Endothelium : journal of endothelial cell research最新文献

The authors hypothesized that matrix metalloproteinase (MMP)-2, -9, and tissue inhibitor metalloproteinase (TIMP)-1, -2 would be abnormal in acute coronary syndromes (ACSs). MMP-2, -9, and TIMP-1, -2 plasma levels were measured in diabetic patients with ACSs compared to nondiabetic patients with ACSs. A total of 46 diabetic and 78 nondiabetic patients with ACSs were enrolled. The following parameters were measured: body mass index (BMI), glycosylated hemoglobin (HbA1c), fasting plasma glucose (FPG), fasting plasma insulin (FPI), homeostasis model assessment index (HOMA index), systolic blood pressure (SBP), diastolic blood pressure (DBP), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglycerides (Tg), lipoprotein(a) [Lp(a)], plasminogen activator inhibitor-1 (PAI-1), homocysteine (Hct), fibrinogen (Fg), high-sensitivity C-reactive protein (hs-CRP), and plasma levels of MMP-2, MMP-9, TIMP-1, and TIMP-2. Significant HbA1c, FPG, FPI, HOMA index, DBP, Tg, Hct, and Fg increases were present in the diabetic group with ACSs, whereas hs-CRP was lower in these patients compared to nondiabetic patients with ACSs. MMP-9, TIMP-1, and TIMP-2 plasma levels were higher in diabetic patients with ACSs compared to nondiabetic patients with ACSs. MMP-9, TIMP-1, and TIMP-2 plasma levels were increased in diabetic patients with ACSs, which may reflect abnormal extracellular matrix metabolism in diabetes during acute event.

Impaired vasoactive release of opposing vasodilator and vasoconstrictor mediators due to endothelial dysfunction is integral to the pathogenesis of diabetic retinopathy. The aim of this study was to determine the effect of hyperglycemia on the expression of endothelial nitric oxide synthase (eNOS) and the release of nitric oxide (NO) in bovine microvascular retinal endothelial cells (BRECs) under both static (basal and acetylcholine stimulated) and flow (laminar shear stress [10 dynes/cm2 and pulsatile flow 0.3 to 23 dynes/cm2) conditions using a laminar shear apparatus and an in vitro perfused transcapillary culture system. The activity and expression of eNOS, measured by nitrate levels and immunoblot, respectively, were determined following exposure of BRECs to varying concentrations of glucose and mannitol (0 to 25 mM). Under static conditions the expression of eNOS decreased significantly following exposure to increasing concentrations of glucose when compared to osmotic mannitol controls and was accompanied by a significant dose-dependent decrease in nitrate levels in conditioned medium. The acetylcholine stimulated increase in NO release (2.0 +/- 0.3-fold) was significantly reduced by 55% +/- 5% and 65% +/- 4.5% following exposure to 16 and 25 mM glucose, respectively, when compared to osmotic controls. In parallel studies, glucose significantly inhibited both laminar shear stress and pulsatile flow-induced activity when compared to mannitol. We conclude that hyperglycemia impairs agonist- and flow-dependent release of NO in retinal microvascular endothelial cells and may thus contribute to the vascular endothelial dysfunction and impaired autoregulation of diabetic retinopathy.

Abundant evidence documents the highly proinflammatory actions of lysophosphatidylcholine (LPC). Further, LPC, found in high amounts in oxidized low-density lipoprotein (LDL), is implicated as an atherogenic factor. In endothelial cells, LPC impairs endothelial barrier function through GPR4, a novel receptor hypothesized to be sensitive to LPC and protons. The authors investigated the stimulation by LPC or low pH of GPR4 in human brain microvascular endothelial cells (HBMECs) and whether the activated GPR4 regulates in vitro monocyte transmigration. The results indicated that HBMECs stimulated by LPC (5 microM), but not low pH, showed a twofold increase in monocyte transmigration. Using retroviruses containing siRNA to GPR4, a > 60% reduction of GPR4 expression resulted in blockade of the LPC-stimulated transmigration. The inhibited response was restored by co-expression with an small interference RNA (siRNA)-resistant, but functional, GPR4 mutant construct. To investigate potential signaling mechanisms, the siRNA-mediated knockdown of GPR4 also prevented LPC-induced RhoA activation. C3 transferase, a Rho inhibitor, prevented approximately approximately 65% of the LPC-stimulated transmigration. LPC also increased MLC phosphorylation by 5 min, which was inhibited by the Rho kinase inhibitor, Y-27632 (10 microM) or ML-7 (myosin light chain kinase (MLCK) inhibitor). The findings indicate that the proinflammatory and atherogenic LPC stimulated endothelial GPR4, which promoted monocyte transmigration through a RhoA-dependent pathway.

Endocardial endothelial cells (EECs), which form the inner lining of the cavities of the heart, are a distinct cell population whose dysfunction can be critical in pathological conditions of heart. Insights into the role and organization of these cells in pathological states of the heart are limited mainly due to a dearth of experimental models. To date no endocardial endothelial cell line is available. The authors attempted to immortalize porcine ventricular EECs by transfecting the cells with human telomerase reverse transcriptase (hTERT). EECs immortalized by ectopic expression of hTERT exhibit phenotypic and functional characteristics similar to primary EECs. The EE cell line could be useful for the study of mechanisms involved in the interaction of EECs with the underlying myocardium and cardiac interstitium and as useful tools in understanding their role in diseased states of heart.

The endothelium is crucial in the maintenance of normal vascular function and disturbance of this balance is a key early event in the development of vascular disease. A wide range of techniques currently exists for assessment of endothelial function in both the coronary and the peripheral vasculature. Many of these in vivo tests have concentrated on measuring nitric oxide bioavailability, however more recently methods for measuring other vascular parameters, such as tissue-plasminogen activator release, have been used. Furthermore indirect systemic measurements of endothelial function and endothelial progenitor cells have been investigated. These methods have given great insights into the pathophysiology of atherosclerosis and have aided in the development of a number of antiatherosclerotic therapies. Importantly the methods used to date for assessing endothelial function in vivo are accurate, reproducible and correlate with the future risk of cardiovascular events. The development of new techniques and the constant refinement of established techniques suggest that many more insights are to be gained from clinical assessment of endothelial function.

The focal development of atherosclerosis in the vascular tree may be explained in part by the local nature of blood flow. Bifurcations and branching points, prone to early atherogenesis, experience disturbed and oscillatory flow, whereas straight vascular regions, resistant to atherosclerosis, are exposed to steady laminar flow. A large number of studies suggest that the antiatherosclerotic effects of laminar flow are in part due to the ability of flow to modulate endothelial cell phenotype. Under steady laminar flow, endothelial cells generate molecules that promote a vasoactive, anticoagulant, anti-inflammatory, and growth-inhibitory surface. In contrast, disturbed flow induces a proliferative, prothrombotic, and adhesive phenotype. Endothelial cells are able to sense the variations of flow via mechanosensitive cell surface proteins and to transduce these signals via intracellular pathways to transcription factors in the nucleus leading to phenotypic changes. This review summarizes the "outside-in" signaling events initiated by flow that modulate endothelial cell phenotype.

The vascular endothelium contributes to and is affected by inflammatory processes. Disturbance of the endothelium's morphologic and functional integrity in response to mechanical, immunologic, and chemical injuries reflects the first step in the pathophysiological cascade of atherosclerotic disorders. At the site of an endothelial injury, invading inflammatory cells producing numerous proinflammatory factors promote and amplify both local and systemic inflammation. These early changes on a cellular and subcellular level that precede the clinical manifestation of atherosclerosis are associated with loss of profound physiological functions of the endothelium. One pivotal function of the endothelium is nitric oxide-mediated regulation of vessel tone and blood flow according to the local requirements. The assessment of nitric oxide-mediated endothelial function by different methods revealed a close relation between inflammatory activation and endothelial dysfunction in healthy volunteers, patients at risk, and patients with established cardiovascular disease. Moreover, anti-inflammatory therapeutic interventions do not only have a positive impact on disease progression, but also on endothelial function, thus further providing an indirect line of evidence linking inflammation with endothelial dysfunction.

This article discusses the importance of the endothelium for successful vascular grafts derived from both native arteries and synthetic materials. It also discusses the fundamental strategies to endothelialize synthetic grafts in animal experiments and in the clinic, as well as the use of endothelial progenitor cells (EPCs), bone marrow-derived cells, and mesothelium as endothelial substitutes.

The identification of circulating endothelial progenitor cells (EPCs) has prompted an explosion of interest in postnatal vasculogenesis and the role of this mechanism in human health and disease. Previously considered restricted to the embryonic phase, the differentiation in situ of progenitor cells to vascular endothelium is now known to occur in the adult. A role for EPCs in the modulation of angiogenesis has also been recognized. These cells are enriched in the mononuclear cell fraction of peripheral blood but have also been isolated from bone marrow, the vessel wall, and a number of other organs and tissues. Accumulating data suggest an important vasculoprotective function for EPCs, although a maladaptive role underpinning a variety of angiogenesis-dependent diseases is also being investigated. Encouraging results observed with experimental and early human trials of EPC-based regenerative therapies have further underscored the significance of this recently discovered cell type. Notwithstanding the scope and pace of these developments, a number of challenges remain: the precise ontogeny and lineage of these cells is unknown, the true extent to which EPCs participate in neovascularization and vascular repair is still uncertain, and the efficacy of EPC-based regenerative therapies has yet to be proven in randomized controlled trials.

Technologies in interventional Cardiology have evolved from balloon to mechanical ablation, atherectomy, stenting, and brachytherapy to current drug eluting interventional strategies. New challenges are to develop techniques that not only prevent restenosis, but also promote vascular and endothelial healing after (balloon) injury. Endothelial healing approaches range from preventing endothelial injury to restoring endothelial function and reendothelialization by pharmacotherapy and cell therapy. These novel healing strategies warrant further exploration as they may represent an alternative to drug-eluting stent approaches.