A hiperemia reativa correlaciona-se com presença de sepse e degradação de glicocálix na unidade de terapia intensiva: um estudo de coorte prospectiva / Reactive hyperemia correlates with the presence of sepsis and glycocalyx degradation in the intensive care unit: a prospective cohort study

RESUMO Objetivo: Investigar se a hiperemia reativa correlaciona-se com marcadores de disfunção endotelial e pode ser utilizada para identificar sepse na doença crítica. Métodos: Trata-se de estudo prospectivo em uma coorte de pacientes críticos. A disfunção endotelial foi avaliada quando da admissão, por meio da quantificação de hiperemia por tonometria arterial periférica e níveis plasmáticos de endotelina 1, E-selectina solúvel, endocana e sindecano 1. Os pacientes sépticos foram comparados com pacientes sem evidência de infecção. Resultados: Cinquenta e oito pacientes sépticos foram comparados com 28 controle. O logaritmo natural da tonometria arterial periférica teve correlação negativa com comorbidades cardiovasculares, severidade da doença e níveis plasmáticos de E-selectina solúvel (p = 0,024) e sindecano 1 (p < 0,001). O logaritmo natural da tonometria arterial periférica foi mais baixo nos pacientes sépticos quando comparado com os de pacientes controle (0,53 ± 0,48 versus 0,69 ± 0,42, respectivamente) e, quando ajustado à idade, o modelo multivariado predisse que cada 0,1 de diminuição em unidades de logaritmo natural da tonometria arterial periférica levou a aumento de 14,6% na probabilidade de infecção. Conclusão: A hiperemia reativa avaliada por tonometria arterial periférica tem estreita relação com E-selectina solúvel e sindecano 1, o que sugere associação entre ativação endotelial, degradação de glicocálix e reatividade vascular. A hiperemia reativa por tonometria arterial periférica parece estar comprometida em pacientes críticos, especialmente os com sepse.

Abstract Objective: To investigate whether reactive hyperemia measured by peripheral arterial tonometry correlates with markers of endothelial dysfunction and may be used to identify sepsis in critical illness. Methods: A prospective study was performed using a cohort of critically ill patients. Endothelial dysfunction was assessed on admission by quantifying reactive hyperemia-peripheral arterial tonometry and plasma levels of endothelin-1, soluble E-selectin, endocan and syndecan-1. Septic patients were compared to patients without evidence of infection. Results: Fifty-eight septic patients were compared to 28 controls. The natural logarithm of reactive hyperemia-peripheral arterial tonometry was negatively correlated with cardiovascular comorbidities, disease severity and plasma levels of soluble E-selectin (p = 0.024) and syndecan-1 (p < 0.001). The natural logarithm of reactive hyperemia-peripheral arterial tonometry was lower in septic patients than in controls (0.53 ± 0.48 versus 0.69 ± 0.42, respectively). When adjusted for age, the multivariable model predicted that each 0.1-unit decrease in natural logarithm of reactive hyperemia-peripheral arterial tonometry increased the odds for infection by 14.6%. m. Conclusion: Reactive hyperemia-peripheral arterial tonometry is closely related to soluble E-selectin and syndecan-1, suggesting an association between endothelial activation, glycocalyx degradation and vascular reactivity. Reactive hyperemia-peripheral arterial tonometry appears to be compromised in critically ill patients, especially those with sepsis.

Role of vitamin K-dependent protein Gas6 in the expression of endothelial cell adhesion molecule-1 and chemokines induced by Porphyromonas gingivalis lipopolysaccharide / 北京大学学报(医学版)

OBJECTIVE@#Growth-arrest-specific protein 6 (Gas6) is a vitamin K-dependent protein and involved in cell proliferation, survival, adhesion and migration . Also it has been shown to play an important role in the inflammatory response .The aim of present study was to investigate the role of Gas6 in the process of the expression of adhesion molecules and chemokines of human umbilical vein endothelial cells (HUVECs) induced by Porphyromonas gingivalis lipopolysaccharide(P.g-LPS).@*METHODS@#After up-regulation and down-regulation of the expression of Gas6, the vascular endothelial cells were stimulated with 1 mg/L P.g-LPS for 3 h and 24 h. Real-time quantitative polymerase chain reaction(real-time PCR) was taken to detect the expression of the cell adhesion molecules:intercellular adhesion molecule-1 (ICAM-1) and E-selectin, as well as chemokines:interleukin-8 (IL-8) and monocyte chemoattractant protein 1 (MCP-1). Wound healing assay was taken to observe the migration ability of endothelium cells in different groups.@*RESULTS@#After 3 h of P.g-LPS stimulation, the expression of adhesion molecules and chemokine in the down-regulation group was not significantly different from that in the control group,while in the up-regulation group the decrease of E-selectin, ICAM-1, IL-8 and MCP-1 was 81%±0%, 47%±3%, 76% ± 3%, 26% ± 6% respectively. After 24 h of P.g-LPS stimulation, the expression of adhesion molecules and chemokine in down-regulation group was significantly higher than that in control group (2.06±0.07, 1.99±0.11, 3.14±0.15, 1.84±0.03 flod), while these molecules in the down-regulation group was significantly lower than in the control group (29%±1%, 62%±3%, 69%±1%, 41%±2%). Differences were statistically significant (P<0.01). Wounding healing assay showed that down-regulation of Gas6 enhanced migration ability of endothelial cells while up-regulation of Gas6 weakened this ability,which was consistent with the trend of real-time PCR result.@*CONCLUSION@#Down-regulation of the Gas6 gene enhanced the expression of ICAM-1, E-selectin, IL-8 and MCP-1 in HUVECs after P.g- LPS stimulating, while up-regulaiton of the Gas6 gene weakened the expression of ICAM-1, E-selectin, IL-8 and MCP-1 in HUVECs after P.g-LPS stimulating,suggesting that Gas6 may play a role in the process of endothelial cell adhesion.

Circulatory and Ventilatory Power: Characterization in Patients with Coronary Artery Disease / Potência Circulatória e Ventilatória: Caracterização em Pacientes com Doença Arterial Coronariana

Background: Circulatory power (CP) and ventilatory power (VP) are indices that have been used for the clinical evaluation of patients with heart failure; however, no study has evaluated these indices in patients with coronary artery disease (CAD) without heart failure. Objective: To characterize both indices in patients with CAD compared with healthy controls. Methods: Eighty-seven men [CAD group = 42 subjects and healthy control group (CG) = 45 subjects] aged 40–65 years were included. Cardiopulmonary exercise testing was performed on a treadmill and the following parameters were measured: 1) peak oxygen consumption (VO2), 2) peak heart rate (HR), 3) peak blood pressure (BP), 4) peak rate-pressure product (peak systolic HR x peak BP), 5) peak oxygen pulse (peak VO2/peak HR), 6) oxygen uptake efficiency (OUES), 7) carbon dioxide production efficiency (minute ventilation/carbon dioxide production slope), 8) CP (peak VO2 x peak systolic BP) and 9) VP (peak systolic BP/carbon dioxide production efficiency). Results: The CAD group had significantly lower values for peak VO2 (p < 0.001), peak HR (p < 0.001), peak systolic BP (p < 0.001), peak rate-pressure product (p < 0.001), peak oxygen pulse (p = 0.008), OUES (p < 0.001), CP (p < 0.001), and VP (p < 0.001) and significantly higher values for peak diastolic BP (p = 0.004) and carbon dioxide production efficiency (p < 0.001) compared with CG. Stepwise regression analysis showed that CP was influenced by group (R2 = 0.44, p < 0.001) and VP was influenced by both group and number of vessels with stenosis after treatment (interaction effects: R2 = 0.46, p < 0.001). Conclusion: The indices CP and VP were lower in men with CAD than healthy controls. .

Fundamento: Os índices da Potência Circulatória (PC) e Potência Ventilatória (PV) têm sido utilizados para avaliação clínica de pacientes com insuficiência cardíaca, mas nenhum estudo avaliou esses índices em pacientes com Doença Arterial Coronariana (DAC). Objetivo: Caracterizar ambos os índices em pacientes com DAC comparados a indivíduos saudáveis. Métodos: Oitenta e sete homens [grupo DAC = 42 sujeitos e, grupo controle (GC) = 45 sujeitos] com idade entre 45 e 65 anos foram incluídos. Um Teste de Exercício Cardiopulmonar (TECP) foi realizado em esteira e as seguintes variáveis foram obtidas: 1) consumo de oxigênio (VO2) pico; 2) Frequência Cardíaca (FC) pico; 3) Pressão Arterial (PA) pico; 4) duplo produto pico (PA sistólica pico x FC pico); 5) pulso de oxigênio pico (VO2 pico dividido pela FC pico); 6) eficiência ventilatória para o consumo de oxigênio (OUES); 7) eficiência ventilatória para a produção de dióxido de carbono (VE/VCO2 slope); 8) PC (VO2 pico x PA sistólica pico); e 9) PV (PA sistólica pico dividido pelo VE/VCO2 slope). Resultados: O grupo DAC apresentou valores significativamente menores das seguintes variáveis no pico do exercício: VO2 (p < 0,001), FC (p < 0,001), PA sistólica (p < 0,001), duplo produto (p < 0,001), pulso de oxigênio (p = 0,008), OUES (p < 0,001), PC (p < 0,001) e PV (p < 0,001), e valores significativamente maiores de PA diastólica (p = 0,004) e VE/VCO2 slope (p < 0,001) em relação ao GC. Uma análise de regressão pelo método stepwise mostrou que a PC foi influenciada pelo grupo (R2 = 0,44, p < 0,001) e a PV tanto pelo grupo quanto pelo número de vasos com estenose pós tratamento (efeito de interação: R2 = 0,46, p < 0,001). Conclusion: Os índices da PC e PV foram menores em homens com DAC comparados ao GC, podendo dessa forma ser utilizados na caracterização dessa população. .

Angiogenic response of advanced glycation end products (AGEs) involves PPAR

Diabetes is associated with increased formation of advanced glycation end products (AGEs), which have been implicated in micro and macrovascular complications of diabetes. Our earlier reports showed proangiogenic effect of AGE-bovine serum albumin (BSA). In order to understand the mechanism of AGE-mediated angiogenesis, the possibility of involvement of peroxisome prolifeator activated receptor (PPAR) , a ligand activated transcription factor was examined. The angiogenic effect was studied in chick chorio allantoic membrane (CAM) and by analyzing angiogenic markers in human umbilical vein endothelial cells (HUVECs) in culture. The involvement of PPAR was investigated using synthetic PPAR agonist GW 1929 and antagonist GW 9662 and by RT-PCR. In CAM assay, PPAR antagonist GW 9662 reversed the AGE-induced effect on vascularity. In HUVECs in culture, GW 9662 reversed the effect of AGE-BSA and decreased the expression of CD 31, E-Selectin and VEGF. RT-PCR analysis showed that treatment with AGE-BSA caused upregulation of PPAR mRNA levels. The reversal of the effect of AGE on angiogenesis by treatment with PPAR antagonists and up-regulation of PPAR gene in HUVECs treated with AGE-BSA suggested the possible involvement of PPAR -dependent downstream pathway in mediating the angiogenic effect of AGE.

Morphine Attenuates Endothelial Cell Adhesion Molecules Induced by the Supernatant of LPS-Stimulated Colon Cancer Cells

A large reservoir of bacterial lipopolysaccharide (LPS) is available in the colon and this could promote colon cancer metastasis by enhancing tumor cell adhesion, intravasation, and extravasation. Furthermore, adhesion molecules like ICAM-1, VCAM-1, and E-selectin play important roles in the adhesion of tumor cells to endothelium. This study was designed to determine whether morphine can attenuate the expressions of adhesion molecules up-regulated by the supernatant of LPS-stimulated HCT 116 colon cancer cells (LPS-Sup). In this study, we divided to three groups by cell-growth medium of human umbilical vascular endothelial cells (HUVECs): the control group was incubated in growth factor-free endothelial medium, the Sup group was incubated in the supernatant of HCT 116 cells (Sup), and the LPS-Sup group was incubated in LPS-Sup. To observe effect of morphine to the adhesion molecules expressions in the LPS-Sup group, we co-treated morphine with LPS or added it to LPS-Sup. Adhesion molecule expressions on HUVECs in all three groups were measured during incubation period. Consquentially, ICAM-1, VCAM-1, and E-selectin expressions on HUVECs were significantly lower when morphine was co-treated with LPS than not co-treated. Thus, we suggest that morphine affects the expressions of adhesion molecules primarily by attenuating LPS stimuli on tumor cells.

Transmigração endotelial do fungo patogênico Sporothrix schenckii: modulação da via de p38MAPK/MLCK, Src e de moléculas de adesão celular / Endothelial transmigration of the pathogenic fungus, Sporothrix schenckii: pathway modulation of p38MAPK/MLCK, Src and cell adhesion molecules

Sporothrix schenckii é um fungo dimórfico, agente etiológico da esporotricose, uma micose subaguda ou crônica que pode eventualmente evoluir para complicações sistêmicas, principalmente em pacientes imunocomprometidos. O endotélio exerce um papel crucial durante infecções disseminadas já que, juntamente com as células epiteliais, representa uma barreira a ser ultrapassada por microorganismos invasores. Em estudos anteriores, observamos que S. schenckii transmigra preferencialmente pela rota paracelular (passagem entre células endoteliais adjacentes), interagindo em seguida com componenetes da matriz subendotelial. Também foram identificadas algumas vias de sinalização relacionadas à diferentes fases da interação de leveduras de S. schenckii com o endotélio in vitro (associação/endocitose, transmigração). No entanto, a correlação entre tais vias de sinalização e os mecanismos celulares da invasão do endotélio pelo fungo não foram efetivamente demonstrados. No presente trabalho, a análise do perfil de proteínas endoteliais totais fosforiladas em resíduos de tirosina mostrou que S. schenckii induz fosforilações em tempos curtos (< 15 minutos), em proteínas de massas moleculares 20, 13, 12 e 6KDa, enquanto alunas proteínas de mais alto peso molecular (83, 123, 136, 140 e 193 KDa) persistem fosforiladas em tempos mais longos durante a infecção (6 horas). As vias de transdução de sinais disparadas pela interação do fungo com o endotélio foram investigadas através do uso de inibidores da ativação de MAPKs p38 (SB 203580) e ERK (PD 98059), MLCK (W7) e de um quelante de Ca2+ intracelular (BAPTA). A transmigração de S. schenckii através de monocamadas de HUVECs por 6 horas mostrou ser dependente da ativação de ERK e p38, ions Ca2+ intracelular e MLCK. Estas vias estão também envolvidas nos rearranjos do citoesqueleto de actina que levam à contratilidade celular e aumento da permeabilidade endotelial. A interação do fungo com HUVECs induziu ativação de Src...

Sporothrix schenckii, a dimorphic fungus, is the causative agent of sporotrichosis, a cutaneous/subcutaneous mycosis which can eventually evolve to systemic complications, mainly in immunocompromised patients. The primary interaction of pathogenic fungi with endothelial cells (EC) is throught to be essential for the development of systemic infections. We have previously shown that S. schenckii cross endothelial monolayers through a paracellular pathway, in a process also modulated by the subendothelial matrix, and that the fungus is able to alter host signaling pathways. We observed that the interaction of S. schenckii with human umbilical vein endothelial cells (HUVECs) was regulated by tyrosine-phosphorylation of EC proteins. In the present work, we observed that S. schenckii stimulates the early increase (<15 minutes) in tyrosine-phorphorylation of 20, 13, 12 e 6 KDa endothelial proteins, whereas tyrosine-phosphorylation of higher molecular weight proteins (83, 123, 136, 140 e 193 KDa) persists up to 6 hours of endothelial infection. Selective signal transduction inhibitors (SB203580 and W7, for blocking p38 MAPK and MLCK activation, respectively) were able to inhibit transendothelial migration of S. schenckii. The process was also modulated by Ca++ions. These signaling pathways are crucial for the actin rearrangement associated to impairment of endothelial permeability. Long-term (3 hours) interaction of S. schenckii with HUVECs lead to increase of MLC2 phosphorylation and Src activation. Src was shown by others to be involved in the phosphorylation of VE-cadherin, thus provoking adherent junctions (AJs) disassembly. We found that S. schenckii induces tyrosine-phosphorylation of endothelial VE-cadherin up to 3 hours of interaction with endothelial cells. VE-cadherin phosphorylation can be triggered by the activation of E-selectin in endothelial cells. Since the time-course of the major signaling events correlated with the time needed...