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T Kooistra

Publications and source records attributed to T Kooistra.

At least 55 records · Page 3Linked to original sources
PubMed1995

Stimulation of tissue-type plasminogen activator gene expression by sodium butyrate and trichostatin A in human endothelial cells involves histone acetylation.

We have previously shown that the pleiotropic agent sodium butyrate strongly stimulates tissue-type plasminogen activator (t-PA) expression in human umbilical vein endothelial cells (HUVEC). Here we provide the following evidence that the butyrate-induced t-PA expression in HUVEC involves histone H4 acetylation. (1) t-PA induction by butyrate occurs at the transcriptional level and does not require new protein synthesis, indicating a direct effect. (2) t-PA induction by butyrate can be fully mimicked by a specific, structurally unrelated, histone deacetylase inhibitor, trichostatin A. (3) At optimally stimulatory conditions, a combination of butyrate and trichostatin A does not enhance t-PA production more than each of the compounds alone, indicating that both compounds act through a common regulatory mechanism. (4) Induction of t-PA transcription by butyrate and trichostatin A was found to be preceded by histone H4 acetylation; at suboptimal inducing concentrations of butyrate and trichostatin A, the degree of acetylation of histone H4 caused by each agent was similarly reduced. These results are consistent with a role for histone H4 acetylation in t-PA induction by butyrate in HUVEC.

Acetylation↗
PubMed1995

Studies on the acute release of tissue-type plasminogen activator from human endothelial cells in vitro and in rats in vivo: evidence for a dynamic storage pool.

The process of acute release of tissue-type plasminogen activator (tPA) is important in locally speeding up fibrinolysis. Using a sensitive enzyme-linked immunosorbent assay for tPA, we investigated the acute release of tPA from cultured human umbilical vein endothelial cells. The addition of thrombin (0.003 to 3 NIH U/mL) caused the dose-dependent release of noncomplexed, enzymatically active tPA into the medium. The amount of tPA released into the medium by thrombin was similar to the difference in the amounts of tPA present in extracts from thrombin-treated cells and control cells. The process of acute release of tPA was complete in 1 minute, whereas the concomitant release of von Willebrand factor into the medium was slightly slower (maximum after 3 minutes). By increasing (c.q. decreasing) tPA synthesis, it was found that the amount of tPA constitutively secreted, the amount acutely released, and the amount in cell extracts were increased (c.q. decreased) to the same extent. The same relation was found in vivo. When rats were pretreated with cholera toxin or retinoic acid to increase tPA synthesis, plasma levels of tPA were increased, whereas acute release of tPA, as induced by bradykinin, was increased to the same extent. Acutely released tPA and constitutively secreted tPA were liberated from different pathways in human umbilical vein endothelial cells; tPA had, relative to the in vivo situation, a short residence time in the acutely releasable pathway.

Animals↗
PubMed1995

Tumor necrosis factor alpha induces endothelial galactosyl transferase activity and verocytotoxin receptors. Role of specific tumor necrosis factor receptors and protein kinase C.

Infections with verocytotoxin (VT) producing Escherichia coli have been strongly implicated in the epidemic form of hemolytic uremic syndrome (HUS). Endothelial damage plays a central role in the pathogenesis of HUS. In vitro studies have shown that VT can damage endothelial cells after interaction with its cellular receptor globotriaosylceramide (GbOse3cer). Cytokines, such as tumor necrosis factor alpha (TNF alpha) and interleukin-1 (IL-1) can potentiate the toxic effect of VT by inducing a protein-synthesis dependent increase in VT receptors on endothelial cells. In this study, the mechanisms underlying the increase in endothelial VT receptors induced by TNF alpha were studied in more detail. To investigate which proteins were involved in this induction, endothelial cells were incubated with and without TNF alpha in the presence of 14C-galactose or 14C-glucose. Thin-layer chromatography (TLC) analysis of the glycolipid extracts of these cells demonstrated a markedly enhanced incorporation of 14C-galactose in GbOse3cer and other galactose-containing glycolipids, suggesting that TNF alpha enhanced galactosyl-transferase activity. To examine the role of the two recently cloned TNF-receptors (TNFR-p75 and TNFR-p55) in the TNF alpha-induced increase in GbOse3cer in human endothelial cells, cells were incubated with TNF alpha, the TNFR-p55 selective R32W-S86T-TNF alpha-mutant, or the TNFR-p75 selective D143N-A145R-TNF alpha-mutant. The effect of TNF alpha activation, determined by binding-experiments with 125I-VT-1, could be largely, but not completely mimicked by R32W-S86T-TNF alpha. Although incubation of cells with D143N-A145R-TNF alpha did not show an increase in VT-1 binding, the monoclonal antibody utr-1, which prevents binding to TNFR-p75, decreased the TNF alpha-induced VT-1 binding. Activation of protein kinase C (PKC) by phorbol ester increases the expression of VT-1 receptors; this effect was prevented by the PKC inhibitor Ro31-8220 and by homologous desensitization by pretreatment with phorbol ester. In contrast, the presence of the protein kinase inhibitor Ro31-8220 or desensitization of PKC activity reduced the TNF alpha-induced increase in VT-1 receptors maximally by 50% and 24%, respectively. Comparable reductions in overall protein synthesis and the synthesis of E-selectin and plasminogen activator inhibitor-1 (PAI-1) were observed. This suggests an effect on general protein synthesis rather than a specific effect of PKC in the signal transduction pathway, by which TNF alpha induces VT-1 receptors. Our results indicate that TNF alpha can increase the VT-1 receptors on endothelial cells by inducing galactosyl-transferase activity, that this action of TNF alpha mainly occurs via the TNFR-p55; and that PKC activation increases expression of VT-1 receptors by a separate mechanism that acts additively to the TNF alpha-induced increase in VT-1 receptors.

Autoradiography↗
PubMed1995

Imbalance between intraperitoneal coagulation and fibrinolysis during peritonitis of CAPD patients: the role of mesothelial cells.

We compared peritoneal dialysis effluents from 18 CAPD patients who had not suffered from peritonitis during the last 6 months (group 1) with the effluents from five patients with acute peritonitis (group 2), measuring activation markers of coagulation and fibrinolysis. These markers included prothrombin fragment F1 + 2 (F1 + 2), thrombin-antithrombin III complex (TAT), fibrin monomer (FM), and fibrin degradation products (FbDP). In the dialysate of group 1 we found remarkably high levels of F1 + 2, TAT and FM concomitant with a high concentration of FbDP, indicating a high rate of intraperitoneal fibrin turnover. The balance between peritoneal generation and degradation of fibrin was disturbed in untreated patients of group 2, who had significantly higher levels of coagulation markers and a higher ratio between FM and FbDP. Seven days after treatment with intraperitoneal administration of antibiotics and heparin, F1 + 2, TAT, FM and FbDP decreased significantly. To evaluate the role of mesothelial cells (MC) in the high peritoneal fibrin turnover we investigated the expression of tissue-type plasminogen activator (t-PA), urokinase-type plasminogen activator (u-PA), plasminogen activator inhibitor type-1 (PAI-1), and tissue factor in cultured human peritoneal MC under basal conditions and after exposure to tumour necrosis factor alpha (TNF alpha), interleukin-1 alpha (IL-1 alpha), or bacterial lipopolysaccharide (LPS). The exposure of MC to TNF alpha or to a lesser extent IL-1 alpha or LPS reduced their fibrinolytic activity by decreasing t-PA production and increasing PAI-1 synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute Disease↗
PubMed1994

Genistein reduces tumor necrosis factor alpha-induced plasminogen activator inhibitor-1 transcription but not urokinase expression in human endothelial cells.

The plasminogen activator inhibitor PAI-1 is markedly elevated in vivo and in vitro upon exposure to the inflammatory mediators tumor necrosis factor alpha (TNF alpha), interleukin-1 (IL-1), and bacterial lipopolysaccharide. Here we report that the isoflavone compound genistein prevents the increase in synthesis of PAI-1 induced by these inflammatory mediators in human endothelial cells in vitro, and partially reduces the basal PAI-1 production by these cells. These effects of genistein were accompanied by a decrease in PAI-1 mRNA and in a suppression of the PAI-1 transcription rate as shown by run-on assay. A specific action of genistein, probably by inhibiting a tyrosine protein kinase, is likely, because the structural genistein analogue daidzein, which has a low tyrosine protein kinase inhibitor activity, did not inhibit PAI-1 synthesis. Vanadate, a tyrosine protein phosphatase inhibitor, increased PAI-1 production. The effect of genistein on PAI-1 synthesis was rather selective. Herbimycin A also reduced PAI-1 synthesis, but several other tyrosine protein kinase inhibitors, namely tyrphostin A47, methyl-2,5-dihydroxy-cinnamate, and compound 5, were unable to do so. All these tyrosine protein kinase inhibitors reduced basic fibroblast growth factor (b-FGF)-induced [3H]thymidine incorporation in endothelial cells. This indicates that the effect of genistein on PAI-1 transcription proceeds independently of its effect on mitogenesis. In contrast to TNF-alpha-induced PAI-1 production, the transcription and synthesis of urokinase-type plasminogen activator (u-PA) was not inhibited by genistein. A TNF-alpha-mutant (Trp32Thr86TNF alpha) that specifically recognizes the 55-kD TNF-receptor, mimicked the effects of TNF alpha on both PAI-1 and u-PA. Because genistein affected PAI-1, but not u-PA induced by this mutant, involvement of different TNF-receptors cannot underlie the difference in the effects of genistein on PAI-1 and u-PA synthesis. Because genistein also inhibited PAI-1 induction by thrombin and IL-4, it is likely that genistein does not act on a TNF alpha-receptor-coupled protein kinase but on the signal transduction pathway enhancing PAI-1 transcription. Our results suggest that the TNF alpha-induced signal transduction pathway of PAI-1 transcription involves a genistein-sensitive step that is not involved in the induction of u-PA by TNF alpha. Given the limited sensitivity to several other tyrosine protein kinase inhibitors, this genistein-sensitive step may be a potential target for pharmacologic intervention to reduce elevated plasma PAI-1 levels.

Cells, Cultured↗
PubMed1994

Regulation of endothelial cell t-PA synthesis and release.

The fibrinolytic activity of blood is to a large extent determined by the plasma level of tissue-type plasminogen activator (t-PA). Changes in the plasma level of t-PA are mainly achieved by the endothelium by two mechanisms: (a) a rapid, short-term release of t-PA, which occurs within minutes (acute release, regulated secretion) and (b) a long-term change in the rate of synthesis and constitutive secretion of t-PA. The rapid t-PA release response of the endothelium upon stimulation may play an important role in the dissolution of fibrin at the initial event of fibrin formation, and hence in the prevention of thrombus formation. The rate of constitutive t-PA secretion is an important determinant of the actual levels of t-PA under basal and stimulated conditions. Our insight into the regulation of the synthesis and release of t-PA has extended markedly in the last decade and still adapts by the continuous stream of new experimental data. In this review we summarize our present knowledge about the factors, including intracellular signalling pathways, promoter elements and transcription factors involved in the modulation of t-PA gene expression, with particular reference to the regulation in human endothelial cells. We also discuss the mechanisms underlying acute release of t-PA from the endothelium, and provide evidence that the regulated and constitutive secretion of t-PA are interrelated in several aspects.

Animals↗
PubMed1994

A sensitive ELISA for human tissue-type plasminogen activator applicable to the study of acute release from cultured human endothelial cells.

The development of a highly sensitive, enzyme-linked immunosorbent assay (ELISA) for tissue-type plasminogen activator (tPA) is described. The use of a biotin-avidin system resulted in a detection limit of 10 pg of tPA per ml, which is 50 to 150 times more sensitive than commercially-available ELISAs. Free tPA and tPA complexed to PA inhibitor type-1 were detected with equal efficiency. The ELISA proved suitable for measuring tPA antigen in conditioned media and in cell extracts. The intra-assay coefficient of variation varied in six different experiments from 1.9% to 3.4% in the range 0 to 250 pg of tPA antigen per ml. The inter-assay coefficient of variation was 7% (n = 6). The ELISA was used to study the acute release of tPA from human umbilical vein endothelial cells in vitro. Upon addition of thrombin (1 NIH U/ml) to endothelial cells, tPA was rapidly released into the medium, the highest release occurring during the first minute. Concomitantly, the tPA concentration in the cell extracts decreased. Evidence is presented that tPA is released from an intracellular source.

Avidin↗
PubMed1993

Effect of isotretinoin on endogenous tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor 1 (PAI-1) in humans.

The effect of isotretinoin on fibrinolysis was investigated in 10 healthy, male volunteers in a randomized, double-blind, crossover-designed study. Isotretinoin (40 mg) was administered in the morning and in the evening for 5 days. t-PA, u-PA and PAI-1 antigen and activity in plasma were measured every morning at 9 a.m. on days 1 to 4 and every 3 hours over 24 hours on day 5. Isotretinoin treatment had no significant stimulatory effect on endogenous t-PA antigen and activity in morning plasma samples nor on their circadian variation. Also, u-PA antigen levels did not change after isotretinoin treatment. Mean PAI-1 antigen and PAI activity in 9 a.m. plasma samples were non-significantly higher during isotretinoin than during placebo treatment. After treatment with isotretinoin a significant rise of fasting triglyceride plasma levels was observed as compared to placebo. The study shows that isotretinoin has no clinically significant effect on endogenous fibrinolysis.

Adult↗
PubMed1993

Triazolobenzodiazepines: a new class of stimulators of tissue-type plasminogen activator synthesis in human endothelial cells.

In our search for compounds that can stimulate endogenous fibrinolysis, we have found that certain triazolobenzodiazepines enhance the production of tissue-type plasminogen activator (t-PA) by vascular endothelial cells maintained in vitro, with no or even a lowering effect on plasminogen activator inhibitor type-1 (PAI-1) production. The most active compounds tested, U-34599, U-46195 and U-51477, were studied in more detail and showed a time- and dose-dependent increase in the production of t-PA by human umbilical vein endothelial cells. At optimal stimulatory concentrations (about 10 microM), the three compounds stimulated t-PA expression about 2-fold after 24 hr and maximally about 4-fold after 48 hr of incubation; this maximal increase in t-PA synthesis was sustained at prolonged incubations of 72 or 96 hr. The triazolobenzodiazepine effects on t-PA production were accompanied by parallel increases in t-PA mRNA levels, without marked changes in PAI-1 or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA concentrations. Numerous analogues of the three lead compounds were then tested to determine the relationship between benzodiazepine structure and the ability to stimulate t-PA production. No positive correlation was found between the ability of the various triazolobenzodiazepines to stimulate t-PA production and their affinity for the benzodiazepine receptor. In agreement with this, no specific binding of [3H]flunitrazepam, a specific ligand for benzodiazepine receptors, to endothelial cell membrane preparations was observed. Thus, it is unlikely that the triazolobenzodiazepines act through central-type benzodiazepine receptors to stimulate t-PA production. Similarly, no evidence was found for the presence of peripheral-type benzodiazepine receptors on endothelial cell membranes. The ability of the benzodiazepines to stimulate t-PA production, however, appeared to be related to their platelet-activating factor (PAF) antagonist activity. Despite this finding, several non-benzodiazepine PAF antagonists did not stimulate t-PA production. While the precise mechanism of action is not yet clear, selected benzodiazepine analogues possessing PAF antagonist activity stimulate the production of t-PA by endothelial cells in vitro.

Benzodiazepines↗
PubMed1993

Modulation of tissue-type plasminogen activator by retinoids in rat plasma and tissues.

In this study the effect of vitamin A status and retinoid treatment on the activity of tissue-type plasminogen activator (TPA) and its inhibitor, plasminogen activator inhibitor-type 1 (PAI-1), in plasma and in several tissues was investigated in BN/BiRij rats. Hypervitaminosis A and retinoic acid treatment increased plasma TPA activity by approximately 50%, but PAI-1 activity was not affected. The effect of retinyl palmitate treatment on plasma TPA activity was already significant after 5 days and continued for at least up to 8 wk. Isotretinoin treatment affected neither plasma TPA nor PAI-1 activity. In plasma of vitamin A-deficient rats, TPA activity was decreased by a factor of three, whereas PAI-1 activity was increased twofold. Modulation of plasma TPA activity by vitamin A status and retinoic acid treatment was associated with similar changes in tissue TPA activity. This suggests that the changes in plasma TPA activities might be the result of changes in tissue TPA synthesis.

Animals↗
PubMed1992

On the fibrinolytic system in aged rats, and its reactivity to endotoxin and cytokines.

Aged rats are more susceptible to endotoxin-induced effects, including microthrombosis and platelet aggregation, than are young rats. To investigate whether changes in the fibrinolytic system might be involved, we investigated the fibrinolytic activity in plasma euglobulin fractions and tissues (lung and heart) of young (6-months old) and aged (24-months old) rats under baseline conditions and after challenge with endotoxin. Aged rats had lower plasma levels of tissue-type plasminogen activator (t-PA) and of urokinase-type PA (u-PA) activity. PA inhibitor (PAI) activity was higher in the plasma of aged rats, as was t-PA activity in lung and heart. Rats were treated with either a low dose (1 microgram/kg) or a high dose (10 mg/kg) of endotoxin. Both treatments induced a transient phase of increased blood fibrinolytic activity, as evidenced by higher levels of tissue-type plasminogen activator (t-PA) activity and decreased levels of PA inhibitor (PAI) activity. Over time, the fibrinolytic activity decreased, probably due to increased levels of PA inhibitor. Both the early increase in t-PA activity, and the subsequent increase in PAI activity, were more pronounced in the aged rats, as compared with the younger rats, after the high dose of endotoxin. The aged rats also responded to an injection of interleukin-1 beta or tumor necrosis factor-alpha with a larger increase of PAI activity than did the younger rats. Together the data suggest that, compared to young rats, aged rats have a decreased base-line plasma fibrinolytic activity, while their fibrinolytic system is more responsive to challenge by endotoxin and cytokines.

Aging↗
PubMed1991

Different induction of two plasminogen activator inhibitor 1 mRNA species by phorbol ester in human hepatoma cells.

In man, the plasminogen activator inhibitor 1 (PAI-1) gene codes for two mRNA species, one of 3.2 kilobases (kb) and the other of 2.4 kb. We report that the protein kinase C activating phorbol ester, phorbol 12-myristate-13-acetate (PMA), causes a different induction of the two PAI-1 mRNA species in the human hepatoma cell line, HepG2. Upon addition of 100 nM PMA, the level of the 3.2-kb PAI-1 mRNA species increased to 25-fold after 3 h, and then declined rapidly. The level of the 2.4-kb species increased more slowly and reached a maximal 18-fold stimulation after 6 h, followed by a gradual decrease towards control levels. Run-on analysis showed that PMA induces a transient 40-fold increase in PAI-1 gene transcription rate. The relative concentration of the two PAI-1 mRNA species in the nuclei of PMA-treated HepG2 cells shifted towards the 2.4-kb form, suggesting that changes in transcription termination site and/or post-transcriptional nuclear processing might contribute to their different accumulation. Also, the two mRNAs differ in turnover rate, with a half-life of about 0.85 h for the 3.2-kb form and a half-life of about 2.5 h for the 2.4-kb form. By itself, cycloheximide had no effect on PAI-1 gene transcription rate or PAI-1 mRNA levels in HepG2. When added 1 h prior to PMA, however, cycloheximide prevented the induction of PAI-1 mRNA, which suggests that PMA exerts its stimulating transcriptional activity through a newly synthesized regulatory protein. When cycloheximide was added 2 h after PMA, when the PAI-1 gene transcription rate was maximally increased, the two PAI-1 mRNAs reached even higher levels than with PMA alone and maximal mRNA levels were maintained for a much longer period (up to 8 h). Thus, ongoing protein synthesis is required for both the induction and the transient nature of the PMA-induced PAI-1 mRNA accumulation. We conclude that the differential accumulation of the two PAI-1 mRNAs by PMA in serum-starved HepG2 cells is due both to changes in transcription termination and/or post-transcriptional nuclear processing and to differences in half-life between the two mRNAs in a process that requires ongoing protein synthesis.

Blotting, Northern↗
PubMed1991

Stimulation of tissue-type plasminogen activator synthesis by retinoids in cultured human endothelial cells and rat tissues in vivo.

Tissue-type plasminogen activator (t-PA) and its inhibitor, plasminogen activator inhibitor 1 (PAI-1), play an important role in regulating the fibrinolytic capacity of plasma. Both t-PA and PAI-1 are synthesized by the endothelium. We report that retinoic acid (vitamin A acid) and other retinoids rather specifically stimulate the production of t-PA by cultured human umbilical vein endothelial cells. Effective retinoids induced a dose-dependent (range: 0.01-50 microM) increase in the production of t-PA of maximally about six-fold, while simultaneously causing no or only a small increase (less than two-fold) of PAI-1. The effects on t-PA synthesis were apparent by 4-8 h, and reached maximal values after about 24-48 h of incubation with retinoid. The retinoid effect on t-PA production was accompanied by increased t-PA mRNA levels, without any parallel change in PAI-1 or GAPDH mRNA concentrations. The study also shows that modifications at the carboxyl group of retinoic acid are associated with a decrease in stimulatory potency. The stimulatory pathway appears to be identical for all retinoids but distinct from a pathway by which another strong inducer, sodium butyrate, induces t-PA synthesis in endothelial cells. The induction of t-PA by retinoids might involve protein kinase C (PKC) as judged by an experiment using a specific PKC inhibitor. The effect of retinoids on the fibrinolytic system in vivo was assessed by feeding rats with a vitamin A deficient diet or a diet with excess of vitamin A or other retinoids.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Further characterization of the 5'-flanking DNA of the gene encoding human plasminogen activator inhibitor-1.

Previous nucleotide (nt) sequence analysis of the 5'-flanking DNA of the gene (PAI-1) encoding plasminogen activator inhibitor-1 revealed an extensive region of shared nt sequence identity with the 5'-flanking region of the gene (t-PA) encoding tissue-type plasminogen activator [Bosma et al., J. Biol. Chem. 263 (1988) 9129-9141]. Additional sequence (1642 bp) from the PAI-1 gene 5'-flanking region reveals that these 'PAI-1/t-PA' sequence elements share an alignment that contains a total of 575 positions. This additional PAI-1 5'-flanking sequence also contains two Alu elements that form inverted repeats. Southern-blot analysis using the PAI-1/t-PA element as a probe indicates that this element is repeated in the human genome. which supports the classification of this element as a medium reiteration frequency sequence [Jurka, Nucleic Acids Res. 18 (1990) 137-141].

Amino Acid Sequence↗
PubMed1990

Progress of fibrinolysis during tumor necrosis factor infusions in humans. Concomitant increase in tissue-type plasminogen activator, plasminogen activator inhibitor type-1, and fibrin(ogen) degradation products.

Several investigators have reported that tumor necrosis factor (TNF) can alter the production of plasminogen activator type-1 (PAI-1) and plasminogen activators (PAs) by endothelial cells in vitro. We have examined the in vivo effects of recombinant human TNF administration on fibrinolysis as assessed by parameters in plasma during a 24-hour period of continuous TNF infusion to 17 cancer patients with active disease. The plasma levels of PAI activity increased sevenfold after 3 and 24 hours of TNF infusion. This was the result of an increase of PAI-1 antigen; PAI-2 antigen was not detectable. Plasma concentrations of tissue-type PA (t-PA) antigen increased twofold to fivefold after 3 and 24 hours of TNF infusion, whereas urokinase-type PA antigen levels in plasma remained unaltered. After 3 hours of TNF infusion the plasma levels of alpha 2-antiplasmin were slightly decreased, 5% on average, suggesting that fibrinolysis continued. After 24 hours of TNF infusion a highly significant increase in fibrin- plus fibrinogen-degradation products, and separately of fibrin degradation products and fibrinogen degradation products, was found. This indicates that fibrinolysis persisted, at least partly, in the presence of high levels of PAI activity. Whereas PAI-1 production increased, t-PA production by human endothelial cells in vitro remains unaltered or even decreases on TNF addition. It has been shown previously that TNF infusion in our patients results in thrombin and fibrin generation. Therefore, it is possible that thrombin, not TNF, is the actual stimulus for t-PA production in our patients. We speculate that fibrin is formed during TNF infusions and that plasmin is generated by t-PA action immediately on the initial formation of (soluble) fibrin molecules. Such a process may explain the generation of degradation products of both fibrin and fibrinogen during infusion of TNF in patients.

Fibrin Fibrinogen Degradation Products↗