Nitric oxide down-regulates MKP-3 mRNA levels: involvement in endothelial cell protection from apoptosis.

MAP kinase-dependent phosphorylation processes have been shown to interfere with the degradation of the antiapoptotic protein Bcl-2. The cytosolic MAP kinase phosphatase MAP kinase phosphatase-3 (MKP-3) induces apoptosis of endothelial cells in response to tumor necrosis factor alpha (TNFalpha) via dephosphorylation of the MAP kinase ERK1/2, leading to Bcl-2 proteolysis. Here we report that the endothelial cell survival factor nitric oxide (NO) down-regulated MKP-3 by destabilization of MKP-3 mRNA.

Reactive oxygen species and vascular cell apoptosis in response to angiotensin II and pro-atherosclerotic factors.

Reactive oxygen species (ROS) are known to induce apoptotic cell death in various cell types. In the vessel wall, ROS can be formed by macrophages within the atherosclerotic plaque or can act on the endothelium after adhesion of monocytes or leucocytes. Moreover, ROS are endogenously synthesized by endothelial and vascular smooth muscle cells by NAD(P)H oxidase.

Allele-specific regulation of matrix metalloproteinase-12 gene activity is associated with coronary artery luminal dimensions in diabetic patients with manifest coronary artery disease.

Both the processes of atherosclerosis and plaque rupture are indicated to be influenced by matrix metalloproteinase (MMP) activity. We therefore searched for common functional variation in the matrix metalloelastase (MMP-12) gene locus that may be implicated in coronary artery disease. Single-strand conformation polymorphism analysis of DNA from healthy individuals detected a common polymorphism within the MMP-12 gene promoter (an A-to-G substitution at position -82).

Ubiquitin-mediated degradation of the proapoptotic active form of bid. A functional consequence on apoptosis induction.

Under basal conditions, the proapoptotic protein Bid is a long-lived protein. Pro-apoptotic stimuli such as tumor necrosis factor-alpha (TNFalpha) or Fas induce its caspase-8-mediated cleavage into two fragments. The COOH-terminal cleavage fragment of Bid (tBid) becomes localized to mitochondrial membranes and triggers the release of cytochrome c.

Inhibition of endogenous nitric oxide synthase potentiates ischemia-reperfusion-induced myocardial apoptosis via a caspase-3 dependent pathway.

Objective: Apoptosis of cardiomyocytes may contribute to ischemia-reperfusion injury. The role of nitric oxide (NO) in apoptosis is controversial. Therefore, we investigated the effect of NO synthase inhibition on apoptosis of cardiomyocytes during ischemia and reperfusion and elucidated the underlying mechanisms.

Phosphorylation and activation of the endothelial nitric oxide synthase by fluid shear stress.

Fluid shear stress activates the endothelial nitric oxide (NO) synthase (eNOS) by a mechanism which does not require an increase in the intracellular concentration of free Ca2+ ([Ca2+]i), and is sensitive to several kinase inhibitors. Although phosphorylation of eNOS has been suggested to regulate enzyme activity, the mechanism of eNOS activation is still unclear. Here we demonstrate that fluid shear stress elicits the phosphorylation of eNOS on tyrosine and serine residues.

Insulin-mediated stimulation of protein kinase Akt: A potent survival signaling cascade for endothelial cells.

Insulin exerts potent antiapoptotic effects in neuronal cells and has been suggested to promote angiogenesis. Therefore, we investigated whether insulin inhibits tumor necrosis factor-alpha (TNF-alpha)-induced apoptosis in human umbilical vein endothelial cells (HUVECs). Because insulin has been shown to stimulate the protein kinase Akt, we investigated whether activation of Akt contributes to the apoptosis-suppressive effect of insulin and characterized the downstream signaling pathway.

Posttranslational modification of Bcl-2 facilitates its proteasome-dependent degradation: molecular characterization of the involved signaling pathway.

The ratio of proapoptotic versus antiapoptotic Bcl-2 members is a critical determinant that plays a significant role in altering susceptibility to apoptosis. Therefore, a reduction of antiapoptotic protein levels in response to proximal signal transduction events may switch on the apoptotic pathway. In endothelial cells, tumor necrosis factor alpha (TNF-alpha) induces dephosphorylation and subsequent ubiquitin-dependent degradation of the antiapoptotic protein Bcl-2.

Laminar shear stress upregulates the complement-inhibitory protein clusterin : a novel potent defense mechanism against complement-induced endothelial cell activation.

Background: The complement system is implicated in the pathogenesis of atherosclerosis. Complement has been shown to activate endothelial cells (ECs) by inducing a proinflammatory response. Physiological levels of shear stress exert potent antiatherosclerotic effects.

Preoperative high dose methylprednisolone improves patients outcome after abdominal surgery.

Objective: To assess the effect of preoperative high dose methylprednisolone on stress response and outcome.

Design: Randomised, placebo-controlled, double-blind study.

Setting: University hospital, Germany.

Nitric oxide-an endothelial cell survival factor.

Due to its unique position in the vessel wall, the endothelium acts as a barrier and thereby controls adhesion, aggregation and invasion of immune competent cells. Apoptosis of endothelial cells may critically disturb the integrity of the endothelial monolayer and contribute to the initiation of proinflammatory events. Endothelial cell apoptosis is counteracted by nitric oxide synthesised by the endothelium nitric oxide synthase (eNOS).

Oxidative stress mediates apoptosis induced by oxidized low-density lipoprotein and oxidized lipoprotein(a).

We measured oxidative stress caused by lipoproteins in human umbilical vein endothelial cells and rabbit aorta. Oxidized lipoprotein(a) was the more potent stimulus over oxidized low density lipoprotein, and we believe that both influence the pathogenesis of atherosclerosis.

Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation.

Nitric oxide (NO) produced by the endothelial NO synthase (eNOS) is a fundamental determinant of cardiovascular homesotasis: it regulates systemic blood pressure, vascular remodelling and angiogenesis. Physiologically, the most important stimulus for the continuous formation of NO is the viscous drag (shear stress) generated by the streaming blood on the endothelial layer. Although shear-stress-mediated phosphorylation of eNOS is thought to regulate enzyme activity, the mechanism of activation of eNOS is not yet known.

Dephosphorylation targets Bcl-2 for ubiquitin-dependent degradation: a link between the apoptosome and the proteasome pathway.

Injury of the endothelial cells by the induction of apoptotic cell death may play an important role in the pathophysiology of atherosclerosis and the progression of inflammatory diseases. Here, we demonstrate an essential role for the ubiquitin-dependent proteasome complex in stimulus-induced degradation of the antiapoptotic protein Bcl-2. Bcl-2 is specifically degraded after stimulation of human endothelial cells with tumor necrosis factor (TNF)-alpha in a process that is inhibited by specific proteasome inhibitors.

Nitric oxide and apoptosis.

Apoptosis occurs during normal development of an organism but also plays an important role under pathophysiological conditions. Nitric oxide (NO) exhibits a double-edged role in apoptosis induction. Both pro- and antiapoptotic effects of NO have been demonstrated.

Lp(a) and LDL induce apoptosis in human endothelial cells and in rabbit aorta: role of oxidative stress.

Background: Atherogenic lipoproteins cause injury to the vascular wall in the early phase of atherogenesis. We assessed the effects of native (nLDL) and oxidized (oxLDL) low-density lipoprotein (LDL) and lipoprotein (a) [Lp(a)] on O2- formation and cell death in cultured human umbilical vein endothelial cells (HUVECs) and rabbit aorta (RA).

Methods And Results: O2- formation of HUVECs and RA segments was not influenced by nLDL, but was dose dependently increased by oxLDL and was moderately increased by nLp(a).

Upregulation of superoxide dismutase and nitric oxide synthase mediates the apoptosis-suppressive effects of shear stress on endothelial cells.

Physiological levels of laminar shear stress completely abrogate apoptosis of human endothelial cells in response to a variety of stimuli and might therefore importantly contribute to endothelial integrity. We show here that the apoptosis-suppressive effects of shear stress are mediated by upregulation of Cu/Zn SOD and NO synthase. Shear stress-mediated inhibition of endothelial cell apoptosis in response to exogenous oxygen radicals, oxidized LDL, and tumor necrosis factor-alpha was associated with complete inhibition of caspase-3-like activity, the central effector arm executing the apoptotic cell death program in endothelial cells.

Nitric oxide inhibits caspase-3 by S-nitrosation in vivo.

In cultured human endothelial cells, physiological levels of NO prevent apoptosis and interfere with the activation of the caspase cascade. In vitro data have demonstrated that NO inhibits the activity of caspase-3 by S-nitrosation of the enzyme. Here we present evidence for the in vivo occurrence and functional relevance of this novel antiapoptotic mechanism.

Immunohistochemical detection of fibronectin in postmortem incised wounds of porcine skin.

Fibronectin plays an important role in tissue repair and wound healing. Previous literature reports indicated that fibronectin could be a marker of vitality for wounds with a survival time of more than a few minutes. In order to verify these findings were performed immunohistochemical investigations on the expression of fibronectin in incised wounds of porcine skin inflicted into various anatomical regions in the early postmortem interval (0-5 min after circulatory arrest).

Cyclosporin A inhibits apoptosis of human endothelial cells by preventing release of cytochrome C from mitochondria.

Background: Several experimental and clinical studies suggest that cyclosporin A (CSA) treatment reduces transplant atherosclerosis. Because oxidized LDL (oxLDL) is believed to play a key role in the development of atherogenesis, causing injury to the endothelium, and has been shown to induce apoptosis of endothelial cells, we investigated whether CSA inhibits oxLDL-induced apoptosis.

Methods And Results: Apoptosis was induced in human umbilical venous endothelial cells (HUVECs) by incubation of 10 microg/mL oxLDL for 18 hours.

Fluid shear stress stimulates phosphorylation of Akt in human endothelial cells: involvement in suppression of apoptosis.

Fluid shear stress alters the morphology and function of the endothelium by activating several kinases. Furthermore, shear stress potently inhibits apoptosis of endothelial cells. Since activation of Akt kinase has been shown to prevent cell death, we investigated the effects of shear stress on Akt phosphorylation.

Platelet glycoprotein IIIa polymorphisms and risk of coronary stent thrombosis.

Background: Coronary stents are an effective treatment for selected coronary stenoses. However, thrombosis of the stented segment is a major adverse complication. Platelet aggregation has a key role in stent thrombosis.

Nitric oxide inhibits APO-1/Fas-mediated cell death.

Activation of the cysteine protease caspases, which are homologous to the product of Caenorhabditis elegans cell-death gene ced 3, is required to mediate APO-1/Fas-induced apoptosis. We report here that nitric oxide (NO) released by exogenous NO donors, as well as NO endogenously derived by transfection with the inducible NO synthase, substantially suppresses APO-1/Fas-triggered cell death of Jurkat cells. The inhibitory NO effect was independent of cGMP, because 8-bromo-cGMP did not influence APO-1/Fas-mediated apoptosis.