RESUMEN
For decades, lipids were confined to the field of structural biology and energetics as they were considered only structural constituents of cellular membranes and efficient sources of energy production. However, with advances in our understanding in lipidomics and improvements in the technological approaches, astounding discoveries have been made in exploring the role of lipids as signaling molecules, termed bioactive lipids. Among these bioactive lipids, sphingolipids have emerged as distinctive mediators of various cellular processes, ranging from cell growth and proliferation to cellular apoptosis, executing immune responses to regulating inflammation. Recent studies have made it clear that sphingolipids, their metabolic intermediates (ceramide, sphingosine-1-phosphate, and N-acetyl sphingosine), and enzyme systems (cyclooxygenases, sphingosine kinases, and sphingomyelinase) harbor diverse yet interconnected signaling pathways in the central nervous system (CNS), orchestrate CNS physiological processes, and participate in a plethora of neuroinflammatory and neurodegenerative disorders. Considering the unequivocal importance of sphingolipids in CNS, we review the recent discoveries detailing the major enzymes involved in sphingolipid metabolism (particularly sphingosine kinase 1), novel metabolic intermediates (N-acetyl sphingosine), and their complex interactions in CNS physiology, disruption of their functionality in neurodegenerative disorders, and therapeutic strategies targeting sphingolipids for improved drug approaches.
Asunto(s)
Sistema Nervioso Central/fisiopatología , Inflamación/fisiopatología , Lípidos de la Membrana/fisiología , Modelos Biológicos , Degeneración Nerviosa/fisiopatología , Enfermedades Neurodegenerativas/fisiopatología , Esfingolípidos/fisiología , Enfermedad de Alzheimer/metabolismo , Enfermedad de Alzheimer/fisiopatología , Ceramidas/fisiología , Eicosanoides/fisiología , Predicción , Homeostasis , Humanos , Inflamación/patología , Lipooxigenasa/fisiología , Lisofosfolípidos/fisiología , Degeneración Nerviosa/patología , Enfermedades Neurodegenerativas/patología , Neuroglía/metabolismo , Neuronas/metabolismo , Enfermedad de Parkinson/metabolismo , Enfermedad de Parkinson/fisiopatología , Fosfotransferasas (Aceptor de Grupo Alcohol)/fisiología , Prostaglandina-Endoperóxido Sintasas/fisiología , Esfingosina/análogos & derivados , Esfingosina/fisiologíaRESUMEN
Ferroptosis is an iron-dependent form of cell death characterized by intracellular lipid peroxide accumulation and redox imbalance. Ferroptosis shows specific biological and morphological features when compared to the other cell death patterns. The loss of lipid peroxide repair activity by glutathione peroxidase 4 (GPX4), the presence of redox-active iron and the oxidation of polyunsaturated fatty acid (PUFA)-containing phospholipids are considered as distinct fingerprints of ferroptosis. Several pathways, including amino acid and iron metabolism, ferritinophagy, cell adhesion, p53, Keap1/Nrf2 and phospholipid biosynthesis, can modify susceptibility to ferroptosis. Through the decades, various diseases, including acute kidney injury; cancer; ischemia-reperfusion injury; and cardiovascular, neurodegenerative and hepatic disorders, have been associated with ferroptosis. In this review, we provide a comprehensive analysis of the main biological and biochemical mechanisms of ferroptosis and an overview of chemicals used as inducers and inhibitors. Then, we report the contribution of ferroptosis to the spectrum of liver diseases, acute or chronic. Finally, we discuss the use of ferroptosis as a therapeutic approach against hepatocellular carcinoma, the most common form of primary liver cancer.
Asunto(s)
Ferroptosis , Hepatopatías/patología , Animales , Autofagia/fisiología , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Ciclohexilaminas/farmacología , Cisteína/metabolismo , Ferroptosis/efectos de los fármacos , Glutatión/biosíntesis , Hemo/metabolismo , Humanos , Hierro/metabolismo , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Lipooxigenasa/fisiología , Neoplasias Hepáticas/patología , Estrés Oxidativo , Fenilendiaminas/farmacología , Fosfolípido Hidroperóxido Glutatión Peroxidasa/fisiología , Piperazinas/farmacología , Quinoxalinas/farmacología , Especies Reactivas de Oxígeno/metabolismo , Daño por Reperfusión/patología , Transducción de Señal , Sorafenib/farmacología , Compuestos de Espiro/farmacología , Sulfasalazina/farmacología , Proteína p53 Supresora de Tumor/metabolismo , alfa-Tocoferol/farmacologíaRESUMEN
We previously reported that intracerebroventricularly (ICV) injected arachidonic acid (AA) could produce pressor and bradycardic responses on the cardiovascular system and hyperventilation effect on the respiratory system by activating cyclooxygenase (COX). We also demonstrated that centrally injected AA-induced cardiovascular and respiratory responses were mediated by COX-metabolites, such as thromboxane A2 (TXA2), prostaglandin (PG) D, PGE, and PGF2α. Brain tissue is also able to express the lipoxygenase (LOX) enzyme and LOX-induced AA-metabolites. The current study was designed to investigate the possible mediation of the central LOX pathway in AA-induced cardiorespiratory effects in anesthetized rats. Central pretreatment with different doses of a non-selective LOX inhibitor, nordihydroguaiaretic acid (NDGA) (500 and 1000 µg; ICV) partially blocked the AA (0.5 µmol; ICV)-evoked pressor and bradycardic cardiovascular responses in male anesthetized Sprague Dawley rats. Pretreatment with different doses of NDGA (500 and 1000 µg; ICV) also reduced AA-induced hyperventilation responses, with an increase in tidal volume, respiratory rate and minute ventilation, in the rats. Moreover, AA-induced increasing pO2 and decreasing pCO2 responses were diminished by central NDGA pretreatment. In summary, our findings show that the central LOX pathway might mediate, at least in part, centrally administered AA-evoked cardiorespiratory and blood gases responses.
Asunto(s)
Ácido Araquidónico/farmacología , Presión Arterial/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Lipooxigenasa/fisiología , Frecuencia Respiratoria/efectos de los fármacos , Volumen de Ventilación Pulmonar/efectos de los fármacos , Animales , Análisis de los Gases de la Sangre , Dióxido de Carbono/sangre , Inyecciones Intraventriculares , Inhibidores de la Lipooxigenasa/farmacología , Masoprocol/farmacología , Oxígeno/sangre , Presión Parcial , RatasRESUMEN
Lipoxygenases (LOXs) are dioxygenases that catalyze the peroxidation of linoleic acid (LA) or arachidonic acid (AA), in the presence of molecular oxygen. The existence of inflammatory component in the tumor microenvironment intimately links the LOXs to gastrointestinal (GI) cancer progression. Amongst the six-different human LOX-isoforms, 5-LOX is the most vital enzyme for leukotriene (LT) biosynthesis, which is the main inflammation intermediaries. As recent investigations have shown the association of 5-LOX with tumor metastasis, there has also been significant progress in discovering the function of 5-LOX pathway in GI cancer. Studies on GI cancer cells using the pharmacological drugs targeting 5-LOX pathway have shown antiproliferative and proapoptotic effects. Pharmacogenetic discoveries in other diseases have revealed strong heritable basis for the leukotriene pathway, which helps in exploring the mechanistic source of genetic alteration within the leukotriene pathway and offer insights into GI cancer pathogenesis and future prospects for treatment and prevention. This review recapitulates the current research status of 5-LOX activity in GI malignancies.
Asunto(s)
Araquidonato 5-Lipooxigenasa/fisiología , Neoplasias Gastrointestinales/etiología , Neoplasias Gastrointestinales/patología , Metabolismo de los Lípidos/fisiología , Animales , Progresión de la Enfermedad , Neoplasias Gastrointestinales/metabolismo , Humanos , Isoenzimas/fisiología , Lipooxigenasa/fisiologíaRESUMEN
Arachidonic acid (AA) plays a pivotal role in the development of edema via its oxidized metabolites derived from cyclooxygenase (COX) and lipoxygenase (LOX), and is recently recognized as an activator of TRPV3. However, it is not clear whether AA plays some TRPV3-mediated pathological roles in the development of edema. Pharmacological and histological studies using ICRTRPV3+/+ and ICRTRPV3-/- mice indicated that higher ear edema responses to topical application of AA were observed in ICRTRPV3+/+ mice compared with ICRTRPV3-/- mice. However, there was no difference in the ear edema response to 12-O-tetradecanoylphorbol 13-acetate, skin histology, and skin barrier function between these mouse strains. Furthermore, oxidized fatty acids from the lesional site were analyzed to elucidate the TRPV3-mediated pathological roles of AA, and the results revealed that there were no differences in the level of COX or LOX metabolites derived from AA between both mouse strains. We concluded that AA plays a role in the development of TRPV3-mediated ear edema and that this result may contribute to better understanding of the pathophysiological mechanisms involved in the development of a certain type of edema.
Asunto(s)
Ácidos Araquidónicos/efectos adversos , Ácidos Araquidónicos/fisiología , Enfermedades del Oído/etiología , Edema/etiología , Canales Catiónicos TRPV/fisiología , Animales , Ácidos Araquidónicos/metabolismo , Femenino , Lipooxigenasa/fisiología , Masculino , Ratones Endogámicos C57BL , Ratones Endogámicos ICR , Prostaglandina-Endoperóxido Sintasas/fisiología , Canales Catiónicos TRPV/metabolismoRESUMEN
Urinary tract infection (UTI) is an extremely common infectious disease. Uropathogenic Escherichia coli (UPEC) is the predominant etiological agent of UTI. Asymptomatic bacteriuric E. coli (ABEC) strains successfully colonize the urinary tract resulting in asymptomatic bacteriuria (ABU) and do not induce symptoms associated with UTI. Oxylipids are key signaling molecules involved in inflammation. Based on the distinct clinical outcomes of E. coli colonization, we hypothesized that UPEC triggers the production of predominantly proinflammatory oxylipids and ABEC leads to production of primarily anti-inflammatory or proresolving oxylipids in the urinary tract. We performed quantitative detection of 39 oxylipid mediators with proinflammatory, anti-inflammatory, and proresolving properties, during UTI and ABU caused by genetically distinct E. coli strains in the murine urinary bladder. Our results reveal that infection with UPEC causes an increased accumulation of proinflammatory oxylipids as early as 6 h postinoculation, compared to controls. To the contrary, ABEC colonization leads to decreased accumulation of proinflammatory oxylipids at the early time point compared to UPEC infection but does not affect the level of proresolving oxylipids. This report represents the first comprehensive investigation on the oxylipidome during benign ABEC colonization observed in ABU and acute inflammation triggered by UPEC leading to UTI.
Asunto(s)
Infecciones por Escherichia coli/etiología , Mediadores de Inflamación/fisiología , Lípidos/fisiología , Vejiga Urinaria/microbiología , Infecciones Urinarias/etiología , Animales , Ácidos Grasos/análisis , Femenino , Lípidos/análisis , Lipooxigenasa/fisiología , Ratones , Ratones Endogámicos CBA , Prostaglandina-Endoperóxido Sintasas/fisiología , Vejiga Urinaria/químicaRESUMEN
Mutations in polycystin-1 (PC1) give rise to autosomal dominant polycystic kidney disease, an important and common cause of kidney failure. Despite its medical importance, the function of PC1 remains poorly understood. Here, we investigated the role of the intracellular polycystin-1, lipoxygenase, and α-toxin (PLAT) signature domain of PC1 using nuclear magnetic resonance, biochemical, cellular, and in vivo functional approaches. We found that the PLAT domain targets PC1 to the plasma membrane in polarized epithelial cells by a mechanism involving the selective binding of the PLAT domain to phosphatidylserine and L-α-phosphatidylinositol-4-phosphate (PI4P) enriched in the plasma membrane. This process is regulated by protein kinase A phosphorylation of the PLAT domain, which reduces PI4P binding and recruits ß-arrestins and the clathrin adaptor AP2 to trigger PC1 internalization. Our results reveal a physiological role for the PC1-PLAT domain in renal epithelial cells and suggest that phosphorylation-dependent internalization of PC1 is closely linked to its function in renal development and homeostasis.
Asunto(s)
Lipooxigenasa/fisiología , Canales Catiónicos TRPP/fisiología , Humanos , Lipooxigenasa/genética , Mutación , Estructura Terciaria de Proteína , Canales Catiónicos TRPP/genéticaRESUMEN
Myocardial infarction (MI) is the main cause for the progression of the left ventricle towards congestive heart failure. The optimal healing after MI requires timely induction and resolution of inflammation. Primarily, there have been a number of strategies applied to inhibit the post-MI inflammation but approaches that focus on the resolution of inflammation have sparsely been used in the treatment of heart failure. The early attempts to inhibit post-MI inflammation resulted in adverse outcomes that were realized in heart failure trials. We provide here an overview on the cyclooxygenase (COX)- and lipoxygenase (LOX)-derived lipid mediators that are either impairing or resolving the post-MI inflammation. With the evolution of lipidomics there has been emerging novel bioactive-specialized lipid mediators that promise to resolve chronic inflammation rather than promoting inhibition. The current review is focused on post-MI immune cells kinetics and the unexplored array of lipid mediators that are coordinated by COX and LOX. Thus, an emphasis on COX and LOX poses key questions and potential for the development of novel targets in the heart failure treatment strategy. This updated dynamic approach aims to fuse basic pre-clinical discoveries and translational bioactive lipid-based resolvin discoveries that could be potentially used in the clinic for the treatment of heart failure.
Asunto(s)
Inflamación/etiología , Infarto del Miocardio/complicaciones , Animales , Insuficiencia Cardíaca/etiología , Insuficiencia Cardíaca/prevención & control , Humanos , Inflamación/complicaciones , Inflamación/fisiopatología , Inflamación/terapia , Lípidos/fisiología , Lipooxigenasa/fisiología , Infarto del Miocardio/inmunología , Prostaglandina-Endoperóxido Sintasas/fisiología , Remodelación VentricularRESUMEN
Breast cancer cells facilitate distant metastasis through the induction of immunosuppressive regulatory B cells, designated tBregs. We report in this study that, to do this, breast cancer cells produce metabolites of the 5-lipoxygenase pathway such as leukotriene B4 to activate the peroxisome proliferator-activated receptor α (PPARα) in B cells. Inactivation of leukotriene B4 signaling or genetic deficiency of PPARα in B cells blocks the generation of tBregs and thereby abrogates lung metastasis in mice with established breast cancer. Thus, in addition to eliciting fatty acid oxidation and metabolic signals, PPARα initiates programs required for differentiation of tBregs. We propose that PPARα in B cells and/or tumor 5-lipoxygenase pathways represents new targets for pharmacological control of tBreg-mediated cancer escape.
Asunto(s)
Subgrupos de Linfocitos B/inmunología , Subgrupos de Linfocitos B/metabolismo , Lipooxigenasa/fisiología , Melanoma Experimental/inmunología , Melanoma Experimental/metabolismo , PPAR alfa/fisiología , Animales , Subgrupos de Linfocitos B/enzimología , Línea Celular Tumoral , Células Cultivadas , Femenino , Lipooxigenasa/genética , Lipooxigenasa/metabolismo , Melanoma Experimental/genética , Ratones , Ratones de la Cepa 129 , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Metástasis de la Neoplasia/genética , Metástasis de la Neoplasia/inmunología , Metástasis de la Neoplasia/patología , PPAR alfa/deficiencia , PPAR alfa/genética , Escape del Tumor/genética , Escape del Tumor/inmunologíaRESUMEN
Loss-of-function mutations in the lipoxygenase (LOX) genes ALOX12B and ALOXE3 are the second most common cause of autosomal recessive congenital ichthyosis. The encoded proteins, 12R-LOX and epidermal LOX-3 (eLOX-3), act in sequence to convert fatty acid substrates via R-hydroperoxides to specific epoxyalcohol derivatives and have been proposed to operate in the same metabolic pathway during epidermal barrier formation. Here, we show that eLOX-3 deficiency in mice results in early postnatal death, associated with similar but somewhat less severe barrier defects and morphological changes than reported earlier for the 12R-LOX-knockout mice. Skin lipid analysis demonstrated that the severity of barrier failure is related to the loss of covalently bound ceramides in both 12R-LOX- and eLOX-3-null mice, confirming a proposed functional linkage of the LOX pathway to ceramide processing and formation of the corneocyte lipid envelope. Furthermore, analysis of free oxygenated fatty acid metabolites revealed strongly reduced levels of hepoxilin metabolites in eLOX-3-deficient epidermis, indicating an additional function of eLOX-3 in mammalian skin as a hepoxilin synthase linked to the 12S-LOX pathway.
Asunto(s)
Araquidonato 12-Lipooxigenasa/fisiología , Epidermis/fisiología , Oxidorreductasas Intramoleculares/fisiología , Lipooxigenasa/fisiología , Animales , Araquidonato 12-Lipooxigenasa/genética , Ceramidas/análisis , Ceramidas/metabolismo , Modelos Animales de Enfermedad , Epidermis/química , Epidermis/metabolismo , Epidermis/patología , Ácidos Grasos no Esterificados/análisis , Ácidos Grasos no Esterificados/metabolismo , Ictiosis/genética , Ictiosis/metabolismo , Oxidorreductasas Intramoleculares/genética , Lípidos/análisis , Lipooxigenasa/genética , Ratones , Ratones Noqueados , Mutación , Índice de Severidad de la EnfermedadRESUMEN
The most significant threat to potato production worldwide is the late blight disease, which is caused by the oomycete pathogen Phytophthora infestans. Based on previous cDNA microarrays and cDNA-amplified fragment length polymorphism analysis, 63 candidate genes that are expected to contribute to developing a durable resistance to late blight were selected for further functional analysis. We performed virus-induced gene silencing (VIGS) to these candidate genes on both Nicotiana benthamiana and potato, subsequently inoculated detached leaves and assessed the resistance level. Ten genes decreased the resistance to P. infestans after VIGS treatment. Among those, a lipoxygenase (LOX; EC 1.13.11.12) and a suberization-associated anionic peroxidase affected the resistance in both N. benthamiana and potato. Our results identify genes that may play a role in quantitative resistance mechanisms to late blight.
Asunto(s)
Resistencia a la Enfermedad/genética , Genes de Plantas , Phytophthora infestans/fisiología , Enfermedades de las Plantas/microbiología , Solanum tuberosum/genética , Agrobacterium tumefaciens , Silenciador del Gen , Estudios de Asociación Genética , Interacciones Huésped-Patógeno , Lipooxigenasa/genética , Lipooxigenasa/fisiología , Anotación de Secuencia Molecular , Peroxidasas/genética , Peroxidasas/fisiología , Hojas de la Planta/genética , Hojas de la Planta/inmunología , Hojas de la Planta/microbiología , Proteínas de Plantas/genética , Proteínas de Plantas/fisiología , Virus de Plantas/genética , Solanum tuberosum/inmunología , Solanum tuberosum/microbiología , Nicotiana/genética , Nicotiana/microbiologíaRESUMEN
Interactions between cancer cells and stromal cells, including blood vessel endothelial cells (BECs), lymphatic vessel endothelial cells (LECs), bone marrow-derived angiogenic cells (BMDACs) and other bone marrow-derived cells (BMDCs) play important roles in cancer progression. Intratumoral hypoxia, which affects both cancer and stromal cells, is associated with a significantly increased risk of metastasis and mortality in many human cancers. Recent studies have begun to delineate the molecular mechanisms underlying the effect of intratumoral hypoxia on cancer progression. Reduced O2 availability induces the activity of hypoxia-inducible factors (HIFs), which activate the transcription of target genes encoding proteins that play important roles in many critical aspects of cancer biology. Included among these are secreted factors, including angiopoietin 2, angiopoietin-like 4, placental growth factor, platelet-derived growth factor B, stem cell factor (kit ligand), stromal-derived factor 1, and vascular endothelial growth factor. These factors are produced by hypoxic cancer cells and directly mediate functional interactions with BECs, LECs, BMDACs and other BMDCs that promote angiogenesis, lymphangiogenesis, and metastasis. In addition, lysyl oxidase (LOX) and LOX-like proteins, which are secreted by hypoxic breast cancer cells, remodel extracellular matrix in the lungs, which leads to BMDC recruitment and metastatic niche formation.
Asunto(s)
Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/fisiología , Neoplasias de la Mama/patología , Factor 1 Inducible por Hipoxia/fisiología , Linfangiogénesis , Neovascularización Patológica/etiología , Células del Estroma/fisiología , Animales , Células de la Médula Ósea/fisiología , Neoplasias de la Mama/irrigación sanguínea , Comunicación Celular , Movimiento Celular , Proliferación Celular , Células Endoteliales/fisiología , Humanos , Lipooxigenasa/fisiología , Metástasis Linfática , Metástasis de la NeoplasiaRESUMEN
In the present study, we investigated the mediators involved in the potentiation of antigen-induced contractions by indomethacin in tracheas isolated from ovalbumin (OA)-sensitized guinea-pigs. Indomethacin-induced potentiation of OA contraction was mimicked by prostaglandin DP/EP(1) /EP(2) receptor antagonist, AH-6809 but not by phospholipase A(2) enzyme inhibitor mepacrine. The lipoxygenase inhibitor AA-861 did not affect the contraction response to OA but prevented its potentiation by indomethacin, while the leukotriene receptor antagonist cinalukast inhibited both the OA response and its potentiation. However, the antagonists of platelet-activating factor (PAF) (BN-52021), adenosine (CGS-15943), endothelin ET(A) and ET(B) receptors (BQ-123, BQ-788), and the neutral endopeptidase inhibitor phosphoramidon did not alter the OA-induced contraction and its potentiation by indomethacin. Furthermore, capsaicin and neuropeptide receptor NK1, NK2, and NK3 antagonists (L-732128, MEN-10376, and SB-218795, respectively) also did not affect the OA-induced contractions and its potentiation. On the other hand, the 'transient receptor potential vanilloid 1' (TRPV1) antagonist capsazepine inhibited the potentiation response, while it did not alter the OA contraction itself. In conclusion, the potentiation of OA-induced contraction by indomethacin is more likely due to the increase in lipoxygenase products by the shift of arachidonic acid towards lipoxygenase pathway. Because some of the lipoxygenase products are potent vanilloid agonists, the stimulation of TRPV1 receptors besides leukotriene receptors seems to participate in the potentiation of contraction response in sensitized guinea-pig tracheas. PAF, adenosine, endothelins, and the neuropeptides present in the afferent neurons do not contribute to the potentiation of OA-induced contraction by indomethacin.
Asunto(s)
Antígenos/fisiología , Indometacina/farmacología , Contracción Muscular/fisiología , Ovalbúmina/farmacología , Tráquea/fisiología , Animales , Ácido Araquidónico/fisiología , Sinergismo Farmacológico , Cobayas , Lipooxigenasa/fisiología , Masculino , Técnicas de Cultivo de Órganos , Ovalbúmina/inmunología , Tráquea/inmunologíaRESUMEN
A European Respiratory Society research seminar on "Metabolic alterations in obstructive sleep apnoea (OSA)" was jointly organised in October 2009 together with two EU COST actions (Cardiovascular risk in the obstructive sleep apnoea syndrome, action B26, and Adipose tissue and the metabolic syndrome, action BM0602) in order to discuss the interactions between obesity and OSA. Such interactions can be particularly significant in the pathogenesis of metabolic abnormalities and in increased cardiovascular risk in OSA patients. However, studying the respective role of OSA and obesity is difficult in patients, making it necessary to refer to animal models or in vitro systems. Since most OSA patients are obese, their management requires a multidisciplinary approach. This review summarises some aspects of the pathophysiology and treatment of obesity, and the possible effects of sleep loss on metabolism. OSA-associated metabolic dysfunction (insulin resistance, liver dysfunction and atherogenic dyslipidaemia) is discussed from the perspective of both obesity and OSA in adults and children. Finally, the effects of treatment for obesity or OSA, or both, on cardio-metabolic variables are summarised. Further interdisciplinary research is needed in order to develop new comprehensive treatment approaches aimed at reducing sleep disordered breathing, obesity and cardiovascular risk.
Asunto(s)
Tejido Adiposo/fisiopatología , Obesidad/fisiopatología , Apnea Obstructiva del Sueño/fisiopatología , Animales , Dislipidemias/enzimología , Dislipidemias/fisiopatología , Femenino , Humanos , Hipoxia/fisiopatología , Inflamación/fisiopatología , Resistencia a la Insulina/fisiología , Lipooxigenasa/fisiología , Masculino , Ratones , Estrés Oxidativo/fisiología , RatasRESUMEN
Shear stress, especially low shear stress (LowSS), plays an important role in vascular remodeling during atherosclerosis. Endothelial cells (ECs), which are directly exposed to shear stress, convert mechanical stimuli into intracellular signals and interact with the underlying vascular smooth muscle cells (VSMCs). The interactions between ECs and VSMCs modulate the LowSS-induced vascular remodeling. With the use of proteomic analysis, the protein profiles of rat aorta cultured under LowSS (5 dyn/cm(2)) and normal shear stress (15 dyn/cm(2)) were compared. By using Ingenuity Pathway Analysis to identify protein-protein association, a network was disclosed that involves two secretary molecules, PDGF-BB and TGF-ß1, and three other linked proteins, lamin A, lysyl oxidase, and ERK 1/2. The roles of this network in cellular communication, migration, and proliferation were further studied in vitro by a cocultured parallel-plate flow chamber system. LowSS up-regulated migration and proliferation of ECs and VSMCs, increased productions of PDGF-BB and TGF-ß1, enhanced expressions of lysyl oxidase and phospho-ERK1/2, and decreased Lamin A in ECs and VSMCs. These changes induced by LowSS were confirmed by using PDGF-BB recombinant protein, siRNA, and neutralizing antibody. TGF-ß1 had similar influences on ECs as PDGF-BB, but not on VSMCs. Our results suggest that ECs convert the LowSS stimuli into up-regulations of PDGF-BB and TGF-ß1, but these two factors play different roles in LowSS-induced vascular remodeling. PDGF-BB is involved in the paracrine control of VSMCs by ECs, whereas TGF-ß1 participates in the feedback control from VSMCs to ECs.
Asunto(s)
Endotelio Vascular/metabolismo , Músculo Liso/metabolismo , Factor de Crecimiento Derivado de Plaquetas/fisiología , Estrés Mecánico , Factor de Crecimiento Transformador beta1/fisiología , Animales , Becaplermina , Movimiento Celular , Proliferación Celular , Células Cultivadas , Técnicas de Cocultivo , Endotelio Vascular/citología , Quinasas MAP Reguladas por Señal Extracelular/fisiología , Lamina Tipo A/fisiología , Lipooxigenasa/fisiología , Músculo Liso/citología , Proteómica , Proteínas Proto-Oncogénicas c-sis , Ratas , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
Atherosclerosis is recognized as an inflammatory condition of the vessel wall, characterized by accumulation of inflammatory cells such as macrophages and T cells. There are accumulating evidences that chemokines, cytokines, and lipid mediators coordinately modulate platelet- or leukocyte-endothelial cell interactions, and contribute to the maintenance of vascular homeostasis. This review focuses on the role of lipid mediators, especially those derived from polyunsaturated fatty acids, in controlling vascular inflammation and the progression of atherosclerosis.
Asunto(s)
Aterosclerosis/etiología , Mediadores de Inflamación/fisiología , Inflamación/etiología , Lipooxigenasa/fisiología , Prostaglandina-Endoperóxido Sintasas/fisiología , Prostaglandinas/fisiología , Animales , Aterosclerosis/patología , Aterosclerosis/prevención & control , Comunicación Celular , Quimiocinas/fisiología , Citocinas/fisiología , Progresión de la Enfermedad , Células Endoteliales/citología , Células Endoteliales/patología , Ácidos Grasos Omega-3/farmacología , Ácidos Grasos Omega-3/uso terapéutico , Ácidos Grasos Insaturados , Humanos , Leucotrienos/fisiología , Macrófagos , Linfocitos TRESUMEN
OBJECTIVE: The Mexican axolotl (Ambystoma mexicanum) is a well-characterized example for intrinsic regeneration. As lipoxygenase signaling is of crucial importance to scarless mammalian wound healing, we postulated that lipoxygenases might be expressed during amphibian regeneration and they might also influence human cells under appropriate conditions. In this study we identified an amphibian lipoxygenase and evaluated its impact on human cells in an in vitro wound model. METHODS: cDNA encoding for amphibian epidermal lipoxygenase (AmbLOXe) was polymerase chain reaction amplified and sequenced followed by phylogenic classification based on T-coffee alignment. Distribution of AmbLOXe was examined in various Ambystoma tissues, using polymerase chain reaction and in situ hybridization. Lipoxgenase influence was investigated using an outgrowth model of amphibian epidermal cells. Human osteosarcoma, as well as keratinocyte cell lines expressing AmbLOXe, were tested concerning in vitro wound closure in a monolayer scratch model. RESULTS: We isolated AmbLOXe from Ambystoma limb bud blastema identified as a homologue of human epidermal lipoxygenase. Amphibian epidermal lipoxygenase is expressed in Axolotl limb blastema and in epidermal cells which show decreased cell migration and proliferation rates when treated with LOX inhibitors. Furthermore, human osteosarcoma and keratinocyte cells showed increased rates of cell migration if transfected with AmbLOXe. CONCLUSION: In this study, AmbLOXe, a new effector of amphibian regeneration is described. In consideration of the presented data, AmbLOXe is important for amphibian epidermal cell proliferation and migration. As AmbLOXe expressing human osteosarcoma and keratinocyte cell lines showed increased rates of in vitro wound closure, an influence of amphibian mediators on human cells could be described for the first time.
Asunto(s)
Ambystoma mexicanum/fisiología , Extremidades/fisiología , Lipooxigenasa/fisiología , Regeneración/fisiología , Cicatrización de Heridas/fisiología , Ambystoma mexicanum/metabolismo , Animales , Línea Celular , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Epidermis/enzimología , Humanos , Lipooxigenasa/genética , Lipooxigenasa/farmacología , Transfección , Cicatrización de Heridas/efectos de los fármacosRESUMEN
Atherogenesis is an inflammatory process with leukocytes infiltrating the arterial intima. The lipoxygenase pathways play a role in leukocyte recruitment through the generation of two classes of arachidonic acid lipid mediators, the leukotrienes and the lipoxins, and one class of omega-3 fatty acid metabolites, the resolvins. There is evidence from animal studies and human genetic studies that the leukotrienes and the enzymes necessary for their generation play a role in atherosclerosis, and possibly even in the development of the vulnerable plaque. Less is known about the effect of the anti-inflammatory lipid mediators in atherosclerosis, the lipoxins and the resolvins. Studies modulating the activity of an enzyme necessary for the production of these lipid mediators, 12/15-lipoxygenase, showed discrepant results in several animal models. Also, human genetic studies have not clearly dissected the effect of the enzyme on atherosclerosis. However, stable forms of the lipoxins and the resolvins protect animals from inflammatory diseases. Whether blocking the leukotrienes or applying anti-inflammatory lipoxins and resolvins will be effective in attenuating human atherosclerosis needs to be demonstrated in future studies. In this review, the biosynthesis of these lipid mediators, their biological effects and the evidence for their possible role in atherosclerosis are discussed with an emphasis on human disease.
Asunto(s)
Aterosclerosis/enzimología , Leucotrienos/biosíntesis , Lipoxinas/biosíntesis , Lipooxigenasa/fisiología , Ácidos Docosahexaenoicos/metabolismo , Ácido Eicosapentaenoico/metabolismo , Ácidos Grasos Omega-3/metabolismo , Humanos , Leucotrienos/metabolismo , Lipoxinas/metabolismo , Lipooxigenasa/metabolismoRESUMEN
Oxidative stress has been suggested as a mechanism contributing to neuronal death induced by hypoglycemia, and an early production of reactive species (RS) during the hypoglycemic episode has been observed. However, the sources of reactive oxygen (ROS) and nitrogen (RNS) species have not been fully identified. In the present study we have examined the contribution of various enzymatic pathways to RS production and neuronal death induced by glucose deprivation (GD) in hippocampal cultures. We have observed a rapid increase in RS during GD, which depends on the activation of NMDA and non-NMDA receptors and on the influx of calcium from the extracellular space. Accordingly, intracellular calcium concentration [Ca(2+)](i) progressively increases more than 30-fold during the GD period. It was observed that superoxide production through the activation of the calcium-dependent enzymes, phospholipase A(2) (cPLA(2)) and xanthine oxidase (XaO), contributes to neuronal damage, while nitric oxide synthase (NOS) is apparently not involved. Inhibition of cPLA(2) decreased RS at early times of GD whereas inhibition of XaO diminished RS at more delayed times. The antioxidants trolox and ebselen also showed a protective effect against neuronal death and diminished RS generation. Inhibition of NADPH oxidase also contributed to the early generation of superoxide. Taking together, the present results suggest that the early activation of calcium-dependent ROS producing pathways is involved in neuronal death associated with glucose deprivation.
Asunto(s)
Glucosa/metabolismo , Hipocampo/metabolismo , Neuronas/metabolismo , Especies de Nitrógeno Reactivo/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Animales , Antioxidantes/farmacología , Calcio/metabolismo , Muerte Celular , Células Cultivadas , Hipocampo/citología , Lipooxigenasa/fisiología , Inhibidores de la Lipooxigenasa/farmacología , NADPH Oxidasas/antagonistas & inhibidores , NADPH Oxidasas/fisiología , Neuronas/citología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxido Nítrico Sintasa/fisiología , Estrés Oxidativo , Fosfolipasas A2 Citosólicas/antagonistas & inhibidores , Ratas , Ratas Wistar , Xantina Oxidasa/antagonistas & inhibidores , Xantina Oxidasa/fisiologíaRESUMEN
Herbivore-induced plant defences influence the behaviour of insects associated with the plant. For biting-chewing herbivores the octadecanoid signal-transduction pathway has been suggested to play a key role in induced plant defence. To test this hypothesis in our plant-herbivore-parasitoid tritrophic system, we used phenidone, an inhibitor of the enzyme lipoxygenase (LOX), that catalyses the initial step in the octadecanoid pathway. Phenidone treatment of Brussels sprouts plants reduced the accumulation of internal signalling compounds in the octadecanoid pathway downstream of the step catalysed by LOX, i.e. 12-oxo-phytodienoic acid (OPDA) and jasmonic acid. The attraction of Cotesia glomerata parasitoids to host-infested plants was significantly reduced by phenidone treatment. The three herbivores investigated, i.e. the specialists Plutella xylostella, Pieris brassicae and Pieris rapae, showed different oviposition preferences for intact and infested plants, and for two species their preference for either intact or infested plants was shown to be LOX dependent. Our results show that phenidone inhibits the LOX-dependent defence response of the plant and that this inhibition can influence the behaviour of members of the associated insect community.