RESUMO
The Forkhead family of transcription factors comprises numerous members and is implicated in various cellular functions, including cell growth, apoptosis, migration, and differentiation. In this study, we identified the Forkhead factor FoxQ1 as increased in expression during TGF-ß1 induced changes in epithelial differentiation, suggesting functional roles of FoxQ1 for epithelial plasticity. The repression of FoxQ1 in mammary epithelial cells led to a change in cell morphology characterized by an increase in cell size, pronounced cell-cell contacts, and an increased expression of several junction proteins (e.g., E-cadherin). In addition, FoxQ1 knock-down cells revealed rearrangements in the actin-cytoskeleton and slowed down cell cycle G1-phase progression. Furthermore, repression of FoxQ1 enhanced the migratory capacity of coherent mammary epithelial cells. Gene expression profiling of NM18 cells indicated that FoxQ1 is a relevant downstream mediator of TGF-ß1-induced gene expression changes. This included the differential expression of transcription factors involved in epithelial plasticity, for example, Ets-1, Zeb1, and Zeb2. In summary, this study has elucidated the functional impact of FoxQ1 on epithelial differentiation.
Assuntos
Diferenciação Celular , Células Epiteliais/citologia , Fatores de Transcrição Forkhead/metabolismo , Actinas/metabolismo , Animais , Comunicação Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Tamanho Celular/efeitos dos fármacos , Quinases Ciclina-Dependentes/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Fatores de Transcrição Forkhead/genética , Fase G1/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Camundongos , Proteínas dos Microfilamentos/metabolismo , Transporte Proteico/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Fator de Crescimento Transformador beta1/farmacologiaRESUMO
Postprandial hyperlipidaemia has been associated with coronary artery disease (CAD). We investigated which of the generally used methods to test postprandial lipaemia differentiated best between patients with premature CAD (50+/-4 years, n=20) and healthy controls. Furthermore, the effects of rosuvastatin 40 mg/day on postprandial parameters were assessed. Standardised oral fat-loading tests (OFLT) and ambulant self-measurements of daylong capillary triglycerides (TGc) were performed. Total responses of individual lipoproteins, plasma TG (TGp) and remnant-like particle cholesterol (RLP-C) were estimated as area under the curve (AUC). Most AUCs were highest in untreated patients and reached control levels after rosuvastatin. From all AUCs, RLP-C-AUC was best associated to TGp-AUC in untreated patients and controls (adjusted R2=0.84, beta=0.92, p.
Assuntos
Doença da Artéria Coronariana/diagnóstico , Hiperlipidemias/diagnóstico , Período Pós-Prandial , Área Sob a Curva , Doença da Artéria Coronariana/tratamento farmacológico , Doença da Artéria Coronariana/fisiopatologia , Feminino , Fluorbenzenos/uso terapêutico , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Hiperlipidemias/tratamento farmacológico , Hiperlipidemias/fisiopatologia , Lipídeos/sangue , Masculino , Pessoa de Meia-Idade , Pirimidinas/uso terapêutico , Análise de Regressão , Rosuvastatina Cálcica , Sulfonamidas/uso terapêutico , Triglicerídeos/sangueRESUMO
Activation of leukocytes is obligatory for inflammation and atherogenesis by adhering to the endothelium via specific ligands. Although in vitro studies have shown that triglycerides (TG) can activate leukocytes, it is unknown whether this occurs in vivo. Using flowcytometry, we studied the expression of leukocyte activation markers CD11A, CD11B, CD62L (all involved in endothelium adhesion) and CD66B (a neutrophil degranulation marker) during a 6 h fat challenge (50 g/m2) and a water test in 10 healthy males (52 +/- 3 years). After fat, neutrophil counts were increased between t=1 and t =6 h, with a maximum at t=3 h (+32% versus t=0, P <0.05), while they remained unchanged after water. Both tests showed gradual lymphocyte count increments. The expression of activation markers on lymphocytes was low and showed comparable responses after both tests. After fat, a significant increase up to a maximum at t=6 h was seen for CD11B on monocytes and on neutrophils for CD11B, CD62L and CD66B. Postprandial activation of monocytes and neutrophils was higher after fat than after water. The maximal postprandial TG increment was significantly related to the increase of CD11B on monocytes (Pearson's R=0.64, P <0.05). In conclusion, postprandially there is a TG-specific increase of neutrophil counts and increased activation of monocytes and neutrophils. These results are suggestive of a pro-inflammatory situation that may correspond with increased adhesive capacity of these cells contributing to the inflammatory component of atherosclerosis.
Assuntos
Ácidos Graxos/sangue , Leucócitos/fisiologia , Período Pós-Prandial , Triglicerídeos/sangue , Triglicerídeos/fisiologia , Adulto , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Activation of leukocytes is obligatory for adherence to the endothelium and atherogenesis. Since leukocyte activation by triglycerides (TG) and glucose has been described in vitro, we hypothesised higher leukocyte activation in patients with type 2 diabetes. METHODS: Using flow cytometry, we studied the expression of the leukocyte activation markers CD11A, CD11B, CD62L and CD66B in 15 patients with type 2 diabetes without clinical evidence of atherosclerosis (55+/-7 years) and in 15 healthy controls (53+/-2 years). All patients were on oral antidiabetic treatment (glyHb 6.3+/-0.9%) and not taking statins or anti-inflammatory drugs. RESULTS: In comparison with controls, the patients had a higher waist circumference (1.08+/-0.09 vs 0.94+/-0.11 m, p<0.005) and higher fasting glucose (8.4+/-2.3 vs 5.3+/-0.7 mM, p<0.005), whereas fasting plasma lipids were not statistically different. The leukocyte count was higher in the patients (6.55+/-1.55 vs 5.07+/-1.10 x 10(9) cells/l, p<0.005) due to higher neutrophils and lymphocytes (+34% and +24%, p<0.05 for each). CD11B on monocytes and CD11B and CD66B on neutrophils were higher in the patients (+30%, +52% and +43%, P<0.05 for each). Fasting glucose, waist circumference, body mass index and systolic blood pressure were positively associated with the leukocyte and neutrophil count. The expressions of CD11B and CD66B on monocytes and neutrophils were strongly positively interrelated, but unrelated to TG and glucose. CONCLUSION: In patients with type 2 diabetes, the expression of activation markers on monocytes and neutrophils is enhanced and not correlated to fasting glucose or TG. These results suggest a proinflammatory situation in type 2 diabetes and most likely represent increased adhesive capacity of neutrophils and monocytes to the endothelium.
Assuntos
Diabetes Mellitus Tipo 2/fisiopatologia , Monócitos/imunologia , Ativação de Neutrófilo , Neutrófilos/imunologia , Biomarcadores/sangue , Estudos de Casos e Controles , Diabetes Mellitus Tipo 2/complicações , Feminino , Citometria de Fluxo , Imunofluorescência , Humanos , Contagem de Leucócitos , Masculino , Pessoa de Meia-IdadeRESUMO
Atherosclerosis is a low-grade inflammatory disease involving leukocytes, lipids, and glucose leading to endothelial dysfunction. Since activation of neutrophils by triglycerides and glucose has been described in vitro, we hypothesized that the postprandial phase is an inflammatory state affecting leukocytes, possibly contributing to endothelial dysfunction. We measured postprandial blood leukocyte counts, cytokines, hydroperoxides (HPOs), and flow-mediated vasodilation (FMD) in eight healthy males (age 23 +/- 2 years) after a FAT (50 g/m2) and GLUCOSE challenge (37.5 g/m2), a combination of both (MIXED test), and after WATER. All tests, except WATER, resulted in significantly impaired FMD (10% reduction) between t = 1 h and t = 3 h, accompanied by a significant increase of neutrophils (59% after FAT and 28% after GLUCOSE and MIXED), total plasma HPOs (15 to 31% increase), and plasma interleukin-8 (IL-8) (50-130% increase). WATER did not affect FMD, neutrophils, HPOs, or IL-8. Lymphocytes increased gradually in all tests (40-70% increase at t = 10 h compared with t = 0; P < 0.005), paralleling a gradual 3- to 5-fold interleukin-6 increase. Monocyte and erythrocyte counts did not change in any test. In conclusion, the neutrophil increment during postprandial lipemia and glycemia with concomitant IL-8 and HPO increases may contribute to endothelial dysfunction. Lymphocyte increment is a nonspecific diurnal process. Postprandial intravascular inflammatory changes may be relevant for the pathogenesis of atherosclerosis.
Assuntos
Arteriosclerose/etiologia , Arteriosclerose/patologia , Endotélio Vascular/patologia , Neutrófilos/metabolismo , Período Pós-Prandial/fisiologia , Adulto , Arteriosclerose/induzido quimicamente , Arteriosclerose/metabolismo , Glicemia/análise , Gorduras na Dieta/administração & dosagem , Gorduras na Dieta/sangue , Gorduras na Dieta/farmacologia , Endotélio Vascular/efeitos dos fármacos , Jejum , Ácidos Graxos não Esterificados/sangue , Glucose/administração & dosagem , Glucose/farmacologia , Humanos , Peróxido de Hidrogênio/sangue , Inflamação/metabolismo , Insulina/sangue , Interleucina-6/sangue , Interleucina-8/sangue , Contagem de Leucócitos , Masculino , Modelos Biológicos , Neutrófilos/efeitos dos fármacos , Fatores de Tempo , Triglicerídeos/sangueRESUMO
Atherosclerosis is an inflammatory disorder involving leukocytes and lipids. To study the relationship between leukocytes and lipids in vivo, leukocyte changes were determined in 14 healthy males (age, 23 +/- 3 years; body mass index [BMI], 21.9 +/- 1.5 kg/m(2)) after an 8-hour oral fat load (50 g/m(2)) and after water. The postprandial triglyceride (TG) increment after fat was paralleled by a leukocyte increment, due to an increase in neutrophils in the first 2 hours (142% +/- 69% higher than baseline, P =.04). Neutrophil counts did not return to baseline at the end of the test. Water ingestion did not induce significant neutrophil changes. Blood lymphocytes increased gradually in both tests (142% +/- 30% higher than baseline, P <.001 after fat, and 128% +/- 36%, P =.02 after water). The total leukocyte increment after fat ingestion was related to the postprandial TG increase (Spearman's r = 0.73, P =.003). An early postprandial, lipid-specific, neutrophil increment is a new characteristic of the postprandial phase. Future studies will elucidate the role of postprandial leukocyte changes in the pathogenesis of atherosclerosis.