endothelin-1 and Hemolysis

Oxidative stress and haemolysis-associated nitric oxide (NO) depletion plays a crucial role in the development of vasculopathy in sickle cell anaemia (SS). However it remains unknown whether oxidative stress and haemolysis levels influence vascular function in patients with sickle haemoglobin C disease (SC). Microvascular response to heat (using Laser Doppler flowmetry on finger), oxidative stress biomarkers, NO metabolites, endothelin-1 and haematological parameters were compared between patients with SS and SC. Vascular function, oxidative and nitrosative markers were also measured in healthy (AA) children. SS and SC had increased plasma advanced oxidation protein products (AOPP), malondialdehyde, plasma antioxidant activities and NO end products, compared to AA. SC had lower catalase activity compared to AA and SS. Haemolytic rate, glutathione peroxidase and nitrotyrosine concentrations were significantly increased in children with SS compared to SC and AA. SS and SC had impaired microvascular reactivity compared to AA. In SS, the plateau phase of the response to local thermal heating was negatively associated with nitrotyrosine and AOPP. No association between vascular function parameters and oxidative stress markers was observed in SC. Mild haemolysis in SC, compared to SS, may limit oxidative and nitrosative stress and could explain the better preserved microvascular function in this group.

Topics: Adolescent; Advanced Oxidation Protein Products; Anemia, Sickle Cell; Antioxidants; Blood Viscosity; Case-Control Studies; Child; Endothelin-1; Female; Fingers; Hemoglobin SC Disease; Hemolysis; Humans; Laser-Doppler Flowmetry; Male; Malondialdehyde; Microcirculation; Nitric Oxide; Oxidative Stress

Drug-eluting stents (DESs) can effectively control the harmful effects of coronary artery disease, because of their excellent ability to reduce in-stent restenosis. However, delayed re-endothelialization and late stent thrombosis have caused concern over the safety of DESs. In this study, according to time-ordered pathological responses after stent implantation, a hierarchical multiple drug-eluting stent was designed and prepared to overcome the existing DES limitations. A platelet membrane glycoprotein IIIa monoclonal antibody (SZ-21) and a vascular endothelial growth factor (VEGF

Topics: Animals; Cell Movement; Cell Proliferation; Chitosan; Drug Design; Drug-Eluting Stents; Endothelin-1; Hemolysis; Human Umbilical Vein Endothelial Cells; Humans; Male; Materials Testing; Nitric Oxide Synthase Type III; Platelet Adhesiveness; Polyesters; Rabbits; Vascular Endothelial Growth Factor A

Placenta growth factor (PlGF) is released by immature erythrocytes and is elevated in sickle cell disease (SCD). Previous data generated in vitro suggest that PlGF may play a role in the pathophysiology of SCD-associated pulmonary hypertension (PHT) by inducing the release of the vasoconstrictor, endothelin-1. In this cross-sectional study of 74 patients with SCD, we confirm that PlGF is significantly elevated in SCD compared with healthy control subjects. We found significantly higher levels of PlGF in SCD patients with PHT but observed no association of PlGF with the frequency of acute pain episodes or history of acute chest syndrome. The observed correlation between PlGF and various measures of red cell destruction suggests that hemolysis, and the resultant erythropoietic response, results in the up-regulation of PlGF. Although relatively specific, PlGF, as well as N-terminal pro-brain natriuretic peptide and soluble vascular cell adhesion molecule, has low predictive accuracy for the presence of PHT. Prospective studies are required to conclusively define the contribution of PlGF to the pathogenesis of PHT and other hemolytic complications in SCD.

Topics: Adult; Anemia, Sickle Cell; Case-Control Studies; Cross-Sectional Studies; Endothelin-1; Female; Hemolysis; Humans; Hypertension, Pulmonary; Inflammation; Male; Middle Aged; Placenta Growth Factor; Predictive Value of Tests; Pregnancy Proteins; Prognosis; Young Adult

Studies on pulsatile and nonpulsatile perfusion have long been performed. However, investigators have not reached a conclusion on which is more effective. In the present study, pulsatile cardiopulmonary bypass (CPB) was investigated in terms of the effects on cytokines, endothelin, catecholamine, and pulmonary and renal functions. Twenty-four patients who underwent coronary artery bypass grafting were divided into a pulsatile CPB group and a nonpulsatile CPB group. Parameters examined were hemodynamics, interleukin-8 (IL-8), endothelin-1 (ET-1), epinephrine, norepinephrine, lactate, arterial ketone body ratio, urine volume, blood urea nitrogen, creatinine, renin activity, angiotensin-II, lactate dehydrogenase, plasma-free hemoglobin, tracheal intubation time, and respiratory index. The IL-8 at 0.5, 3, and 6 h after CPB, and ET-1 at 3, 6, 9, and 18 h after CPB were significantly lower in the pulsatile group. Both epinephrine and norepinephrine were significantly lower in the pulsatile group. The respiratory index was significantly higher in the pulsatile group. In the present study, inhibitory effects on cytokine activity, edema in pulmonary alveoli, and endothelial damage were shown in addition to the favorable effects on catecholamine level, renal function, and peripheral circulation that have already been documented.

Topics: Aged; Cardiopulmonary Bypass; Catecholamines; Coronary Artery Bypass; Cytokines; Endothelin-1; Female; Hemolysis; Humans; Interleukin-8; Kidney Function Tests; Male; Middle Aged; Perfusion; Pulsatile Flow; Respiratory Function Tests

We investigated the effects of sickle erythrocytes on the production of vasotone mediators in endothelial cells (ECs) using an in vitro recirculating flow system. Sickle erythrocytes increased the EC production of two important vasoactivators, prostacyclin and endothelin-1, under venous wall shear stress conditions of 1dyncm2. The presence of interleukin-1 in the perfusion system, as a model for inflammatory cytokine effects, enhanced the overall amounts of released prostacyclin but did not affect the production of endothelin-1. This study demonstrates the effects of sickle erythrocytes on the function and metabolism of ECs under vascular flow environments. The altered production of vasoactivators may contribute to the vasotone instability and vasoocclusive crises in sickle cell anemia.

Topics: 6-Ketoprostaglandin F1 alpha; Adult; Anemia, Sickle Cell; Cells, Cultured; Endothelin-1; Endothelium, Vascular; Epoprostenol; Erythrocytes, Abnormal; Hemolysis; Hemorheology; Humans; Interleukin-1; Kinetics

Hemoglobin released from hemolysed erythrocytes has been postulated to be responsible for delayed cerebral vasospasm after subarachnoid hemorrhage (SAH). However, the evidence is indirect and the mechanisms of action are unclear. Cerebrovascular tone is regulated by a dynamic balance of relaxing and contracting factors. Loss of the endothelium-derived relaxing factor-nitric oxide in the presence of oxyhemoglobin and overproduction of endothelin-1 stimulated by oxyhemoglobin have been postulated as causes of delayed cerebral vasospasm after SAH.. The authors aimed to investigate this hypothesis using in vivo microdialysis to examine time-dependent changes in the perivascular concentrations of oxyhemoglobin, deoxyhemoglobin, and methemoglobin in a primate model of SAH.. Nine cynomolgus monkeys underwent right-sided frontotemporal craniectomy and placement of a semipermeable microdialysis catheter adjacent to the right middle cerebral artery (MCA). Saline (control group, three animals) or an arterial blood clot (SAH group, six animals) was then placed around the MCA and the catheter. Arteriographically confirmed vasospasm had developed in all animals with SAH but in none of the control animals on Day 7. The dialysate was collected daily for 12 days. Levels of oxyhemoglobin, deoxyhemoglobin, and methemoglobin were measured by means of spectrophotometry. Perivascular concentrations of oxyhemoglobin, deoxyhemoglobin, and methemoglobin peaked on Day 2 in the control monkeys and could not be detected on Days 5 to 12. Perivascular concentrations of oxyhemoglobin and deoxyhemoglobin peaked on Day 7 in the SAH group, at which time the concentrations in the dialysate were 100-fold higher than in any sample obtained from the control animals. Methemoglobin levels increased only slightly, peaking between Days 7 and 12, at which time the concentration in the dialysate was 10-fold higher than in samples from the control animals.. This study provides in vivo evidence that the concentrations of oxyhemoglobin and deoxyhemoglobin increase in the cerebral subarachnoid perivascular space during the development of delayed cerebral vasospasm. The results support the hypothesis that oxyhemoglobin is involved in the pathogenesis of delayed cerebral vasospasm after SAH and implicate deoxyhemoglobin as a possible vasospastic agent.

Topics: Analysis of Variance; Animals; Catheterization; Cerebral Angiography; Cerebral Arteries; Craniotomy; Disease Models, Animal; Endothelin-1; Exudates and Transudates; Hemoglobins; Hemolysis; Ischemic Attack, Transient; Macaca fascicularis; Methemoglobin; Microdialysis; Nitric Oxide; Oxyhemoglobins; Sodium Chloride; Spectrophotometry; Subarachnoid Hemorrhage; Time Factors

Although phorbol esters, synthetic activators of protein kinase C (PKC), can stimulate large increases in the binding of cytosolic PKC to form membrane-bound PKC (PKCm, an indicator of PKC activation), the authors report that even small increases in PKCm induced by phorbol esters (8-12% of total PKC content) can be associated with significant PKC-mediated contractions in vitro (50-85% of maximum) in normal canine cerebral arteries. Increases in PKCm of similarly small magnitude were found in vitro when control artery segments were exposed to hemolysate, but only if the arterial smooth-muscle cells were first slightly depolarized by increased extracellular potassium to values of membrane potential similar to those observed in canine cerebral arteries during chronic cerebral vasospasm. These increases in PKCm (6-8% of total PKC content) coincided with a greatly augmented contractile response to hemolysate. These results show that the previous observation of only a small increase in PKCm (approximately 7% of total PKC content) after experimental subarachnoid hemorrhage in the canine model does not preclude a potentially important role for PKC-mediated contraction in the pathogenesis of cerebral vasospasm.

Topics: Animals; Cell Membrane; Cerebral Arteries; Chronic Disease; Cytosol; Dogs; Endothelin-1; Enzyme Activation; Enzyme Inhibitors; Hemolysis; Ischemic Attack, Transient; Membrane Potentials; Muscle, Smooth, Vascular; Naphthalenes; Phorbol 12,13-Dibutyrate; Phorbol Esters; Potassium; Protein Binding; Protein Kinase C; Stress, Mechanical; Subarachnoid Hemorrhage; Vasoconstriction; Vasoconstrictor Agents

The release of endothelin from various blood cell fractions was investigated. Human as well as rat blood cell fractions homogenized by sonification were incubated in buffer for up to 60 min. Neither in platelet nor leukocyte homogenates from either species could immunoreactive endothelin be detected. In contrast, homogenates of red blood cells from both species showed a rapid and time-dependent rise of immunoreactive endothelin levels, reaching a peak at 15 min and decreasing thereafter. However, at time point 0 no immunoreactive endothelin could be detected. Reverse phase high performance liquid chromatography showed immunoreactive endothelin to consist of endothelin-1 as well as big endothelin-1. The release of immunoreactive endothelin in human and rat homogenates was concentration-dependently inhibited by the protease inhibitors, leupeptin, phosphoramidon, chymostatin and pepstatin A in order of increasing potency. Intact red blood cells did not incorporate [125I]endothelin-1 nor did they transform exogenous big endothelin-1 to endothelin-1. However, haemolysis of red blood cells with hypotonic saline (0.2%) or incubation with pore-forming staphylococcal alpha-toxin induced the release of immunoreactive endothelin into the buffer samples. Thus, apart from the indirect vasoconstrictor, haemoglobin, red blood cells can also liberate the direct vasoconstrictor, endothelin, a finding expected to be of considerable pathophysiological significance.

Topics: Animals; Bacterial Toxins; Chromatography, High Pressure Liquid; Endothelin-1; Endothelins; Erythrocytes; Hemolysin Proteins; Hemolysis; Humans; In Vitro Techniques; Male; Protease Inhibitors; Protein Precursors; Radioimmunoassay; Rats; Rats, Wistar