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1.
AIDS Care ; 28(7): 834-41, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-26885765

RESUMO

Depression is a common but frequently undiagnosed feature in individuals with HIV infection. To find a strategy to detect depression in a non-specialized clinical setting, the overall performance of the Hospital Anxiety and Depression Scale (HADS) and the depression identification questions proposed by the European AIDS Clinical Society (EACS) guidelines were assessed in a descriptive cross-sectional study of 113 patients with HIV infection. The clinician asked the two screening questions that were proposed under the EACS guidelines and requested patients to complete the HADS. A psychiatrist or psychologist administered semi-structured clinical interviews to yield psychiatric diagnoses of depression (gold standard). A receiver operating characteristic (ROC) analysis for the HADS-Depression (HADS-D) subscale indicated that the best sensitivity and specificity were obtained between the cut-off points of 5 and 8, and the ROC curve for the HADS-Total (HADS-T) indicated that the best cut-off points were between 12 and 14. There were no statistically significant differences in the correlations of the EACS (considering positive responses to one [A] or both questions [B]), the HADS-D ≥ 8 or the HADS-T ≥ 12 with the gold standard. The study concludes that both approaches (the two EACS questions and the HADS-D subscale) are appropriate depression-screening methods in HIV population. We believe that using the EACS-B and the HADS-D subscale in a two-step approach allows for rapid, assumable and accurate clinical diagnosis in non-psychiatric hospital settings.


Assuntos
Depressão , Infecções por HIV , Programas de Rastreamento/métodos , Adulto , Assistência Integral à Saúde/métodos , Estudos Transversais , Depressão/diagnóstico , Depressão/epidemiologia , Depressão/etiologia , Feminino , Infecções por HIV/complicações , Infecções por HIV/epidemiologia , Infecções por HIV/psicologia , Humanos , Entrevista Psicológica/métodos , Masculino , Pessoa de Meia-Idade , Curva ROC , Sensibilidade e Especificidade , Espanha/epidemiologia
2.
FEBS Lett ; 581(13): 2394-400, 2007 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-17481611

RESUMO

Oxidative stress and low grade chronic inflammation are increased in accumulating fat. Our objective was to test whether 4-hydroxynonenal (4-HNE), an end-product of lipid peroxidation, affects cyclooxygenases in 3T3-L1 adipose cells. 4-HNE increased COX-2 mRNA and protein expression and p38MAP-kinase phosphorylation in a dose-dependent manner. Pretreatment of 3T3-L1 cells by a selective inhibitor of p38MAPK (PD 169316) abolished 4-HNE and glucose oxidase induced COX-2 expression. Our results show that oxidative stress induces COX-2 expression through the production of 4-HNE which activates p38MAPKinase, suggesting that 4-HNE links oxidative stress and chronic inflammation through the activation of cyclooxygenase.


Assuntos
Adipócitos/enzimologia , Aldeídos/farmacologia , Ciclo-Oxigenase 2/genética , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Estresse Oxidativo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Células 3T3 , Adipócitos/efeitos dos fármacos , Animais , Inibidores de Cisteína Proteinase/farmacologia , Dinoprosta/biossíntese , Dinoprostona/biossíntese , Ativação Enzimática , Peroxidação de Lipídeos , Camundongos , Prostaglandina-Endoperóxido Sintases/genética , RNA Mensageiro/genética
3.
Radiat Res ; 156(5 Pt 1): 476-87, 2001 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11604060

RESUMO

The monohydroxylated fatty acid content of peripheral blood mononuclear cells from 23 cleanup workers and 16 unexposed individuals was studied in relation to their immune status after the Chernobyl accident. Men with absorbed doses below 0.32 Gy showed higher levels of free and esterified 12-hydroxyeicosatetraenoic acid (12-HETE) than unexposed men, whereas 15-HETE and the 17-hydroxy derivative of C22 fatty acid (17-OH 22), either free or esterified in phospholipids, were increased in a dose-dependent manner. The percentage of CD4-positive cells was also increased significantly in heavily irradiated men, whereas the percentage of CD8-positive cells tended to decrease with dose. Furthermore, the absolute count of CD4-positive cells was correlated positively with the amount of esterified 15-HETE in the phospholipid fraction of the mononuclear cells and with the total 15-HETE. These results show for the first time that the accumulation of autoxidized/lipoxygenase products of polyunsaturated fatty acids in the mononuclear cells of irradiated individuals was associated with immune imbalance. This may be the basis for certain late effects of radiation such as autoimmune disorders, somatic and neoplastic diseases, and early aging.


Assuntos
Ácido 12-Hidroxi-5,8,10,14-Eicosatetraenoico/sangue , Ácidos Hidroxieicosatetraenoicos/sangue , Imunidade/efeitos da radiação , Leucócitos Mononucleares/efeitos da radiação , Centrais Elétricas , Liberação Nociva de Radioativos , Adulto , Idoso , Antígenos CD4/análise , Antígenos CD8/análise , Antígenos HLA-DR/análise , Humanos , Pessoa de Meia-Idade , Ucrânia
5.
J Biol Chem ; 275(43): 33379-87, 2000 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-10938092

RESUMO

Hormones and growth factors induce in many cell types the production of phosphatidic acid (PA), which has been proposed to play a role as a second messenger. We have previously shown in an acellular system that PA selectively stimulates certain isoforms of type 4 cAMP-phosphodiesterases (PDE4). Here we studied the effect of endogenous PA on PDE activity of transiently transfected MA10 cells overexpressing the PA-sensitive isoform PDE4D3. Cell treatment with inhibitors of PA degradation, including propranolol, induced an accumulation of endogenous PA accompanied by a stimulation of PDE activity and a significant decrease in both cAMP levels and protein kinase A activity. Furthermore, in FRTL5 cells, which natively express PDE4D3, pretreatment with compounds inducing PA accumulation prevented both cAMP increase and cAMP-responsive element-binding protein phosphorylation triggered by thyroid-stimulating hormone. To determine the mechanism of PDE stimulation by PA, endogenous phospholipids were labeled by preincubating MA10 cells overexpressing PDE4D3 with [(32)P]orthophosphate. Immuno- precipitation experiments showed that PA was specifically bound to PDE4D3, supporting the hypothesis that PDE4D3 activation occurs through direct binding of PA to the protein. PA binding site on PDE4D3 was characterized by engineering deletions of selected regions in the N-terminal regulatory domain of the enzyme. Deletion of amino acid residues 31-59 suppressed both PA-activating effect and PA binding, suggesting that this region rich in basic and hydrophobic residues contains the PA binding site. These observations strongly suggest that endogenous PA can modulate cAMP levels in intact cells, through a direct activation of PDE4D3.


Assuntos
3',5'-AMP Cíclico Fosfodiesterases/metabolismo , AMP Cíclico/fisiologia , Isoenzimas/metabolismo , Ácidos Fosfatídicos/metabolismo , 3',5'-AMP Cíclico Fosfodiesterases/química , Sequência de Aminoácidos , Animais , Sítios de Ligação , Células Cultivadas , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4 , Isoenzimas/química , Dados de Sequência Molecular , Fosforilação , Propranolol/farmacologia , Ratos , Tireotropina/farmacologia
6.
Mol Biol Cell ; 10(12): 4355-67, 1999 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-10588663

RESUMO

Myogenic cell differentiation is induced by Arg(8)-vasopressin, whereas high cAMP levels and protein kinase A (PKA) activity inhibit myogenesis. We investigated the role of type 4 phosphodiesterase (PDE4) during L6-C5 myoblast differentiation. Selective PDE4 inhibition resulted in suppression of differentiation induced by vasopressin. PDE4 inhibition prevented vasopressin-induced nuclear translocation of the muscle-specific transcription factor myogenin without affecting its overall expression level. The effects of PDE4 inhibition could be attributed to an increase of cAMP levels and PKA activity. RNase protection, reverse transcriptase PCR, immunoprecipitation, Western blot, and enzyme activity assays demonstrated that the PDE4D3 isoform is the major PDE4 expressed in L6-C5 myoblasts and myotubes, accounting for 75% of total cAMP-hydrolyzing activity. Vasopressin cell stimulation caused a biphasic increase of PDE4 activity, which peaked at 2 and 15 min and remained elevated for 48 h. In the continuous presence of vasopressin, cAMP levels and PKA activity were lowered. PDE4D3 overexpression increased spontaneous and vasopressin-dependent differentiation of L6-C5 cells. These results show that PDE4D3 plays a key role in the control of cAMP levels and differentiation of L6-C5 cells. Through the modulation of PDE4 activity, vasopressin inhibits the cAMP signal transduction pathway, which regulates myogenesis possibly by controlling the subcellular localization of myogenin.


Assuntos
3',5'-AMP Cíclico Fosfodiesterases/metabolismo , Diferenciação Celular/fisiologia , Músculo Esquelético/enzimologia , 3',5'-AMP Cíclico Fosfodiesterases/antagonistas & inibidores , Animais , Linhagem Celular , AMP Cíclico/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4 , Camundongos , Músculo Esquelético/citologia , Miosinas/metabolismo , Inibidores de Fosfodiesterase/farmacologia , Testes de Precipitina , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rolipram/farmacologia , Vasopressinas/metabolismo
7.
Eur J Cell Biol ; 78(6): 398-406, 1999 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-10430021

RESUMO

Fatty acids have been involved in the proliferation and differentiation of numerous cells, as mediated via peroxisome proliferator-activated receptors (PPARs) or lipid metabolites (prostaglandins, diacylglycerol). In the present study, we have investigated the effect of arachidonic acid (AA), docosahexaenoic acid (DHA) and its precursor eicosapentaenoic acid (EPA) on the differentiation of a rat uterine stromal cell line, UIII. As markers of decidualization, we have investigated morphological changes, monitored by inverted light and scanning electron microscopy. The induction of 3 proteins, desmin, hsp-25 and prolactin, which are all considered to be markers of decidualization, were analyzed by immunocytochemistry or Western blotting. Addition of AA (30 microM) to the medium of cultured cells for 48h induced cell spreading and flattening. Cells became enlarged (x 2.5) and some of them were binucleated. Using scanning electron microscopy, we confirmed these morphological changes and showed that the enlargement of the cells was followed by numerous extracellular processes, leading to an increase in cell surface area and intercellular communications. Immunocytochemistry showed that this treatment also induced the expression of desmin, which seems to direct morphological changes, beginning as a perinuclear ring and extending to the cell membrane. The time course of desmin expression was studied by Western blotting. No desmin expression was present before 4h of AA treatment. Desmin induction was maximum at 24h of treatment and plateaued thereafter. DHA and EPA (30 microM), added to the medium, failed to induce any change. However, in cells previously differentiated with AA and expressing desmin, treatment with DHA or EPA (30microM) reversed partially the action of AA, EPA being the most effective. AA also induced hsp-25, though all cells did not express this protein. A prolactin (PRL)-like factor was induced by AA, as recognized by an antibody against pituitary rPRL, and migrated as the standard. Moreover, a fragment of 16 kDa was also revealed by this antibody, suggesting that the PRL-like factor cleaved, was similar to PRL and that the PRL-like factor could be identical to PRL. In conclusion, these results show that AA is able to specifically induce the decidualization of uterine stromal cells in vitro.


Assuntos
Ácido Araquidônico/farmacologia , Decídua/citologia , Proteínas de Choque Térmico , Células Estromais/citologia , Útero/citologia , Animais , Western Blotting , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Desmina/biossíntese , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Proteínas de Choque Térmico HSP27 , Proteínas de Neoplasias/biossíntese , Ratos , Células Estromais/metabolismo
10.
J Leukoc Biol ; 65(3): 381-90, 1999 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10080543

RESUMO

We have previously shown that mitogenic activation of human PBMC rapidly increases both the intracellular phosphatidic acid (PA) level and cyclic nucleotide phosphodiesterase (PDE) activity, with time-course responses, suggesting a causative relationship between the two events. PA also directly stimulated cAMP-PDE activity in acellular systems. Thus the mitogenic properties of PA night be due to its ability to lower the level of cAMP, a negative effector of lymphocyte activation, through PDE activation. In this study, human PBMC were stimulated either with the mitogenic lectin ConA, the anti-CD3 mAb OKT3, or the phorbol ester TPA. All three agonists increased the radiolabeled PA level and the PA mass in treated cells and simultaneously increased cytosolic and particulate cAMP- and cGMP-PDE activities, with significant positive correlations between PA accumulation and PDE activities. Furthermore, the ConA-induced PDE activation was dose-dependently reduced by treatment of PBMC with the diacylglycerol-kinase inhibitor R59022. This compound also dose-dependently lowered the PA level and inhibited the proliferative response to ConA. In addition, TPA-induced PDE activation was totally abolished by ethanol, which strongly reduced PA accumulation in response to the phorbol ester. These data suggest that PA increase may be linked to mitogen-induced PDE activation. Experiments performed in the presence of rolipram indicated that ConA and TPA stimulated both the rolipram-sensitive PDE4 and the rolipram-insensitive PDE activities, OKT3 being more active on PDE4. All three agonists stimulated the cGMP-specific PDE5. These results suggest that PA is an important component of the mechanisms that maintain a low level of cyclic nucleotides, which is a prerequisite for an optimal lymphoproliferative response.


Assuntos
3',5'-AMP Cíclico Fosfodiesterases/metabolismo , Leucócitos Mononucleares/metabolismo , Ácidos Fosfatídicos/metabolismo , Divisão Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Humanos , Imunossupressores/farmacologia , Leucócitos Mononucleares/citologia , Mitógenos/farmacologia , Muromonab-CD3/farmacologia , Pirimidinonas/farmacologia , Tiazóis/farmacologia
11.
Biochem Pharmacol ; 57(9): 1021-30, 1999 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-10796072

RESUMO

Oxidatively stressed lymphocytes exhibit decreased proliferative response to mitogenic stimulation. Although several sensitive targets involved in lymphocyte suppression have already been identified, little is known about the influence of oxidative stress on cyclic nucleotide phosphodiesterases (PDE) (EC 3.1.4.17), thought to play a major role in the control of cyclic AMP (cAMP) level, a well-recognized negative effector of lymphoproliferation. Although the polyunsaturated fatty acid content of membrane phospholipids is thought to be directly related to the extent of oxidant-induced lipid peroxidation, some n-3 fatty acids also seem to have antioxidant effects, depending on the concentration used and the overall redox status of the cells in question. Results of the present study showed that human peripheral blood mononuclear cells (PBMC) as well as rat thymocytes were relatively resistant to a short-term exposure (10 min) to hydrogen peroxide (H2O2). Indeed, H2O2-induced lipid peroxidation, estimated by malondialdehyde (MDA) production, was only 2-fold increased by H2O2 concentrations lower than 2 mM, whereas a larger increase (10-fold) could be observed in PBMC at the highest dose (5 mM). Previous enrichment of PBMC with 5 microM docosahexaenoic acid (22:6n-3), brought to the cells as a fatty acid-albumin complex (ratio 1), significantly reduced MDA production induced by low doses of H2O2, the protective effect no longer being observed at the highest doses. In contrast, eicosapentaenoic acid (20:5n-3) did not have any protective effect. Cytosolic PDE activities of both human PBMC and rat thymocytes were significantly inhibited (40-50%) after H2O2 treatment of the cells, whereas particulate PDE activities were not modified. Different responses of PDE activities to H2O2 treatment were observed when PBMC were first enriched with 22:6n-3 prior to H2O2 addition. In 22:6n-3-treated cells, the H2O2-induced inhibition of both cAMP- and cGMP-PDE cytosolic activities was abolished, whereas the particulate activities were increased by the highest H2O2 concentration used (5 mM). At the same time, the glutathione peroxidase (glutathione: oxidoreductase, EC 1.11.1.9) (GSH-Px) activity of PBMC and thymocytes was only marginally inhibited by H2O2 addition (20%), and pretreatment of the cells with 22:6n-3 did not modify the slight inhibitory effect of H2O2. Collectively, these results suggest that lymphocytes are relatively resistant to H2O2-induced lipid peroxidation due to their high GSH-Px content, and that low doses of 22:6n-3 are able to prevent some of the H2O2-induced alterations such as lipid peroxidation and PDE inhibition. Docosahexaenoic acid might thus offer some protection against oxidant-induced lymphocyte suppression.


Assuntos
Ácidos Docosa-Hexaenoicos/farmacologia , Peróxido de Hidrogênio/farmacologia , Linfócitos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Animais , Interações Medicamentosas , Ácidos Graxos Ômega-3/farmacologia , Óleos de Peixe/farmacologia , Glutationa Peroxidase/efeitos dos fármacos , Glutationa Peroxidase/metabolismo , Humanos , Técnicas In Vitro , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Linfócitos/metabolismo , Diester Fosfórico Hidrolases/efeitos dos fármacos , Diester Fosfórico Hidrolases/metabolismo , Ratos
12.
Cell Biochem Biophys ; 29(1-2): 1-17, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9631235

RESUMO

Phosphatidic acid (PA) has been previously shown to activate specifically some of the isoforms of type 4 cylic nucleotide phosphodiesterases (PDE-4) in an acellular system. In the present work, we have investigated the mechanism of PA-activating effect by using a recombinant PA-sensitive isoform, PDE-4D3. The enzyme was specifically activated by acidic phospholipids, but not by zwitterionic phospholipids or anionic detergents. The importance of the role of PA acidic groups in the activation process was confirmed by studying the influence of pH and ionic strength on activation. Crosslinking experiments suggested that PA might influence the ability of PDE-4D3 to form dimers. Binding studies performed with radiolabeled PA showed that PA binds to a PDE-4D3 preparation in a saturable manner. Specifically bound PA was displaced by anionic, but not by zwitterionic phospholipids. With a preparation of PDE-4B2, a PDE-4 isoform insensitive to PA activation, PA binding was only displaced by high concentrations of unlabeled PA, suggesting that high-affinity PA binding sites are only present on PDE-4D3. These data support the hypothesis that PA-activating effect depends on direct binding of the effector on specific sites carried by the PDE-4D3 protein.


Assuntos
3',5'-AMP Cíclico Fosfodiesterases/efeitos dos fármacos , 3',5'-AMP Cíclico Fosfodiesterases/metabolismo , AMP Cíclico/metabolismo , Ácidos Fosfatídicos/farmacologia , 3',5'-AMP Cíclico Fosfodiesterases/genética , Animais , Ânions , Linhagem Celular , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4 , Detergentes/farmacologia , Ativação Enzimática/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Concentração Osmolar , Ácidos Fosfatídicos/metabolismo , Ligação Proteica/efeitos dos fármacos , Conformação Proteica/efeitos dos fármacos , Ratos , Spodoptera/genética , Especificidade por Substrato
13.
J Lipid Res ; 39(4): 873-83, 1998 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-9555950

RESUMO

N-3 polyunsaturated fatty acids from marine oil have been shown to decrease T cell-mediated immune function both in animals and humans, and to inhibit the mitogen-induced lymphoproliferative response when added to lymphocyte culture medium. As phosphatidic acid (PA) is a key mediator of the mitogenic process, the present study aims to investigate whether docosahexaenoic (DHA) and eicosapentaenoic (EPA) acids, the main n-3 fatty acids from fish oil, are able to alter the mitogen-induced synthesis of PA, when added to the culture medium of human peripheral blood mononuclear cells (PBMC). Incubation of PBMC in a medium containing 5 microM DHA bound to 5 microM human delipidated serum albumin induced a 2-fold increase in the basal PA mass whereas incubation with EPA, in the same conditions, had no effect. In contrast, both fatty acids markedly reduced the concanavalin A (ConA)-induced production of PA as compared with untreated cells. Paradoxically, phospholipase D (PLD) activity, evidenced by the synthesis of phosphatidylbutanol, was only detected in DHA-treated cells further stimulated by ConA, indicating that both DHA and ConA are required for PLD activation. Similarly, an increased diacylglycerol (DAG) mass was only observed in DHA-treated cells stimulated by ConA, whereas no modification occurred in control or EPA-treated cells stimulated or not by ConA. Furthermore, 1-butanol suppressed the ConA-induced increase of DAG mass observed in DHA-treated cells, indicating that phosphatidate was the source of the newly synthesized diacylglycerol. Altogether, these results show that, in concanavalin A-activated human peripheral blood mononuclear cells, docosahexaenoate stimulates both phospholipase D and phosphatidate phosphohydrolase activities, which ultimately results in an increased diacylglycerol production at the expense of phosphatidate.


Assuntos
Ácidos Docosa-Hexaenoicos/farmacologia , Imunossupressores/farmacologia , Linfócitos/efeitos dos fármacos , Ácidos Fosfatídicos/análise , Fosfolipase D/metabolismo , Concanavalina A/farmacologia , Diglicerídeos/análise , Ácido Eicosapentaenoico/farmacologia , Ativação Enzimática , Óleos de Peixe/farmacologia , Humanos , Ativação Linfocitária , Mitógenos/farmacologia , Fosfatidato Fosfatase/metabolismo
14.
Pulm Pharmacol Ther ; 11(4): 245-52, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-10101740

RESUMO

Leukotriene B4 (LTB4), an inflammatory mediator, is a potent chemoattractant for neutrophils (PMN) that plays an important role in the late reaction in asthma. Human airway epithelial cells (HAEC) can interact with PMN to increase LTB4 production. The aim of this study was to determine the influence of loratadine, an antihistaminic drug, on the production of LTB4 by PMN either alone or during interaction with transformed HAEC. The effect of tumour necrosis factor-alpha (TNF-alpha) was also examined. LTB4 production was measured by RP-HPLC after cell stimulation with calcium ionophore. Loratadine (0.25-25 microM) induced a significant and dose-dependent decrease of LTB4 production by PMN alone whereas it was up-regulated by TNF-alpha. As reported by others, we confirmed the increase of LTB4 release when PMN were cocultured with HAEC as compared to PMN alone. Addition of loratadine to HAEC before co-culture with PMN induced a significant decrease of LTB4 formation by cell interaction. This effect was noted when HAEC were washed following incubation with loratadine, demonstrating a direct action of the drug on this cell type. Moreover, the TNF-alpha-induced stimulation of LTB4 release that we demonstrated in PMN-HAEC interaction was also inhibited by loratadine. These results indicate that loratadine might reduce inflammatory reaction by a direct effect on PMN LTB4 production but also through an influence on HAEC during interaction with PMN.


Assuntos
Antialérgicos/farmacologia , Antagonistas dos Receptores Histamínicos H1/farmacologia , Leucotrieno B4/metabolismo , Loratadina/farmacologia , Pulmão/metabolismo , Neutrófilos/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologia , Células Cultivadas/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Técnicas de Cocultura , Relação Dose-Resposta a Droga , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Citometria de Fluxo , Humanos , Molécula 1 de Adesão Intercelular/metabolismo , Pulmão/citologia , Pulmão/efeitos dos fármacos , Neutrófilos/metabolismo , Regulação para Cima
15.
Eur J Pharmacol ; 312(3): 379-84, 1996 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-8894621

RESUMO

This work showed that docosahexaenoic (22:6n-3) and eicosapentaenoic (20:5n-3) acid supplementation for 48 h have opposite effects on the norepinephrine-stimulated cyclic AMP accumulation in rat pinealocytes. We found that 22:6n-3 supplementation of pineal cells, done by increasing specifically 22:6n-3 in phospholipid and triacylglycerol pools, led to inhibition of norepinephrine-stimulated cyclic AMP production whereas 20:5n-3 supplementation, by increasing 20:5n-3, and 22:5n-3 and 22:6n-3 in the same pools, stimulated it. In contrast, direct treatment of pinealocytes with each fatty acid (50 microM) did not affect cyclic AMP production in the presence of (0.1-10 microM) norepinephrine. The results indicate that, using pharmacological agents such as forskolin or prazosin: (a) neither basal nor forskolin-stimulated cyclic AMP levels were modified in fatty acid-supplemented cells compared to control cells; (b) in the presence of 1 microM prazosin, the activation by 20:5n-3 was still effective whereas no additional inhibition of norepinephrine stimulation was observed in 22:6n-3-supplemented cells. Taken together our results suggest that 22:6n-3 or 20:5n-3 supplementation modulates specifically the alpha 1- or beta-adrenoceptors in the rat pineal gland.


Assuntos
AMP Cíclico/metabolismo , Ácidos Graxos/farmacologia , Norepinefrina/farmacologia , Glândula Pineal/efeitos dos fármacos , Antagonistas Adrenérgicos alfa/farmacologia , Agonistas Adrenérgicos beta/farmacologia , Animais , Colforsina/farmacologia , Sinergismo Farmacológico , Masculino , Fosfolipídeos/metabolismo , Glândula Pineal/citologia , Glândula Pineal/metabolismo , Prazosina/farmacologia , Ratos , Ratos Sprague-Dawley , Triglicerídeos/metabolismo
16.
J Mol Cell Cardiol ; 28(10): 2151-61, 1996 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-8930810

RESUMO

The authors have previously resolved four forms of cyclic nucleotide phosphodiesterase (PDE) in neonatal rat cultured cardiomyocytes by use of high-performance liquid chromatography (HPLC) and have shown that the response of the cGMP-stimulated PDE to the effector cGMP was markedly reduced as compared to that of the corresponding isoform present in the heart ventricle of adult rat (70% v 350%). When neonatal rat ventricular myocytes were grown in a medium enriched in docosahexaenoic acid (DHA) or eicosapentaenoic acid (EPA), an increase of the basal level of cAMP and mainly cGMP was observed. The cGMP-PDE specific activity in the unfractionated cytosol of DHA-, EPA- or 8-bromo-cGMP-enriched cardiomyocytes was lower than that observed in control cells. Whereas the cAMP-PDE specific activity remained unchanged whatever the treatment used. At the same time, the response of the cGMP-stimulated PDE to cGMP was substantially increased in n-3 fatty acid-enriched cardiomyocytes and reached the same level as in the whole ventricle (340%). The treatment of neonatal cardiomyocytes with 8-bromo-cGMP also re-established the sensitivity of this cGMP-stimulated isozyme to cGMP (320%). These results suggest a common mechanism for polyunsaturated fatty acids and 8-bromo-cGMP in the regulation of the cGMP-stimulated PDE activity in cardiomyocytes from new-born rats.


Assuntos
3',5'-AMP Cíclico Fosfodiesterases/metabolismo , 3',5'-GMP Cíclico Fosfodiesterases/metabolismo , GMP Cíclico/análogos & derivados , Ácidos Graxos Ômega-3/farmacologia , Miocárdio/enzimologia , 3',5'-AMP Cíclico Fosfodiesterases/antagonistas & inibidores , Animais , Animais Recém-Nascidos , Células Cultivadas , AMP Cíclico/metabolismo , AMP Cíclico/farmacologia , GMP Cíclico/metabolismo , GMP Cíclico/farmacologia , Ácidos Graxos/análise , Cinética , Milrinona , Inibidores de Fosfodiesterase/farmacologia , Piridonas/farmacologia , Pirrolidinonas/farmacologia , Ratos , Ratos Wistar , Rolipram
17.
Prostaglandins ; 51(4): 263-73, 1996 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-8935186

RESUMO

12-HETE, the major lipoxygenase end-product of platelets and macrophages, may be released in contact of bronchial epithelium in inflammatory diseases of the lung. We have studied the outcome of 12-HETE in presence of human bronchial epithelial cells (HBEC). When HBEC were incubated with [3H]12-HETE for 30 minutes, 27.5% of total radioactivity was found in HBEC and 72.5% in supernatants. Unesterified 12-HETE accounted for 22.4% of total radioactivity, 4.5% being recovered in phospholipids, preferentially in phosphatidylcholine and phosphatidylethanolamine. No incorporation in neutral lipids was detected. 72.9% of the incubated radioactivity was recovered in un identified metabolites. As 12-HETE has been shown to modulate the expression and production of various proteins, the consequence of the 12-HETE uptake on the release of GM-CSF and IL8 by HBEC was assessed. HBEC from control subjects were cultured for 24 hours with 12-HETE (10(-9) to 10(-7)M) in the presence or absence of TNF alpha. Detectable amounts of both cytokines were released in the supernatant in basal conditions at 24hr, and TNF alpha increased significantly the release of GM-CSF. 12-HETE at 10(-7)M weakly but significantly decreased the TNF-induced release of GM-CSF from HBEC. Thus the uptake of 12-HETE could affect the epithelial cell function in some situations.


Assuntos
Brônquios/metabolismo , Fator Estimulador de Colônias de Granulócitos e Macrófagos/biossíntese , Ácidos Hidroxieicosatetraenoicos/metabolismo , Interleucina-8/biossíntese , Lipídeos/química , Adulto , Brônquios/citologia , Células Cultivadas , Células Epiteliais , Epitélio/metabolismo , Feminino , Humanos , Ácidos Hidroxieicosatetraenoicos/análise , Masculino , Pessoa de Meia-Idade
18.
Gen Pharmacol ; 27(2): 269-72, 1996 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-8919641

RESUMO

1. In this study, we compared the effects of two antihistamine drugs on the production of granulocyte-macrophage colony-stimulating factor and interleukin-8 by human bronchial epithelial cells in vitro. 2. Cytokine production was assessed by the use of an enzyme-linked immunosorbent assay. 3. Epithelial cells spontaneously released both cytokines and tumor necrosis factor alone induced a significant increase in this production but loratadine and cetirizine had no effect at the various concentrations studied. 4. The antihistamines have no effect and this suggests that histamine plays no role in cytokine production under these conditions.


Assuntos
Brônquios/efeitos dos fármacos , Brônquios/metabolismo , Cetirizina/farmacologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/biossíntese , Antagonistas dos Receptores Histamínicos H1/farmacologia , Interleucina-8/biossíntese , Loratadina/farmacologia , Adulto , Idoso , Células Cultivadas , Ensaio de Imunoadsorção Enzimática , Epitélio/efeitos dos fármacos , Epitélio/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fator de Necrose Tumoral alfa/farmacologia
19.
Lipids ; 31 Suppl: S109-15, 1996 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-8729104

RESUMO

The exchange of docosahexaenoic acid (22:6n-3) within lipid pools in rat and human has been followed as a function of time after the ingestion of triglycerides (TG) containing 22:6n-3 labeled with 13C(13C 22:6n-3). The 13C abundance in the fatty acid was measured by gas-chromatography-combustion isotope ratio mass spectrometry which allowed the detection of 0.001 atom 13C percent 12C. The 13C 22:6n-3 appearance was rapid in the TG of very low density lipoprotein plus chylomicron fraction, in which the maximal labeling was observed at 3 and 2 h after ingestion in rat and human, respectively. Concomitant with the TG utilization of this fraction by lipoprotein lipase from tissues, unesterified 13C 22:6n-3 appeared in the plasma albumin. 13C 22:6n-3 bound to albumin was mostly present in unesterified form before 12 h post-ingestion while after that period, lysophosphatidylcholine (lysoPC) bound to albumin carried higher 13C 22:6n-3 concentrations. These lyso-PC were mostly from hepatic origin and might represent a potential source of 22:6n-3 redistribution to tissues. The 13C 22:6n-3 uptake into rat brain PC and phosphatidylethanolamine was still increasing when the concentration of plasma unesterified 13C 22:6n-3 had already dropped to a minimal plateau value and during the period of maximal plasma circulation of 13C 22:6n-3-lysoPC bound to albumin. In contrast, the uptake of 13C 22:6n-3 into blood platelet PC occurred during the phase of important circulation of 13C-22:6n-3 bound to albumin, suggesting the in vivo efficiency of the Lands pathway for this fatty acid. It is concluded that 13C 22:6n-3 esterified in TG is rapidly absorbed and redistributed within plasma lipoproteins and that its redistribution within the two lipid species bound to albumin might influence its uptake by platelets and rat brain.


Assuntos
Plaquetas/metabolismo , Ácidos Docosa-Hexaenoicos/metabolismo , Cromatografia Gasosa-Espectrometria de Massas/métodos , Metabolismo dos Lipídeos , Lipoproteínas/sangue , Animais , Isótopos de Carbono , Humanos , Masculino , Ratos , Ratos Sprague-Dawley , Valores de Referência
20.
Clin Exp Allergy ; 25(8): 729-36, 1995 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-7584684

RESUMO

BACKGROUND: Mucosal inflammatory processes in late phase of allergic diseases involve cytokine production, cell adhesion molecule overexpression and release of inflammatory mediators with chemotactic activity, such as leukotriene B4 (LTB4). We had previously observed increased production of LTB4 by neutrophils in patients with allergic rhinitis and discussed the role of granulocyte macrophage-colony stimulating factor (GM-CSF) priming. Some antihistaminic compounds were shown to diminish the production of leukotrienes by neutrophils. OBJECTIVES: In a first step, we evaluated in ex vivo and in vitro studies, the effects of cetirizine on LTB4 production by blood neutrophils from allergic and healthy subjects. In a second step, we studied the in vitro effect of cetirizine on LTB4 production by neutrophils from healthy subjects during GM-CSF priming of these cells. METHODS: Neutrophils from both populations were purified from venous blood and LTB4 production was measured using high performance liquid cromatography (HPLC) method. RESULTS: In ex vivo studies, cetirizine treatment induced a decreased LTB4 production by neutrophils in allergic rhinitis. This effect of decreased LTB4 production was reproduced in vitro with 10(-8)-10(-6)M cetirizine. Nevertheless, this anti-H1 compound had no effect on neutrophil priming with GM-CSF. CONCLUSION: As LTB4 is an important chemotactic factor, Cetirizine could act on inflammatory cell recruitment by inhibiting LTB4 production by neutrophils.


Assuntos
Antialérgicos/farmacologia , Cetirizina/farmacologia , Antagonistas dos Receptores Histamínicos H1/farmacologia , Leucotrieno B4/biossíntese , Neutrófilos/metabolismo , Rinite Alérgica Perene/metabolismo , Adulto , Asma/metabolismo , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Humanos , Masculino , Pessoa de Meia-Idade
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