RESUMEN
BACKGROUND: DBA/1 mice arthritis models have contributed to our understanding of human rheumatoid arthritis (RA) and spondyloarthritis (SpA) pathogenesis, as well as the exploration of therapeutic targets for treatment. Quantitative polymerase chain reaction (qPCR) is an indispensable tool in molecular research, which requires reference gene validation to obtain consistent and reliable results. OBJECTIVE: To determine the stability of candidate reference genes for qPCR in the joint of collagen-induced arthritis (CIA) and spontaneous arthritis (SpAD) DBA/1 mice. METHODS: The expression of eleven commonly used reference genes (ACTB, B2M, EF1a, GAPDH, HMBS, HPRT, PPIB, RPL13A, SDHA, TBP, and YWHAZ) was assessed by qPCR and the data were compared using delta-Ct methods and the geNorm, NormFinder, and RefFinder software packages. Genes identified as stable in each model were used for the quantification of inflammatory cytokines RESULTS: The gene stabilities differed between the two arthritis models in the DBA/1 mice. EF1a and RPL13A were the best reference genes for SpAD, while RPL13A and TBP were the best for the CIA. These genes allowed the data normalization for the quantification of the inflammatory cytokines in both models; these results showed an increase in the expression of IL-1B, IL-12B, IL-17A, and IL-6 in the inflamed joints. The use of different primer sequences for the same reference gene resulted in different relative quantification values. CONCLUSION: This study demonstrates that commonly used reference genes may not be suitable for arthritic tissues from DBA/1 mice, and strengthening the principle that meticulous validation of reference genes is essential before each experiment to obtain valid and reproducible qPCR data for analysis or interpretation.
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Artritis Experimental/genética , Articulaciones del Pie , Genes Esenciales , Animales , Artritis Experimental/inmunología , Citocinas/genética , Citocinas/inmunología , Articulaciones del Pie/inmunología , Expresión Génica , Masculino , Ratones Endogámicos DBA , Reacción en Cadena en Tiempo Real de la Polimerasa , Reproducibilidad de los ResultadosRESUMEN
Physical exercise (PE) is recommended for Rheumatoid Arthritis (RA), but the molecular and biological mechanisms that impact the inflammatory process and joint destruction in RA remain unknown. The objective of this study was to evaluate the effect of PE on the histological and transcriptional changes in the joints of adjuvant-induced arthritis (AIA) rat model. AIA rats were subjected to PE on a treadmill for eight weeks. The joints were subjected to histological and microarray analysis. The differentially expressed genes (DEGs) by PE in the arthritic rats were obtained from the microarray. The bioinformatic analysis allowed the association of these genes in biological processes and signaling pathways. PE induced the differential expression of 719 genes. The DEGs were significantly associated with pathogenic mechanisms in RA, including HIF-1, VEGF, PI3-Akt, and Jak-STAT signaling pathways, as well as response to oxidative stress and inflammatory response. At a histological level, PE exacerbated joint inflammatory infiltrate and tissue destruction. The PE exacerbated the stressed joint environment aggravating the inflammatory process, the hypoxia, and the oxidative stress, conditions described as detrimental in the RA joints. Research on the effect of PE on the pathogenesis process of RA is still necessary for animal models and human.
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Artritis Experimental/genética , Hipoxia/genética , Inflamación/genética , Estrés Oxidativo/genética , Condicionamiento Físico Animal , Animales , Artritis Experimental/inducido químicamente , Modelos Animales de Enfermedad , Adyuvante de Freund/administración & dosificación , Perfilación de la Expresión Génica , Inflamación/patología , Masculino , Ratas , Ratas WistarRESUMEN
Healthy and adjuvant-induced arthritic rats were treated for 23 days with daily doses of 400 and 800 mg kg-1Ilex paraguariensis extract. This treatment (a) diminished the ROS levels in the liver and brain, (b) decreased oxidative protein and lipid damage in liver and brain, (c) increased the plasma antioxidant capacity, (d) increased the GSH levels and the GSH/GSSH ratio in both the liver and the brain, (e) almost restored the enzymatic activities linked to the metabolism of GSH-GSSG, and (f) reversed the modified activities of xanthine oxidase, superoxide dismutase and catalase. The anti-inflammatory actions (firstly) and the antioxidant actions (in the second place) of the yerba mate constituents (e.g., chlorogenic acid derivatives) are the causes of these beneficial effects. Daily ingestion of traditional yerba mate beverages may be effective in attenuating the symptoms of inflammatory diseases, especially in older adults.
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Antiinflamatorios/administración & dosificación , Antioxidantes/administración & dosificación , Artritis Experimental/tratamiento farmacológico , Ilex paraguariensis/química , Extractos Vegetales/administración & dosificación , Animales , Artritis Experimental/genética , Artritis Experimental/inmunología , Artritis Experimental/metabolismo , Bebidas/análisis , Catalasa/metabolismo , Humanos , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno/metabolismo , Superóxido Dismutasa/metabolismoRESUMEN
Mouse lines selected for maximal (AIRmax) or minimal acute inflammatory reaction (AIRmin) were used to characterize the immune response and the influence of genetic background during pristane-induced arthritis (PIA). Susceptible AIRmax mice demonstrated exacerbated cellular profiles during PIA, with intense infiltration of lymphocytes, as well as monocytes/macrophages and neutrophils, producing higher levels of IL-1ß, IFN-γ, TNF-α, IL-10, total IgG3, and chemokines. Resistant AIRmin mice controlled cell activation more efficiently than the AIRmax during arthritis progression. The weight alterations of the spleen and thymus in the course of PIA were observed. Our data suggest that selected AIRmax cellular and genetic immune mechanisms contribute to cartilage damage and arthritis severity, evidencing many targets for therapeutic actions.
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Artritis Experimental/inmunología , Citocinas/inmunología , Inmunoglobulina G/inmunología , Terpenos/efectos adversos , Enfermedad Aguda , Animales , Artritis Experimental/inducido químicamente , Artritis Experimental/genética , Artritis Experimental/patología , Citocinas/genética , Inmunoglobulina G/genética , Inflamación , Ratones , Bazo/inmunología , Bazo/patología , Terpenos/farmacología , Timo/inmunología , Timo/patologíaRESUMEN
BACKGROUND: Epidemiologic studies have highlighted the association of environmental factors with the development and progression of autoimmune and chronic inflammatory diseases. Among the environmental factors, smoking has been associated with increased susceptibility and poor prognosis in rheumatoid arthritis (RA). However, the immune and molecular mechanism of smoking-induced arthritis aggravation remains unclear. The transcription factor aryl hydrocarbon receptor (AHR) regulates the generation of Th17 cells, CD4 T cells linked the development of autoimmune diseases. AHR is activated by organic compounds including polycyclic aromatic hydrocarbons (PAHs), which are environmental pollutants that are also present in cigarette smoke. In this study, we investigated the role of AHR activation in the aggravation of experiment arthritis induced by exposure to cigarette smoke. METHODS: Mice were exposed to cigarette smoke during the developmental phase of antigen-induced arthritis and collagen-induced arthritis to evaluate the effects of smoking on disease development. Aggravation of articular inflammation was assessed by measuring neutrophil migration to the joints, increase in articular hyperalgesia and changes in the frequencies of Th17 cells. In vitro studies were performed to evaluate the direct effects of cigarette smoke and PAH on Th17 differentiation. We also used mice genetically deficient for AHR (Ahr KO) and IL-17Ra (Il17ra KO) to determine the in vivo mechanism of smoking-induced arthritis aggravation. RESULTS: We found that smoking induces arthritis aggravation and increase in the frequencies of Th17 cells. The absence of IL-17 signaling (Il17ra KO) conferred protection to smoking-induced arthritis aggravation. Moreover, in vitro experiments showed that cigarette smoke can directly increase Th17 differentiation of T cells by inducing AHR activation. Indeed, Ahr KO mice were protected from cigarette smoke-induced arthritis aggravation and did not display increase in TH17 frequencies, suggesting that AHR activation is an important mechanism for cigarette smoke effects on arthritis. Finally, we demonstrate that PAHs are also able to induce arthritis aggravation. CONCLUSIONS: Our data demonstrate that the disease-exacerbating effects of cigarette smoking are AHR dependent and environmental pollutants with AHR agonist activity can induce arthritis aggravation by directly enhancing Th17 cell development.
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Artritis Experimental/metabolismo , Receptores de Hidrocarburo de Aril/metabolismo , Humo/efectos adversos , Células Th17/metabolismo , Animales , Artritis Experimental/etiología , Artritis Experimental/genética , Compuestos Azo/farmacología , Masculino , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Ratones Noqueados , Pirazoles/farmacología , Receptores de Hidrocarburo de Aril/antagonistas & inhibidores , Receptores de Hidrocarburo de Aril/genética , Receptores de Interleucina-17/genética , Receptores de Interleucina-17/metabolismo , Nicotiana/químicaRESUMEN
Anti-inflammatory and antioxidant properties have been attributed to constituents of Rosmarinus officinalis. Considering the inflammatory nature of arthritis and the oxidative stress induced by the disease, this work aimed to investigate if R. officinalis is able to attenuate oxidative and inflammatory injuries caused by the disease. Rats with complete Freund's adjuvant induced arthritis were used. An aqueous extract of the rosemary leaves was administered orally. The parameters related to oxidative stress were measured in the liver, brain and plasma. The administration of the rosemary extract (RE) to arthritic rats diminished oxidative damage (e.g., less carbonylated proteins), improved the oxidative state (e.g., less reactive oxygen species) and also increased the antioxidant capacity by increasing the GSH level and the GSH/GSSG ratio and by almost normalizing the activity of several antioxidant enzymes. The treatment of arthritic rats with the extract also diminished paw edema, the number of leukocytes recruited in the femoro-tibial joint cavities and the weight of the lymph nodes and delayed the appearance of secondary lesions. Twelve phenolic compounds were identified and quantified in the aqueous RE, rosmarinic acid being the most abundant one. In addition, the extract also contains polysaccharides and proteins. The anti-inflammatory and antioxidant activities of the RE can be attributed, partly at least, to its content of polyphenolics with the strong possibility of synergistic interactions as well as metabolic activations. The results corroborate and amplify the general notion that rosemary aqueous extracts possess efficient anti-inflammatory agents and suggest that they are able to attenuate the oxidative stress inherent to arthritis.
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Antiinflamatorios/administración & dosificación , Antioxidantes/administración & dosificación , Artritis Experimental/tratamiento farmacológico , Extractos Vegetales/administración & dosificación , Rosmarinus/química , Animales , Antiinflamatorios/química , Antiinflamatorios/aislamiento & purificación , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Artritis Experimental/genética , Artritis Experimental/inmunología , Artritis Experimental/metabolismo , Humanos , Masculino , Estrés Oxidativo/efectos de los fármacos , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Hojas de la Planta/química , RatasRESUMEN
Pristane-induced arthritis (PIA) in mice is an experimental model that resembles human rheumatoid arthritis, a chronic autoimmune disease that affects joints and is characterized by synovial inflammation and articular cartilage and bone destruction. AIRmax and AIRmin mouse lines differ in their susceptibility to PIA, and linkage analysis in this model mapped arthritis severity QTLs in chromosomes 5 and 8. miRNAs are a class of small RNA molecules that have been extensively studied in the development of arthritis. We analyzed miRNA and gene expression profiles in peritoneal cells of AIRmax and AIRmin lines, in order to evaluate the genetic architecture in this model. Susceptible AIRmax mice showed higher gene (2025 vs 1043) and miRNA (240 vs 59) modulation than resistant AIRmin mice at the onset of disease symptoms. miR-132-3p/212-3p, miR-106-5p, miR-27b-3p, and miR-25-3p were among the miRNAs with the highest expression in susceptible animals, showing a negative correlation with the expression of predicted target genes (Il10, Cd69, and Sp1r1). Our study showed that global gene and miRNA expression profiles in peritoneal cells of susceptible AIRmax and resistant AIRmin lines during pristane-induced arthritis are distinct, evidencing interesting targets for further validation.
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Artritis Experimental/genética , Artritis Reumatoide/genética , MicroARNs/genética , Peritoneo/fisiología , Animales , Artritis Experimental/inducido químicamente , Células Cultivadas , Susceptibilidad a Enfermedades , Femenino , Humanos , Interleucina-10/genética , Masculino , Ratones , Ratones Mutantes , Peritoneo/patología , Sitios de Carácter Cuantitativo/genética , Terpenos/administración & dosificación , TranscriptomaRESUMEN
OBJECTIVE AND DESIGN: Pristane-induced arthritis (PIA) in AIRmax mice homozygous for Slc11a1 R and S alleles was used to characterize the influence of Slc11a1 gene polymorphism on immune responses during disease manifestation. Previous reports demonstrated that the presence of the Slc11a1 S allele increased the incidence and severity of PIA in AIRmax SS , suggesting that this gene could interact with inflammatory loci to modulate PIA. We investigated the effects of Slc11a1 alleles on the activation of phagocytes during PIA. TREATMENT: Mice were injected intraperitoneally with two doses of 0.5 mL of mineral oil pristane at 60-day intervals. Arthritis development was accompanied for 180 days. RESULTS: AIRmax SS mice showed differential peritoneal macrophage gene expression profiles during PIA, with higher expression and production of H2O2, NO, IL-1ß, IL-6, TNF-α, and several chemokines. The presence of the Slc11a1 R allele, on the other hand, diminished the intensity of macrophage activation, restricting arthritis development. CONCLUSION: Our data demonstrated the fine-tuning roles of Slc11a1 alleles modulating macrophage activation, and consequent PIA susceptibility, in those mouse lines.
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Artritis Experimental/genética , Artritis Experimental/inmunología , Artritis Reumatoide/genética , Artritis Reumatoide/inmunología , Proteínas de Transporte de Catión/genética , Proteínas de Transporte de Catión/inmunología , Macrófagos Peritoneales/inmunología , Animales , Artritis Experimental/patología , Artritis Reumatoide/patología , Citocinas/sangre , Citocinas/inmunología , Femenino , Peróxido de Hidrógeno/inmunología , Articulaciones/patología , Masculino , Ratones , Óxido Nítrico/inmunología , Terpenos , TranscriptomaRESUMEN
BACKGROUND: Increasing evidences indicate that an unbalance between tryptases and their endogenous inhibitors, leading to an increased proteolytic activity, is implicated in the pathophysiology of rheumatoid arthritis. The aim of the present study was to evaluate the impact of tryptase inhibition on experimental arthritis. METHODS: Analysis of gene expression and regulation in the mouse knee joint was performed by RT-qPCR and in situ hybridization. Arthritis was induced in male C57BL/6 mice with mBSA/IL-1ß. Tryptase was inhibited by two approaches: a lentivirus-mediated heterologous expression of the human endogenous tryptase inhibitor, sperm-associated antigen 11B isoform C (hSPAG11B/C), or a chronic treatment with the synthetic tryptase inhibitor APC366. Several inflammatory parameters were evaluated, such as oedema formation, histopathology, production of IL-1ß, -6, -17A and CXCL1/KC, myeloperoxidase and tryptase-like activities. RESULTS: Spag11c was constitutively expressed in chondrocytes and cells from the synovial membrane in mice, but its expression did not change 7 days after the induction of arthritis, while tryptase expression and activity were upregulated. The intra-articular transduction of animals with the lentivirus phSPAG11B/C or the treatment with APC366 inhibited the increase of tryptase-like activity, the late phase of oedema formation, the production of IL-6 and CXCL1/KC. In contrast, neutrophil infiltration, degeneration of hyaline cartilage and erosion of subchondral bone were not affected. CONCLUSIONS: Tryptase inhibition was effective in inhibiting some inflammatory parameters associated to mBSA/IL-1ß-induced arthritis, notably late phase oedema formation and IL-6 production, but not neutrophil infiltration and joint degeneration. These results suggest that the therapeutic application of tryptase inhibitors to rheumatoid arthritis would be restrained to palliative care, but not as disease-modifying drugs. Finally, this study highlighted lentivirus-based gene delivery as an instrumental tool to study the relevance of target genes in synovial joint physiology and disease.
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Técnicas de Transferencia de Gen , Inflamación/metabolismo , Articulación de la Rodilla/metabolismo , Triptasas/metabolismo , Animales , Antígenos de Superficie/genética , Antígenos de Superficie/metabolismo , Artritis Experimental/genética , Artritis Experimental/metabolismo , Artritis Experimental/terapia , Artritis Reumatoide/genética , Artritis Reumatoide/metabolismo , Artritis Reumatoide/terapia , Condrocitos/metabolismo , Citocinas/metabolismo , Dipéptidos/farmacología , Células HEK293 , Humanos , Inflamación/genética , Inflamación/terapia , Articulación de la Rodilla/efectos de los fármacos , Articulación de la Rodilla/patología , Lentivirus/genética , Masculino , Ratones Endogámicos C57BL , Membrana Sinovial/metabolismo , Triptasas/antagonistas & inhibidores , Triptasas/genéticaRESUMEN
Anti-inflammatory property of low-level laser therapy (LLLT) has been widely described in literature, although action mechanisms are not always clarified. Thus, this study aimed to evaluate apoptosis mechanisms in the LLLT anti-inflammatory effects on the arthritis experimental model in vivo at two different energy densities (3 and 30 Jcm-2). Arthritis was induced in mice by zymosan solution, animals were distributed into five groups, and morphological analysis, immunocytochemistry and gene expressions for apoptotic proteins were performed. Data showed an anti-inflammatory effect, DNA fragmentation in polymorphonuclear (PMN) cells and alteration in gene expression of proteins related to apoptosis pathways after LLLT. p53 gene expression increased at both energy densities, Bcl2 gene expression increased at 3 Jcm-2, and Bcl2 tissue expression decreased at 30 Jcm-2. In addition, apoptosis was restricted to PMN cells. Results suggest that apoptosis in PMN cells comprise part of LLLT anti-inflammatory mechanisms by disbalance promotion between expression of pro-apoptotic (Bax and p53) and anti-apoptotic (Bcl-2) proteins, with pro-apoptotic gene expression selectively in PMN cells.
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Apoptosis/efectos de la radiación , Inflamación/patología , Articulaciones/patología , Terapia por Luz de Baja Intensidad , Neutrófilos/patología , Neutrófilos/efectos de la radiación , Enfermedad Aguda , Animales , Apoptosis/efectos de los fármacos , Apoptosis/genética , Artritis Experimental/genética , Artritis Experimental/patología , Artritis Experimental/radioterapia , Fragmentación del ADN/efectos de la radiación , Regulación de la Expresión Génica/efectos de la radiación , Inflamación/genética , Masculino , Ratones Endogámicos C57BL , Neutrófilos/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , ZimosanRESUMEN
INTRODUCTION: Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by neutrophil articular infiltration, joint pain and the progressive destruction of cartilage and bone. IL-22 is a key effector molecule that plays a critical role in autoimmune diseases. However, the function of IL-22 in the pathogenesis of RA remains controversial. In this study, we investigated the role of IL-22 in the early phase of antigen-induced arthritis (AIA) in mice. METHODS: AIA was induced in C57BL/6, IL-22(-/-), ASC(-/-) and IL-1R1(-/-) immunized mice challenged intra-articularly with methylated bovine serum albumin (mBSA). Expression of IL-22 in synovial membranes was determined by RT-PCR. Articular hypernociception was evaluated using an electronic von Frey. Neutrophil recruitment and histopathological analyses were assessed in inflamed knee joint. Joint levels of inflammatory mediators and mBSA-specific IgG concentration in the serum were measured by ELISA. RESULTS: The IL-22 mRNA expression and protein levels in synovial tissue were increased during the onset of AIA. In addition, pharmacological inhibition (anti-IL-22 antibody) and genetic deficiency (IL-22(-/-) mice) reduced articular pain and neutrophil migration in arthritic mice. Consistent with these findings, recombinant IL-22 joint administration promoted articular inflammation per se in WT mice, restoring joint nociception and neutrophil infiltration in IL-22(-/-) mice. Moreover, IL-22-deficient mice showed reduced synovitis (inflammatory cell influx) and lower joint IL-1ß levels, whereas the production of IL-17, MCP-1/CCL2, and KC/CXCL1 and the humoral immune response were similar, compared with WT mice. Corroborating these results, the exogenous administration of IL-22 into the joints induced IL-1ß production in WT mice and reestablished IL-1ß production in IL-22(-/-) mice challenged with mBSA. Additionally, IL-1R1(-/-) mice showed attenuated inflammatory features induced by mBSA or IL-22 challenge. Articular nociception and neutrophil migration induced by IL-22 were also reduced in ASC(-/-) mice. CONCLUSIONS: These results suggest that IL-22 plays a pro-inflammatory/pathogenic role in the onset of AIA through an ASC-dependent stimulation of IL-1ß production.
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Artritis Experimental/inmunología , Interleucina-1beta/inmunología , Interleucinas/inmunología , Articulación de la Rodilla/inmunología , Animales , Antígenos/inmunología , Proteínas Reguladoras de la Apoptosis/genética , Proteínas Reguladoras de la Apoptosis/inmunología , Proteínas Reguladoras de la Apoptosis/metabolismo , Artralgia/genética , Artralgia/inmunología , Artralgia/metabolismo , Artritis Experimental/genética , Artritis Experimental/metabolismo , Proteínas Adaptadoras de Señalización CARD , Movimiento Celular/inmunología , Ensayo de Inmunoadsorción Enzimática , Expresión Génica/inmunología , Interleucina-1beta/metabolismo , Interleucinas/biosíntesis , Interleucinas/genética , Articulación de la Rodilla/metabolismo , Ratones Endogámicos C57BL , Ratones Noqueados , Neutrófilos/citología , Neutrófilos/inmunología , Neutrófilos/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Membrana Sinovial/inmunología , Membrana Sinovial/metabolismo , Sinovitis/genética , Sinovitis/inmunología , Sinovitis/metabolismo , Zimosan/inmunología , Interleucina-22RESUMEN
BACKGROUND AND OBJECTIVE: To investigate the influence of intraoperative and preoperative positive pressure in the time of extubation in patients undergoing bariatric surgery. METHOD: Randomized clinical trial, in which 40 individuals with a body mass index between 40 and 55 kg/m2, age between 25 and 55 years, nonsmokers, underwent bariatric surgery type Roux-en-Y gastric bypass by laparotomy and with normal preoperative pulmonary function were randomized into the following groups: G-pre (n = 10): individuals who received treatment with noninvasive positive pressure before surgery for 1 h; G-intra (n = 10): individuals who received positive end-expiratory pressure of 10 cm H2O throughout the surgical procedure; and G-control (n = 20): not received any preoperative or intraoperative intervention. Following were recorded: time between induction of anesthesia and extubation, between the end of anesthesia and extubation, duration of mechanical ventilation, and time between extubation and discharge from the post-anesthetic recovery. RESULTS: There was no statistical difference between groups. However, when applied to the Cohen coefficient, the use of positive end-expiratory pressure of 10 cm H2O during surgery showed a large effect on the time between the end of anesthesia and extubation. About this same time, the treatment performed preoperatively showed moderate effect. CONCLUSION: The use of positive end-expiratory pressure of 10 cm H2O in the intraoperative and positive pressure preoperatively, influenced the time of extubation of patients undergoing bariatric surgery. .
JUSTIFICATIVA E OBJETIVO: investigar a influência do uso da pressão positiva nas vias aéreas intraoperatória e pré-operatória no tempo de extubação de pacientes submetidos à cirurgia bariátrica. MÉTODO: Trata-se de ensaio clínico randomizado, no qual 40 indivíduos com índice de massa corporal entre 40 e 55 kg/m2, idade entre 25 e 55 anos, não tabagistas, submetidos à cirurgia bariátrica do tipo derivação gástrica em Y de Roux por laparotomia e com prova de função pulmonar pré-operatória dentro da normalidade foram randomizados nos seguintes grupos: G-pré (n = 10): indivíduos que receberam tratamento com pressão positiva não invasiva antes da cirurgia, durante uma hora, G-intra (n = 10): indivíduos que receberam Positive End-expiratory Pressure de 10 cm H2O durante todo o procedimento cirúrgico e G-controle (n = 20): não receberam qualquer tipo de intervenção pré ou intraoperatória. foram anotados os seguintes tempos: tempo decorrido entre a indução anestésica e a extubação, entre o término da anestesia e extubação, tempo de ventilação mecânica, e tempo entre a extubação e a alta da Recuperação Pós-Anestésica. RESULTADOS: Não houve diferença estatística entre os grupos, porém quando aplicado ao Coeficiente de Cohen, o uso da Positive End-expiratory Pressure de 10 cm H2O no intraoperatório mostrou um efeito grande sobre o tempo entre o término da anestesia e a extubação. Sobre este mesmo tempo, o tratamento realizado no pré-operatório apresentou efeito moderado. CONCLUSÃO: O uso da Positive End-expiratory Pressure de 10 cm H2O no intraoperatório e da pressão positiva no pré-operatório, pode influenciar o tempo de extubação de pacientes submetidos à cirurgia bariátrica. .
JUSTIFICACIÓN Y OBJETIVO: Investigar la influencia del uso de la presión positiva en las vías aéreas intraoperatoria y preoperatoria en el tiempo de extubación de pacientes sometidos a la cirugía bariátrica. MÉTODO: Se trata de un ensayo clínico aleatorizado, en el cual 40 individuos con IMC entre 40 y 55 kg/m2, edad entre 25 y 55 años, no fumadores, sometidos a cirugía bariátrica del tipo derivación gástrica en Y de Roux por laparotomía y con prueba de función pulmonar preoperatoria dentro de la normalidad fueron aleatorizados en los siguientes grupos: G-pre (n = 10): individuos que recibieron tratamiento con presión positiva no invasiva antes de la cirugía durante una hora; G-intra (n = 10): individuos que recibieron PEEP de 10 cm H2O durante todo el procedimiento quirúrgico y G-control (n = 20): no recibieron ningún tipo de intervención pre- o intraoperatoria. Fueron anotados los siguientes tiempos: tiempo trascurrido entre la inducción anestésica y la extubación, entre el fin de la anestesia y la extubación, tiempo de ventilación mecánica, y tiempo entre la extubación y el alta de la sala de recuperación postanestésica. RESULTADOS: No hubo diferencia estadística entre los grupos, sin embargo cuando se aplicó el coeficiente de Cohen, el uso de la PEEP de 10 cm H2O en el intraoperatorio mostró un efecto importante sobre el tiempo entre el término de la anestesia y la extubación. Sobre ese mismo tiempo, el tratamiento realizado en el preoperatorio presentó un efecto moderado. CONCLUSIÓN: El uso de la PEEP de 10 cm H2O en el intraoperatorio y de la presión positiva en el preoperatorio puede influir en el tiempo de extubación de pacientes sometidos a cirugía bariátrica. .
Asunto(s)
Animales , Femenino , Humanos , Masculino , Ratones , Artritis Experimental/inmunología , Subgrupos de Linfocitos B/inmunología , Proteína del Síndrome de Wiskott-Aldrich/inmunología , Artritis Experimental/genética , Artritis Experimental/patología , Enfermedades Autoinmunes/genética , Enfermedades Autoinmunes/inmunología , Enfermedades Autoinmunes/patología , Subgrupos de Linfocitos B/patología , /genética , /inmunología , Ratones Noqueados , Linfocitos T Reguladores/inmunología , Linfocitos T Reguladores/patología , /inmunología , /patología , Proteína del Síndrome de Wiskott-Aldrich/genéticaRESUMEN
BACKGROUND: Due to recent studies indicating that the deregulation of microRNAs (miRNAs) in T cells contributes to increased severity of rheumatoid arthritis, we hypothesized that deregulated miRNAs may interact with key mRNA targets controlling the function or differentiation of these cells in this disease. METHODOLOGY/PRINCIPAL FINDINGS: To test our hypothesis, we used microarrays to survey, for the first time, the expression of all known mouse miRNAs in parallel with genome-wide mRNAs in thymocytes and naïve and activated peripheral CD3(+) T cells from two mouse strains the DBA-1/J strain (MHC-H2q), which is susceptible to collagen induced arthritis (CIA), and the DBA-2/J strain (MHC-H2d), which is resistant. Hierarchical clustering of data showed the several T cell miRNAs and mRNAs differentially expressed between the mouse strains in different stages of immunization with collagen. Bayesian statistics using the GenMir(++) algorithm allowed reconstruction of post-transcriptional miRNA-mRNA interaction networks for target prediction. We revealed the participation of miR-500, miR-202-3p and miR-30b*, which established interactions with at least one of the following mRNAs: Rorc, Fas, Fasl, Il-10 and Foxo3. Among the interactions that were validated by calculating the minimal free-energy of base pairing between the miRNA and the 3'UTR of the mRNA target and luciferase assay, we highlight the interaction of miR-30b*-Rorc mRNA because the mRNA encodes a protein implicated in pro-inflammatory Th17 cell differentiation (Rorγt). FACS analysis revealed that Rorγt protein levels and Th17 cell counts were comparatively reduced in the DBA-2/J strain. CONCLUSIONS/SIGNIFICANCE: This result showed that the miRNAs and mRNAs identified in this study represent new candidates regulating T cell function and controlling susceptibility and resistance to CIA.
Asunto(s)
Artritis Experimental/genética , Colágeno/administración & dosificación , Susceptibilidad a Enfermedades , MicroARNs/metabolismo , Procesamiento Postranscripcional del ARN , ARN Mensajero/metabolismo , Linfocitos T/metabolismo , Algoritmos , Animales , Artritis Experimental/inmunología , Citometría de Flujo , Perfilación de la Expresión Génica , Masculino , Ratones , Ratones Endogámicos DBA , Análisis de Secuencia por Matrices de Oligonucleótidos , Linfocitos T/inmunologíaRESUMEN
Propolis is a resinous substance collected by honeybees from leaf buds and cracks in the bark of various plants. Propolis has been reported to have immunomodulatory activity. We hypothesized that propolis would be able to reduce the disease severity of rheumatoid arthritis. We evaluated the effect of Brazilian propolis ethanolic extract on the pathogenesis of collagen-induced arthritis (CIA) in mice. Mice fed propolis exhibited significant lower clinical arthritis scores than those fed the control diet. To investigate the mechanism of the effect of propolis on CIA mice, we examined interleukin-17 (IL-17) production in CIA mice fed propolis using an enzyme-linked immunospot assay and flow cytometric analysis. The numbers of IL-17-producing cells in the CIA mice fed propolis were significantly decreased. To determine direct influence of propolis on cytokine production, splenocytes were stimulated with phorbol myristate acetate in the presence of propolis extract in vitro. Concentration-dependent declines in IL-17 expression were observed by ELISA and real-time PCR methods. We further found that propolis significantly inhibited the differentiation of Th17 cells from murine splenocytes in a concentration-dependent manner. Taken together, our results may provide a new light on the potential mechanism of the immunosuppressive and anti-inflammatory effects of propolis.
Asunto(s)
Artritis Experimental/tratamiento farmacológico , Artritis Experimental/inmunología , Interleucina-17/antagonistas & inhibidores , Interleucina-17/biosíntesis , Própolis/farmacología , Animales , Antiinflamatorios/farmacología , Artritis Experimental/genética , Artritis Experimental/metabolismo , Brasil , Diferenciación Celular/efectos de los fármacos , Femenino , Inmunosupresores/farmacología , Interleucina-17/genética , Interleucina-17/inmunología , Ratones , Ratones Endogámicos DBA , ARN Mensajero/genética , ARN Mensajero/metabolismo , Bazo/inmunología , Bazo/metabolismo , Células Th17/efectos de los fármacos , Células Th17/inmunología , Células Th17/metabolismoRESUMEN
Arthritis of the knee is the most common type of joint inflammatory disorder and it is associated with pain and inflammation of the joint capsule. Few studies address the effects of the 810-nm laser in such conditions. Here we investigated the effects of low-level laser therapy (LLLT; infrared, 810-nm) in experimentally induced rat knee inflammation. Thirty male Wistar rats (230-250 g) were anesthetized and injected with carrageenan by an intra-articular route. After 6 and 12 h, all animals were killed by CO(2) inhalation and the articular cavity was washed for cellular and biochemical analysis. Articular tissue was carefully removed for real-time PCR analysis in order to evaluate COX-1 and COX-2 expression. LLLT was able to significantly inhibit the total number of leukocytes, as well as the myeloperoxidase activity with 1, 3, and 6 J (Joules) of energy. This result was corroborated by cell counting showing the reduction of polymorphonuclear cells at the inflammatory site. Vascular extravasation was significantly inhibited at the higher dose of energy of 10 J. Both COX-1 and 2 gene expression were significantly enhanced by laser irradiation while PGE(2) production was inhibited. Low-level laser therapy operating at 810 nm markedly reduced inflammatory signs of inflammation but increased COX-1 and 2 gene expression. Further studies are necessary to investigate the possible production of antiinflammatory mediators by COX enzymes induced by laser irradiation in knee inflammation.
Asunto(s)
Artritis Experimental/terapia , Terapia por Luz de Baja Intensidad/métodos , Animales , Artritis Experimental/genética , Artritis Experimental/metabolismo , Carragenina/toxicidad , Ciclooxigenasa 1/genética , Ciclooxigenasa 2/genética , Dinoprostona/metabolismo , Azul de Evans , Extravasación de Materiales Terapéuticos y Diagnósticos , Regulación de la Expresión Génica , Interleucina-1/metabolismo , Interleucina-6/metabolismo , Articulación de la Rodilla/metabolismo , Articulación de la Rodilla/efectos de la radiación , Recuento de Leucocitos , Masculino , Proteínas de la Membrana/genética , Peroxidasa/metabolismo , Ratas , Ratas WistarRESUMEN
Considering that imbalance of central tolerance in the thymus contributes to aggressive autoimmunity, we compared the expression of peripheral tissue autoantigens (PTA) genes, which are involved in self-representation in the thymic stroma, of two mouse strains; DBA-1/J (MHC-H2(q)) susceptible and DBA-2/J (MHC-H2(d)) resistant to collagen induced arthritis (CIA). We evaluate whether these strains differ in their thymic gene expression, allowing identification of genes that might play a role in susceptibility/resistance to CIA. Microarray profiling showed that 1093 PTA genes were differentially modulated between collagen immunized DBA-1/J and DBA-2/J mice. These genes were assigned to 17 different tissues/organs, including joints/bone, characterizing the promiscuous gene expression (PGE), which is implicated in self-representation. Hierarchical clustering of microarray data and quantitative RT-PCR analysis showed that Aire (autoimmune regulator), an important regulator of the PGE process, Aire-dependent (insulin), Aire-independent (Col2A1 and Gad67), and other 22 joint/bone autoantigen genes were down-regulated in DBA-1/J compared with DBA-2/J in the thymus. Considering the importance of MHC-H2 in peptide-self presentation and autoimmunity susceptibility, we reconstructed transcriptional networks of both strains based on actual microarray data. The networks clearly demonstrated different MHC-H2 transcriptional interactions with PTAs genes. DBA-1/J strain featured MHC-H2 as a node influencing downstream genes. Differently, in DBA-2/J strain network MHC-H2 was exclusively self-regulated and does not control other genes. These findings provide evidence that CIA susceptibility in mice may be a reflex of a cascade-like transcriptional control connecting different genes to MHC-H2 in the thymus.
Asunto(s)
Artritis Experimental/genética , Artritis Experimental/inmunología , Autoantígenos/metabolismo , Redes Reguladoras de Genes , Timo/metabolismo , Animales , Artritis Experimental/metabolismo , Autoantígenos/genética , Autoantígenos/inmunología , Células Cultivadas , Biología Computacional , Perfilación de la Expresión Génica , Redes Reguladoras de Genes/inmunología , Antígenos de Histocompatibilidad/metabolismo , Ratones , Ratones Endogámicos DBA , Análisis por Micromatrices , Autotolerancia , Especificidad de la Especie , Timo/inmunología , Timo/patología , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Proteína AIRERESUMEN
Mice obtained by bidirectional selective breeding for high (HIII) or low (LIII) antibody (Ab) production are resistant or extremely susceptible to pristane-induced arthritis (PIA), respectively. Several quantitative trait loci regulating Ab production (Ab QTL) have been mapped in these lines, which were used to investigate the influence of these Ab QTL in PIA. Parental HIII and LIII mice and their F1 and F2 intercrosses were injected twice with pristane, and arthritis was observed for 200 days. In LIII mice PIA was more severe and incidence was 100% at day 105, while F1 and F2 mice showed intermediate values. HIII mice were totally resistant. Microsatellite polymorphisms of Ab QTL were analysed and D3Mit100 alleles cosegregated significantly with PIA incidence, severity and onset in F2 intercross mice, while the other four markers showed suggestive values. Results indicate colocalization of QTL for Ab production and PIA susceptibility. Moreover, the different cytokine and IgG isotype profiles observed in HIII and LIII lines after PIA induction are useful to candidate genes endowed with the regulation of the Ab production and arthritis phenotypes.
Asunto(s)
Artritis Experimental/genética , Artritis Experimental/inmunología , Autoanticuerpos/biosíntesis , Predisposición Genética a la Enfermedad , Sitios de Carácter Cuantitativo , Animales , Artritis Experimental/inducido químicamente , Cruzamientos Genéticos , Modelos Animales de Enfermedad , Femenino , Masculino , Ratones , Ratones Endogámicos , Repeticiones de Microsatélite , Terpenos/toxicidadRESUMEN
As part of a program researching the synthesis and immunopharmacological evaluation of novel synthetic compounds, we have described the immune modulatory profile of the new achiral thalidomide analogue LASSBio-468 in the present work. This compound was planned as an N-substituted phthalimide derivate, structurally designed as a hybrid of thalidomide and aryl sulfonamides, which were previously described as tumor necrosis factor-alpha (TNF-alpha) and PDE4 inhibitors. LASSBio-468 was recently demonstrated to inhibit the TNF-alpha production induced by lipopolysaccharide (LPS), in vivo. Here, we investigated whether this compound would affect chronic inflammation processes associated with the production of this pro-inflammatory cytokine. Treatment with LASSBio-468 before a lethal dose injection of LPS in animals greatly inhibited endotoxic shock. This effect seems to be mediated by a specific down regulation of TNF-alpha and nitric oxide production, regulated mainly at the RNA level. In another model, histopathological analysis indicated that this compound also inhibited adjuvant-induced arthritis in rats. Taken together, our data demonstrated a potent anti-inflammatory effect of LASSBio-468, suggesting its use as a potential drug against chronic inflammatory diseases.
Asunto(s)
Artritis Experimental/tratamiento farmacológico , Indoles/farmacología , Óxido Nítrico/sangre , Choque Séptico/tratamiento farmacológico , Talidomida/análogos & derivados , Factor de Necrosis Tumoral alfa/metabolismo , Animales , Antiinflamatorios/farmacología , Artritis Experimental/genética , Artritis Experimental/patología , Expresión Génica/efectos de los fármacos , Isoindoles , Lipopolisacáridos/farmacología , Activación de Linfocitos/efectos de los fármacos , Macrófagos Peritoneales/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos BALB C , Óxido Nítrico/metabolismo , Pentoxifilina/farmacología , Ratas , Ratas Wistar , Talidomida/farmacología , Factor de Necrosis Tumoral alfa/genética , Vasculitis del Sistema Nervioso Central/tratamiento farmacológico , Vasculitis del Sistema Nervioso Central/patologíaRESUMEN
Galectin-1 (GAL-1), a member of a family of conserved beta-galactoside-binding proteins, has been shown to induce in vitro apoptosis of activated T cells and immature thymocytes. We assessed the therapeutic effects and mechanisms of action of delivery of GAL-1 in a collagen-induced arthritis model. A single injection of syngeneic DBA/1 fibroblasts engineered to secrete GAL-1 at the day of disease onset was able to abrogate clinical and histopathological manifestations of arthritis. This effect was reproduced by daily administration of recombinant GAL-1. GAL-1 treatment resulted in reduction in anticollagen immunoglobulin (Ig)G levels. The cytokine profile in draining lymph node cells and the anticollagen IgG isotypes in mice sera at the end of the treatment clearly showed inhibition of the proinflammatory response and skewing towards a type 2-polarized immune reaction. Lymph node cells from mice engaged in the gene therapy protocol increased their susceptibility to antigen-induced apoptosis. Moreover, GAL-1-expressing fibroblasts and recombinant GAL-1 revealed a specific dose-dependent inhibitory effect in vitro in antigen-dependent interleukin 2 production to an A(q)-restricted, collagen type 2-specific T cell hybridoma clone. Thus, a correlation between the apoptotic properties of GAL-1 in vitro and its immunomodulatory properties in vivo supports its therapeutic potential in the treatment of T helper cell type 1-mediated autoimmune disorders.