THROMBOSIS RESEARCH 63; 661-666,199l 0049-3848/91 $3.00 + .OOPrinted in the USA. Copyright (c) 1991 Pergamon Press pk. All rights reserved.
BRIEF
FIBRINOLYTIC
COMMUNICATION
ACTIVITY IN DEPRESSION
AND NEUROSIS.
Michal H. Pietraszek#, Yumiko Takada, Masahiko Koichi Oharal, Kenshiro Oharal and Akikazu Departments of Psychiatry’ School of Medicine,
and Physiology, Hamamatsu-shi, 431-31, Japan.
Nishimotol, Takada*
Hamamatsu University Shizuoka-ken,
.
(Received 5.1 .1991; accepted in revised form 1.7.1991 by Editor M.B. Donati)
INTRODUCTION. a long-established clinical and epidemiological It has been observation that patients with affective disorders have a higher than expected rate of mortality from cardiovascular disease (1,2). Further evidence suggests that major depression has a direct negative impact on the outcome of cardiovascular disease (3’1. The mechanism through which mental disorder is a cause of cardiovascular death in depressed patients and affects outcome in cardiac disease is far from clear. The fibrinolytic system plays a key role in defence against fibrin deposition on the vessel wall and is thus an essential protection against blood vessel occlusion. Decreased fibrinolytic activity is considered to be a factor involved in the pathogenesis of myocardial infarction, cerebral thrombosis and deep vein thrombosis. It seemed, therefore, of interest to evaluate the function of the fibrinolytic system in patients suffering from depression and neurosis developed assay for the determination of plasma using a newly plasminogen activators and inhibitors (4). Key
words:
plasminogen
tissue-type activator
plasminogen inhibitor
#On leave of absence from Medical Academy, Bialystok, *Author for correspondence.
activator,
1, depression, Department Poland.
661
urokinase, neurosis.
of Pharmacodynamics.
662
Flf3RlNOLYSlS IN CNS DISORDERS
MATERIALS
Vol. 63, No 6
AND METHODS.
Subjects; Twenty seven patients. who were diagnosed at the Department :)I Psychiatry, Hamamatsu University School of Medicine participated in the present investigation. Patients were divided into 2 groups according to The first group was with major unipolar examinations. psychiatric 10 males, 7 females) and the second group was with depression (n=l7; depressive neurosis (dysthymia) (n=lO; 8 males, 2 females). The mean ages of these groups were SO.8 + 8.5 and 46.0 + 6.6 years, respectively. The mean Hamilton depression rating score on the 24-item scale was 18.7 (range 12-24) and 16.5 (range 1 l-20), respectively. Diagnosis was based on the Diagnostic and Statistical Manual of Mental Disorders (DSM-III-R) which requires for the diagnosis of depressive neurosis at least two-year numerous periods of depressive symptoms of continued or history characteristic of major depression that do not meet full severity and/or duration criteria for the syndrome. There were IO smokers, and 17 nonsmokers. All subjects were drug free for one week before the experiment. Patients were matched in pairs according to sex and age with the control group composed of 27 apparently healthy volunteers. Blood
samples; Blood was drawn from the antecubital vein with minimal stasis mixed with 3.8% sodium citrate in proportion of 9:1, v/v. The blood centrifugated at 3000 rpm for 15 min at 4’ C to obtain plasma. Plasma kept frozen at -70” C until assayed.
and was was
The
preparation of euglobulin fraction; The plasma euglobulin fraction was prepared by 20 times dilution of titrated plasma (0.5 ml) and acidificated at pH 5.2. After leaving for one hour at 4” C followed by the centrifugation for 10 min at 3000 rpm at 4” C, the precipitate was dissolved by 0.5 ml of 0.1 M Tris-HCI buffer, pH 7.4. All samples were used for assay immediately after preparation.
Euglobulin clot lysis time; 50 p1 of human thrombin (100 W/ml) was placed in individual wells of the microtiter plate and the clot formation was initiated by the addition of 150 pl of the plasma euglobulin fraction. The turbidity of the wells was measured as a function of the absorbance at 340 nm every 30 min employing an automatic microtiter plate reader (Sigma Seiki Inc., Japan). All the samples were assayed in duplicate. Tissue plasminogen Plasma levels WA) (5).
activator of t-PA
(t-PA) and urokinase (UK) ; and UK were assayed by enzyme-immunoassay
Vol. 63, No. 6
Total
FIBRINOLYSIS IN CNS DISORDERS
PAI-1, free PAIand Plasma levels of t-PA by EIA (4). The concentration by the subtraction of the complex from that of total was expressed by the order
663
t-PA - PAIcomplex ; - PAIcomplex and total PAIwere measured of free PAIin the plasma was calculated concentration of all forms of t-PA - PAIPAI-1. The concentration of all forms of PAIof ng/ml of PAI-1.
Statistical analysis; Group differences were tested for significance using a Wilcoxon rank sum test, A value of ~~0.05 was taken as the level of significance. Results were expressed as a mean f. standard deviation. RESULTS As shown in Table 1 there were significant differences in euglobulin lysis time as well as in t-PA, UK, and PAIplasma concentrations between controls and psychiatric patients. The euglobulin lysis time was slightly shorter in the group of depressed subjects. A group of neurotic patients showed the shortest euglobulin lysis time. It was significant at p < 0.01. The group of patients with depression showed significantly lower t-PA concentrations than normal persons (~~0.05). The lowest concentration of tPA was observed in depressive neurosis. It was significant at ~~0.01. A different pattern was shown in plasma urokinase level. In patients with almost two fold higher compared to depression plasma urokinase was that of normal persons (~~0.01). Plasma urokinase levels in depressive neurosis were also higher (~~0.01). Both groups I3f patients with affective disorders had significantly lower total PAIthan controls (p
BRIEF
FIBRINOLYTIC
COMMUNICATION
ACTIVITY IN DEPRESSION
AND NEUROSIS.
Michal H. Pietraszek#, Yumiko Takada, Masahiko Koichi Oharal, Kenshiro Oharal and Akikazu Departments of Psychiatry’ School of Medicine,
and Physiology, Hamamatsu-shi, 431-31, Japan.
Nishimotol, Takada*
Hamamatsu University Shizuoka-ken,
.
(Received 5.1 .1991; accepted in revised form 1.7.1991 by Editor M.B. Donati)
INTRODUCTION. a long-established clinical and epidemiological It has been observation that patients with affective disorders have a higher than expected rate of mortality from cardiovascular disease (1,2). Further evidence suggests that major depression has a direct negative impact on the outcome of cardiovascular disease (3’1. The mechanism through which mental disorder is a cause of cardiovascular death in depressed patients and affects outcome in cardiac disease is far from clear. The fibrinolytic system plays a key role in defence against fibrin deposition on the vessel wall and is thus an essential protection against blood vessel occlusion. Decreased fibrinolytic activity is considered to be a factor involved in the pathogenesis of myocardial infarction, cerebral thrombosis and deep vein thrombosis. It seemed, therefore, of interest to evaluate the function of the fibrinolytic system in patients suffering from depression and neurosis developed assay for the determination of plasma using a newly plasminogen activators and inhibitors (4). Key
words:
plasminogen
tissue-type activator
plasminogen inhibitor
#On leave of absence from Medical Academy, Bialystok, *Author for correspondence.
activator,
1, depression, Department Poland.
661
urokinase, neurosis.
of Pharmacodynamics.
662
Flf3RlNOLYSlS IN CNS DISORDERS
MATERIALS
Vol. 63, No 6
AND METHODS.
Subjects; Twenty seven patients. who were diagnosed at the Department :)I Psychiatry, Hamamatsu University School of Medicine participated in the present investigation. Patients were divided into 2 groups according to The first group was with major unipolar examinations. psychiatric 10 males, 7 females) and the second group was with depression (n=l7; depressive neurosis (dysthymia) (n=lO; 8 males, 2 females). The mean ages of these groups were SO.8 + 8.5 and 46.0 + 6.6 years, respectively. The mean Hamilton depression rating score on the 24-item scale was 18.7 (range 12-24) and 16.5 (range 1 l-20), respectively. Diagnosis was based on the Diagnostic and Statistical Manual of Mental Disorders (DSM-III-R) which requires for the diagnosis of depressive neurosis at least two-year numerous periods of depressive symptoms of continued or history characteristic of major depression that do not meet full severity and/or duration criteria for the syndrome. There were IO smokers, and 17 nonsmokers. All subjects were drug free for one week before the experiment. Patients were matched in pairs according to sex and age with the control group composed of 27 apparently healthy volunteers. Blood
samples; Blood was drawn from the antecubital vein with minimal stasis mixed with 3.8% sodium citrate in proportion of 9:1, v/v. The blood centrifugated at 3000 rpm for 15 min at 4’ C to obtain plasma. Plasma kept frozen at -70” C until assayed.
and was was
The
preparation of euglobulin fraction; The plasma euglobulin fraction was prepared by 20 times dilution of titrated plasma (0.5 ml) and acidificated at pH 5.2. After leaving for one hour at 4” C followed by the centrifugation for 10 min at 3000 rpm at 4” C, the precipitate was dissolved by 0.5 ml of 0.1 M Tris-HCI buffer, pH 7.4. All samples were used for assay immediately after preparation.
Euglobulin clot lysis time; 50 p1 of human thrombin (100 W/ml) was placed in individual wells of the microtiter plate and the clot formation was initiated by the addition of 150 pl of the plasma euglobulin fraction. The turbidity of the wells was measured as a function of the absorbance at 340 nm every 30 min employing an automatic microtiter plate reader (Sigma Seiki Inc., Japan). All the samples were assayed in duplicate. Tissue plasminogen Plasma levels WA) (5).
activator of t-PA
(t-PA) and urokinase (UK) ; and UK were assayed by enzyme-immunoassay
Vol. 63, No. 6
Total
FIBRINOLYSIS IN CNS DISORDERS
PAI-1, free PAIand Plasma levels of t-PA by EIA (4). The concentration by the subtraction of the complex from that of total was expressed by the order
663
t-PA - PAIcomplex ; - PAIcomplex and total PAIwere measured of free PAIin the plasma was calculated concentration of all forms of t-PA - PAIPAI-1. The concentration of all forms of PAIof ng/ml of PAI-1.
Statistical analysis; Group differences were tested for significance using a Wilcoxon rank sum test, A value of ~~0.05 was taken as the level of significance. Results were expressed as a mean f. standard deviation. RESULTS As shown in Table 1 there were significant differences in euglobulin lysis time as well as in t-PA, UK, and PAIplasma concentrations between controls and psychiatric patients. The euglobulin lysis time was slightly shorter in the group of depressed subjects. A group of neurotic patients showed the shortest euglobulin lysis time. It was significant at p < 0.01. The group of patients with depression showed significantly lower t-PA concentrations than normal persons (~~0.05). The lowest concentration of tPA was observed in depressive neurosis. It was significant at ~~0.01. A different pattern was shown in plasma urokinase level. In patients with almost two fold higher compared to depression plasma urokinase was that of normal persons (~~0.01). Plasma urokinase levels in depressive neurosis were also higher (~~0.01). Both groups I3f patients with affective disorders had significantly lower total PAIthan controls (p
