators could be beneficial for the treatment of several inflammatory and immune disorders. As a result, attractive compounds targeting LTB4 receptors have been developed by several pharmaceutical companies. This review aims to understand the potential of BLT1 and BLT2 as therapeutic targets for the treatment of various inflammatory diseases. In addition, recent topics are discussed with major focuses on the structure and post-translational modifications of BLT1 and BLT2. Collectively, current evidence on modulating LTB4 receptor functions provides new strategies for the treatment of various diseases.
Asunto(s)
Asma , Psoriasis , Animales , Leucocitos/metabolismo , Leucotrieno B4/genética , Leucotrieno B4/metabolismo , Leucotrieno B4/farmacología , Ratones , Receptores de Leucotrieno B4/genética , Receptores de Leucotrieno B4/metabolismoRESUMEN
Diabetes is a known risk factor for severe coronavirus disease 2019 (COVID-19), the disease caused by the new coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, there is a lack of knowledge about the mechanisms involved in the evolution of COVID-19 in individuals with diabetes. We aimed to evaluate whether the chronic low-grade inflammation of diabetes could play a role in the development of severe COVID-19. We collected clinical data and blood samples of patients with and without diabetes hospitalized for COVID-19. Plasma samples were used to measure inflammatory mediators and peripheral blood mononuclear cells, for gene expression analysis of the SARS-CoV-2 main receptor system (ACE2/TMPRSS2), and for the main molecule of the leukotriene B4 (LTB4) pathway (ALOX5). We found that diabetes activates the LTB4 pathway and that during COVID-19 it increases ACE2/TMPRSS2 as well as ALOX5 expression. Diabetes was also associated with COVID-19-related disorders, such as reduced oxygen saturation as measured by pulse oximetry/fraction of inspired oxygen (FiO2) and arterial partial pressure of oxygen/FiO2 levels, and increased disease duration. In addition, the expressions of ACE2 and ALOX5 are positively correlated, with increased expression in patients with diabetes and COVID-19 requiring intensive care assistance. We confirmed these molecular results at the protein level, where plasma LTB4 is significantly increased in individuals with diabetes. In addition, IL-6 serum levels are increased only in individuals with diabetes requiring intensive care assistance. Together, these results indicate that LTB4 and IL-6 systemic levels, as well as ACE2/ALOX5 blood expression, could be early markers of severe COVID-19 in individuals with diabetes.
Asunto(s)
Enzima Convertidora de Angiotensina 2/metabolismo , Araquidonato 5-Lipooxigenasa/metabolismo , COVID-19/patología , Diabetes Mellitus/patología , Leucotrieno B4/metabolismo , SARS-CoV-2 , Enzima Convertidora de Angiotensina 2/genética , Araquidonato 5-Lipooxigenasa/genética , COVID-19/metabolismo , Regulación de la Expresión Génica , Humanos , Inflamación/metabolismo , Leucotrieno B4/genética , Factores de Riesgo , Transducción de SeñalRESUMEN
The eicosanoid leukotriene B4 (LTB4) relays chemotactic signals to direct neutrophil migration to inflamed sites through its receptor BLT1. However, the mechanisms by which the LTB4-BLT1 axis relays chemotactic signals during intravascular neutrophil response to inflammation remain unclear. Here, we report that LTB4 produced by neutrophils acts as an autocrine/paracrine signal to direct the vascular recruitment, arrest, and extravasation of neutrophils in a sterile inflammation model in the mouse footpad. Using intravital subcellular microscopy, we reveal that LTB4 elicits sustained cell polarization and adhesion responses during neutrophil arrest in vivo. Specifically, LTB4 signaling coordinates the dynamic redistribution of non-muscle myosin IIA and ß2-integrin, which facilitate neutrophil arrest and extravasation. Notably, we also found that neutrophils shed extracellular vesicles in the vascular lumen and that inhibition of extracellular vesicle release blocks LTB4-mediated autocrine/paracrine signaling required for neutrophil arrest and extravasation. Overall, we uncover a novel complementary mechanism by which LTB4 relays extravasation signals in neutrophils during early inflammation response.
Asunto(s)
Inflamación/genética , Leucotrieno B4/genética , Neutrófilos/metabolismo , Receptores de Leucotrieno B4/genética , Animales , Comunicación Autocrina/genética , Antígenos CD18/genética , Movimiento Celular/genética , Factores Quimiotácticos/metabolismo , Humanos , Inflamación/metabolismo , Inflamación/patología , Ratones , Miosina Tipo IIA no Muscular/genética , Comunicación Paracrina/genéticaRESUMEN
Leukotriene B4 (LTB4) is a major type of lipid mediator that is rapidly generated from arachidonic acid through sequential action of 5-lipoxygenase (5-LO), 5-lipoxygenase-activating protein (FLAP) and LTA4 hydrolase (LTA4H) in response to various stimuli. LTB4 is well known to be a chemoattractant for leukocytes, particularly neutrophils, via interaction with its high-affinity receptor BLT1. Extensive attention has been paid to the role of the LTB4-BLT1 axis in acute and chronic inflammatory diseases, such as infectious diseases, allergy, autoimmune diseases, and metabolic disease via mediating recruitment and/or activation of different types of inflammatory cells depending on different stages or the nature of inflammatory response. Recent studies also demonstrated that LTB4 acts on non-immune cells via BLT1 to initiate and/or amplify pathological inflammation in various tissues. In addition, emerging evidence reveals a complex role of the LTB4-BLT1 axis in cancer, either tumor-inhibitory or tumor-promoting, depending on the different target cells. In this review, we summarize both established understanding and the most recent progress in our knowledge about the LTB4-BLT1 axis in host defense, inflammatory diseases and cancer.
Asunto(s)
Leucotrieno B4/fisiología , Receptores de Leucotrieno B4/fisiología , Animales , Enfermedad , Salud , Humanos , Inflamación/fisiopatología , Leucotrieno B4/genética , Neoplasias/fisiopatología , Receptores de Leucotrieno B4/efectos de los fármacos , Receptores de Leucotrieno B4/genéticaRESUMEN
Activation of the leukotriene A4 hydrolase (LTA4H) aminopeptidase (AP) activity with 4-methoxydiphenylmethane (4MDM) promoted resolution of neutrophil infiltration in a murine cigarette smoke-induced model for emphysematous chronic obstructive pulmonary disease. Recently, 4-(4-benzylphenyl)thiazol-2-amine (ARM1) was published as a ligand for LTA4H with potential anti-inflammatory properties. To investigate the effect of modifier structure on enzyme kinetics of LTA4H, a series of analogues bearing structural features of ARM1 and 4MDM were synthesized using trifluoroborate Suzuki coupling reactions. Following, the 2.8 Å X-ray crystal structure of LTA4H complexed with 4-OMe-ARM1, a 4MDM-ARM1 hybrid molecule, was determined. Kinetic analysis showed that ARM1 and related analogues lowered affinity for the enzyme-substrate complex, resulting in a change of mechanism from hyperbolic mixed predominately catalytic activation (HMx(Sp < Ca)A) as observed for 4MDM to a predominately specific activation (HMx(Sp > Ca)A) mechanism. 4-OMe-ARM1 was then shown to dose responsively reduce LTB4 production in human neutrophils.
Asunto(s)
Aminopeptidasas/metabolismo , Antiinflamatorios/farmacología , Activación Enzimática/efectos de los fármacos , Epóxido Hidrolasas/metabolismo , Antiinflamatorios/química , Células Cultivadas , Epóxido Hidrolasas/genética , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Mediadores de Inflamación/metabolismo , Leucotrieno B4/genética , Leucotrieno B4/metabolismo , Estructura Molecular , Neutrófilos/efectos de los fármacos , Neutrófilos/metabolismoRESUMEN
Leishmaniasis is a neglected tropical disease affecting millions of individuals worldwide. P2X7 receptor has been linked to the elimination of Leishmania amazonensis. Biological responses evoked by P2X7 receptor activation have been well-documented, including apoptosis, phagocytosis, cytokine release, such as IL-1ß. It was demonstrated that NLRP3 inflammasome activation and IL-1ß signaling participated in resistance against L. amazonensis. Furthermore, our group has shown that L. amazonensis elimination through P2X7 receptor activation depended on leukotriene B4 (LTB4) production and release. Therefore, we investigated whether L. amazonensis elimination by P2X7 receptor and LTB4 involved NLRP3 inflammasome activation and IL-1ß signaling. We showed that macrophages from NLRP3-/-, ASC-/-, Casp-1/11-/-, gp91phox-/- , and IL-1R-/- mice treated with ATP or LTB4 did not decrease parasitic load as was observed in WT mice. When ASC-/- macrophages were treated with exogenous IL-1ß, parasite killing was noted, however, we did not see parasitic load reduction in IL-1R-/- macrophages. Similarly, macrophages from P2X7 receptor-deficient mice treated with IL-1ß also showed decreased parasitic load. In addition, when we infected Casp-11-/- macrophages, neither ATP nor LTB4 were able to reduce parasitic load, and Casp-11-/- mice were more susceptible to L. amazonensis infection than were WT mice. Furthermore, P2X7-/- L. amazonensis-infected mice locally treated with exogenous LTB4 showed resistance to infection, characterized by lower parasite load and smaller lesions compared to untreated P2X7-/- mice. A similar observation was noted when infected P2X7-/- mice were treated with IL-1ß, i.e., lower parasite load and smaller lesions compared to P2X7-/- mice. These data suggested that L. amazonensis elimination mediated by P2X7 receptor and LTB4 was dependent on non-canonical NLRP3 inflammasome activation, ROS production, and IL-1ß signaling.
Asunto(s)
Inflamasomas/inmunología , Interleucina-1beta/inmunología , Leishmania/inmunología , Leishmaniasis/inmunología , Leucotrieno B4/inmunología , Macrófagos/inmunología , Proteína con Dominio Pirina 3 de la Familia NLR/inmunología , Receptores Purinérgicos P2X7/inmunología , Transducción de Señal/inmunología , Animales , Inflamasomas/genética , Interleucina-1beta/genética , Leishmaniasis/genética , Leishmaniasis/patología , Leucotrieno B4/genética , Macrófagos/parasitología , Ratones , Ratones Noqueados , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Receptores Purinérgicos P2X7/genética , Transducción de Señal/genéticaRESUMEN
BACKGROUND: Genetic variation in the genes ALOX5 (arachidonate 5-lipoxygenase), ALOX5AP (arachidonate 5-lipoxygenase-activating protein) and LTA4H (leukotriene A4 hydrolase) has previously been shown to contribute to the risk of MI (myocardial infarction) in Caucasian and African American populations. All genes encode proteins playing a role in the synthesis of the pro-inflammatory leukotriene B mediators, possibly providing a link between MI and inflammation. The aim of the present study was to investigate whether these associations could be confirmed in the study of China MI patients. The study included 401 Han Chinese MI patients and 409 controls. Six tag single nucleotide polymorphisms (SNPs)-ALOX5 rs12762303 and rs12264801, ALOX5AP rs10507391, LTA4H rs2072512, rs2540487 and rs2540477-were selected. SNP genotyping was performed by an improved multiplex ligation detection reaction assay. RESULTS: The rs2540487 genotype was associated with the risk of MI in overdominant model (P = 0.008). rs12762303 and rs10507391 SNPs were significantly associated with lipid levels in MI patients (P < 0.006-0.008). Several SNPs interacted with alcohol consumption, cigarette smoking, and hypertension to modify TC, TG, LDL-C and CRE levels, and the risk of MI (P < 0.0017 for all). No association between the SNPs of LT pathway and susceptibility to MI was found (P > 0.05 for all). CONCLUSIONS: Taken together, this study provides additional evidence that functional genetic variation of the LT pathway can mediate atherogenic processes and the risk of MI in Chinese.
Asunto(s)
Proteínas Activadoras de la 5-Lipooxigenasa/genética , Araquidonato 5-Lipooxigenasa/genética , Aterosclerosis/genética , Epóxido Hidrolasas/genética , Infarto del Miocardio/genética , Consumo de Bebidas Alcohólicas/genética , Consumo de Bebidas Alcohólicas/fisiopatología , Aterosclerosis/fisiopatología , China , Femenino , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Genotipo , Haplotipos/genética , Humanos , Inflamación/genética , Inflamación/fisiopatología , Leucotrieno B4/genética , Leucotrieno B4/metabolismo , Masculino , Redes y Vías Metabólicas/genética , Persona de Mediana Edad , Infarto del Miocardio/fisiopatología , Polimorfismo de Nucleótido Simple/genética , Factores de Riesgo , Población BlancaRESUMEN
Poorly controlled diabetes leads to comorbidities and enhanced susceptibility to infections. While the immune components involved in wound healing in diabetes have been studied, the components involved in susceptibility to skin infections remain unclear. Here, we examined the effects of the inflammatory lipid mediator leukotriene B4 (LTB4) signaling through its receptor B leukotriene receptor 1 (BLT1) in the progression of methicillin-resistant Staphylococcus aureus (MRSA) skin infection in 2 models of diabetes. Diabetic mice produced higher levels of LTB4 in the skin, which correlated with larger nonhealing lesion areas and increased bacterial loads compared with nondiabetic mice. High LTB4 levels were also associated with dysregulated cytokine and chemokine production, excessive neutrophil migration but impaired abscess formation, and uncontrolled collagen deposition. Both genetic deletion and topical pharmacological BLT1 antagonism restored inflammatory response and abscess formation, followed by a reduction in the bacterial load and lesion area in the diabetic mice. Macrophage depletion in diabetic mice limited LTB4 production and improved abscess architecture and skin host defense. These data demonstrate that exaggerated LTB4/BLT1 responses mediate a derailed inflammatory milieu that underlies poor host defense in diabetes. Prevention of LTB4 production/actions could provide a new therapeutic strategy to restore host defense in diabetes.
Asunto(s)
Diabetes Mellitus Experimental/metabolismo , Leucotrieno B4/metabolismo , Piel/inmunología , Piel/metabolismo , Infecciones Cutáneas Estafilocócicas/inmunología , Absceso/inmunología , Absceso/patología , Animales , Carga Bacteriana , Movimiento Celular , Quimiocinas/metabolismo , Citocinas/metabolismo , Femenino , Inflamación , Leucotrieno B4/genética , Leucotrieno B4/inmunología , Macrófagos/inmunología , Masculino , Staphylococcus aureus Resistente a Meticilina/patogenicidad , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neutrófilos/inmunología , Receptores de Leucotrieno B4/efectos de los fármacos , Receptores de Leucotrieno B4/genética , Receptores de Leucotrieno B4/metabolismo , Transducción de Señal , Piel/patología , Infecciones Cutáneas Estafilocócicas/patologíaRESUMEN
Age-related macular degeneration (AMD), a progressive chronic disease of the central retina, is associated with aging and is a leading cause of blindness worldwide. Here, we demonstrate that leukotriene B4 (LTB4) receptor 1 (BLT1) promotes laser-induced choroidal neovascularization (CNV) in a mouse model for wet-type AMD. CNV was significantly less in BLT1-deficient (BLT1-KO) mice compared with BLT1-WT controls. Expression of several proangiogenic and profibrotic factors was lower in BLT1-KO eyes than in BLT1-WT eyes. LTB4 production in the eyes was substantially increased in the early phase after laser injury. BLT1 was highly expressed in M2 macrophages in vitro and in vivo, and ocular BLT1+ M2 macrophages were increased in the aged eyes after laser injury. Furthermore, M2 macrophages were rapidly attracted by LTB4 and subsequently produced VEGF-A- through BLT1-mediated signaling. Consequently, intravitreal injection of M2 macrophages augmented CNV formation, which was attenuated by BLT1 deficiency. Thus, laser-induced injury to the retina triggered LTB4 production and attracted M2 macrophages via BLT1, leading to development of CNV. A selective BLT1 antagonist (CP105696) and 3 LTB4 inhibitors (zileuton, MK-886, and bestatin) reduced CNV in a dose-dependent manner. CP105696 also inhibited the accumulation of BLT1+ M2 macrophages in the laser-injured eyes of aged mice. Together, these results indicate that the LTB4-BLT1 axis is a potentially novel therapeutic target for CNV of wet-type AMD.
Asunto(s)
Leucotrieno B4/metabolismo , Macrófagos/metabolismo , Degeneración Macular/metabolismo , Neovascularización Patológica/metabolismo , Receptores de Leucotrieno B4/metabolismo , Animales , Benzopiranos/farmacología , Ácidos Carboxílicos/farmacología , Neovascularización Coroidal , Modelos Animales de Enfermedad , Ojo/efectos de la radiación , Lesiones Oculares , Hidroxiurea/análogos & derivados , Hidroxiurea/farmacología , Indoles/farmacología , Rayos Láser/efectos adversos , Leucina/análogos & derivados , Leucina/farmacología , Leucotrieno B4/antagonistas & inhibidores , Leucotrieno B4/genética , Macrófagos/efectos de los fármacos , Degeneración Macular/patología , Masculino , Ratones , Ratones Noqueados , Neovascularización Patológica/tratamiento farmacológico , Receptores de Leucotrieno B4/genética , Transducción de SeñalRESUMEN
BACKGROUND: Recognizing patients at risk for pulmonary complications (PC) is of high clinical relevance. Migration of polymorphonuclear leukocytes (PMN) to inflammatory sites plays an important role in PC, and is tightly regulated by specific chemokines including interleukin (IL)-8 and other mediators such as leukotriene (LT)B4. Previously, we have reported that LTB4 indicated early patients at risk for PC after trauma. Here, the relevance of LTB4 to indicating lung integrity in a newly established long-term porcine severe trauma model (polytrauma, PT) was explored. METHODS: Twelve pigs (3 months old, 30 ± 5kg) underwent PT including standardized femur fracture, lung contusion, liver laceration, hemorrhagic shock, subsequent resuscitation and surgical fracture fixation. Six animals served as controls (sham). After 72h lung damage and inflammatory changes were assessed. LTB4 was determined in plasma before the experiment, immediately after trauma, and after 2, 4, 24 or 72h. Bronchoalveolar lavage (BAL)-fluid was collected prior and after the experiment. RESULTS: Lung injury, local gene expression of IL-8, IL-1ß, IL-10, IL-18 and PMN-infiltration into lungs increased significantly in PT compared with sham. Systemic LTB4 increased markedly in both groups 4h after trauma. Compared with declined plasma LTB4 levels in sham, LTB4 increased further in PT after 72h. Similar increase was observed in BAL-fluid after PT. CONCLUSIONS: In a severe trauma model, sustained changes in terms of lung injury and inflammation are determined at day 3 post-trauma. Specifically, increased LTB4 in this porcine long-term model indicated a rapid inflammatory alteration both locally and systemically. The results support the concept of LTB4 as a biomarker for PC after severe trauma and lung contusion.
Asunto(s)
Inflamación/sangre , Leucotrieno B4/sangre , Lesión Pulmonar/sangre , Heridas y Lesiones/sangre , Animales , Líquido del Lavado Bronquioalveolar , Modelos Animales de Enfermedad , Humanos , Inflamación/etiología , Inflamación/genética , Inflamación/fisiopatología , Interleucinas/sangre , Leucotrieno B4/genética , Pulmón/metabolismo , Pulmón/fisiopatología , Lesión Pulmonar/etiología , Lesión Pulmonar/genética , Lesión Pulmonar/fisiopatología , Neutrófilos/metabolismo , Porcinos , Heridas y Lesiones/complicaciones , Heridas y Lesiones/fisiopatologíaRESUMEN
Dioxin and related chemicals alter the expression of a number of genes by activating the aryl hydrocarbon receptors (AHR) to produce a variety of disorders including hepatotoxicity. However, it remains largely unknown how these changes in gene expression are linked to toxicity. To address this issue, we initially examined the effect of 2,3,7,8-tetrachrolodibenzo-p-dioxin (TCDD), a most toxic dioxin, on the hepatic and serum metabolome in male pubertal rats and found that TCDD causes many changes in the level of fatty acids, bile acids, amino acids, and their metabolites. Among these findings was the discovery that TCDD increases the content of leukotriene B4 (LTB4), an inducer of inflammation due to the activation of leukocytes, in the liver of rats and mice. Further analyses suggested that an increase in LTB4 comes from a dual mechanism consisting of an induction of arachidonate lipoxygenase-5, a rate-limiting enzyme in LTB4 synthesis, and the down-regulation of LTC4 synthase, an enzyme that converts LTA4 to LTC4. The above changes required AHR activation, because the same was not observed in AHR knock-out rats. In agreement with LTB4 accumulation, TCDD caused the marked infiltration of neutrophils into the liver. However, deleting LTB4 receptors (BLT1) blocked this effect. A TCDD-produced increase in the mRNA expression of inflammatory markers, including tumor-necrosis factor and hepatic damage, was also suppressed in BLT1-null mice. The above observations focusing on metabolomic changes provide novel evidence that TCDD accumulates LTB4 in the liver by an AHR-dependent induction of LTB4 biosynthesis to cause hepatotoxicity through neutrophil activation.
Asunto(s)
Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Dioxinas/toxicidad , Leucotrieno B4/biosíntesis , Infiltración Neutrófila/efectos de los fármacos , Neutrófilos/metabolismo , Dibenzodioxinas Policloradas/toxicidad , Receptores de Hidrocarburo de Aril/metabolismo , Animales , Enfermedad Hepática Inducida por Sustancias y Drogas/genética , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Leucotrieno B4/genética , Activación Neutrófila/efectos de los fármacos , Infiltración Neutrófila/genética , Neutrófilos/patología , Ratas , Ratas Mutantes , Receptores de Hidrocarburo de Aril/genética , Receptores de Leucotrieno B4/genética , Receptores de Leucotrieno B4/metabolismoRESUMEN
Tissue inflammation is a key component of obesity-induced insulin resistance, with a variety of immune cell types accumulating in adipose tissue. Here, we have demonstrated increased numbers of B2 lymphocytes in obese adipose tissue and have shown that high-fat diet-induced (HFD-induced) insulin resistance is mitigated in B cell-deficient (Bnull) mice. Adoptive transfer of adipose tissue B2 cells (ATB2) from wild-type HFD donor mice into HFD Bnull recipients completely restored the effect of HFD to induce insulin resistance. Recruitment and activation of ATB2 cells was mediated by signaling through the chemokine leukotriene B4 (LTB4) and its receptor LTB4R1. Furthermore, the adverse effects of ATB2 cells on glucose homeostasis were partially dependent upon T cells and macrophages. These results demonstrate the importance of ATB2 cells in obesity-induced insulin resistance and suggest that inhibition of the LTB4/LTB4R1 axis might be a useful approach for developing insulin-sensitizing therapeutics.
Asunto(s)
Tejido Adiposo/inmunología , Subgrupos de Linfocitos B/inmunología , Resistencia a la Insulina/inmunología , Leucotrieno B4/inmunología , Receptores de Leucotrieno B4/inmunología , Transducción de Señal/inmunología , Tejido Adiposo/patología , Animales , Subgrupos de Linfocitos B/patología , Grasas de la Dieta/efectos adversos , Grasas de la Dieta/farmacología , Resistencia a la Insulina/genética , Leucotrieno B4/genética , Macrófagos/inmunología , Macrófagos/patología , Ratones , Ratones Noqueados , Obesidad/inducido químicamente , Obesidad/genética , Obesidad/inmunología , Obesidad/patología , Receptores de Leucotrieno B4/genética , Transducción de Señal/genética , Linfocitos T/inmunología , Linfocitos T/patologíaRESUMEN
Unresolved experimental Lyme arthritis in C3H 5-lipoxygenase (5-LOX)-/- mice is associated with impaired macrophage phagocytosis of Borrelia burgdorferi In the present study, we further investigated the effects of the 5-LOX metabolite, leukotriene (LT)B4 on phagocytosis of B. burgdorferi Bone marrow-derived macrophages (BMDMs) from 5-LOX-/- mice were defective in the uptake and killing of B. burgdorferi from the earliest stages of spirochete internalization. BMDMs from mice deficient for the LTB4 high-affinity receptor (BLT1-/-) were also unable to efficiently phagocytose B. burgdorferi Addition of exogenous LTB4 augmented the phagocytic capability of BMDMs from both 5-LOX-/- and BLT1-/- mice, suggesting that the low-affinity LTB4 receptor, BLT2, might be involved. Blocking BLT2 activity with the specific antagonist, LY255283, inhibited phagocytosis in LTB4-stimulated BLT1-/- BMDMs, demonstrating a role for BLT2. However, the lack of a phagocytic defect in BLT2-/- BMDMs suggested that this was a compensatory effect. In contrast, 12(S)-hydroxyheptadeca-5Z,8E,10E-trienoic acid, a natural BLT2-specific high-affinity ligand, and resolvin E1, a BLT1 agonist, were both unable to boost phagocytosis in BMDMs from either 5-LOX-/- or BLT1-/- mice, suggesting a specific role for LTB4 in mediating phagocytosis in murine macrophages. This study demonstrates that LTB4 promotes macrophage phagocytosis of bacteria via BLT1, and that BLT2 can fulfill this role in the absence of BLT1.
Asunto(s)
Araquidonato 5-Lipooxigenasa/genética , Enfermedad de Lyme/genética , Receptores de Leucotrieno B4/genética , Animales , Araquidonato 5-Lipooxigenasa/metabolismo , Borrelia burgdorferi/genética , Borrelia burgdorferi/patogenicidad , Modelos Animales de Enfermedad , Humanos , Leucotrieno B4/administración & dosificación , Leucotrieno B4/genética , Leucotrieno B4/metabolismo , Enfermedad de Lyme/metabolismo , Enfermedad de Lyme/microbiología , Enfermedad de Lyme/patología , Macrófagos/metabolismo , Macrófagos/patología , Ratones , Ratones Transgénicos , Fagocitosis/genética , Receptores de Leucotrieno B4/antagonistas & inhibidores , Tetrazoles/administración & dosificaciónRESUMEN
Future feed for farmed fish are based on untraditional feed ingredients, which will change nutrient profiles compared to traditional feed based on marine ingredients. To understand the impact of oils from different sources on fish health, n-6 and n-3 polyunsaturated fatty acids (PUFAs) were added to salmon head kidney cells, in a fully crossed design, to monitor their individual and combined effects on gene expression. Exposing salmon head kidney cells to single fatty acids, arachidonic acid (AA) or decosahexaenoic acid (DHA), resulted in down-regulation of cell signaling pathway genes and specific fatty acid metabolism genes as well as reduced prostaglandin E2 (PGE2) secretion. Eicosapentaenoic acid (EPA) had no impact on gene transcription in this study, but reduced the cell secretion of PGE2. The combined effect of AA + EPA resulted in up-regulation of eicosanoid pathway genes and the pro-inflammatory cytokine, tumor necrosis factor alpha (TNF-α), Bclx (an inducer of apoptosis) and fatty acid translocase (CD36) as well as increased cell secretion of PGE2 into the media. Adding single fatty acids to salmon head kidney cells decreased inflammation markers in this model. The combination AA + EPA acted differently than the rest of the fatty acid combinations by increasing the inflammation markers in these cells. The concentration of fatty acid used in this experiment did not induce any lipid peroxidation responses.
Asunto(s)
Ácido Araquidónico/farmacología , Ácidos Docosahexaenoicos/farmacología , Ácido Eicosapentaenoico/farmacología , Riñón Cefálico/citología , Leucocitos/efectos de los fármacos , Salmón/metabolismo , Alprostadil/análogos & derivados , Alprostadil/metabolismo , Animales , Antígenos CD36/genética , Células Cultivadas , Ciclooxigenasa 2/genética , Dinoprostona/metabolismo , Ácido Eicosapentaenoico/análogos & derivados , Ácido Eicosapentaenoico/genética , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Oxidorreductasas Intramoleculares/genética , Leucocitos/metabolismo , Leucotrieno B4/análogos & derivados , Leucotrieno B4/genética , Masculino , Salmón/genética , Factor de Necrosis Tumoral alfa/genética , Proteína bcl-X/genéticaRESUMEN
Leukotriene B4 (LTB4) is a potent inflammatory mediator derived from arachidonic acid. Two G protein-coupled receptors for LTB4 have been identified: a high-affinity receptor, BLT1, and a low-affinity receptor, BLT2. Both receptors mainly couple to pertussis toxin-sensitive Gi-like G proteins and induce cell migration. 12(S)-hydroxy-5Z,8E,10E-heptadecatrienoic acid (12-HHT) was identified to bind BLT2 with higher affinity than LTB4. Expression of BLT1 was confirmed in type 1 helper T cells, type 2 helper T cells, type 17 helper T cells, effector CD8(+) T cells, dendritic cells and osteoclasts in addition to granulocytes, eosinophils and macrophages, and BLT1-deficient mice showed greatly reduced phenotypes in models of various inflammatory diseases, such as peritonitis, bronchial asthma, rheumatoid arthritis, atherosclerosis and osteoporosis. In mice, BLT2 expression is restricted to intestinal epithelial cells and epidermal keratinocytes. BLT2-deficient mice showed enhanced colitis after administration of dextran sulfate, possibly due to reduced intestinal barrier function. An aspirin-dependent reduction in 12-HHT production was responsible for delayed skin wound healing, showing that the 12-HHT/BLT2 axis also plays an important role in skin biology. BLT1 and BLT2 are therefore potential targets for the development of novel drugs.
Asunto(s)
Inmunidad Innata/genética , Inflamación/genética , Receptores de Leucotrieno B4/genética , Animales , Aspirina/administración & dosificación , Linfocitos T CD8-positivos/metabolismo , Movimiento Celular/efectos de los fármacos , Movimiento Celular/genética , Clonación Molecular , Humanos , Inflamación/tratamiento farmacológico , Inflamación/patología , Leucotrieno B4/genética , Leucotrieno B4/metabolismo , Ratones , Receptores de Leucotrieno B4/biosíntesis , Receptores de Leucotrieno B4/metabolismo , Cicatrización de Heridas/genéticaRESUMEN
BACKGROUND: We investigated the relationships between the ALOX5AP gene rs10507391 and rs4769874 polymorphisms, serum levels of leukotriene (LT) B4, and risk of acute coronary syndrome (ACS). METHODS: A total of 709 participants, comprising 508 ACS patients (ACS group) and 201 noncoronary artery disease patients with chest pain (control group) were recruited from the Han population of the Changwu region in China. Two polymorphic loci were genotyped using polymerase chain reaction and restriction fragment length polymorphism analysis. Serum LTB4 level was determined by enzyme-linked immunosorbent assay. RESULTS: Serum LTB4 levels were significantly higher (P<0.001) in the ACS group (median/interquartile range, 470.27/316.32 pg/ml) than in the control group (233.05/226.82 pg/ml). No statistical differences were observed between genotype, allele and haplotype frequencies for the tested loci in either the ACS group or the control group, even after adjustments were made for conventional risk factors by multivariate logistic regression. This suggests there is no association between the ALOX5AP rs10507391 and rs4769874 polymorphisms and ACS risk. Elevated serum LTB4 level was closely linked to ACS risk, and may be independent of traditional risk factors as a risk factor for ACS (P<0.001). There was no significant association between serum LTB4 levels and the two variants in either the ACS group or the control group. CONCLUSIONS: Rs10507391, rs4769874 and its haplotypes in ALOX5AP are unrelated to ACS risk in the Chinese Han population of Changwu, but elevated serum LTB4 level is strongly associated with ACS risk. Serum LTB4 level is not subject to the influence of either the rs10507391, rs4769874 or the haplotype.