RESUMO
Vascular endothelial growth factor (VEGF) is not only an important factor for angiogenesis but also lung development and homeostasis. VEGF-A binds three tyrosine kinase (TK) receptors VEGFR1-3. Idiopathic pulmonary fibrosis (IPF) is one of the poor prognoses of lung diseases. The relationship of VEGF and IPF remains to be clarified. Treatment with nintedanib used for the treatment of IPF reduced fibroblast proliferation, inhibited TK receptors, platelet-derived growth factor receptor (PDGFR), fibroblast growth factor receptor (FGFR), and VEGFR. Because the effect of that treatment is still not satisfactory, the emergence of new therapeutic agents is needed. This review describes the enhancement of pulmonary fibrosis by VEGFR1-TK signal and suggests that the blocking of the VEGFR1-TK signal may be useful for the treatment of pulmonary fibrosis.
RESUMO
Abscess formation is a classic host response to infection by many pathogenic microorganisms. Here, we studied the role of prostaglandins (PGs) and their signal transduction in abscess formation. Zymosan was injected into the pleural cavity of rats. Expression of enzymes involved in PG synthesis, their receptors, and cytokines in exudate leukocytes and abscesses were analyzed by polymerase chain reaction, Western blotting, and immunohistochemistry. Treatment with ketorolac, a cyclooxygenase (COX)-1 inhibitor, or N-[2-cyclohexyloxy-4-nitrophenyl] methanesulfonamide (NS-398), a COX-2 inhibitor, reduced the size of abscesses and the number of cells recovered from the abscess. COX-2 was detected in leukocytes of the exudate and a marginal area of abscesses. Among detected terminal PG synthases, the major one was cytosolic PGE synthase. Membrane-bound PGE synthase (mPGES)-1 was detected in cells that were similar to the COX-2-expressing cells in morphology and localization. A high level of the E-prostanoid (EP)(2) receptor and a low level of the EP(4) receptor were detected. The expression pattern of the EP(2) receptor paralleled that of COX-2 and mPGES-1. 11,15-O-Dimethyl PGE(2) (ONO-AE1-259), an EP(2) receptor agonist, and rolipram, a phosphodiesterase type-4 inhibitor, reversed the effects of COX inhibitors on abscess formation. In contrast, 16-(3-methoxymethyl) phenyl-omega-tetranor-3,7-dithia PGE(1) (ONO-AE1-329), an EP(4) receptor agonist, did not reverse the effects of NS-398. Moreover, NS-398 reduced the mRNA levels in exudate leukocytes of some proinflammatory and fibrogenic cytokines, which was reversed by ONO-AE1-259. These results suggest that PGE(2) generated via COX-1 and COX-2 may interact with the EP(2) receptor and may up-regulate in cAMP-dependent fashion the production of cytokines that promote abscess formation.
Assuntos
Abscesso/etiologia , Dinoprostona/biossíntese , Pleurisia/complicações , Receptores de Prostaglandina E/biossíntese , Transdução de Sinais , Abscesso/enzimologia , Abscesso/metabolismo , Abscesso/prevenção & controle , Animais , Western Blotting , Ciclo-Oxigenase 2/biossíntese , Inibidores de Ciclo-Oxigenase 2/farmacologia , Inibidores de Ciclo-Oxigenase 2/uso terapêutico , Imuno-Histoquímica , Masculino , Pleurisia/induzido quimicamente , Pleurisia/enzimologia , Pleurisia/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores de Prostaglandina E/agonistas , Receptores de Prostaglandina E Subtipo EP2 , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/efeitos dos fármacos , ZimosanRESUMO
Prostanoid production depends on the activity of two cyclooxygenase (COX) isoforms. It is appreciated that COX-1 plays a role in physiological processes, whereas COX-2 acts in pathological conditions. However their roles, particularly roles of COX-1, have not yet been fully established in inflammation. Here, we examined the effects of COX inhibitors, having differential isoform selectivity, on the late phase of rat carrageenin-induced pleurisy to elucidate the role of COX-2 expressed in the draining lymph nodes and found substantial contribution of COX-1-product(s). Protein and mRNA of COX-2 were detectable with Western blotting analysis and reverse-transcription polymerase chain reaction (RT-PCR) analysis in parathymic lymph nodes, peaking at 48 h after induction of pleurisy. Microsomal prostaglandin E synthase (mPGES)-1 was detectable by immunohistochemical analysis in cells with dendritic processes, a morphological characteristic similar to that of COX-2 expressing cells. Although aspirin, indomethacin and a COX-1 inhibitor, ketorolac, significantly decreased the volume of pleural exudate, they did not affect the levels of COX-2 and mPGES-1 in the lymph node 24 h after induction of pleurisy. In contrast, COX-2 inhibitors, nimesulide and NS-398, had no effect on the exudate volume, but they increased the number of COX-2- and mPGES-1-expressing cells and extension of their dendritic processes with significant increase in the COX-2 level, which were antagonised by ketorolac. These results suggest that COX-2-expressing cells may negatively self-regulate their functions by producing PGE2 via mPGES-1: migration into the draining lymph node and their differentiation. Moreover, COX-1- and COX-2-derived prostanoids may play differential or sometimes antagonistic roles in the late phase of acute inflammation.
Assuntos
Ciclo-Oxigenase 1/metabolismo , Ciclo-Oxigenase 2/biossíntese , Linfonodos/enzimologia , Proteínas de Membrana/metabolismo , Derrame Pleural/enzimologia , Pleurisia/enzimologia , Prostaglandinas/metabolismo , Doença Aguda , Animais , Western Blotting , Carragenina , Inibidores de Ciclo-Oxigenase 2/farmacologia , Inibidores de Ciclo-Oxigenase 2/uso terapêutico , Inibidores de Ciclo-Oxigenase/farmacologia , Inibidores de Ciclo-Oxigenase/uso terapêutico , Dinoprostona/metabolismo , Modelos Animais de Doenças , Indução Enzimática/efeitos dos fármacos , Linfonodos/efeitos dos fármacos , Linfonodos/metabolismo , Linfonodos/patologia , Masculino , Proteínas de Membrana/antagonistas & inibidores , Derrame Pleural/induzido quimicamente , Derrame Pleural/tratamento farmacológico , Derrame Pleural/patologia , Pleurisia/induzido quimicamente , Pleurisia/tratamento farmacológico , Pleurisia/patologia , Prostaglandina-E Sintases , Prostaglandina-Endoperóxido Sintases/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de TempoRESUMO
Granulocyte apoptosis and subsequent clearance by phagocytes are critical for the resolution of inflammation. However, no studies have addressed how the resolution proceeds in the inflammatory site. We studied the time course of neutrophil apoptosis and the following ingestion by mononuclear leukocytes in rat carrageenin-induced pleurisy, detecting DNA fragmentation by the deoxyuridine triphosphate-biotin nick-end labeling (TUNEL) method, by acridine orange staining, and from the DNA ladder pattern on electrophoresis. Neutrophil accumulation started 3-5 h after carrageenin injection and then maintained a plateau until 24 h. Neutrophils decreased steeply between days 1 and 3. Mononuclear leukocytes started to accumulate at 5 h and reached a peak at day 2. TUNEL-positive bodies and acridine orange-positive bodies first became detectable in the cytoplasm of the mononuclear leukocytes from 24 h and 9 h, respectively. Both methods indicated that mononuclear leukocytes containing fragmented DNA increased rapidly on days 1 and 2 and reached a peak at day 3. The characteristic ladder pattern of neutrophil DNA was observed from 5 h. Tumor necrosis factor alpha was detectable on the start, and the levels of interleukin-10 and transforming growth factor-beta1 rose together with signs of neutrophil apoptosis and the following ingestion by mononuclear leukocytes. These results indicate that neutrophils start to undergo apoptosis just after the beginning of their accumulation in the inflammation site. Thus, evolution and resolution processes may proceed concurrently in acute inflammation.
Assuntos
Apoptose , Neutrófilos/patologia , Pleurisia/fisiopatologia , Laranja de Acridina , Animais , Carragenina/toxicidade , DNA/metabolismo , Marcação In Situ das Extremidades Cortadas , Interleucina-10/metabolismo , Leucócitos Mononucleares/fisiologia , Masculino , Fagocitose/fisiologia , Pleurisia/induzido quimicamente , Ratos , Ratos Sprague-Dawley , Fator de Crescimento Transformador beta/metabolismo , Fator de Crescimento Transformador beta1 , Fator de Necrose Tumoral alfa/metabolismoRESUMO
We showed in Drosophila that nuclear migration was reduced all through cleavage stages in embryos with any one of the maternal-effect mutations, gs(1)N441 and gs(1)N26, in which F-actin reorganization in cleavage embryos is disordered. Moreover, we determined nuclear positions in embryos at cycle 1 and 2 in the wild type and two mutants, gs(1)N441 and gs(1)N26, in order to test if the nuclear migration is regulated within a nuclear cycle. At cycle 1, there was no difference in nuclear position among the strains that we observed. At cycle 2 the two sister nuclei had already migrated posteriorly in the wild type. However, migration was not detectable at cycle 2 in the mutants. Besides, the two sister nuclei were less-separated from each other, and orientation of the two nuclei with regard to the anteroposterior axis was random, different from the wild type. These results support the hypothesis that F-actin is involved in the regulation to separate cleavage nuclei from each other and from the egg cortex. This regulation is apparently required for posteriorward nuclear migration, and for synchronous nuclear arrival in the whole egg cortex.
RESUMO
Expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) is up-regulated in response to inflammatory stimuli. To evaluate the extent to which local pleural inflammation involves additional site in the pleural cavity and elsewhere, we investigated the time course of the levels of iNOS and its product in the inflammatory and other sites, and compared those with a level of COX-2 in rat carrageenin-induced pleurisy. The exudate and plasma NOx levels rose, reaching peaks at 9 and 14 h, respectively. Both COX-2 and iNOS became detectable in exudate leukocytes, their levels reaching peaks at 3 and 9 h after irritation, respectively. COX-2 was detectable mainly in neutrophils, but iNOS was detectable in both neutrophils and mononuclear leukocytes. Furthermore, iNOS became detectable in neutrophils and mononuclear leukocytes in enlarged parathymic lymph nodes from 3h in addition to those in peripheral blood and Kupffer cells from 3 to 14 h, respectively. The gene product is also detectable in thymic large dendritic cells of pleurisy-induced rats as well as normal control rats. COX-2 became detectable in stellar dendritic cells of the enlarged draining lymph nodes from 14 h. Thus, these gene products were induced in the immediate proximity of regional lymph nodes, and even at a considerable distance of liver by the local inflammatory stimulus. Although their expression pattern was quite different from each other, these gene products were detectable in phagocytic cells.