RESUMO
Chronic thromboembolic pulmonary hypertension (CTEPH) and pulmonary arterial hypertension (PAH) are two forms of pulmonary hypertension (PH) characterized by obstructive vasculopathy. Endothelial dysfunction along with metabolic changes towards increased glycolysis are important in PAH pathophysiology. Less is known about such abnormalities in endothelial cells (ECs) from CTEPH patients. This study provides a systematic metabolic comparison of ECs derived from CTEPH and PAH patients. Metabolic gene expression was studied using qPCR in cultured CTEPH-EC and PAH-EC. Western blot analyses were done for HK2, LDHA, PDHA1, PDK and G6PD. Basal viability of CTEPH-EC and PAH-EC with the incubation with metabolic inhibitors was measured using colorimetric viability assays. Human pulmonary artery endothelial cells (HPAEC) were used as healthy controls. Whereas PAH-EC showed significant higher mRNA levels of GLUT1, HK2, LDHA, PDHA1 and GLUD1 metabolic enzymes compared to HPAEC, CTEPH-EC did not. Oxidative phosphorylation associated proteins had an increased expression in PAH-EC compared to CTEPH-EC and HPAEC. PAH-EC, CTEPH-EC and HPAEC presented similar HOXD macrovascular gene expression. Metabolic inhibitors showed a dose-dependent reduction in viability in all three groups, predominantly in PAH-EC. A different metabolic profile is present in CTEPH-EC compared to PAH-EC and suggests differences in molecular mechanisms important in the disease pathology and treatment.
Assuntos
Células Endoteliais/metabolismo , Hipertensão Arterial Pulmonar/genética , Hipertensão Arterial Pulmonar/metabolismo , Embolia Pulmonar/genética , Embolia Pulmonar/metabolismo , Adulto , Idoso , Células Cultivadas , Doença Crônica , Feminino , Expressão Gênica , Glutamato Desidrogenase/genética , Glutamato Desidrogenase/metabolismo , Glicólise/genética , Hexoquinase/genética , Hexoquinase/metabolismo , Humanos , L-Lactato Desidrogenase/genética , L-Lactato Desidrogenase/metabolismo , Masculino , Pessoa de Meia-Idade , Fosforilação Oxidativa , Artéria Pulmonar/citologia , Piruvato Desidrogenase (Lipoamida)/genética , Piruvato Desidrogenase (Lipoamida)/metabolismoRESUMO
Cardiovascular disease is the leading cause of human mortality and disability worldwide, primarily due to myocardial infarction (MI) and the resultant heart failure. To address this, animal models of MI have been developed to better understand the pathophysiological process and to enable the discovery and development of new therapies. The most commonly used small and large mammal models of MI accurately reproduce histopathologically the four characteristic post-MI phases: cardiac cell death, inflammation, myocardial repair and remodelling. However, differences between the time of onset of each characteristic phase and the kinetics of various cellular reactions between human MI and animal models, and between animal models, require careful consideration when defining the variables to be analysed and the timepoints of assessment in experimental studies. Typically, the progression of the different phases post-MI occur more rapidly in rodent models compared with large-animal models and man, suggesting the use of large-animal models is more translational for studying human MI. This review provides an overview of the main anatomopathological features of small and large animal models of MI and discusses the key species-specific histopathological similarities and differences.
Assuntos
Modelos Animais de Doenças , Infarto do Miocárdio , Animais , HumanosRESUMO
Previous work has shown that optimal ex vivo expansion and differentiation of CD34(+) progenitor cells into neutrophils is by addition of Flt3-L, SCF and G-CSF. Here we report that a variety of genes involved in the WNT pathway are transcriptionally active in both undifferentiated and differentiated umbilical cord blood CD34(+) cells, however statistically significant changes in gene expression are not always consistent across UCB samples.
Assuntos
Diferenciação Celular/genética , Sangue Fetal/citologia , Receptores Frizzled/genética , Perfilação da Expressão Gênica , Células-Tronco Hematopoéticas/metabolismo , Neutrófilos/citologia , Proteínas Wnt/genética , Antígenos CD34/análise , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas/citologia , Células Cultivadas/metabolismo , DNA Complementar/genética , Fatores de Crescimento de Células Hematopoéticas/farmacologia , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/efeitos dos fármacos , Humanos , Recém-Nascido , Reação em Cadeia da Polimerase/métodos , RNA Mensageiro/genética , Transcrição GênicaRESUMO
BACKGROUND: Endothelial progenitor cells (EPCs) are circulating mononuclear cells that participate in angiogenesis. The aim of this study was to determine the influence of the menstrual cycle on the number and function of EPCs, and to investigate their relationship with circulating concentrations of sex steroids and inflammatory mediators. METHODS: Ten healthy nulliparous, premenopausal, non-smoking women with regular menses were studied over a single menstrual cycle. Venepuncture was performed in the menstrual, follicular, peri-ovulatory and luteal phases. EPCs were quantified by flow cytometry (CD133(+)CD34(+)KDR(+) phenotype) and the colony-forming unit (CFU-EPC) functional assay. Circulating concentrations of estradiol, progesterone and inflammatory mediators (TNF-alpha, IL-6, sICAM-1 and VEGF) were measured by immunoassays. RESULTS: The numbers of CD133(+)CD34(+)KDR(+) cells were higher in the follicular phase (0.99 +/- 0.3 x 10(6) cells/l) compared with the peri-ovulatory phase (0.29 +/- 0.1 x 10(6) cells/l; P < 0.05). In contrast, the numbers of CFU-EPCs did not vary over the menstrual cycle. There were no correlations between EPCs and concentrations of either circulating sex steroids or inflammatory mediators. CONCLUSIONS: CD133(+)CD34(+)KDR(+) cells but not CFU-EPCs vary during the menstrual cycle. Our findings suggest a potential role for circulating EPCs in the normal cycle of physiological angiogenesis and repair of the uterine endometrium that is independent of circulating sex steroids or inflammatory mediators.
