Full text of DOI:10.1007/BF03343806

J. Endocrinol. Invest. 24: 37-41, 2001
Administration of glutathione in patients with type 2
diabetes mellitus increases the platelet constitutive nitric
oxide synthase activity and reduces PAI-1
V. Martina*, G.A. Bruno**, E. Zumpano*, C. Origlia*, L. Quaranta*, and
G.P. Pescarmona**
*Department of Internal Medicine, Division of Endocrinology; **Department of Genetic, Biology e
Chemical Medicine, University of Torino, Italy
ABSTRACT. Several studies suggest that nitric ox-
ide (NO) production is impaired in diabetes melli-
tus. Reduced levels of NO could contribute to car-
diovascular mortality. Furthermore, NO synthesis
is impaired in glutathione (GSH)-depleted human
umbilical vein endothelial cells and GSH is reduced
in patients with type 2 diabetes mellitus (T2DM).
We tested the hypothesis that treatment with GSH
may improve platelet constitutive NO sinthase
(cNOS) activity in patients with T2DM. Fifteen pa-
tients with T2DM underwent a treatment with GSH
600 mg/day im for 10 days. With respect to the
basal values on the 10th day of treatment, the red
blood cell GSH concentration and platelets cNOS
increased (1.4 0.1 vs 1.9 0.1 μmol/1010 RBC,
p<0.001 and 0.7 0.1 vs 2.9 0.2 fmol·min-1·10-9
PLTs, p<0.001, respectively) and the plasma PAI-1
levels diminished (81.4 3.7 vs 68.7 4.0 ng/ml,
p<0.002). A negative correlation between the
cNOS and the PAI-1 was found on the basal values.
After a wash-out of 30 days the values of red blood
cell GSH concentration, platelet cNOS activity and
PAI-1 Ag returned to the basal levels. These data
suggest that the administration of GSH, in patients
with T2DM, is able to improve platelet cNOS ac-
tivity together with a reduction of PAI-1.
(J. Endocrinol. Invest. 24: 37-41, 2001)
©2001, Editrice Kurtis
INTRODUCTION
(cNOS) activity shows a reduction in patients with type
1 and type 2 diabetes mellitus when compared with
that of normal subjects. On the other hand it has been
reported that the lack of some cofactors essential for
cNOS activity (namely NADPH, BH4, GSH) can im-
pair endothelial cell function: NADPH and BH4
(tetrahydrobiopterin) deficiency can be responsible
for an attenuation of endothelial dependent relax-
ation in diabetes (13) and glutathione (GSH) avail-
ability can modulate cNOS activity in the same cells
(14). GSH level is often decreased in the erythrocytes
(15-17) and platelets (18) of diabetic patients: the
GSH decrease is secondary both to an increased free
radical production induced by hyperglycaemia (19,
20) and to a decreased NADPH level due to polyol
pathway activation (15).
Cardiovascular complications represent 80% of the
cause of death in patients with type 2 diabetes mel-
litus (1). Among the several causes which may explain
this excess of mortality, the role of nitric oxide (NO)
has increasingly gained credit (2-7). NO, which is syn-
thesized from L-arginine by nitric oxide synthase, is a
potent vasodilator (8) and inhibitor of platelet adhe-
sion and aggregation (9, 10). Therefore, reduced lev-
els of NO may contribute to vascular alteration facil-
itating platelet aggregation, platelet-vascular wall in-
teraction, adhesion of monocytes to endothelium,
vascular smooth muscle proliferation and decreasing
endothelium-dependent vasodilation. Our recent da-
ta (11) together with the data of Rabini et al. (12) doc-
ument that the platelet constitutive NO synthase
In this study we tested the hypothesis that a treatment
with GSH given im may improve platelet cNOS activ-
ity in patients with type 2 diabetes mellitus.
Key-words: Nitric oxide, cNOS activity, PAI-1, glutathione, type 2 diabe-
tes mellitus.
Correspondence: Dr. V. Martina, Dipartimento di Medicina Interna,
Divisione di Endocrinologia, Ospedale Molinette, Corso A.M. Dogliotti
14, 10126 Torino, Italy.
SUBJECTS AND METHODS
Fifteen patients with type 2 diabetes mellitus (7 males
and 8 females), recruited from the out-patients of the
E-mail: camanni@pianeta.net
Accepted July 26, 2000.
37

V. Martina, G.A. Bruno, E. Zumpano, et al.
Diabetic Center of the Division of Endocrinology of
the University of Torino, Italy, were studied. The study
had been approved by the Ethical Committee of our
Department and informed consent had been ob-
tained from all patients. The mean age was 59.0 1.1
years (range 49-65), BMI 26.8 0.4 kg/m2 (range 24-
30) and the known duration of diabetes 11.4 0.7
years (range 3-22). The fasting plasma glucose levels
were 10.5 0.6 mmol/l (range 7.5-16.5) and of HbA1c
7.6 0.3% (range 6.2-9.8) (n.v. 5.1-5.8). Four of them
presented background retinopathy, six microalbu-
minuria and none-clinical macrovascular disease. All
the patients were treated by diet plus sulphonylureas.
They underwent a full medical history and physical
examination. Subjects were excluded from the study
if there was a history of cerebrovascular disease, is-
chaemic heart disease, peripheral vascular disease,
hypertension or if they were taking any cardiovascu-
lar drugs. All patients were treated with 600 mg/day
of GSH (Tationil, Boehringer Mannheim), given im at
18:00 h, for 10 days. The day before the first admin-
istration, during the administration (10th day) and 30
days after the last administration, the patients un-
derwent at 08:00 h a blood sample collection for
evaluation of plasma glucose, platelet cNOS activi-
ty, red blood cell GSH concentration, PAI-1 antigen
(PAI-1 Ag) and t-PA activity (t-PA-a).
Plasma glucose was measured by a glucose oxidase
method (Beckman, Palo Alto, U.S.A.). The red blood
cell and platelet quantification was performed by
Coulter S.T.S. Microdiff 18. The evaluation of red
blood cell GSH concentration was performed by a
modification of the method of Beutler et al. (21).
Platelet cNOS was extracted and assayed according
to the method previously described in more detail
(11). cNOS extraction was performed from platelet
rich plasma (PRP) using a buffer containing Hepes-
Na 20 mM, EDTA 0.5 mM, Triton-X-100, Leupeptin
50 μg/ml, pepstatin 50 μg/ml and phenylmethylsul-
fonylfluoride 1 mM. Ph 7.2. PRP (ph7.6) was prepared
by centrifugation at room temperature for 15 min at
200xg. The platelet PRP qualification was performed
by impedance method with Coulter S.T.S. Microdiff
18. The activity of the cNOS was measured by fol-
lowing the conversion of radiolabelled L-arginine to
L-citrulline. In each test tube the following reagents
were added to 100 μl lysate at the final concentra-
tions: 2 mM of NADPH, dithiothreitol 1 mM, 1.5 mM
of CaCl2 and 2.5 Ci L-(3H) arginine (=0.4 μM). In some
preliminary experiments the activity was also mea-
sured without CaCl2. After 15 min incubation at 37
C, the reaction was stopped by adding 2 ml Hepes-
Na ph 6 containing 2 mM EDTA; the whole reaction
mixture was passed through a Dowex column. The
radioactivity corresponding to L-(3H)citrulline content
in 6 ml eluate was measured by liquid scintillation
counting. The coefficient of variation was 10.2-18.1%.
In preliminary experiments NO synthesis was evalu-
ated in the absence or presence of CaCl2. Since no
detectable activity was found in 10 normal subjects
and in 5 type 2 diabetic patients studied when CaCl2
was omitted, next experiments (presented here) were
therefore performed in the presence of CaCl2 and
the activity found referred to as cNOS. Plasma PAI-1
Ag was measured by commercially available ELISA
method from Biopool (22). tPA-a was measured ac-
cording to Verheijeen et al. (23). KABI reagents were
employed for tPA-a. Fourteen age and sex matched
normal volunteers were evaluated as a control group
for basal plasma values of platelet cNOS activity
(1.54 0.3 fmol·min-1·10-9), red blood cell GSH con-
centration (1.99 0.1 μmol/1010 RBC), PAI-1 Ag
(30.6 15.4 ng/ml) and t-PA activity (0.46 0.2 UI/ml).
Statistical analysis
Results are expressed by mean SE. Statistical analy-
sis was carried out by t-test for paired data and by
linear regression analysis.
RESULTS
During the treatment, compared to the basal values
red blood cell GSH concentration increased (1.4 0.1
vs 1.9 0.1 mol/1010 RBC, p<0.001) as well as platelet
cNOS activity (0.7 0.1 vs 2.9 0.2 fmol·min-1·10-9
PLTs, p<0.001) (Fig. 1). Furthermore, PAI-1 Ag de-
5.0
4.0
3.0
2.0
1.0
0
Before
treatment
During
treatment
30 days
after treatment
Fig. 1 - Mean (bars) of the cNOS activity in 15 patients (ꢀ) be-
fore and during the treatment with GSH and after a wash-out of
30 days.
38

GSH increases cNOS activity in T2DM
110
100
90
99% in the reduced form (GSH) and 1% in the oxi-
dized form (GSSG); the high ratio GSH/GSSG is driv-
en by a similar ratio of NADPH/NADP+ redox cou-
ple. GSSG is reduced by GSH reductase, a FAD en-
zyme, using NADPH as a cofactor; NADPH, in turn, is
constantly regenerated in the hexose monophos-
phate shunt (24). In diabetes mellitus there is both an
increased rate of GSH oxidation to GSSG, produced
by hyperglycemia via ROS enhanced production and
GSH peroxidase activity, and also an impaired re-
duction of GSSG to GSH, which is secondary to the
decreased availability of NADPH. As GSSG is active-
ly pumped out from the cells and de novo GSH syn-
thesis rate is low, any long standing oxidative stress
will lead to a steady state of low total GSH (24).
In platelets low GSH is associated with higher sensi-
tivity to agonists (24), spontaneous aggregation (25)
and higher agonists induced calcium influx (26). Ca2+
influx is down regulated by NO via an increase of
cGMP, that inhibits L-Ca-channels, or via a cGMP-in-
dependent pathway (27). We have previously demon-
strated that the cNOS activity depends on GSH (14)
and that in platelets of diabetic patients it is low (11);
furthermore, Schaeffer demonstrated that the intra-
cellular Ca2+ signalling is increased in the platelets of
diabetic patients and that the platelet preincubation
with GSH was able to normalize them (28); similar ef-
fects have been shown by Graier et al. in endothelial
cells during hyperglycemia (29). In our study GSH ad-
ministration has been able to reversibly normalize in-
tracellular GSH. The enhanced availability of GSH may
explain the increase of the cNOS activity; NOS is ex-
tremely sensitive to inhibition by its own product NO
which strongly binds cNOS SH groups, forming stable
nitroso-thiols. GSH or other thiols like α-mercapto-
propionylglycine or β-mercaptoethanol show an ef-
fective scavenging effect on any kind of radicals in-
cluding NO and ROS and prevent early inactivation
of cNOS by radical intermediates (30); higher doses of
these reducing agents can even induce reversible in-
hibition of aggregation (31, 32). On this basis, we sug-
gest that GSH could normalize Ca2+ signalling, as al-
ready demonstrated by Schaeffer, who showed how
calcium influx measured on platelets loaded with fu-
ra-2 was upregulated by treatment with BSO (that de-
creases GSH) and downregulated by treatment with
GSH (28), via the increase of NO production.
80
70
60
50
40
0
0.5
1.0
1.5
2.0
cNOS activity (fmol·min-1 10-9 PLTs)
Fig. 2 - Negative correlation between the basal levels of PAI-1
Ag and cNOS activity.
creased (81.4 3.7 vs 68.7 4.0 ng/ml, p<0.002) and
fasting plasma glucose levels did not change
(10.5 0.6 vs 10.8 0.5 mmol/l, NS) and neither did
plasma t-PA-a (0.8 0.2 vs 0.5 0.1 UI/ml, NS).
A negative correlations between the cNOS activity
and the PAI-1 Ag plasma levels (r=-0.613, p<0.02)
(Fig. 2) was found on the basal values.
No correlation were found between the duration or
severity of the diabetes and the GSH red blood cell
concentration, PAI-1 Ag and platelet cNOS activity.
Thirty days after the last administration of GSH red
blood cell GSH concentration and platelet cNOS
activity returned to the basal values (1.9 0.1 vs
1.3 0.1 mol/1010 RBC and 2.9 0.2 vs 0.7 0.1
fmol·min-1·10-9 PLTs, p<0.001 and p<0.001 re-
spectively) and PAI-1 Ag correspondingly increased
(68.7 4.0 vs 75.4 3.6 ng/ml, p<0.05) (Fig. 1).
Fasting plasma glucose levels (10.5 0.6 vs 10.4 0.6
mmol/l, NS) and plasma t-PA-a (0.8 0.2 vs 0.6 0.2
UI/ml, NS) remained unchanged. During the treat-
ment there was no correlation between the cNOS
activity and the PAI-1 Ag plasma levels.
DISCUSSION
The main result of the present study is that the GSH
administration is able to enhance the platelet cNOS
activity in patients with type 2 diabetes mellitus. The
reversibility of the effects of GSH administration (in-
crease of platelets cNOS activity and of red blood
cells GSH), 30 days after the last administration, rea-
sonably excludes a placebo effect and confirms the
role of GSH as a cofactor of NO-synthase (14).
Low total GSH has been demonstrated in red blood
cells by many authors (15-17) and a similar decrease
has also been shown in platelets, paralleling those of
red blood cells (18). Intracellular glutathione is about
As far as the PAI-1 Ag is concerned it is well known
that PAI-1 Ag is elevated in patients with type 2 dia-
betes mellitus (33-35) and it has been demonstrated
that hypofibrinolysis, via increased levels of plas-
minogen activator inhibitor, leading to decreased re-
moval of fibrin deposit, would be a prime candidate
for a role in the development of atherothrombosis
(36). The negative correlation between the basal
39

V. Martina, G.A. Bruno, E. Zumpano, et al.
6. Pieper G.M.
cNOS activity and the basal PAI-1 Ag plasma levels
and the increase of the cNOS activity together with
the reduction of PAI-1 levels during the treatment
with GSH, confirming our previous findings (33), sup-
port the hypothesis of Korbut et al. that the NO lev-
els can modulate PAI-1 Ag levels (37, 38). t-PA plas-
ma activity is not affected by GSH treatment, as its
release is not calcium-dependent (39).
Taken together these data suggest that one of the
possible pathways leading to the increased mor-
tality for cardiovascular diseases in patients with
type 2 diabetes mellitus could be: decreased GSH
→ higher peroxides + lower NOS activity → high-
er Ca2+ influx → hyperaggregability + higher gran-
ules release → higher PAI-1 release → higher car-
diovascular accidents rate. Based on the foregoing
consideration, the development of a pharmaco-
logical approach to enhance the cNOS activity and
to reduce the PAI-1 plasma concentration in type
2 diabetes mellitus would be of relevant interest.
In conclusion these preliminary data suggest, for
the first time in vivo, that GSH administration in pa-
tients with type 2 diabetes mellitus is able to im-
prove platelet cNOS activity and simultaneously to
reduce plasma PAI-1 levels confirming the rela-
tionship between the two parameters. Further stud-
ies are necessary, above all a double blind study,
in order to confirm these results.
Review of alterations in endothelial nitric oxide pro-
duction in diabetes: protective role of arginine on
endothelial disfunction.
Hypertension 1998, 31: 1047-1060.
7. Veves A., Akbari C.M., Primavera J., Donaghue V.M.
Endothelial dysfunction and the expression of en-
dothelial nitric oxide synthetase in diabetic neu-
ropathy, vascular disease, and foot ulceration.
Diabetes 1998, 47: 457-463.
8. Palmer R.M.J., Moncada S.
A novel citrulline-forming enzyme implicated in the
formation of nitric oxide by vascular endothelial
cells.
Biochem. Biophys. Res. Commun. 1989, 158: 348-352.
9. Moncada S., Radomski M.W., Palmer R.M.J.
Endothelium derived relaxing factor: identification
as nitric oxide and role in the control of vascular tone
and platelet function.
Biochem. Pharmacol. 1988, 37: 2495-2502.
10. Yang Z., von Segesser L., Bauer Estulz P., Turina M.,
Luscher F.T.
Different activation of the endothelial L-arginine and
ciclooxygenase pathway in the human internal mam-
mary artery and saphenous vein.
Circ. Res. 1991, 68: 52-60.
11. Martina V., Bruno G.A., Trucco F., Zumpano E.,
Tagliabue M., Pescarmona G.P.
Platelet cNOS activity is reduced in patients with
IDDM and NIDDM.
Thromb. Haemost. 1998, 79: 520-522.
ACKNOWLEDGMENTS
This work was supported by a grant of MURST.
12. Rabini R.A., Staffolani R., Fumelli P., Mutus B.,
Curatola G., Mazzanti L.
Decreased nitric oxide synthase activity in platelets
from IDDM and NIDDM patients.
Diabetologia 1998, 41: 101-104.
REFERENCES
13. Pieper G.M.
1. Stamler J., Vaccaro O., Neaton J.D.
Diabetes, other risk factors, and 12 yr cardiovascu-
lar mortality for men screened in the multiple risk fac-
tor intervention trial.
Diabetes Care 1993, 16: 434-444.
2. Cosentino F., Luscher T.F.
Endothelial disfunction in diabetes mellitus.
J. Cardiovasc. Pharmacol. 1998, 32 (Suppl. 3): S54-S61.
3. Hayoz D., Ziegler T., Brunner H.R., Ruiz J.
Diabetes mellitus and vascular lesions.
Metabolism 1998, 47 (Suppl. 1): 16-19.
Acute amelioration of diabetic endothelial disfunc-
tion with a derivative of nitric oxide sinthase cofactor,
tetrahydrobiopterin.
J. Cardiovasc. Pharmacol. 1997, 29: 8-15.
14. Ghigo D., Alessio P., Foco A., Bosia A.
A Nitric oxide synthesis is impaired in GSH depleted
human umbilical vein endothelial cells.
Am. J. Phisiol. Endocrinol. Metab. 1993, 265 (Cell
Physiol. 34): C728-C732.
15. Pescarmona G.P., Bosia A., Ghigo D.
Shortened red cell life span in diabetes: mechanism
of haemolysis.
4. Watts G.F., Playford D.A.
In: Weatherall D.J., Fiorelli G., Gorinio S. (Eds),
Advances in red cell biology.
Raven, New York, 1992, p. 391.
Dyslipoproteinemia and hyperoxidative stress in the
pathogenesis of endothelial dysfunction in non-in-
sulin dependent diabetes mellitus: an hypothesis.
Atherosclerosis 1998, 14: 17-30.
16. Murakami K., Kondo T., Ohtsuda Y., Fujiwara Y.,
Shimada M., Kawakami Y.
5. Honing M.L., Morrison P.J., Banga J.D., Stroes E.S.,
Rabelink T.J.
Impairment of GSH metabolism in erytrocytes from
patients with diabetes mellitus.
Metabolism 1989, 38: 753-758.
Nitric oxide availability in diabetes mellitus.
Diabetes Metab. Rev. 1998, 14: 241-249.
40

GSH increases cNOS activity in T2DM
17. Yoshida K., Hirokawa J., Tagami S., Kawakami Y.,
Urzata Y., Kondo T.
28. Schaeffer G., Wascher T.C., Kostner G.M., Graier W.F.
Alterations in Ca2+ signalling in diabetic patients is
due to increased formation of superoxide anions and
reduced nitric oxide production.
Weakened cellular scavenging activity against ox-
idative stress in diabetes mellitus: regulation of GSH
synthesis and efflux.
Diabetologia 1999, 42: 167-176.
Diabetologia 1985, 38: 201-210.
29. Graier W.F., Simecek S., Hoebel B., Wascher T.C.,
Dittrich P., Kostner G.M.
18. Thomas G., Skrinska V., Lucas F.V., Schumacher O.P.
Platelet glutathione and thromboxane synthesis in
diabetes.
Antioxidant prevent high D-glucose-enhanced en-
dothelial Ca2+/cGMP response scavenging superox-
ide anions.
Diabetes 1985, 34: 951-954.
Eur. J. Pharmacol. 1997, 322: 113-122.
19. Graier W.F., Simecek S., Kukovctz W.R., Kostner G.M.
High-D-glucose-induced changes in endothelial
Ca2+/EDRF signalling is due to generation of super-
oxide anions.
Diabetes 1996, 45: 1386-1395.
20. Graier W.F., Posch K., Wascher T.C., Kukovetz W.R.,
Kostner G.M.
30. Schmidt K., Graier W.F., Kostner G.M., Mayer B.,
Kukovetz W.R.
Activation of soluble guanylate cyclase by nitrovaso-
dilators is inhibited by oxidized low-density lipoprotein.
Biochem. Biophys. Res. Commun. 1990, 172: 614-619.
31. Losche W., Breddin K., Pescarmona G.P.
Inhibition of platelet activation by 2-mercaptopro-
pionylglycin in vitro and in vivo.
Role of superoxide anions in changes of endothelial
vasoactive response during acute hyperglycaemia.
Horm. Metab. Res. 1997, 29: 622-629.
Folia Haematol. 1998, 115: 185-188.
21. Beutler E.
32. Pacchiarini L., Tua A., Grignani G.
In vitro effect of reduced glutathione on platelet
function.
Red cell metabolism.
In: Grune & Stratton (Eds.), A manual of biochemical
methods.
New York and London, 1973, p. 103.
Haematologica 1996, 81: 497-502.
33. Martina V., Bruno G.A., Pannocchia A., Zumpano E.,
Tagliabue M., Stella G., Pescarmona G.P.
22. DeClerk P.J., Alessi M.C., Vestreken M., Kruithoff
E.K.O., Vague J., Collen D.
PAI-1 reduction after treatment with GSH in NIDDM.
Fibrinolysis 1996, 10 (Suppl. 2): 63-65.
34. Auwerx J., Bouillon R., Collen D., Geboers D.
Tissue-type plasminogen activator antigen and plas-
minogen activator inhibitor in diabetes mellitus.
Arteriosclerosis 1988, 8: 68-72.
Measurement of plasminogen activator inhibitor-1 in
biological fluids with a murine monoclonal antibody-
hosed enzyme linked immunosorbent assay.
Blood 1988, 71: 220-225.
23. Verheijen J.H., Mullaart E., Chang G.T.G., Kluft C.,
Wijn Gaards G.
35. Garcia Frade L.J., De La Calle H., Torrado M., Lara
J.I., Cuellar L., Garcia A.
A simple sensitive spectrophotometric assay for ex-
trinsic (tissue-type) plasminogen activator applicable
to measurements in plasma.
Hypofibrinolysis associated with vasculopathy in non-
insulin-dependent diabetes mellitus.
Thromb. Res. 1990, 59: 51-59.
Thromb. Haemost. 1982, 48: 266-269.
24. Bosia A., Losche W., Spangenberg P., Pescarmona G.P.
Role of glutathione in blood platelet function.
In: Dolphin D., Poulson R., Avramovic O., (Eds.), Glu-
tathione. Chemical, biochemical and medical aspects.
J. Wiley and Sons, Inc. Publ., New York, London,
Sidney, Toronto, 1989, p. 235.
36. Bachmann F.
Fibrinolysis.
In: Verstraete M., Vermeylen J., Lijnen H.R., Arnout J.
(Eds.), Thrombosis and haemostasis.
Leuven University Press, Leuven, 1987, p. 227.
37. Korbut R., Warner T.D., Gryglewski R.J., Vane J.R.
The effect of nitric oxide synthase inhibition on the
plasma fibrinolytic system in septic shock in rats.
Br. J. Pharmacol. 1994, 112: 289-291.
25. Bosia A., Spangenberg P., Ghigo D.
Effect of GSH depletion by 1-chloro-2,4-dinitroben-
zene on human platelet aggregation, arachidonic
acid oxidative metabolism and cytoskeletal proteins.
Thromb. Res. 1985, 37: 423-434.
38. Korbut R., Marcinkiewicz W., Cieslik K., Gryglevski R.J.
The effect of nitric oxide donors on the release of
plasminogen activator inhibitor (PAI) from rabbit
platelets in vitro.
26. Bosia A., Treves S., Pannocchia A.
Influence of platelet GSH level on arachidonic acid-
dependent Ca++ influx.
J. Phys. Pharm. 1995, 46: 37-44.
Ital. J. Biochem. 1986, 35: 59-61.
39. van den Eijnden-Schraunen Y., Atsma D.E., Lupu F.,
de Vries R.E., Kooistra T., Emeis J.J.
27. Tsikas D., Ikic M., Tewes K.S., Raida M., Frolich J.C.
Inhibition of platelet aggregation by S-nitroso-cys-
teine via cGMP-independent mechanisms: evidence
of inhibition of thromboxane A2 synthesis in human
blood platelets.
Involvement of calcium and G proteins in the acute re-
lease of tissue-type plasminogen activator and von
Willebrand factor from cultured human endothelial cells.
Arterioscler. Thromb. Vasc. Biol. 1997, 17: 2177-2187.
FEBS Lett. 1999, 442:162-166.
41