Navegando por Palavras-chave "Lipopolysaccharide"

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    Adenosine A(2A) receptor agonist (CGS-21680) prevents endotoxin-induced effects on nucleotidase activities in mouse lymphocytes
    (Elsevier B.V., 2011-01-25) Vuaden, Fernanda Cenci; Savio, Luiz Eduardo Baggio; Bastos, Carolina Maria Alves [UNIFESP]; Bogo, Mauricio Reis; Bonan, Carla Denise; Pontificia Univ Catolica Rio Grande do Sul; Univ Fed Rio Grande do Sul; Universidade Federal de São Paulo (UNIFESP)
    Adenosine 5'-triphosphate (ATP) released during inflammation presents proinflammatory properties. Adenosine, produced by catabolism of ATP, is an anti-inflammatory compound. Considering the role of ATP and adenosine in inflammation and the importance of ectonucleotidases in the maintenance of their extracellular levels, we investigated the effect of a selective agonist of the adenosine A(2A) receptor (CGS-21680) on ectonucleotidase activities and gene expression patterns in lymphocytes from mice submitted to an endotoxemia model. Animals were injected intraperitoneally with 12 mg/kg Lipopolyssacharide (LPS) and/or 0.5 mg/kg CGS-21680 or saline. Nucleotidase activities were determined in lymphocytes from mesenteric lymph nodes and analysis of ectonucleotidase expression was carried out by a semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) assay. Exposure to endotoxemia promoted an increase in nucleotide hydrolysis. When CGS-21680 was administered concomitantly with LPS, this increase was prevented for ATP, adenosine 5'-monophosphate (AMP), and p-Nitrophenyl thymidine 5'-monophosphate (p-Nph-5'-TMP) hydrolysis. However, when CGS-21680 was administered 24 h after LPS injection, the increase was not reversed. the expression pattern of ectonucleotidases was not altered between LPS and LPS plus CGS-21680 groups, indicating that the transcriptional control was not involved on the effect exerted for CGS-21680. These results showed an enhancement of extracellular nucleotide catabolism in lymphocytes after induction of endotoxemia, which was prevented, but not reversed by CGS-21680 administration. These findings suggest that the control of nucleotide and nucleoside levels exerted by CGS-21680 could contribute to the modulation of the inflammatory process promoted by adenosine A(2A) agonists. (C) 2010 Elsevier B.V. All rights reserved.
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    The anti-inflammatory action of the analgesic kyotorphin neuropeptide derivatives: insights of a lipid-mediated mechanism
    (Springer wien, 2016) Conceicao, Katia [UNIFESP]; Magalhaes, Pedro R.; Campos, Sara R. R.; Domingues, Marco M.; Ramu, Vasanthakumar G.; Michalek, Matthias; Bertani, Philippe; Baptista, Antonio M.; Heras, Montserrat; Bardaji, Eduard R.; Bechinger, Burkhard; Ferreira, Monica Lopes; Castanho, Miguel A. R. B.
    Recently, a designed class of efficient analgesic drugs derived from an endogenous neuropeptide, kyotorphin (KTP, Tyr-Arg) combining C-terminal amidation (KTP-NH2) and N-terminal conjugation to ibuprofen (Ib), IbKTP-NH2, was developed. The Ib moiety is an enhancer of KTP-NH2 analgesic action. In the present study, we have tested the hypothesis that KTP-NH2 is an enhancer of the Ib anti-inflammatory action. Moreover, the impact of the IbKTP-NH2 conjugation on microcirculation was also evaluated by a unified approach based on intravital microscopy in the murine cremasteric muscle. Our data show that KTP-NH2 and conjugates do not cause damage on microcirculatory environment and efficiently decrease the number of leukocyte rolling induced by lipopolysaccharide (LPS). Isothermal titration calorimetry showed that the drugs bind to LPS directly thus contributing to LPS aggregation and subsequent elimination. In a parallel study, molecular dynamics simulations and NMR data showed that the IbKTP-NH2 tandem adopts a preferential "stretched" conformation in lipid bilayers and micelles, with the simulations indicating that the Ib moiety is anchored in the hydrophobic core, which explains the improved partition of IbKTP-NH2 to membranes and the permeability of lipid bilayers to this conjugate relative to KTP-NH2. The ability to bind glycolipids concomitant to the anchoring in the lipid membranes through the Ib residue explains the analgesic potency of IbKTP-NH2 given the enriched glycocalyx of the blood-brain barrier cells. Accumulation of IbKTP-NH2 in the membrane favors both direct permeation and local interaction with putative receptors as the location of the KTP-NH2 residue of IbKTP-NH2 and free KTP-NH2 in lipid membranes is the same.
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    Both adiponectin and interleukin-10 inhibit LPS-induced activation of the NF-kappa B pathway in 3T3-L1 adipocytes
    (Elsevier B.V., 2012-01-01) Lira, Fábio Santos de [UNIFESP]; Rosa Neto, José Cesar [UNIFESP]; Pimentel, Gustavo Duarte [UNIFESP]; Seelaender, Marilia; Dâmaso, Ana Raimunda [UNIFESP]; Oyama, Lila Missae [UNIFESP]; Nascimento, Claudia Maria da Penha Oller do [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Universidade de São Paulo (USP)
    Adiponectin and interleukin 10 (IL-10) are adipokines that are predominantly secreted by differentiated adipocytes and are involved in energy homeostasis, insulin sensitivity, and the anti-inflammatory response. These two adipokines are reduced in obese subjects, which favors increased activation of nuclear factor kappa B (NF-kappa B) and leads to elevation of pro-inflammatory adipokines. However, the effects of adiponectin and IL-10 on NF-kappa B DNA binding activity (NF-kappa Bp50 and NF-kappa Bp65) and proteins involved with the toll-like receptor (TLR-2 and TLR-4) pathway, such as MYD88 and TRAF6 expression, in lipopolysaccharide-treated 3T3-L1 adipocytes are unknown. Stimulation of lipopolysaccharide-treated 3T3-L1 adipocytes for 24 h elevated IL-6 levels; activated the NF-kappa B pathway cascade; increased protein expression of IL-6R, TLR-4, MYD88, and TRAF6; and increased the nuclear activity of NF-kappa B (p50 and p65) DNA binding. Adiponectin and IL-10 inhibited the elevation of IL-6 levels and activated NF-kappa B (p50 and p65) DNA binding. Taken together, the present results provide evidence that adiponectin and IL-10 have an important role in the anti-inflammatory response in adipocytes. in addition, inhibition of NF-kappa B signaling pathways may be an excellent strategy for the treatment of inflammation in obese individuals. (C) 2011 Elsevier B.V. All rights reserved.
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    Bradykinin inducible receptor is essential to lipopolysaccharide-induced acute lung injury in mice
    (Elsevier B.V., 2010-05-25) Campanholle, Gabriela; Landgraf, Richardt G. [UNIFESP]; Borducchi, Erica; Semedo, Patricia [UNIFESP]; Wang, Pamela H. M. [UNIFESP]; Amano, Mariane T.; Russo, Momtchilo; Pacheco-Silva, Alvaro [UNIFESP]; Jancar, Sonia; Camara, Niels O. S. [UNIFESP]; Universidade de São Paulo (USP); Universidade Federal de São Paulo (UNIFESP)
    Lipopolysaccharides from gram-negative bacteria are amongst the most common causative agents of acute lung injury, which is characterized by an inflammatory response, with cellular infiltration and the release of mediators/cytokines. There is evidence that bradykinin plays a role in lung inflammation in asthma but in other types of lung inflammation its role is less clear. in the present study we evaluated the role of the bradykinin B(1) receptor in acute lung injury caused by lipopolysaccharide inhalation and the mechanisms behind bradykinin actions participating in the inflammatory response. We found that in C57BI/6 mice, the bradykinin B(1) receptor expression was up-regulated 24 h after lipopolysaccharide inhalation. At this time, the number of cells and protein concentration were significantly increased in the bronchoalveolar lavage fluid and the mice developed airway hyperreactivity to methacholine. in addition, there was an increased expression of tumor necrosis factor-alpha, interleukin-1 beta and interferon-gamma and chemokines (monocytes chemotactic protein-1 and KC) in the bronchoalveolar lavage fluid and in the lung tissue. We then treated the mice with a bradykinin B, receptor antagonist, R-954 (Ac-Orn-[Oic(2), alpha-MePhe(5), D-beta Nal(7), Ile(8)]desArg(9)-bradykinin), 30 min after lipopolysaccharide administration. We observed that this treatment prevented the airway hyperreactivity as well as the increased cellular infiltration and protein content in the bronchoalveolar lavage fluid. Moreover, R-954 inhibited the expression of cytokines/chemokines. These results implicate bradykinin, acting through B(1) receptor, in the development of acute lung injury caused by lipopolysaccharide inhalation. (C) 2010 Elsevier B.V. All rights reserved.
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    Changes in the proteomic profile during differentiation and maturation of human monocyte-derived dendritic cells stimulated with granulocyte macrophage colony stimulating factor/interleukin-4 and lipopolysaccharide
    (Wiley-Blackwell, 2005-04-01) Pereira, Sandra Rodrigues [UNIFESP]; Faça, Vitor Marcel; Gomes, Glauce Gaspar; Chammas, Roger; Fontes, Aparecida Maria; Covas, Dimas Tadeu; Greene, Lewis Joel; Universidade de São Paulo (USP); Universidade Federal de São Paulo (UNIFESP)
    Dendritic cells (DCs) are highly specialized antigen-presenting cells that play an essential role in the immune response. We used the proteomic approach based on two-dimensional gel electrophoresis and mass spectrometry to identify the protein changes that occur during differentiation of DCs from monocytes (Mo) stimulated with granulocyte macrophage colony stimulating factor/interleukin-4 (GM-CSF/IL-4) and during the maturation of immature DCs stimulated with lipopolysaccharide. Sixty-three differentially expressed proteins (+/- two-fold) were unambiguously identified with sequence coverage greater than 20%. They corresponded to only 36 different proteins, because 11 were present as 38 electrophoretic forms. Some proteins such as tropomyosin 4 and heat shock protein 71 presented differentially expressed electrophoretic forms, suggesting that many of the changes in protein expression that accompany differentiation and maturation of DCs occur in post-translationally modified proteins. the largest differences in expression were observed for actin (21-fold in Mo), Rho GDP-dissociation inhibitor 2 (20-fold in Mo), vimentin (eight-fold in immature DCs), lymphocyte-specific protein 1 (12-fold in mature DCs) and thioredoxin (14-fold in mature DCs). Several proteins are directly related to functional and morphological characteristics of DCs, such as cytoskeletal proteins (cytoskeleton rearrangement) and chaperones (antigen processing and presentation), but other proteins have not been assigned specific functions in DCs. Only a few proteins identified here were the same as those reported in proteomic studies of DCs, which used different stimuli to produce the cells (GM-CSF/IL-4 and tumor necrosis factor-alpha). These data suggest that the DC protein profile depends on the stimuli used for differentiation and especially for maturation.
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    Collagen XVIII/endostatin expression in experimental endotoxemic acute renal failure
    (Associação Brasileira de Divulgação Científica, 2009-12-01) Cichy, Milene Cristina [UNIFESP]; Rocha, Flavia Gomes de Goes [UNIFESP]; Tristão, Vivian Regina [UNIFESP]; Pessoa, Edson de Andrade [UNIFESP]; Cenedeze, Marcos Antonio [UNIFESP]; Nürmberg Junior, R.; Schor, Nestor [UNIFESP]; Bellini, Maria Helena [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Instituto de Pesquisas Energéticas e Nucleares Centro de Biotecnologia; Faculdades Metropolitanas Unidas Faculdade de Medicina Veterinária
    Acute renal failure (ARF) is a frequent complication of Gram-negative sepsis, with a high risk of mortality. Lipopolysaccharide (LPS)-induced ARF is associated with hemodynamic changes that are strongly influenced by the overproduction of nitric oxide (NO) through the cytokine-mediated up-regulation of inducible NO synthase. LPS-induced reductions in systemic vascular resistance paradoxically culminate in renal vasoconstriction. Collagen XVIII is an important component of the extracellular matrix expressed in basement membranes. Its degradation by matrix metalloproteases, cathepsins and elastases results in the formation of endostatin, claimed to have antiangiogenic activity and to be a prominent vasorelaxing agent. We evaluated the expression of endostatin/collagen XVIII in an endotoxemic ARF model. ARF was induced in C57BL/6 mice by intraperitoneal injection of LPS (10 mg/kg) followed by sacrifice 4 and 12 h later. Kidney tissue was the source of RNA and protein and the subject of histological analysis. As early as 4 h after LPS administration, blood urea, creatinine and NO levels were significantly increased compared to control. Endostatin/collagen XVIII mRNA levels were 0.71 times lower than sham-inoculated mice 4 h after LPS inoculation, returning to normal levels 12 h after LPS inoculation. Immunohistological examination revealed that acute injury caused by LPS leads to an increase of endostatin basement membrane staining in association with the decrease of CD31 endothelial basement membrane staining. These results indicate that in the early phase of endotoxemic ARF the endostatin levels were not regulated by gene expression, but by the metabolism of collagen XVIII.
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    Estudos in vitro da participação da leptina em células endoteliais pulmonares de ratos nutridos e desnutridos intrauterinamente
    (Universidade Federal de São Paulo, 2014-03-02) Santos, Leila Aparecida dos [UNIFESP]; Landgraf, Richardt Gama [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Studies suggest that intrauterine malnutrition can "program" the fetal tissues making them more prone to disorders associated with food, such as type 2 diabetes, metabolic syndrome and other chronic diseases in adulthood , and induce changes in the immune response and inflammatory. Leptin, the main hormone secreted by adipose tissue, has a pleiotropic action, working in various systems such as the regulation of energy balance, reproduction, hematopoiesis, lymphopoiesis and has a modulating action of the immune system. Pulmonary endothelial cells were obtained from Wistar rats, male or intrauterine undernourished fed rats at 12 weeks of age. These cells were stimulated with leptin or LPS or leptin plus LPS, furthermore, cells with no stimulus were used as controls. Two hours after stimulation, production of inflammatory mediators PGE2, LTB4, IL1?, TNF? and the expression of ERK ½, and leptin receptors were analyzed. We observed that expression of the leptin receptor is 63 % lower in primary cultures of endothelial cells derived from intrauterine undernourished rats. Leptin alone did not induce any change in levels of inflammatory mediators evaluated, whereas LPS increased levels of PGE2 (250 %) and LTB4 (29%). Only endothelial cells of rats fed LPS administration + leptin production increased lipid mediators compared to LPS group (PGE 2 - and LTB4 28 % - 18%). The assay for IL1? showed that only cells from the control animals showed expression of IL1?. The expression of TNF and activation of ERK ½ showed a contradictory result because only the endothelial cells from the malnourished animals showed higher expression and activation, respectively. This result may suggest an expression of TNFa stimulation via LPS via another receptor with activation of ERK. These results suggest that the lower expression of the leptin receptor in lung endothelial cells in intrauterine undernourished rats could negatively modulate lipid mediator production in these cells.
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    Immune development in HIV-exposed uninfected children born to HIV-infected women
    (Inst Medicina Tropical Sao Paulo, 2017) Miyamoto, Maristela [UNIFESP]; Gouvêa, Aida de Fátima Thomé Barbosa [UNIFESP]; Ono, Erika [UNIFESP]; Succi, Regina Célia de Menezes [UNIFESP]; Pahwa, Savita; Moraes-Pinto, Maria Isabel de [UNIFESP]
    Immunological and clinical findings suggestive of some immune dysfunction have been reported among HIV-exposed uninfected(HEU) children and adolescents. Whether these defects are persistent or transitory is still unknown. HEU pediatric population at birth, 12 months, 6-12 years were evaluated in comparison to healthy age-matched HIV-unexposed controls. Plasma levels of LPS, sCD14, cytokines, lymphocyte immunophenotyping and T-cell receptor excision circles (TREC) were assessed. HEU and controls had similar LPS levels, which remained low from birth to 6-12 years
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    Immune Outcomes of Paradoxical Sleep Deprivation on Cellular Distribution in Naive and Lipopolysaccharide-Stimulated Mice
    (Karger, 2012-01-01) Zager, Adriano [UNIFESP]; Ruiz, Francieli Silva [UNIFESP]; Tufik, Sergio [UNIFESP]; Andersen, Monica Levy [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Background/Aims: Several lines of evidence indicate that sleep loss imposes significant consequences on the host defense system, including changes in cell number, activity and distribution. However, it is not clear whether cellular alterations after sleep deprivation are caused by redistribution to immune organs or by death of these cells or how the response to a nonspecific immune activator would be affected. Therefore, the aim of this study was to assess the leukocyte distribution after paradoxical sleep deprivation (PSD) in saline-and lipopolysaccharide-treated mice. Methods: Adult inbred mice were paradoxical sleep deprived (72 h), whereas the controls were kept in their home cages. After PSD, both groups received an injection of either saline or lipopolysaccharide (LPS; 1 or 5 mu g/animal, intraperitoneally), 2 h prior to the collection of blood, spleen, lymph nodes and peritoneal wash. Isolated cells were then designated to differential leukocyte count (blood) and flow cytometry analysis of immune cell subsets (immune sites). Results: the data revealed that PSD caused a significant reduction of circulating lymphocytes and a general decrease in all cellular subsets of spleen, mainly T and B cells. However, no alteration in response of PSD was found on other immune sites, such as lymph nodes and peritoneum. of note, immune cell distribution in response to in vivo LPS stimulation remained unchanged after PSD. Conclusions: Our study provided original evidence concerning the immune outcomes of PSD, indicating that cellular decrease caused by PSD is not restricted to circulation, but also to immune sites. Taken together, our results could help shed light on the physiological mechanisms of leukocyte trafficking. Copyright (C) 2012 S. Karger AG, Basel
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    Inflammatory markers are associated with inhibitory avoidance memory deficit induced by sleep deprivation in rats
    (Elsevier B.V., 2011-08-01) Esumi, Lia Assae [UNIFESP]; Palma, B. D. [UNIFESP]; Gomes, V. L. [UNIFESP]; Tufik, S. [UNIFESP]; Hipolide, D. C. [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Sleep deprivation (SD) causes detrimental effects to the body, such as memory impairment and weight loss. SD also changes the concentration of inflammatory mediators such as cytokines, which, in turn, can affect cognitive functioning. Thus, the objective of this study was to investigate the involvement of these inflammatory mediators in inhibitory avoidance memory deficit in sleep-deprived rats. Male Wistar rats were deprived of sleep by the modified multiple platform method for 96 h, while their respective controls remained in their housing cages. To assess memory after SD, all animals underwent training, followed by the inhibitory avoidance task test 24 h later. Also, the weight of each animal was recorded daily. in the first experiment, animals received an acute administration of lipopolysaccharide (LPS, 50 or 75 mu g/kg i.p.) 3 h before the inhibitory avoidance training. in the experiment 2, the animals received acute or chronic administration of anti-IL-6 antibody (Ab, 2 mu g/kg i.p.). the acute administration was performed 3 h before the inhibitory avoidance training, while the chronic treatment administrations were performed daily during the SD period. the 75 mu g/kg dose of LPS, but not the 50 mu g/kg dose, caused a significant attenuation of memory impairment in the sleep-deprived animals. Although the treatments with the anti-IL-6 Ab did not produce any significant changes in cognitive performance, the Ab attenuated weight loss in sleep-deprived animals. Taken together, these results suggest the involvement of inflammatory mediators in the modulation of memory deficit and weight loss that are observed in sleep-deprived rats. (C) 2011 Elsevier B.V. All rights reserved.
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    Leptin Downregulates LPS-Induced Lung Injury: Role of Corticosterone and Insulin
    (Karger, 2014-01-01) Landgraf, Maristella de Almeida [UNIFESP]; Silva, Reinaldo Correia [UNIFESP]; Correa-Costa, Matheus; Hiyane, Meire Ioshie [UNIFESP]; Carvalho, Maria Helena C.; Landgraf, Richardt Gama [UNIFESP]; Câmara, Niels Olsen Saraiva [UNIFESP]; Universidade de São Paulo (USP); Universidade Federal de São Paulo (UNIFESP)
    Background/Aims: We investigated the effects of leptin in the development of lipopolysaccharide (LPS)-induced acute lung inflammation (ALI) in lean mice. Methods: Mice were administered leptin (1.0 mu g/g) or leptin (1.0 mu g/g) followed by LPS (1.5 mu g/g) intranasally. Additionally, some animals were given LPS (1.5 mu g/g) or saline intranasally alone, as a control. Tissue samples and fluids were collected six hours after instillation. Results: We demonstrated that leptin alone did not induce any injury. Local LPS exposure resulted in significant acute lung inflammation, characterized by a substantial increase in total cells, mainly neutrophils, in bronchoalveolar lavages (BAL). We also observed a significant lymphocyte influx into the lungs associated with enhanced lung expression of chemokines and cytokines (KC, RANTES, TNF-alpha, IFN-beta, GM-CSF and VEGF). LPS-induced ALI was characterized by the enhanced expression of ICAM-1 and iNOS in the lungs. Mice that received LPS showed an increase in insulin levels. Leptin, when administered prior to LPS instillation, abolished all of these effects. LPS induced an increase in corticosterone levels, and leptin potentiated this event. Conclusion: These data suggest that exogenous leptin may promote protection during sepsis, and downregulation of the insulin levels and upregulation of corticosterone may be important mechanisms in the amelioration of LPS-induced ALI.Copyright (c) 2014 S. Karger AG, Basel
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    Lipopolysaccharide-induced expression of cell surface receptors and cell activation of neutrophils and monocytes in whole human blood
    (Associação Brasileira de Divulgação Científica, 2010-09-01) Gomes, Natalia Estevam [UNIFESP]; Brunialti, Milena Karina Coló [UNIFESP]; Mendes, Marialice Erdelyi [UNIFESP]; Freudenberg, M.; Galanos, C.; Salomão, Reinaldo [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Max-Planck-Institute for Immunobiology
    Lipopolysaccharide (LPS) activates neutrophils and monocytes, inducing a wide array of biological activities. LPS rough (R) and smooth (S) forms signal through Toll-like receptor 4 (TLR4), but differ in their requirement for CD14. Since the R-form LPS can interact with TLR4 independent of CD14 and the differential expression of CD14 on neutrophils and monocytes, we used the S-form LPS from Salmonella abortus equi and the R-form LPS from Salmonella minnesota mutants to evaluate LPS-induced activation of human neutrophils and monocytes in whole blood from healthy volunteers. Expression of cell surface receptors and reactive oxygen species (ROS) and nitric oxide (NO) generation were measured by flow cytometry in whole blood monocytes and neutrophils. The oxidative burst was quantified by measuring the oxidation of 2',7'-dichlorofluorescein diacetate and the NO production was quantified by measuring the oxidation of 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate. A small increase of TLR4 expression by monocytes was observed after 6 h of LPS stimulation. Monocyte CD14 modulation by LPS was biphasic, with an initial 30% increase followed by a 40% decrease in expression after 6 h of incubation. Expression of CD11b was rapidly up-regulated, doubling after 5 min on monocytes, while down-regulation of CXCR2 was observed on neutrophils, reaching a 50% reduction after 6 h. LPS induced low production of ROS and NO. This study shows a complex LPS-induced cell surface receptor modulation on human monocytes and neutrophils, with up- and down-regulation depending on the receptor. R- and S-form LPS activate human neutrophils similarly, despite the low CD14 expression, if the stimulation occurs in whole blood.
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    Modulation of sickness behavior by sleep: the role of neurochemical and neuroinflammatory pathways in mice
    (Elsevier B.V., 2009-08-01) Zager, Adriano [UNIFESP]; Andersen, Monica L. [UNIFESP]; Lima, Marcelo M. S. [UNIFESP]; Reksidler, Angela B.; Machado, Ricardo B. [UNIFESP]; Tufik, Sergio [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Univ Fed Parana
    Activation of the immune system elicits several behavioral changes that are collectively called sickness behavior and consists in a strategy to overcome infection. Steep deprivation can increase susceptibility to pathogens and to behavioral alterations. Thus, the present study aimed to determine how paradoxical steep deprivation (PSD) affects the behavioral and neurochemical responses to lipopotysaccharide (LPS, potent activator of the immune response). Adult inbred mice were paradoxical steep deprived (72 h), whereas the control group was kept in their home cages. Both groups received either an injection of saline or LPS (5, 10 or 20 mu g/animal ip) before behavioral tasks and tissue collection. During the recovery sleep period, LPS provoked a strong inhibition of steep rebound due to a suppression of paradoxical steep. PSD increased the susceptibility of mice to LPS-induced immobility in the open field, which was capable of affecting the anxiety-like behavior also. These altered behavioral responses to LPS were accompanied by reduction in dopamine turnover within the striatum and increased expression of cyctooxygenase-2 in the cortex. the study provides some insights into how the steep-wake cycle affects the expression of sickness behavior induced by LPS. (C) 2009 Elsevier B.V. and ECNP. All rights reserved.
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    Noncrystalline uric acid inhibits proteoglycan and glycosaminoglycan synthesis in distal tubular epithelial cells (MDCK)
    (Associação Brasileira de Divulgação Científica, 2010-10-01) Borges, Fernanda Teixeira [UNIFESP]; Dalboni, Maria Aparecida [UNIFESP]; Michelacci, Yara Maria [UNIFESP]; Schor, Nestor [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Hyperuricemia is associated with renal stones, not only consisting of uric acid (UrAc) but also of calcium oxalate (CaOx). Glycosaminoglycans (GAGs) are well-known inhibitors of growth and aggregation of CaOx crystals. We analyzed the effect of noncrystalline UrAc on GAG synthesis in tubular distal cells. MDCK (Madin-Darby canine kidney) cells were exposed to noncrystalline UrAc (80 µg/mL) for 24 h. GAGs were labeled metabolically and characterized by agarose gel electrophoresis. The expression of proteoglycans and cyclooxygenase 2 (COX-2) was assessed by real-time PCR. Necrosis, apoptosis and prostaglandin E2 (PGE2) were determined by acridine orange, HOESCHT 33346, and ELISA, respectively. CaOx crystal endocytosis was evaluated by flow cytometry. Noncrystalline UrAc significantly decreased the synthesis and secretion of heparan sulfate into the culture medium (UrAc: 2127 ± 377; control: 4447 ± 730 cpm) and decreased the expression of perlecan core protein (UrAc: 0.61 ± 0.13; control: 1.07 ± 0.16 arbitrary units), but not versican. Noncrystalline UrAc did not induce necrosis or apoptosis, but significantly increased COX-2 and PGE2 production. The effects of noncrystalline UrAc on GAG synthesis could not be attributed to inflammatory actions because lipopolysaccharide, as the positive control, did not have the same effect. CaOx was significantly endocytosed by MDCK cells, but this endocytosis was inhibited by exposure to noncrystalline UrAc (control: 674.6 ± 4.6, CaOx: 724.2 ± 4.2, and UrAc + CaOx: 688.6 ± 5.4 geometric mean), perhaps allowing interaction with CaOx crystals. Our results indicate that UrAc decreases GAG synthesis in MDCK cells and this effect could be related to the formation of UrAc and CaOx stones.
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    Participação do receptor P2x7 nos efeitos de longo prazo da inflamação sistêmica neonatal: estresse oxidativo hipocampal, alteração da nocicepção e ansiedade
    (Universidade Federal de São Paulo (UNIFESP), 2018-03-29) Silva, Clivandir Severino da [UNIFESP]; Mello, Luiz Eugenio Araujo de Moraes [UNIFESP]; http://lattes.cnpq.br/4462750801249231; http://lattes.cnpq.br/8977562964399129; Universidade Federal de São Paulo [UNIFESP]
    Objetivos: Avaliar os efeitos da inflamação sistêmica no período neonatal sobre a taxa de mortalidade, a massa corporal, o comportamento associado à ansiedade, a nocicepção e o estresse oxidativo no giro denteado do hipocampo em longo prazo e se o bloqueio do receptor P2X7 (P2X7R) com Brilliant Blue G (BBG) modula os efeitos da inflamação sobre essas variáveis. Metodologia: Filhotes de ratos da cepa Wistar norvegicus (machos e fêmeas) foram distribuídos nos grupos experimentais NAIVE, salina (SAL)+SAL, SAL+lipopolissacarídeo (LPS) e BBG+LPS. As soluções foram injetadas via intraperitoneal no primeiro, terceiro, quinto e sétimo dia pós-natal (DPN). A massa corporal foi analisada no DPN1, DPN10, DPN21, DPN45 e DPN89. No DPN80, DPN82 e DPN84 foram realizados respectivamente os testes do labirinto em cruz elevado, da placa quente e de retirada da cauda. Os animais foram perfundidos no DPN89 e os encéfalos extraídos. Os encéfalos foram cortados em fatias coronais e algumas, contendo o hipocampo, coradas com etidina. Resultados: Os animais expostos à inflamação sistêmica neonatal (grupo SAL+LPS) apresentaram aumento da taxa de mortalidade nas primeiras semanas de vida, atraso transitório no ganho de massa corporal e aumento da concentração de ânion superóxido no giro denteado do hipocampo na fase adulta quando comparados com os animais dos grupos controles (grupos NAIVE e SAL+SAL) e o bloqueio do P2X7R (grupo BBG+LPS) reduziu a taxa de mortalidade e a concentração de ânion superóxido. A exposição à inflamação não induziu comportamento associado à ansiedade e alteração da nocicepção em longo prazo. Conclusão: A ativação do P2X7R durante o processo inflamatório sistêmico no período neonatal colabora para desencadear eventos fisiopatológicos que podem culminar com a morte dos neonatos e, caso ela não ocorra, promove estresse oxidativo na idade adulta no giro denteado do hipocampo, por induzir aumento da produção de ânion superóxido. Portanto, o bloqueio da ativação do P2X7R, usando BBG, pode ser uma estratégia para reduzir a taxa mortalidade decorrente de complicações da inflamação sistêmica neonatal e prevenir alterações no tecido nervoso associadas ao estresse oxidativo.
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    Pro-inflammatory and oxidative effects of noncrystalline uric acid in human mesangial cells: contribution to hyperuricemic glomerular damage
    (Springer, 2011-02-01) Convento, M. S. [UNIFESP]; Pessoa, E. [UNIFESP]; Dalboni, Maria Aparecida [UNIFESP]; Borges, F. T. [UNIFESP]; Schor, N. [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Hyperuricemia is associated with cardiovascular and renal diseases, as glomerulosclerosis. Noncrystalline uric acid induces deleterious effects on endothelial and vascular smooth muscle cells. in the present study, we analyzed the damage induced by UA on human mesangial cells (HMC), the potential mechanism involved in this injury, and its consequences during infection. HMC were exposed to noncrystalline UA (8 mg/dl) and/or lipopolysaccharide (LPS, 100 mu g/ml) for 24 h. in the experiments of cellular viability, HMC were exposed to 8-50 mg/dl of UA. Necrosis was assessed by acridine orange and ethidium bromide. Reactive oxygen species (ROS) were analyzed by 2',7'-dichlorofluorescein. Prostaglandin E2 (PGE2) was evaluated by ELISA. Cyclooxygenase 2 (COX-2) expression was assessed by real-time PCR. UA induced necrosis only at supraphysiological concentrations. Nevertheless, it significantly increased ROS production at 8 mg/dl. LPS increased necrosis and ROS production. Interestingly, the association between UA and LPS decreased ROS and necrosis. UA associated or not with LPS induced COX-2 expression and PGE2 increases in HMC. Results suggest that UA has pro- and anti-oxidant effects in HMC. During infections, it acts like scavenger increasing cellular viability, but alone it can induce ROS production and cellular death in higher concentrations. Additionally, UA has direct pro-inflammatory effects inducing COX-2 expression and PGE2 synthesis. It is concluded that elevated concentrations of uric acid potentially contributes to glomerular damage.
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    TLR4 and CD14 receptors expressed in rat pineal gland trigger NFKB pathway
    (Wiley-Blackwell, 2010-09-01) Cruz-Machado, Sanseray da Silveira; Carvalho-Sousa, Claudia Emanuele; Tamura, Eduardo Koji; Pinato, Luciana; Cecon, Erika; Magno Fernandes, Pedro Augusto Carlos; Avellar, Maria Christina Werneck [UNIFESP]; Ferreira, Zulma Silva; Markus, Regina Pekelmann; Universidade de São Paulo (USP); Univ Estadual Paulista; Universidade Federal de São Paulo (UNIFESP)
    Nuclear factor-kappa B (NFKB), a pivotal player in inflammatory responses, is constitutively expressed in the pineal gland. Corticosterone inhibits pineal NFKB leading to an enhancement of melatonin production, while tumor necrosis factor (TNF) leads to inhibition of Aa-nat transcription and the production of N-acetylserotonin in cultured glands. the reduction in nocturnal melatonin surge favors the mounting of the inflammatory response. Despite these data, there is no clear evidence of the ability of the pineal gland to recognize molecules that signal infection. This study investigated whether the rat pineal gland expresses receptors for lipopolysaccharide (LPS), the endotoxin from the membranes of Gram-negative bacteria, and to establish the mechanism of action of LPS. Here, we show that pineal glands possess both CD14 and toll-like receptor 4 (TLR4), membrane proteins that bind LPS and trigger the NFKB pathway. LPS induced the nuclear translocation of p50/p50 and p50/RELA dimers and the synthesis of TNF. the maximal expression of TNF in cultured glands coincides with an increase in the expression of TNF receptor 1 (TNFR1) in isolated pinealocytes. in addition, LPS inhibited the synthesis of N-acetylserotonin and melatonin. Therefore, the pineal gland transduces Gram-negative endotoxin stimulation by producing TNF and inhibiting melatonin synthesis. Here, we provide evidence to reinforce the idea of an immune-pineal axis, showing that the pineal gland is a constitutive player in the innate immune response.
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    The wake-promoting drug Modafinil prevents motor impairment in sickness behavior induced by LPS in mice: Role for dopaminergic D1 receptor
    (Pergamon-Elsevier Science Ltd, 2018) Zager, Adriano; Brandao, Wesley Nogueira; Margatho, Rafael Oliveira; Peron, Jean Pierre; Tufik, Sergio [UNIFESP]; Andersen, Monica Levy [UNIFESP]; Kornum, Birgitte Rahbek; Palermo-Neto, Joao
    The wake-promoting drug Modafinil has been used for many years for treatment of Narcolepsy and Excessive Daytime Sleepiness, due to a dopamine-related psychostimulant action. Recent studies have indicated that Modafinil prevents neuroinflammation in animal models. Thus, the aim of the present study was to evaluate the effect of Modafinil pretreatment in the Lipopolysaccharide (LPS)-induced sickness and depressive-like behaviors. Adult male C57BL/6J mice were pretreated with Vehicle or Modafinil (90 mg/Kg) and, 30 min later, received a single saline or LPS (2 mg/Kg) administration, and were submitted to the open field and elevated plus maze test 2 h later. After 24 h, mice were subjected to tail suspension test, followed by either flow cytometry with whole brain for CD11b(+) CD45(+) cells or qPCR in brain areas for cytokine gene expression. Modafinil treatment prevented the LPS-induced motor impairment, anxiety-like and depressive-like behaviors, as well as the increase in brain CD11b(+) CD45(high) cells induced by LPS. Our results indicate that Modafinil pretreatment also decreased the IL-1 beta gene upregulation caused by LPS in brain areas, which is possibly correlated with the preventive behavioral effects. The pharmacological blockage of the dopaminergic D1R by the drug SCH-23390 counteracted the effect of Modafinil on locomotion and anxiety-like behavior, but not on depressive-like behavior and brain immune cells. The dopaminergic D1 receptor signaling is essential to the Modafinil effects on LPS-induced alterations in locomotion and anxiety, but not on depression and brain macrophages. This evidence suggests that Modafinil treatment might be useful to prevent inflammation-related behavioral alterations, possibly due to a neuroimmune mechanism.