Carbon Monoxide-Releasing Molecule-Derived CO Regulates Tissue Factor, Plasminogen Activator Inhibitor Type 1, and Thrombomodulin Production by Human Endothelial Cells.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1146-1146
Author(s):  
Keiko Maruyama ◽  
Eriko Morishita ◽  
Shigeki Ohtake ◽  
Akihiro Yachie ◽  
Shinji Nakao
Keyword(s):  
Plasminogen Activator Inhibitor ◽  
Mapk Signaling ◽  
Mrna Levels ◽  
Protein Levels ◽  
Tnf Α ◽  
Inhibitor Type ◽  
Activator Inhibitor Type ◽  
Activator Inhibitor ◽  
Pai 1

Abstract Abstract 1146 Background and purpose: Heme oxygenase-1 (HO-1) is a cytoprotective and anti-inflammatory protein that catalyzes the conversion of heme into biliverdin, free iron, and carbon monoxide (CO). HO-1 is rapidly induced by various oxidative stresses and inflammation, thereby playing an important role in the self defence system. Recently, Yachie, et al. reported the first human case of HO-1 deficiency. This patient showed prominent signs of intravascular hemolysis, endothelial cell injury, and abnormalities in the coagulation / fibrinolysis system, suggesting the involvement of HO-1 or HO-1 products, such as CO, regulation of coagulation / fibrinolysis system. The current study examined whether tricarbonyldichlororuthenium (II) dimer (CORM-2), which liberated CO in the presence of dimethyl sulfoxide (DMSO), modulates the expression of tissue factor (TF), plasminogen activator inhibitor type 1 (PAI-1) and thrombomodulin (TM) in human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were pretreated with CORM-2 at the concentration of 50 μM for 3h, washed and stimulated with tumor necrosis factor-α(TNF-α, 10 ng/ml) for additional 2 or 5h. The mRNA and protein levels of TF, PAI-1 and TM in the cultured HUVECs were determined by real-time reverse transcriptase polymerase chain reaction and Western blotting. To determine whether CORM-2 affects the MAPK signaling pathways, the phosphorylation of p38 mitogen-activated protein kinase (p38MAPK) and extracellular signal-regulated kinase1/2 (ERK1/2) in the HUVECs were analyzed with Western blotting. Results: After TNF-α stimulation, TF mRNA levels were approximately 6-fold and PAI-1 mRNA levels were approximately 8-fold increased, and TM mRNA levels were decreased by 20% compared to the control. Similarly to the mRNA expression, TF and PAI-1 protein levels were increased while the TM protein level was decreased. On the other hand, pretreatment of HUVECs with CORM-2 significantly decreased TF mRNA levels (approximately 80% suppression), and PAI-1 mRNA levels (approximately 90% suppression) while increased TM mRNA levels by 3-fold as compared to the TNF-α-stimulated group (p<0.05; Figure 1). Similarly, the pretreatment with CORM-2 inhibited TNF-α-induced TF and PAI-1 protein up-regulation and TM protein down-regulation. CORM-2 inhibited TNF-α-induced activation of p38MAPK and ERK1/2 by 50% compared to the control. Conclusions: These results indicate that CO liberated by CORM-2 suppresses the TNF-α-induced TF and PAI-1 up-regulation and prevents the TNF-α-induced TM down-regulation in HUVECs via MAPK signaling pathways. Thus, the modulation of endothelial function by CO may offer a novel antithrombotic option for treatment of the hypercoagulable state associated with inflammation. Disclosures: No relevant conflicts of interest to declare.

Physiological testosterone stimulates tissue plasminogen activator and tissue factor pathway inhibitor and inhibits plasminogen activator inhibitor type 1 release in endothelial cellsThis paper is one of a selection of papers in this Special Issue, entitled International Symposium on Recent Advances in Molecular, Clinical, and Social Medicine, and has undergone the Journal's usual peer-review process.

2007 ◽  
Vol 85 (2) ◽  
pp. 246-251 ◽  
Author(s):  
Hong Jin ◽  
Jijin Lin ◽  
Lu Fu ◽  
Yi-Fang Mei ◽  
Geng Peng ◽  
...  
Keyword(s):  
Plasminogen Activator ◽  
Plasminogen Activator Inhibitor ◽  
Mrna Levels ◽  
Tissue Plasminogen ◽  
Tissue Factor Pathway ◽  
Inhibitor Type ◽  
Activator Inhibitor Type ◽  
Activator Inhibitor ◽  
Pai 1

There is a striking gender difference in atherosclerotic vascular disease. For decades, testosterone was considered detrimental to the cardiovascular system. Recent studies, however, have presented some alternative results. The aim of this study was to evaluate the effect of testosterone, using physiological and supraphysiological concentrations, on antigen and mRNA levels of tissue plasminogen activator (tPA), plasminogen activator inhibitor type 1 (PAI-1), and tissue factor pathway inhibitor (TFPI) released by human umbilical vein endothelial cells and to investigate the cellular mechanism. Cells within 2–3 passages were cultured in 25 cm2 flasks or plated onto 96-well plates with a density of about 1 × 105 cells/mL as recommended. The cells were incubated in the presence or absence of testosterone (3, 30, 3 × 103, 3 × 104 nmol/L) for 48 h. Levels of tPA, PAI-1, and TFPI antigen were assayed with ELISA kits. Reverse transcriptase PCR was carried out to detect tPA, PAI-1, and TFPI mRNA levels. Cells were incubated in androgen-receptor antagonist (flutamide 10 µmol/L) or aromatase inhibitor (aminoglutethimide 50 µmol/L) for 3 h, and then the experiments were repeated. Testosterone at a physiologic concentration (30 nmol/L) increased the antigen levels of tPA and TFPI significantly (P < 0.05). However, tPA and TFPI levels were markedly reduced (P < 0.05) at a larger dose (3 × 104 nmol/L). On the other hand, PAI-1 antigen levels decreased significantly at the testosterone concentrations ranging from 3 to 3 × 104 nmol/L (P < 0.05). The change in the levels of tPA and TFPI were reflected in the corresponding change in mRNA levels. Flutamide attenuated the effect of testosterone at physiological concentration (30 nmol/L). The results demonstrated that testosterone at physiological concentrations may have a beneficial influence on the haemostatic system through enhancement of anticoagulant activity, resulting from stimulation of TFPI and tPA expression and inhibition of PAI-1 secretion by the endothelium.

The Production of Heparin Cofactor li Is Not Regulated by Inflammatory Cytokines in Human Hepatoma Cells: Comparison with Plasminogen Activator Inhibitor Type-1

1996 ◽  
Vol 75 (02) ◽  
pp. 298-302 ◽  
Author(s):  
Chisato Kolke ◽  
Yumiko Hayakawa ◽  
Kenji Niiya ◽  
Nobuo Sakuragawa ◽  
Hideo Sasaki
Keyword(s):  
Plasminogen Activator Inhibitor ◽  
Mrna Levels ◽  
Hep G2 ◽  
Hep G2 Cells ◽  
Tnf Α ◽  
G2 Cells ◽  
Inhibitor Type ◽  
Activator Inhibitor Type ◽  
Activator Inhibitor

SummaryUsing the Northern blot technique, we screened 6 human hepatoma cell lines to investigate the regulation mechanism of heparin cofactor II (HCII) biosynthesis. We found that HuH-7 and Hep G2 cells constitutively expressed the HC II gene. In conditioned medium, HuH-7 cells constantly produced HC II that was functionally active and formed a complex with thrombin in the presence of dermatan sulfate. HC II is thought be an acute phase reactant, and, therefore, we examined the effects of the major inflammatory cytokines, IL-6, IL-1β, and TNF-α, on the regulation of HC II production in HuH-7 and Hep G2 cells. In HuH-7 cells, the antigen and mRNA levels of plasminogen activator inhibitor type-1 (PAI-1), an acute phase protein produced by hepa-tocytes, were increased in response to stimulation with either IL-6 or IL-1 (3 or both, but HC II antigen and mRNA levels were not changed by the same stimulation. Even when Hep G2 cells were treated with a combination of three cytokines, IL-6, IL-1β, and TNF-α, HC II antigen and mRNA levels were not changed; however, PAI-1 antigen and mRNA levels were clearly increased. These results suggest that the production of HC II in hepatoma cells is not regulated by the major inflammatory mediators, IL-6, IL-iβ, and TNF-α.

Limited Proteolysis of Vitronectin by Plasmin Destroys Heparin Binding Activity

1991 ◽  
Vol 66 (03) ◽  
pp. 310-314 ◽  
Author(s):  
David C Sane ◽  
Tammy L Moser ◽  
Charles S Greenberg
Keyword(s):  
Limited Proteolysis ◽  
Plasminogen Activator Inhibitor ◽  
Binding Activity ◽  
Binding Domain ◽  
Heparin Binding ◽  
Inhibitor Type ◽  
Activator Inhibitor Type ◽  
Activator Inhibitor ◽  
Pai 1

SummaryVitronectin (VN) stabilizes plasminogen activator inhibitor type 1 (PAI-1) activity and prevents the fibrin(ogen)-induced acceleration of plasminogen activation by t-PA. These antifibrinolytic activities as well as other functions are mediated by the glycosaminoglycan (GAG) binding domain of VN. Since the GAG binding region is rich in arginyl and lysyl residues, it is a potential target for enzymes such as plasmin. In this paper, the dose and time-dependent proteolysis of VN by plasmin is demonstrated. The addition of urokinase or streptokinase (200 units/ml) to plasma also produced proteolysis of VN. With minimal proteolysis, the 75 kDa band was degraded to a 62-65 kDa form of VN. This minimal proteolysis destroyed the binding of [3H]-heparin to VN and reversed the neutralization of heparin by VN.Thus, the plasmin-mediated proteolysis of the GAG binding activity of VN could destroy the antifibrinolytic activity of VN during physiologic conditions and during thrombolytic therapy. Furthermore, other functions of VN in complement and coagulation systems that are mediated by the GAG binding domain may be destroyed by plasmin proteolysis.

Adiponectin is inversely related to plasminogen activator inhibitor type 1 in patients with stable exertional angina

2004 ◽  
Vol 91 (05) ◽  
pp. 1026-1030 ◽  
Author(s):  
Hidetomo Maruyoshi ◽  
Tohru Funahashi ◽  
Shinzo Miyamoto ◽  
Jun Hokamaki ◽  
Hirofumi Soejima ◽  
...  
Keyword(s):  
Plasminogen Activator ◽  
Plasma Levels ◽  
Plasminogen Activator Inhibitor ◽  
Plasminogen Activator Inhibitor Type ◽  
Exertional Angina ◽  
Inhibitor Type ◽  
Activator Inhibitor Type ◽  
Activator Inhibitor ◽  
Pai 1

SummaryAdipose tissue is a secretory organ producing a variety of bioactive substances, such as adiponectin. Adiponectin has antiatherogenic properties while plasminogen activator inhibitor type 1 (PAI-1) is closely involved in the development of atherosclerosis. The relationship between adiponectin and PAI-1 in patients with coronary artery disease (CAD) has not been clarified. This study examined plasma levels of adiponectin and PAI-1 in 64 patients with stable exertional angina (SEA) and 65 patients with the chest pain syndrome (CPS). Plasma logadiponectin levels were significantly lower in patients with SEA (0.62±0.08 µg/dL) compared to those with CPS (0.86± 0.05 µg/dL) (p<0.0001). The plasma levels of log-PAI-1 were significantly higher in patients with SEA (1.23±0.18 ng/mL) compared to those with CPS (1.15±0.22 ng/mL) (p<0.05). Plasma log-adiponectin levels correlated negatively with diabetes mellitus (DM), body mass index (BMI), log-PAI-1 (r=−0.284, p<0.001), triglyceride (TG), and remnant-like particles cholesterol (RLP-C), and positively with high-density lipoprotein cholesterol (HDL-C) levels. Plasma levels of log-PAI-1 correlated positively with DM, BMI, TG and RLP-C levels, and negatively with HDL-C levels. Multiple logistic regression analysis identified sex, angina pectoris, and PAI-1 as independent determinants of hyperadiponectinemia (p<0.05). Adiponectin is inversely related to PAI-1. DM, BMI, TG, HDL-C, and RLP-C are common mediators between adiponectin and PAI-1, and treatment for common mediators may prevent the development of CAD by reducing PAI-1 and increasing adiponectin levels.

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