RESUMO
OBJECTIVE: Recently, members of the semaphorin family have received major attention in various medical fields, especially autoimmunity. In this study, we selected semaphorin-3A (Sema3A), semaphorin-7A (Sema7A), and their receptors to determine the possible relationship between these molecules and multiple sclerosis (MS). METHOD: We measured the gene expression of Sema3A, Sema7A, neuropilin-1 (NP-1), plexin-C1, and ß1 integrin in the blood samples of relapsing-remitting multiple sclerosis (RRMS) patients, treated with high-dose interferon-ß1a (IFN-ß1a), low-dose IFN-ß1a, IFN-ß1b, and glatiramer acetate (GA) via quantitative real-time polymerase chain reaction (qRT-PCR) assay, and then, compared the results of treatment-naive patients with the healthy controls. RESULTS: The gene expression of Sema3A (P = 0.02), NP-1 (P < 0.001), and plexin-C1 (P < 0.01) significantly decreased in the treatment-naive group, compared to the healthy controls. Sema3A significantly increased in all treated patients, compared to the treatment-naive patients (P < 0.001). However, expression of NP-1 (P < 0.001), plexin-C1 (P < 0.001), and ß1 integrin (P < 0.05) only increased in patients receiving high-dose IFN-ß1a, IFN-ß1b, and GA. Expression of Sema7A increased in only two groups of patients treated with IFN-ß1b (P < 0.001) and GA (P = 0.018), without any significant decrease in the treatment-naive group, compared to the healthy controls (P > 0.05). CONCLUSION: Our findings confirm that the presence of Sema3A, Sema7A, and their receptors can play critical roles in the treatment of MS patients. Therefore, they can be potential target molecules for MS treatment in the future.
Assuntos
Antígenos CD/genética , Expressão Gênica , Integrina beta1/genética , Esclerose Múltipla Recidivante-Remitente/genética , Neuropilina-1/genética , Semaforina-3A/genética , Semaforinas/genética , Adulto , Antígenos CD/sangue , Feminino , Proteínas Ligadas por GPI/sangue , Proteínas Ligadas por GPI/genética , Acetato de Glatiramer/uso terapêutico , Humanos , Integrina beta1/sangue , Interferon beta/uso terapêutico , Masculino , Esclerose Múltipla Recidivante-Remitente/sangue , Esclerose Múltipla Recidivante-Remitente/tratamento farmacológico , Neuropilina-1/sangue , Semaforina-3A/sangue , Semaforinas/sangueRESUMO
Both major subcategories of inflammatory bowel disease (IBD), ulcerative colitis and Crohn's disease are characterized by infiltration of the gut wall by inflammatory effector cells and elevated biomarkers of inflammation in blood and feces. We investigated the phenotypes of circulating lymphocytes in the two types of IBD in treatment-naive pediatric patients by analysis of blood samples by flow cytometry. Multivariate analysis was used to compare the phenotypes of the blood lymphocytes of children with ulcerative colitis (n = 17) or Crohn's disease (n = 8) and non-IBD control children with gastrointestinal symptoms, but no signs of gut inflammation (n = 23). The two IBD subcategories could be distinguished based on the results from the flow cytometry panel. Ulcerative colitis was characterized by activated T cells, primarily in the CD8+ population, as judged by increased expression of human leukocyte antigen D-related (HLA-DR) and the ß1-integrins [very late antigen (VLA)] and a reduced proportion of naive (CD62L+ ) T cells, compared with the non-IBD controls. This T cell activation correlated positively with fecal and blood biomarkers of inflammation. In contrast, the patients with Crohn's disease were characterized by a reduced proportion of B cells of the memory CD27+ phenotype compared to the non-IBD controls. Both the patients with ulcerative colitis and those with Crohn's disease showed increased percentages of CD23+ B cells, which we demonstrate here as being naive B cells. The results support the notion that the two major forms of IBD may partially have different pathogenic mechanisms.
Assuntos
Subpopulações de Linfócitos B/imunologia , Colite Ulcerativa/imunologia , Doença de Crohn/imunologia , Subpopulações de Linfócitos T/imunologia , Adolescente , Estudos de Casos e Controles , Criança , Pré-Escolar , Colite Ulcerativa/sangue , Doença de Crohn/sangue , Feminino , Humanos , Memória Imunológica , Mediadores da Inflamação/sangue , Integrina beta1/sangue , Ativação Linfocitária , Masculino , Modelos Imunológicos , Fenótipo , Receptores de IgE/sangue , Membro 7 da Superfamília de Receptores de Fatores de Necrose Tumoral/sangueRESUMO
Schizophrenia is one of the most severe chronic psychiatric disorders, which lacks of objective and effective diagnosis and observation indicators.In this work, the serum miRNA profiles of schizophrenic patients were analyzed. Targets of abnormal miRNAs, and their regulatory mechanisms were studied. A miRNA array was used to analyze the serum from 3 schizophrenic patients without treatment, 3 clinically cured patients and 3 healthy controls. The findings from the array were confirmed by real-time PCR in a larger cohort, including 59 patients and 60 healthy controls. The candidate miRNAs were analyzed using bioinformatics tools. Their potential targets were studied through in vitro cellular experiments.MiR-320a-3p and miR-320b were found to be down-regulated in patients compared with cured patients and controls in the miRNA array, which was also confirmed by real-time PCR in the larger cohort. Integrin ß1 (ITG ß1) was found to be one of the targets of miR-320a-3p. An enzyme-linked immune sorbent assay demonstrated that the ITG ß1 concentration increased significantly in the patients' serum, and the in vitro study confirmed that miR-320a-3p targeted the 3' UTR of ITG ß1 mRNA and reduced its expression.Our results demonstrated that the regulatory effect of miR-320a-3p on its target ITG ß1 might play an important role in schizophrenia pathogenesis, which could be a potential pathway for schizophrenia diagnosis and therapy.
Assuntos
Integrina beta1/genética , MicroRNAs/genética , Esquizofrenia/diagnóstico , Esquizofrenia/genética , Regiões 3' não Traduzidas , Adulto , Regulação para Baixo , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Integrina beta1/sangue , Masculino , MicroRNAs/sangue , Pessoa de Meia-Idade , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase em Tempo Real , Esquizofrenia/sangue , Esquizofrenia/terapia , Transdução de Sinais , Adulto JovemRESUMO
Extracellular vesicles (EVs) in the plasma mediate important intercellular communications in the pathogenesis of cancer and inflammatory diseases. EVs express integrins that regulate target specificities and programmed cell death ligand 1 and 2 (PD-L1 and 2) that suppress lymphocyte activation. However, the roles of these molecules on EVs in systemic inflammatory response syndrome (SIRS) and sepsis remain little understood. This study aimed to investigate how the EV expression of integrins and PD-1 ligands might differ in SIRS and sepsis, compared with healthy controls, and to correlate their expression with the clinical parameters reflecting pathogenesis. Twenty-seven SIRS patients without sepsis, 27 sepsis patients, and 18 healthy volunteers were included. EVs were isolated from plasma samples. The expression of three major integrins (ß1, ß2, ß3 integrins) and PD-L1 and 2 were measured. The EV expression of ß2 integrin and PD-L2 was significantly increased in sepsis patients compared with healthy controls. EV expression of PD-L1 was not elevated in sepsis and SIRS; however, circulating soluble PD-L1 levels were significantly higher in sepsis. Furthermore, EV expression of ß2 integrin in sepsis patients correlated with hypotension and reduced kidney function. In addition, soluble PD-L1 levels correlated with sepsis severity, impaired kidney function, and impaired central nervous system function. These results suggest the potential involvements of the EV ß2 integrin, as well as EV PD-L2 and soluble PD-L1, in the septic pathogenesis that occurs with the systemic immune activation leading to multiple organ dysfunctions.
Assuntos
Antígeno B7-H1/sangue , Antígenos CD18/sangue , Integrina beta1/sangue , Integrina beta3/sangue , Proteína 2 Ligante de Morte Celular Programada 1/sangue , Sepse/sangue , Síndrome de Resposta Inflamatória Sistêmica/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Voluntários Saudáveis , Humanos , Masculino , Pessoa de Meia-Idade , Sepse/imunologia , Sepse/patologia , Síndrome de Resposta Inflamatória Sistêmica/imunologia , Síndrome de Resposta Inflamatória Sistêmica/patologiaRESUMO
OBJECTIVE: Cardiovascular diseases and depression are the leading causes of disability in Western countries. Clinical data on potential cardiovascular effects of serotonin reuptake inhibitors (SSRIs), the most commonly used antidepressant drugs, are controversial. In addition to blocking serotonin reuptake transporter in the brain, SSRIs deplete the major peripheral serotonin (5-hydroxytryptamine [5-HT]) storage by inhibiting serotonin reuptake transporter-mediated uptake in platelets. In this study, we aimed to investigate the effect of chronic SSRI intake on the development of atherosclerosis. APPROACH AND RESULTS: Treatment of apolipoprotein E-deficient mice with the SSRI fluoxetine for 2, 4, or 16 weeks increased atherosclerotic lesion formation, with most pronounced effect during early plaque development. Intravital microscopy of carotid arteries revealed enhanced myeloid cell adhesion on fluoxetine treatment. Mechanistically, we found that fluoxetine augmented vascular permeability and increased chemokine-induced integrin-binding activity of circulating leukocytes. In vitro stimulation of murine blood demonstrated that fluoxetine, but not 5-HT, could directly promote ß1 and ß2 integrin activation provided C-C motif chemokine ligand 5 was also present. Similar effects were observed with the SSRI escitalopram. Enhanced C-C motif chemokine ligand 5-induced integrin activation by fluoxetine was also confirmed in a human neutrophil-like cell line. In contrast to the proatherogenic properties of fluoxetine, pharmacological inhibition of the peripheral 5-HT synthesizing enzyme tryptophan hydroxylase 1 did not promote atherosclerosis, suggesting that the proatherogenic effect of fluoxetine occurs independent of peripheral 5-HT depletion. CONCLUSIONS: SSRI intake may promote atherosclerosis and therefore potentially increase the risk for acute cardiovascular events by a mechanism that is independent of 5-HT depletion.
Assuntos
Aorta/efeitos dos fármacos , Doenças da Aorta/induzido quimicamente , Aterosclerose/induzido quimicamente , Artérias Carótidas/efeitos dos fármacos , Doenças das Artérias Carótidas/induzido quimicamente , Fluoxetina/toxicidade , Placa Aterosclerótica , Inibidores Seletivos de Recaptação de Serotonina/toxicidade , Animais , Aorta/metabolismo , Aorta/patologia , Doenças da Aorta/sangue , Doenças da Aorta/genética , Doenças da Aorta/patologia , Aterosclerose/sangue , Aterosclerose/genética , Aterosclerose/patologia , Plaquetas/efeitos dos fármacos , Plaquetas/metabolismo , Antígenos CD18/sangue , Permeabilidade Capilar/efeitos dos fármacos , Artérias Carótidas/metabolismo , Artérias Carótidas/patologia , Doenças das Artérias Carótidas/sangue , Doenças das Artérias Carótidas/genética , Doenças das Artérias Carótidas/patologia , Adesão Celular/efeitos dos fármacos , Quimiocina CCL5/sangue , Modelos Animais de Doenças , Progressão da Doença , Esquema de Medicação , Fluoxetina/administração & dosagem , Células HEK293 , Células HL-60 , Humanos , Integrina beta1/sangue , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout para ApoE , Células Mieloides/efeitos dos fármacos , Células Mieloides/metabolismo , Serotonina/sangue , Inibidores Seletivos de Recaptação de Serotonina/administração & dosagem , Transdução de Sinais , Fatores de TempoRESUMO
BACKGROUND AND AIMS: The risk of developing cardiovascular disease (CVD) is twice as high among smoking individuals compared to non-smokers. Monocytes are involved in smoking-related atherosclerotic plaque formation. In this study, we investigated whether smokers with an increased risk of developing CVD can be identified on the basis of monocyte-derived miRNA expression levels. METHODS: We performed a miRNA microarray experiment on isolated monocytes from smoking, former smoking and non-smoking individuals in a cohort of patients with premature CVD and healthy controls (Cohort I, n = 76). RESULTS: We found miR-124-3p to be heterogeneously expressed among all smoking individuals, whereas expression was low in non-smokers. Subsequently, RT-qPCR measurements on whole blood showed that among smoking individuals an increase in miR-124-3p is associated with an increased risk for advanced atherosclerotic disease (cohort II, n = 24) (OR 11.72 95% CI 1.09-126.53) and subclinical atherosclerosis (coronary artery calcium score ≥ 80th percentile, cohort III n = 138) (OR 2.71, 95% CI 1.05-7.01). This was not observed among former smokers or non-smoking individuals. Flow cytometric analysis demonstrated that high miR-124-3p expression was associated with upregulation of the monocyte surface markers CD45RA, CD29 and CD206, indicating an altered monocyte phenotype. Finally, overexpression of miR-124-3p resulted in an upregulation of CD206 surface expression on monocytes. CONCLUSIONS: High miR-124-3p expression is associated with an increased risk of subclinical atherosclerosis in smoking individuals and with an altered monocyte phenotype. This may suggest that miR-124-3p identifies which smoking individuals are susceptible to the atherogenic effects of smoking.
Assuntos
Aterosclerose/genética , MicroRNAs/genética , Monócitos/metabolismo , Fumar/efeitos adversos , Fumar/genética , Adulto , Aterosclerose/sangue , Aterosclerose/diagnóstico , Estudos de Casos e Controles , Feminino , Citometria de Fluxo , Marcadores Genéticos , Predisposição Genética para Doença , Humanos , Integrina beta1/sangue , Lectinas Tipo C/sangue , Antígenos Comuns de Leucócito/sangue , Modelos Logísticos , Masculino , Receptor de Manose , Lectinas de Ligação a Manose/sangue , MicroRNAs/sangue , Pessoa de Meia-Idade , Razão de Chances , Análise de Sequência com Séries de Oligonucleotídeos , Fenótipo , Receptores de Superfície Celular/sangue , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Risco , Fumar/sangue , Regulação para CimaRESUMO
OBJECTIVE: To explore the effects of acupuncture at Baihui (GV 20) and Zusanli (ST 36) on the peripheral serum expression of microRNA 124 (miRNA 124), laminin and integrin ß1 in rats with cerebral ischemia reperfusion injury (CIRI). METHODS: Seventy-two healthy male Sprague-Dawley rats were randomized into a model group, an acupuncture group, and a sham-operated group using a random digits table, with 24 rats per group. Each group was further randomly divided into 1-, 3-, 5-, and 7-day subgroups based on the reperfusion time according to a random digits table, with 6 rats in each subgroup. In the model and acupuncture groups, CIRI was induced using the thread occlusion method. Electroacupuncture stimulation was applied daily to GV 20 and left ST 36 for 20 min at the indicated time points after successful operations. Serum was sampled for detecting laminin and integrin ß1 protein via enzyme-linked immunosorbent assay, and serum miRNA 124 was examined using quantitative polymerase chain reaction. RESULTS: The serum level of miRNA 124 in the cerebral ischemia rats increased significantly, and the peak expression of miRNA 124 in both the model and acupuncture groups occurred at 3 days. The expression of miRNA 124 in the acupuncture group was higher than in the model group at the same time point (5.96±0.01 vs. 3.11±0.04, P <0.05). Laminin expression in serum from the cerebral ischemia group was higher than that in the sham-operated group. Compared with the model group, the level of laminin in the serum of the acupuncture group was significantly lower at each time point, especially at the 3-day, and 7-day time points (589.12±3.57 vs. 793.05±5.28, and 600.53±3.05 vs. 899.06±5.74, P <0.05). The level of integrin ß1 in the serum from the acupuncture group was lower than that in the model group particularly at the 3-day and 7-day time points (208.66±0.95 vs. 280.83±1.77, and 212.36±0.95 vs. 316.77±2.42, P <0.05). Additionally, the model group and the acupuncture group showed dual peaks of integrin ß1 and laminin expression at 3-day and 7-day. CONCLUSIONS: Acupuncture at GV 20 and ST 36 in rats alleviated CIRI and was associated with upregulated expression of miRNA 124 and with downregulated expression of integrin ß1 and laminin in peripheral serum. These changes may represent one of the mechanisms underlying acupuncture's attenuation of CIRI.
Assuntos
Pontos de Acupuntura , Terapia por Acupuntura/métodos , Isquemia Encefálica/terapia , Integrina beta1/genética , Laminina/sangue , MicroRNAs/sangue , Traumatismo por Reperfusão/terapia , Animais , Isquemia Encefálica/sangue , Isquemia Encefálica/complicações , Isquemia Encefálica/genética , Regulação da Expressão Gênica , Integrina beta1/sangue , Masculino , MicroRNAs/genética , Ratos Sprague-Dawley , Traumatismo por Reperfusão/sangue , Traumatismo por Reperfusão/complicações , Traumatismo por Reperfusão/genéticaRESUMO
BACKGROUND: Multiple myeloma (MM) represents a malignancy of B-cells characterized by proliferation of malignant plasma cells in the bone marrow (BM). Versican (VCAN), an extracellular matrix (ECM) protein, appears to be involved in multiple processes in several cancers. Identifying optimum diagnostic markers and delineating its association with disease severity might be important for controlling MM. METHODS: Expression of VCAN and its associated molecules (ß-catenin, ß1 integrin and FAK) were investigated in 60 subjects to evaluate their usefulness as diagnostic marker. Circulatory and molecular levels of above molecules were analyzed in their BM and Blood using ELISA, Q-PCR and western blotting along with their ROC curve analysis. RESULTS: Circulatory levels of VCAN, ß-catenin and FAK were significantly higher in patients with varying significance in each stage. ß-Catenin and FAK intracellular levels were significantly elevated in patients. mRNA levels of all molecules were significantly higher in BMMNCs while VCAN and ß-catenin also showed increase in PBMCs. Upregulation of these molecules was also observed at protein level. ROC curve analysis for VCAN showed absolute combination of sensitivity and specificity for diagnosis in serum. CONCLUSIONS: Significant elevation of VCAN and its associated molecules imply their role in MM. Optimal sensitivity and specificity of VCAN might utilize its importance as potential marker for active disease.
Assuntos
Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/genética , Mieloma Múltiplo/sangue , Mieloma Múltiplo/diagnóstico , Versicanas/sangue , Versicanas/genética , Adulto , Idoso , Biomarcadores Tumorais/metabolismo , Feminino , Quinase 1 de Adesão Focal/sangue , Quinase 1 de Adesão Focal/genética , Quinase 1 de Adesão Focal/metabolismo , Regulação Neoplásica da Expressão Gênica , Humanos , Integrina beta1/sangue , Integrina beta1/genética , Integrina beta1/metabolismo , Espaço Intracelular/metabolismo , Masculino , Pessoa de Meia-Idade , Mieloma Múltiplo/genética , Mieloma Múltiplo/patologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Curva ROC , Versicanas/metabolismo , beta Catenina/sangue , beta Catenina/genética , beta Catenina/metabolismoRESUMO
AIM: To evaluate the role of baicalin in ulcerative colitis (UC) with regard to the CD4(+)CD29(+) T helper cell, its surface markers and serum inflammatory cytokines. METHODS: Flow cytometry was used to detect the percentage of CD4(+)CD29(+) cells in patients with UC. Real time polymerase chain reaction was used to detect expression of GATA-3, forkhead box P3, T-box expressed in T cells (T-bet), and retinoic acid-related orphan nuclear hormone receptor C (RORC). Western blotting was used to analyze expression of nuclear factor-κB (NF-κB) p65, phosphorylation of NF-κB (p-NF-κB) p65, STAT4, p-STAT4, STAT6 and p-STAT6. The concentrations of interferon-γ (IFN-γ), interleukin (IL)-4, IL-5, IL-6, IL-10 and TGF-ß in serum were determined by ELISA assay. RESULTS: The percentages of CD4(+)CD29(+) T cells were lower in treatment with 40 and 20 µmol/L baicalin than in the treatment of no baicalin. Treatment with 40 or 20 µmol/L baicalin significantly upregulated expression of IL-4, TGF-ß1 and IL-10, increased p-STAT6/STAT6 ratio, but downregulated expression of IFN-γ, IL-5, IL-6, RORC, Foxp3 and T-bet, and decreased ratios of T-bet/GATA-3, p-STAT4/STAT4 and p-NF-κB/NF-κB compared to the treatment of no baicalin. CONCLUSION: The results indicate that baicalin regulates immune balance and relieves the ulcerative colitis-induced inflammation reaction by promoting proliferation of CD4(+)CD29(+) cells and modulating immunosuppressive pathways.
Assuntos
Anti-Inflamatórios/farmacologia , Colite Ulcerativa/imunologia , Flavonoides/farmacologia , Imunossupressores/farmacologia , Integrina beta1/sangue , Ativação Linfocitária/efeitos dos fármacos , Linfócitos T Auxiliares-Indutores/efeitos dos fármacos , Adolescente , Adulto , Biomarcadores/sangue , Estudos de Casos e Controles , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Colite Ulcerativa/sangue , Colite Ulcerativa/genética , Citocinas/sangue , Relação Dose-Resposta a Droga , Feminino , Regulação da Expressão Gênica , Humanos , Mediadores da Inflamação/sangue , Integrina beta1/imunologia , Masculino , Pessoa de Meia-Idade , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Auxiliares-Indutores/metabolismo , Fatores de Tempo , Adulto JovemRESUMO
BACKGROUND: Many previous studies demonstrated that cell adhesion molecules CD44v6 and integrin-ß1 had been extensively investigated as potential prognostic markers of various cancers. However, data in PC are scarce. METHODS: We now investigate CD44v6 and integrin-ß1 mRNA expression in PBMC by a triplex real-time RT-PCR assay and protein expression in plasma by ELISA. All specimens were collected from 54 PC patients who received the treatment of cryosurgery as well as 20 healthy individuals (control). RESULTS: The mRNA and protein expression levels of CD44v6 and integrin-ß1 in patients were significantly increased compared with control group (P<0.05). The high CD44v6 mRNA and protein expression were significantly correlated with clinical stage, tumor differentiation, LNM, liver metastasis and decreased median DFS (P<0.05), while the high integrin-ß1 mRNA and protein expression were significantly correlated with clinical stage, LNM, liver metastasis and decreased median DFS (P<0.05). Clinical stage, LNM, liver metastasis, CD44v6 mRNA and protein expression were the independent predictors of survival in PC patients (P<0.05). Moreover, CD44v6 and integrin-ß1 mRNA and protein expression levels were significantly decreased in patients in 3 months after cryosurgery (P<0.05). No significant difference was found in CD44v6 mRNA and protein expression between patients in 3 months after cryosurgery and control group (P>0.05). CONCLUSION: CD44v6 and integrin-ß1 mRNA and protein expression in blood may serve as biomarkers for the development and metastasis of PC, and as prognostic indicators for PC. They may become useful predictors in assessing outcome of PC patients after cryosurgery. VIRTUAL SLIDES: The virtual slides for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/4035308681009006.
Assuntos
Biomarcadores Tumorais/sangue , Criocirurgia , Receptores de Hialuronatos/sangue , Integrina beta1/sangue , Neoplasias Pancreáticas/imunologia , Neoplasias Pancreáticas/cirurgia , Adulto , Idoso , Biomarcadores Tumorais/genética , Estudos de Casos e Controles , Diferenciação Celular , Intervalo Livre de Doença , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Receptores de Hialuronatos/genética , Integrina beta1/genética , Estimativa de Kaplan-Meier , Neoplasias Hepáticas/imunologia , Neoplasias Hepáticas/secundário , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Multiplex , Estadiamento de Neoplasias , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/mortalidade , Neoplasias Pancreáticas/patologia , RNA Mensageiro/sangue , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Risco , Fatores de Tempo , Resultado do Tratamento , Regulação para CimaRESUMO
BACKGROUND: Integrin-mediated platelet function plays an important role in primary hemostasis. Growth-differentiation factor 15 (GDF-15) has been shown to inhibit ß(2) -integrin activation in leukocytes. METHODS: We investigated the effect of GDF-15 on platelet integrin activation in vitro and in different in vivo models of thrombus formation. RESULTS: GDF-15(-/-) mice showed an accelerated thrombus formation and a reduced survival rate after collagen-induced pulmonary thromboembolism. In reconstitution experiments, recombinant GDF-15 decelerated thrombus formation and prolonged the bleeding time. In vitro experiments demonstrated that GDF-15 pretreated, agonist-stimulated platelets showed decreased binding to fibrinogen in flow chamber assays and reduced activation of ß(1) - and ß(3) -integrins in flow cytometry experiments. Pretreating human and mouse platelets with GDF-15 reduced platelet aggregation. Mechanistically, GDF-15 prevents agonist-induced Rap1- dependent α(II) (b) ß(3) activation by activating PKA. Platelet P-selectin expression and dense granule secretion after stimulation were unaffected by GDF-15, indicating a specific effect of GDF-15 on integrin activation. CONCLUSION: GDF-15 specifically inhibits platelet integrin activation. These findings may have profound clinical implications for the treatment of hemostatic conditions involving platelets.
Assuntos
Plaquetas/metabolismo , Fator 15 de Diferenciação de Crescimento/metabolismo , Hemostasia , Integrinas/sangue , Ativação Plaquetária , Embolia Pulmonar/prevenção & controle , Trombose/prevenção & controle , Animais , Tempo de Sangramento , Plaquetas/efeitos dos fármacos , Cloretos , Colágeno , Proteínas Quinases Dependentes de AMP Cíclico/sangue , Modelos Animais de Doenças , Ativação Enzimática , Compostos Férricos , Citometria de Fluxo , Fator 15 de Diferenciação de Crescimento/deficiência , Fator 15 de Diferenciação de Crescimento/genética , Hemostasia/efeitos dos fármacos , Humanos , Integrina beta1/sangue , Integrina beta3/sangue , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Selectina-P/sangue , Ativação Plaquetária/efeitos dos fármacos , Inibidores da Agregação Plaquetária/farmacologia , Complexo Glicoproteico GPIIb-IIIa de Plaquetas/metabolismo , Embolia Pulmonar/sangue , Embolia Pulmonar/induzido quimicamente , Embolia Pulmonar/genética , Proteínas Recombinantes/metabolismo , Trombose/sangue , Trombose/induzido quimicamente , Trombose/genética , Fatores de Tempo , Proteínas rap1 de Ligação ao GTP/sangueRESUMO
INTRODUCTION: Adhesive molecules, particularly selectins and integrins, are critical for the inflammatory cell trafficking from blood to the lungs. Among integrins, the most important for cell infiltration are those containing α4 and ß2 subunits. OBJECTIVES: The aim of this study was to evaluate the expression of α1 and α2 integrin subunits on peripheral blood T cells in asthmatic subjects, because previously we showed evidence that α1ß1 and α2ß1 integrins may be found on peripheral blood eosinophils in these subjects. In this study, we also analyzed the expression of α4 and ß1 subunits as a positive reference. PATIENTS AND METHODS: Expression of α1, α2, α4, and ß1 subunits was analyzed by flow cytometry on CD4+ and CD8+ T lymphocytes obtained from the peripheral blood of 54 clinically stable, asymptomatic, mild-to-moderate persistent asthmatics and 40 healthy controls. RESULTS: The α1 subunit was not present on peripheral blood T cells in the majority of subjects in both study groups. Expression of α2 was detectable on CD8+ cells in both groups and was increased on CD4+ in asthmatics. Both types of T cells showed higher expression of α4 and ß1 in patients with asthma. Expression of α4 was higher on CD8+ T cells both in asthmatics and controls. CONCLUSIONS: Expression of α4 and ß1 integrin subunits is increased on peripheral blood T cells in patients with asthma, which confirms the preactivation of blood lymphocytes even in stable and asymptomatic disease. The biological role of α2 subunit on T cells remains to be elucidated.
Assuntos
Asma/imunologia , Antígenos CD18/sangue , Linfócitos T CD4-Positivos/metabolismo , Integrina alfa4/sangue , Integrina beta1/sangue , Ativação Linfocitária/imunologia , Receptores de Colágeno/imunologia , Adulto , Moléculas de Adesão Celular/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
RATIONALE: Eosinophil ß1-integrin activation correlates inversely with FEV1 and directly with eosinophil-bound P-selectin in subjects with nonsevere allergic asthma. OBJECTIVES: Determine the relationships between ß1-integrin activation and pulmonary function or eosinophil-bound P-selectin in subjects with asthma of varying severity and discern the source of eosinophil-bound P-selectin. METHODS: Blood was assayed by flow cytometry for P-selectin and activated ß1-integrin on eosinophils and platelets. Plasma was analyzed with ELISA for soluble P-selectin, platelet factor 4, and thrombospondin-1. MEASUREMENTS AND MAIN RESULTS: Activated ß1-integrin correlated with eosinophil-bound P-selectin among all subjects with asthma even though activated ß1-integrin was higher in subjects with nonsevere asthma than severe asthma. Activated ß1-integrin correlated inversely with FEV1 corrected for FVC only in younger subjects with nonsevere asthma. Paradoxically, platelet surface P-selectin, a platelet activation marker, was low in subjects with severe asthma, whereas plasma platelet factor 4, a second platelet activation marker, was high. Correlations indicated that P-selectin-positive platelets complexed to eosinophils are the major source of the eosinophil-bound P-selectin associated with ß1-integrin activation. After whole-lung antigen challenge of subjects with nonsevere asthma, a model of asthma exacerbation known to cause platelet activation, circulating eosinophils bearing P-selectin and activated ß1-integrin disappeared. CONCLUSIONS: The relationship between eosinophil ß1-integrin activation and pulmonary function was replicated only for younger subjects with nonsevere asthma. However, we infer that platelet activation and binding of activated platelets to eosinophils followed by P-selectin-mediated eosinophil ß1-integrin activation occur in both nonsevere and severe asthma with rapid movement of platelet-eosinophil complexes into the lung in more severe disease.
Assuntos
Asma/fisiopatologia , Eosinófilos/fisiologia , Integrina beta1/fisiologia , Selectina-P/fisiologia , Ativação Plaquetária/fisiologia , Adulto , Asma/sangue , Estudos de Casos e Controles , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Humanos , Integrina beta1/sangue , Masculino , Selectina-P/sangue , Fator Plaquetário 4/sangue , Trombospondina 1/sangueRESUMO
BACKGROUND: We examined CEACAM6, ITGB1, and cyr61 concentrations from patients with gastric cancers (GCs) to assess their clinical application for diagnosing and monitoring diseases. METHODS: Real-time reverse transcription-polymerase chain reaction was used to detect the expressions of CEA, CEACAM6, ITGB1, IGF1R, CK20, cyr61, and S100A4 in peripheral blood karyocyte from 82 patients with GC, 24 patients with recurrence, and 37 healthy volunteers. Receiver operating characteristics (ROC) curves were constructed. RESULTS: There were significant association between these CEACAM6, ITGB1, and cyr61 and TNM Stages and distant metastasis. The AUC of CEACAM6 was 0.884 ± 0.044 (P = 0.0001), the AUC of cyr61 was 0.833 ± 0.047 (P = 0.0001), and the AUC of ITGB1 was 0.838 ± 0.042 (P = 0.0001) by differentiating preoperative GC patients from healthy volunteers from ROC curve analysis. The AUC of CEACAM6 was 0.761 ± 0.066 (P = 0.001), the AUC of CYR61 was 0.762 ± 0.063 (P = 0.001), and the AUC of ITGB1 was 0.824 ± 0.051 (P = 0.0001), by differentiating recurrence of GC from healthy volunteers from ROC curve analysis. CONCLUSION: The method of detecting the expression of CEACAM6, ITGB1, and CYR61 in peripheral blood of GC patients was more sensitive than CEA, IGF1R, CK20, and S100A4 for the early diagnosis of metastasis and recurrence.
Assuntos
Biomarcadores Tumorais/sangue , Recidiva Local de Neoplasia/sangue , Neoplasias Gástricas/sangue , Neoplasias Gástricas/diagnóstico , Adenocarcinoma/sangue , Adenocarcinoma/secundário , Adenocarcinoma Mucinoso/sangue , Adenocarcinoma Mucinoso/secundário , Adenocarcinoma Papilar/sangue , Adenocarcinoma Papilar/secundário , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos CD/sangue , Área Sob a Curva , Carcinoma de Células em Anel de Sinete/sangue , Carcinoma de Células em Anel de Sinete/secundário , Estudos de Casos e Controles , Moléculas de Adesão Celular/sangue , Proteína Rica em Cisteína 61/sangue , Feminino , Seguimentos , Proteínas Ligadas por GPI/sangue , Mucosa Gástrica/metabolismo , Humanos , Integrina beta1/sangue , Queratina-20/sangue , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/patologia , Prognóstico , RNA Mensageiro/sangue , RNA Mensageiro/genética , Receptor IGF Tipo 1/sangue , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteína A4 de Ligação a Cálcio da Família S100 , Proteínas S100/sangueRESUMO
BACKGROUND AND PURPOSE: Migration of leukocytes into tissue is a key element of innate and adaptive immunity. An animal study showed that liver irradiation, in spite of induction of chemokine gene expression, does not lead to recruitment of leukocytes into the parenchyma. The aim of this study was to analyze gene expression of adhesion molecules, which mediate leukocyte recruitment into organs, in irradiated rat liver in vivo and rat hepatocytes in vitro. MATERIAL AND METHODS: Rat livers in vivo were irradiated selectively at 25 Gy. Isolated hepatocytes in vitro were irradiated at 8 Gy. RNA extracted within 48 h after irradiation in vivo and in vitro was analyzed by real-time PCR (polymerase chain reaction) and Northern blot. Adhesion molecule concentration in serum was measured by ELISA (enzyme-linked immunosorbent assay). Cryostat sections of livers were used for immunohistology. RESULTS: Significant radiation-induced increase of ICAM-1 (intercellular adhesion molecule-1), VCAM-1 (vascular cell adhesion molecule-1), JAM-1 (junctional adhesion molecule-1), beta1-integrin, beta2-integrin, E-cadherin, and P-selectin gene expression could be detected in vivo, while PECAM-1 (platelet-endothelial cell adhesion molecule-1) gene expression remained unchanged. In vitro, beta1-integrin, JAM-1, and ICAM-2 showed a radiation-induced increased expression, whereas the levels of P-selectin, ICAM-1, PECAM-1, VCAM-1, Madcam-1 (mucosal addressin cell adhesion molecule-1), beta2-integrin, and E-cadherin were downregulated. However, incubation of irradiated hepatocytes with either tumor necrosis factor-(TNF-)alpha, interleukin-(IL-)1beta, or IL-6 plus TNF-alpha led to an upregulation of P-selectin, ICAM-1 and VCAM-1. CONCLUSION: The findings suggest that liver irradiation modulates gene expression of the main adhesion molecules in vivo and in cytokine-activated hepatocytes, with the exception of PECAM-1. This may be one reason for the lack of inflammation in the irradiated rat liver.
Assuntos
Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/efeitos da radiação , Regulação da Expressão Gênica/efeitos da radiação , Hepatócitos/efeitos da radiação , Fígado/efeitos da radiação , Infiltração de Neutrófilos/efeitos da radiação , Lesões Experimentais por Radiação/imunologia , Animais , Antígenos CD18/sangue , Antígenos CD18/genética , Caderinas/sangue , Caderinas/genética , Moléculas de Adesão Celular/sangue , Regulação para Baixo/efeitos da radiação , Técnicas In Vitro , Integrina beta1/sangue , Integrina beta1/genética , Molécula 1 de Adesão Intercelular/sangue , Molécula 1 de Adesão Intercelular/genética , Interleucina-1beta/farmacologia , Interleucina-6/farmacologia , Masculino , Selectina-P/sangue , Selectina-P/genética , Molécula-1 de Adesão Celular Endotelial a Plaquetas/sangue , Molécula-1 de Adesão Celular Endotelial a Plaquetas/genética , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Necrose Tumoral alfa/farmacologia , Regulação para Cima/efeitos da radiação , Molécula 1 de Adesão de Célula Vascular/sangue , Molécula 1 de Adesão de Célula Vascular/genéticaRESUMO
The past few years have witnessed a remarkable advance in our understanding of the pathophysiology of coronary heart disease. Myocardial ischemia usually occurs on the basis of coronary atherosclerosis. Although the functional consequences of depriving the myocardium of its blood supply have been appreciated for many years, coronary heart disease is still the leading cause of morbidity and mortality in the western world. This has focused the attention of physicians on restoring blood flow to the ischemic region in order to prevent tissue necrosis and regain organ function. Reperfusion of ischemic tissues is often associated with microvascular dysfunction that is manifested as impaired endothelial-dependent dilatation in arterioles and leukocyte plugging in capillaries. The availability of a broad variety of knockout mice provides important clues about the progression of the ischemia/reperfusion (I/R) injury. Therefore mouse models for I/R are of great importance for the development of new therapeutic strategies for humans.
Assuntos
Modelos Animais de Doenças , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Animais , Endotélio Vascular/fisiopatologia , Integrina beta1/sangue , Integrina beta1/genética , Molécula 1 de Adesão Intercelular/sangue , Molécula 1 de Adesão Intercelular/genética , Isoenzimas/sangue , Isoenzimas/genética , Camundongos , Camundongos Knockout , Microcirculação/fisiopatologia , Infarto do Miocárdio/fisiopatologia , Proteína Quinase C/sangue , Proteína Quinase C/genética , Proteína Quinase C-theta , Troponina T/sangue , Vasodilatação/fisiologiaRESUMO
Mesenchymal stem cells from bone marrow of Rhesus monkey (RhBMSCs) were isolated by density gradient centrifugation, purified by adherence separation, and further identified by phenotype and karyotype analysis. Growth characteristics of RhBMSCs were investigated by observation of cell proliferation and detection of apoptosis. Density gradient centrifugation and adherence separation revealed a simple method to obtain fairly pure RhBMSCs. Fusiform was the most common form of cell under observation, and cell karyotype was normal. Phenotyping assay revealed that the RhBMSCs are highly positive (99.2%) for CD29 when compared with the low positive rates (less than 3.2%) for CD34, CD45 and HLA-DR, which indicated a successful isolation of high purity RhBMSCs and a vigorous activity of cells to proliferate. But the proliferation activity of RhBMSCs gradually decreased following the increasing of cell generations. The methods and results of isolation, expansion, identification and growth characteristics of RhBMSCs were discussed in this paper, which may be helpful to understanding the bone marrow mesenchymal stem cells of human, and may serve as the groundwork for orientated differentiation of RhBMSCs and tissue repairment on experimental animal model of rhesus monkey.
Assuntos
Células da Medula Óssea/citologia , Células-Tronco Mesenquimais/citologia , Animais , Técnicas de Cultura de Células , Separação Celular , Células Cultivadas , Integrina beta1/sangue , Macaca mulatta , FenótipoAssuntos
Corticosteroides/efeitos adversos , Asma/metabolismo , Eosinófilos/metabolismo , Integrina beta1/sangue , Síndrome de Abstinência a Substâncias/imunologia , Síndrome de Abstinência a Substâncias/metabolismo , Administração por Inalação , Corticosteroides/administração & dosagem , Asma/tratamento farmacológico , Asma/patologia , Movimento Celular/efeitos dos fármacos , Movimento Celular/imunologia , Método Duplo-Cego , Eosinófilos/patologia , HumanosRESUMO
AIM: To study the expression levels of E- selectin, integrin beta1 and immunoglobulin superfamily member-intercellular adhesion molecule-1 (ICAM-1) in human gastric carcinoma cells, and to explore the relationship between these three kinds of cell adhesion molecules and gastric carcinoma. METHODS: The serum contents of E-selectin, integrin beta1 and ICAM-1 were detected by enzyme-linked immunosorbent assay (ELISA), in 47 healthy individuals (control group) and in 57 patients with gastric carcinoma (gastric carcinoma group) respectively prior to operation and 7 d after operation. RESULTS: The serum E-selectin, ECAM-1 and integrin beta1 were found to be expressed in both control and gastric carcinoma groups. However, they were highly expressed in patients with gastric carcinoma patients before operation or with unresectable tumours. The expression levels of ICAM-1 and integrin beta1 were significantly higher in gastric carcinoma patients than in controls (P < 0.01). A comparison of the E-selectin levels between the two groups showed statistically insignificant difference (P = 0.64). In addition, the expression levels were all decreased substantially in the postoperative patients subjected to radical resection of the tumours, indicating that the high level expressions of these compounds might be the important factor for predicting the prognosis of these patients. CONCLUSION: Serum E-selectin, ICAM-1 and integrin beta1 expression levels are probably related to the metastasis and relapse of gastric cancer.
Assuntos
Selectina E/genética , Integrina beta1/genética , Molécula 1 de Adesão Intercelular/genética , Neoplasias Gástricas/genética , Adulto , Idoso , Estudos de Casos e Controles , Selectina E/sangue , Ensaio de Imunoadsorção Enzimática , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Integrina beta1/sangue , Molécula 1 de Adesão Intercelular/sangue , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia , Estadiamento de Neoplasias , Valor Preditivo dos Testes , Prognóstico , Neoplasias Gástricas/sangue , Neoplasias Gástricas/patologia , Neoplasias Gástricas/cirurgiaRESUMO
The present study examines serum zinc concentrations in patients with chronic fatigue syndrome (CFS) versus normal volunteers. Serum zinc levels were determined by means of an atomic absorption method. We found that serum zinc was significantly lower in the CFS patients than in the normal controls. There was a trend toward a significant negative correlation between serum zinc and the severity of CFS and there was a significant and negative correlation between serum zinc and the subjective experience of infection. We found that serum zinc was significantly and negatively correlated to the increase in the alpha2 protein fraction and positively correlated to decreases in the expression of mitogen-induced CD69+ (a T cell activation marker) on CD3+ as well as CD3+CD8+ T cells. These results show that CFS is accompanied by a low serum zinc status and that the latter is related to signs of inflammation and defects in early T cell activation pathways. Since zinc is a strong anti-oxidant, the present results further support the findings that CFS is accompanied by increased oxidative stress. The results of these reports suggest that some patients with CFS should be treated with specific antioxidants, including zinc supplements.