National Sanatorium Toneyama Hospital, Toyonaka. Osaka, Japan and of Surgery1), Obstetrics and Gynaecology-), Urology') and Pathology4), Osaka University Medical School, Osaka, Japan
Departments
EFFECTS OF GENERAL ANAESTHESIA AND SEVERITY OF SURGICAL STRESS ON SERUM LH AND TESTOSTERONE IN MALES
By A. Nakashima, K. Koshiyama, T. Uozumi, Y. Monden, Y. Hamanaka, K. Kurachi, T. Aono, S. Mizutani and K. Matsumoto
ABSTRACT
Significantly decreased levels of serum testosterone from the pre-anaesthesia level were found during and up to 7 days following major surgery under general anaesthesia (nitrous oxide, oxygen and halothane following induction with thiopental and succinylcholine chloride) in 18 male patients. On the other hand, in the same patients, the serum luteinizing hormone (LH) increased significantly from the pre-anaesthesia level 30 min and 1 h after the beginning of anaesthesia. A slight increase in LH level was also noted on the 7th post-operative day. The determinations of serum testosterone and LH in fiberoptic bronchoscopy under the same general anaesthesia as that used in surgery or local anaesthesia in 26 male patients, revealed that the change in the serum LH during and following surgery seemed to be mainly induced by the general anaesthesia and that the rate of decrease in the serum testosterone may be related to the severity of surgical stress including the anaesthesia. The rate of increase in serum testosterone following the injection of gonadotrophin in 20 males on the 6th post-operative day was similar to that in 10 pre-operative males. The effects of pulmonary lobectomy on serum
urinary steroids were also studied in 6 males under adrenal suppression with dexamethasone. On the 6th post-operative day, the urinary aetiocholanolone plus androsterone and serum testosterone were found to be half the level of those on the pre-operative day, while the urinary 5\g=b\-pregnane-3\g=a\,17\g=a\,20\g=a\-triolremained unchanged. These observations in human are not inconsistent with the report of Tcholakian & Eik-Nes (1971) in dogs namely that a shift in androgen biosynthetic pathway is present in the testis under surgical stress. testosterone and
In a previous paper (Aono et al 1972), we reported in male patients that the levels of plasma testosterone decreased significantly from the pre-operative level during pulmonary lobectomy or gastrectomy, shortly after the operation and on the 2nd and 6th post-operative days, while the levels of plasma lute¬ inizing hormone (LH) increased significantly from that of the pre-operation level 30 min, 1 and 2 h after the induction of general anaesthesia and also on the 6th post-operative day. The concentration of LH decreased slightly but significantly on the 2nd post-operative day when the plasma testosterone showed a most marked decrease. We postulated that there might be some lack of testicular sensitivity to gonadotrophins during surgical stress. With regard to the plasma gonadotrophins in surgery, Charters et al. (1969) found a decrease in plasma LH on the first 2 post-operative days in 10 females and 3 males and Carstensen et al. (1969, 1972) and (Monden et al. 1972) reported a signi¬ ficant increase in plasma LH in males several days after surgery. However, Charters et al. (1969) found no significant intra-operative increase in the concentration of LH. With regard to plasma testosterone in surgery, a sig¬ nificant intra- and post-operative decrease in male patients has been reported (Carstensen et al. 1968, 1969, 1972, 1973; Matsumoto et al. 1970; Monden et al. 1972; Oyama 8c Kudo 1972). The present study was attempted in order to examine the cause of these
changes.
MATERIALS AND METHODS
Serum
samples
Blood samples were taken from 70 male patients, aged from 19-61 years. None of the patients had any disorders of the endocrine system, liver or kidney functions as judged by routine tests. None of them had received andrenocorticotrophin (ACTH), gonadotrophins, corticosteroids or sex steroids before the studies. The patients had not been exposed to any particular kind of stress before examination. The blood was collected in containers. The serum was separated soon by centrifugation, stored at -20° until required and used within 5 weeks.
The effects of major surgery on the serum levels of testosterone and LH were in¬ vestigated in 15 males with pulmonary tuberculosis or pulmonary cancer who had undergone lobectomy and in 3 males with gastric ulcer who had undergone gastrectomy. One hour before the operation, the patients were injected with 50 mg of hydroxyzinepamoate and 0.5 mg of atropine sulphate. All the operations, which began at 9 a. m..
carried out at room temperature and the duration of surgery from incision to closure ranged from 2-4 h. Anaesthesia was maintained with nitrous oxide, oxygen and halothane following induction with thiopental and succinylcholine chloride. About 500 ml of plasma substitutes and 1000 ml of electrolyte solution were infused into each of the patients during and shortly following the operation. None of the patients showed any complications following the operation. Control serum samples were taken immediately before anaesthesia. Subsequent samples were obtained from the patients 30 min, 1 and 2-3 h after anaesthesia and at 9 a. m. on the 2nd, 4th and 7th post¬ operative days. The interval between the induction of anaesthesia and the beginning of incision was about 30 min. The effects of fiberoptic bronchoscopy under general anaesthesia on the serum levels of testosterone and LH were examined in 16 males with pulmonary tuberculosis or pulmonary cancer. The injection of hydroxyzine-pamoate and atropine sulphate was as described above. Bronchoscopy began at 9 a. m. and the duration ranged from 40-60 min. Anaesthesia was maintained with nitrous oxide and oxygen with or without halothane following induction with thiopental and succinylcholine chloride. Broncho¬ scopy began immediately after the induction of general anaesthesia. About 500 ml of electrolyte solution was infused during the procedure. Control serum samples were taken immediately before the induction of anaesthesia. Subsequent samples were obtained 30 min, 1 and 2.5 h after the anaesthesia and at 9 a. m. on the 2nd and 7th were
post-bronchoscopy days. The effects of the fiberoptic bronchoscopy under local anaesthesia with lidocaine hydrochloride on the serum levels of testosterone and LH were examined in 10 males with pulmonary tuberculosis or pulmonary cancer. One hour before the bronchoscopy, which began at 9 a. m., the patients were treated with hydroxyzine-pamoate and atropine sulphate. The duration of the bronchoscopy under local anaesthesia was about 30 min. None of the patients received thiopental, succinylcholine chloride or transfusion. Serum samples were taken immediately before local anaesthesia, 30 min and 1 h after the beginning of anaesthesia and at 9 a. m. on the 2nd, 4th and 7th post-bronchoscopy days. Bronchoscopy began immediately after the onset of local
anaesthesia. The effect of human chorionic gonadotrophin (HCG) on serum level of testosterone was examined in 10 pre-operative males with pulmonary tuberculosis and in 20 male patients who had undergone lobectomy or gastrectomy 7 days before the test. Two thousand units of HCG was injected intramuscularly at 9 a. m. Serum samples were obtained from the pre- and post-operative patients immediately before the HCG in¬ jection, and 3 and 6 h after the injection.
Serum and urine
samples
The effects of surgical stress on the excretion of urinary steroids and serum levels of testosterone and LH under adrenal suppression were examined in 6 male patients with pulmonary tuberculosis. Serum and urine samples were taken 7 days before lobectomy. The patients were then given dexamethasone, 6 mg daily for 12 days. On the day of operation and on the next day, an additional dose of dexamethasone,
i.e.
3mg was injected. The operation was performed on the 6th day of adrenal suppression, and samples were obtained 1 day before and 6 days after the operation. Serum samples were taken at 9 a. m. and urine samples were collected for 24 h.
Samples
were
Estimation
stored at -20°C until
of serum
Concentrations of
testosterone
required.
and LH
testosterone were measured by the competitive protein binding method of Mayes Se Nugent (1968). Serum levels of LH were measured by the radioimmunoassay method (Aono et al. 1967), with slight modifications as pre¬ viously described (Aono et al. 1972). The evaluation of the LH and testosterone assays including specificity study was satisfactory as previously reported (Aono et al. 1972; Matsumoto et al. 1970). The intra- and inter-assay coefficient of variation in a normal male range obtained from 10 assays, were 8.9 and 9.7%, respectively in the testosterone assay and 8.8 and 13.6%, respectively in the LH assay. serum
Quantitative analyses of urinary steroids The method for the quantitative determination of urinary steroid fractions has been described in previous reports (Matsumoto et al. 1966; Seki Se Matsumoto 1967). One hundred ml of each urine sample was first treated with limpet /J-glucuronidase. After extraction of the steroids with chloroform, steroid sulphate was hydrolyzed by the method of Burstein Se Lieberman (1958). Each of the steroid hydrolysates thus obtained was washed with 1 NaOH, H.,0 and separated by elution chromatography on Amberlite IRC-50 using methanol:ethanol:water (3:9:8 v/v) as eluent. The steroids extracted following hydrolysis with /i-glucuronidase were separated into cortisol + cortisone + tetrahydrocortisol (3«,1 l/?,17a,21-tetrahydroxy-5/i-pregnan-20-one) + tetra¬
hydrocortisone (3ci,l7a,21-trihydroxy-5/?-pregnane-ll,20-dione), allotetrahydrocortisol
(3a,ll/?,17a,21-tetrahydroxy-5a-pregnan-20-one), ll/j-hydroxyaetiocholanolone (3«,ll/idihydroxy-5/?-androstan-17-one), 11-ketoaetiocholanolone (3cc-hydroxy-5/i-androstane11.1
7-dione), 11/J-hydroxyandrosterone (3a,ll/J-dihydroxy-5«-androstan-17-one),
aetio-
cholanolone
(3a-hydroxy-5/?-androstan-17-one), 5/?-pregnane-3a,17a,20a-triol, androsterone and 5/?-pregnane-3a,20a-dioL The other hydrolysate was separated into dehydro¬ epiandrosterone and 5-pregnene-3/i,20a-diol. Steroid fractions thus obtained were quantitatively
determined by Porter-Silber reagent, Zimmermann reagent, acid and Oertel-Eik-Nes reagent (Oertel Se Eik-Nes 1959), respectively.
sulphuric
RESULTS surgery and fiberoptic bronchoscopy under general or local anaesthesia on serum levels of LH and testosterone The pre-, during- and post-anaesthesia levels of serum LH and testosterone in male patients are shown in Tables 1 and 2. Fig. 1 shows the average per¬ centage changes from the pre-anaesthesia levels in the same patients. In sur¬ gical patients, the level of serum LH increased significantly from that of
Effects of major
SERUM LH
0
05
2-3
1
Fig
12
4
DAYS
HOURS
L
major surgery and fiberoptic bronchoscopy under general or local anaesthesia on serum levels of LH and testosterone in males. The results are expressed as the average percentage change from the pre-anaesthesia level. Conditions of experiments are shown in Tables 1 and 2. Differences from "Pre-anaesthesia" (P): I < 0.05, 2 < 0.01. Effects of
the control (pre-anaesthesia) 30 min and 1 h after the beginning of general anaesthesia. In spite of continuing surgical stress and general anaesthesia, the LH concentration then decreased toward the control level 2-3 h after the beginning of anaesthesia. A significant decrease in the level of serum LH was noted on the 2nd post-operative day, but the level increased slightly on the 7th day following surgery under general anaesthesia, thought the increase was not statistically significant. Although the duration of fiberoptic broncho¬ scopy under general anaesthesia ranged from 40-60 min instead of 2-4 h in surgery, the change in the serum levels of LH during and following the bron¬ choscopy under general anaesthesia was similar to that found in surgery (Table 1, Fig. 1). However, the rate of change was lower in the bronchoscopy than in surgery. On the other hand, the level of LH exhibited no significant change in the bronchoscopy under local anaesthesia (Table 1, Fig. 1).
Table i. Effects of major surgery and fiberoptic bronchoscopy under general or local anaesthesia on serum level of LH (Mean + se ug/100 ml) in males.
Before anaesthesia
Surgery
Bronchoscopy
Bronchoscopy
under general anaesthesia
under general anaesthesia
under local anaesthesia
4.5 ± 0.5
(If
.7 ± 0.6
(16)
4.3 ± 0.8
(10)
(10) ( 6)
After onset of anaesthesia 30 min 1 h
9.5 ±
0.8*;;-(12)
8.6 ±
0.8**(15)
4.5 ± 0.8
10.0 ±
1.0**(18)
9.4 ±
1.1**(15)
4.2 ± 0.4
5.0 ± 0.9
2-3 h
Post-anaesthesia 2nd day 4th day 7th day
2.9 ±
(10)
0.4*s-(16) (17) 0.8 (17)
6.1 ± 0.8
4.5 ±
(12)
0.4**(15)
4.6 ± 0.7 6.0 ±
5.1 ± 1.1 5.0 ± 0.8
7.0 ± 1.2
( 9)
4.6 + 0.6
(10) ( 9) ( 9)
Differences from "Before anaesthesia" (P): ** < 0.01 (i-test for paired observations). The duration of major surgery and fiberoptic bronchoscopy under general and local anaesthesia was 2-4 h, 40-60 min and 30 min, respectively. ( ) indicates number of patients at each point.
levels of testosterone progressively decreased during the opera¬ value significantly smaller than the control level was found 2-3 h after the beginning of anaesthesia. The lowest concentration was obtained on the 2nd post-operative day when a significant decrease in the level of LH was noted. Significantly decreased levels of serum testosterone were maintained until the 7th post-operative day when the level of serum LH showed a slight increase. The levels of serum testosterone showed no significant decrease in fiberoptic bronchoscopy under general and local anaesthesia, except for that obtained on the 2nd day following bronchoscopy under general anaesthesia, when the serum LH was found to be slightly but significantly decreased The
serum
tions, and
a
(Table 2, Fig. 1). on serum level of testosterone in pre- and male patients post-operative Since the results shown in Tables 1 and 2 and Fig. 1 suggested some decrease of testicular sensitivity to gonadotrophins under major surgical stress, the ex¬ periment shown in Table 3 was performed. However, the rate of increase in
Effect of HCG
Table 2. Effects of major surgery and fiberoptic bronchoscopy under general or local anaesthesia + se «g/100 on serum level of testosterone (Mean ml) in males.
Surgery
under general anaesthesia
Bronchoscopy
Bronchoscopy
under general anaesthesia
under local anaesthesia
0.77 ± 0.04
(18)
0.57 ± 0.04
(16)
0.73 ± 0.04
(10)
30 min
0.83 ± 0.06
(12)
0.62 ± 0.04
0.72 ± 0.05 (18) 0.47 ± 0.09**(10)
0.58 ± 0.04
(15) (15) (12)
0.67 ± 0.05
1 h
(10) ( 6)
0.22 ± 0.29 ± 0.51 ±
0.47 ±
0.04*(15)
0.68 ± 0.07
( 9)
0.67 ± 0.05 0.74 ± 0.05
Before anaesthesia After onset of anaesthesia
2-3 h
Post-anaesthesia 2nd day 4th day 7 th day
0.03**(16) 0.03**(17) 0.05**(17)
Differences from "Before anaesthesia"
observations). ( ) indicates number
Effect of HCG
of
patients
on serum
0.63 ± 0.04
at
*
(P):
each
0.53 ± 0.04
< 0.05,
**
< 0.01
0.67 ± 0.05
(i-test
for
Table 3. level of testosterone in pre- and
post-operative
Mean ±
After operation 6th day2 6th
day3
Differences from "Before
observations). 1
2 3
se
males.
injection of HCG'
3 h
operation
paired
point.
Alter
Before
(10) ( 9) ( 9)
6 h
(«g/100 ml)
10
0.68 ± 0.12
0.76 ± 0.11
0.91 ± 0.10*
10
0.32 ± 0.04
0.45 ± 0.04**
10
0.49 ± 0.06
0.57
0.46 ± 0.04* 0.65 ± 0.08*
injection" (P):
*
< 0.05,
**
± 0.06*
< 0.01
(¿-test
for
paired
Two thousand units of HCG was injected intramuscularly at 9.00 a. m. Control serum samples were obtained immediately before the HCG injection. Post-operative and pre-operative samples were obtained from the same patients. Post-operative samples were obtained from different groups of patients.
testosterone level similar to that in
post-operative patients was pre-operative patients, though the pre- and post-injection values were significantly lower in the post-operative patients than in the pre¬ operative patients, respectively.
following
Effects of surgical stress
HCG injection in
urinary steroid excretion and serum levels of patients under adrenal suppression Since the results shown in Tables 1-3 and those obtained by Tcholakian Sc Eik-Nes (1971) in dogs seemed to suggest a shift in the androgen biosynthetic pathway of the testis following surgical stress, urinary steroids were measured under adrenal suppression with dexamethasone in 6 male patients before and 6 days after pulmonary lobectomy. Metabolites of cortisol decreased markedly testosterone
fiftieth after dexamethasone administration. Under adrenal suppression, urinary excretion of aetiocholanolone plus androsterone de¬ creased significantly from the pre-operative level on the 6th post-operative to
less than
on
and LH in male
one
the
Effects of
Table 4. stress on urinary steroid excretion and serum levels of testosterone and LH in 6 males under adrenal suppression with dexamethasone.
surgical
No
suppression
Before op.
urinary
steroids
Metabolites of cortisol2
11-Oxy-17-ketosteroids 11-Deoxy-17-ketosteroids 5-Pregnene-3/?,20a-diol 5/LPregnane-3a,20a-diol 5/?-Pregnane-3a,17a,20a-triol Serum testosterone and LH Testosterone LH
Adrenal
Before op.1 Mean ±
se
1
2
After
op.1
(mg/24 h)
3.18 ± 0.75 0.89 ± 0.24 1.92 ± 0.46
0.06 ± 0.02
0.06 ± 0.03
0.14 ± 0.05
0.06
1.24 ± 0.20
0.52 ± 0.08s
< 0.10 < 0.20 0.98 ± 0.20
Departments
EFFECTS OF GENERAL ANAESTHESIA AND SEVERITY OF SURGICAL STRESS ON SERUM LH AND TESTOSTERONE IN MALES
By A. Nakashima, K. Koshiyama, T. Uozumi, Y. Monden, Y. Hamanaka, K. Kurachi, T. Aono, S. Mizutani and K. Matsumoto
ABSTRACT
Significantly decreased levels of serum testosterone from the pre-anaesthesia level were found during and up to 7 days following major surgery under general anaesthesia (nitrous oxide, oxygen and halothane following induction with thiopental and succinylcholine chloride) in 18 male patients. On the other hand, in the same patients, the serum luteinizing hormone (LH) increased significantly from the pre-anaesthesia level 30 min and 1 h after the beginning of anaesthesia. A slight increase in LH level was also noted on the 7th post-operative day. The determinations of serum testosterone and LH in fiberoptic bronchoscopy under the same general anaesthesia as that used in surgery or local anaesthesia in 26 male patients, revealed that the change in the serum LH during and following surgery seemed to be mainly induced by the general anaesthesia and that the rate of decrease in the serum testosterone may be related to the severity of surgical stress including the anaesthesia. The rate of increase in serum testosterone following the injection of gonadotrophin in 20 males on the 6th post-operative day was similar to that in 10 pre-operative males. The effects of pulmonary lobectomy on serum
urinary steroids were also studied in 6 males under adrenal suppression with dexamethasone. On the 6th post-operative day, the urinary aetiocholanolone plus androsterone and serum testosterone were found to be half the level of those on the pre-operative day, while the urinary 5\g=b\-pregnane-3\g=a\,17\g=a\,20\g=a\-triolremained unchanged. These observations in human are not inconsistent with the report of Tcholakian & Eik-Nes (1971) in dogs namely that a shift in androgen biosynthetic pathway is present in the testis under surgical stress. testosterone and
In a previous paper (Aono et al 1972), we reported in male patients that the levels of plasma testosterone decreased significantly from the pre-operative level during pulmonary lobectomy or gastrectomy, shortly after the operation and on the 2nd and 6th post-operative days, while the levels of plasma lute¬ inizing hormone (LH) increased significantly from that of the pre-operation level 30 min, 1 and 2 h after the induction of general anaesthesia and also on the 6th post-operative day. The concentration of LH decreased slightly but significantly on the 2nd post-operative day when the plasma testosterone showed a most marked decrease. We postulated that there might be some lack of testicular sensitivity to gonadotrophins during surgical stress. With regard to the plasma gonadotrophins in surgery, Charters et al. (1969) found a decrease in plasma LH on the first 2 post-operative days in 10 females and 3 males and Carstensen et al. (1969, 1972) and (Monden et al. 1972) reported a signi¬ ficant increase in plasma LH in males several days after surgery. However, Charters et al. (1969) found no significant intra-operative increase in the concentration of LH. With regard to plasma testosterone in surgery, a sig¬ nificant intra- and post-operative decrease in male patients has been reported (Carstensen et al. 1968, 1969, 1972, 1973; Matsumoto et al. 1970; Monden et al. 1972; Oyama 8c Kudo 1972). The present study was attempted in order to examine the cause of these
changes.
MATERIALS AND METHODS
Serum
samples
Blood samples were taken from 70 male patients, aged from 19-61 years. None of the patients had any disorders of the endocrine system, liver or kidney functions as judged by routine tests. None of them had received andrenocorticotrophin (ACTH), gonadotrophins, corticosteroids or sex steroids before the studies. The patients had not been exposed to any particular kind of stress before examination. The blood was collected in containers. The serum was separated soon by centrifugation, stored at -20° until required and used within 5 weeks.
The effects of major surgery on the serum levels of testosterone and LH were in¬ vestigated in 15 males with pulmonary tuberculosis or pulmonary cancer who had undergone lobectomy and in 3 males with gastric ulcer who had undergone gastrectomy. One hour before the operation, the patients were injected with 50 mg of hydroxyzinepamoate and 0.5 mg of atropine sulphate. All the operations, which began at 9 a. m..
carried out at room temperature and the duration of surgery from incision to closure ranged from 2-4 h. Anaesthesia was maintained with nitrous oxide, oxygen and halothane following induction with thiopental and succinylcholine chloride. About 500 ml of plasma substitutes and 1000 ml of electrolyte solution were infused into each of the patients during and shortly following the operation. None of the patients showed any complications following the operation. Control serum samples were taken immediately before anaesthesia. Subsequent samples were obtained from the patients 30 min, 1 and 2-3 h after anaesthesia and at 9 a. m. on the 2nd, 4th and 7th post¬ operative days. The interval between the induction of anaesthesia and the beginning of incision was about 30 min. The effects of fiberoptic bronchoscopy under general anaesthesia on the serum levels of testosterone and LH were examined in 16 males with pulmonary tuberculosis or pulmonary cancer. The injection of hydroxyzine-pamoate and atropine sulphate was as described above. Bronchoscopy began at 9 a. m. and the duration ranged from 40-60 min. Anaesthesia was maintained with nitrous oxide and oxygen with or without halothane following induction with thiopental and succinylcholine chloride. Broncho¬ scopy began immediately after the induction of general anaesthesia. About 500 ml of electrolyte solution was infused during the procedure. Control serum samples were taken immediately before the induction of anaesthesia. Subsequent samples were obtained 30 min, 1 and 2.5 h after the anaesthesia and at 9 a. m. on the 2nd and 7th were
post-bronchoscopy days. The effects of the fiberoptic bronchoscopy under local anaesthesia with lidocaine hydrochloride on the serum levels of testosterone and LH were examined in 10 males with pulmonary tuberculosis or pulmonary cancer. One hour before the bronchoscopy, which began at 9 a. m., the patients were treated with hydroxyzine-pamoate and atropine sulphate. The duration of the bronchoscopy under local anaesthesia was about 30 min. None of the patients received thiopental, succinylcholine chloride or transfusion. Serum samples were taken immediately before local anaesthesia, 30 min and 1 h after the beginning of anaesthesia and at 9 a. m. on the 2nd, 4th and 7th post-bronchoscopy days. Bronchoscopy began immediately after the onset of local
anaesthesia. The effect of human chorionic gonadotrophin (HCG) on serum level of testosterone was examined in 10 pre-operative males with pulmonary tuberculosis and in 20 male patients who had undergone lobectomy or gastrectomy 7 days before the test. Two thousand units of HCG was injected intramuscularly at 9 a. m. Serum samples were obtained from the pre- and post-operative patients immediately before the HCG in¬ jection, and 3 and 6 h after the injection.
Serum and urine
samples
The effects of surgical stress on the excretion of urinary steroids and serum levels of testosterone and LH under adrenal suppression were examined in 6 male patients with pulmonary tuberculosis. Serum and urine samples were taken 7 days before lobectomy. The patients were then given dexamethasone, 6 mg daily for 12 days. On the day of operation and on the next day, an additional dose of dexamethasone,
i.e.
3mg was injected. The operation was performed on the 6th day of adrenal suppression, and samples were obtained 1 day before and 6 days after the operation. Serum samples were taken at 9 a. m. and urine samples were collected for 24 h.
Samples
were
Estimation
stored at -20°C until
of serum
Concentrations of
testosterone
required.
and LH
testosterone were measured by the competitive protein binding method of Mayes Se Nugent (1968). Serum levels of LH were measured by the radioimmunoassay method (Aono et al. 1967), with slight modifications as pre¬ viously described (Aono et al. 1972). The evaluation of the LH and testosterone assays including specificity study was satisfactory as previously reported (Aono et al. 1972; Matsumoto et al. 1970). The intra- and inter-assay coefficient of variation in a normal male range obtained from 10 assays, were 8.9 and 9.7%, respectively in the testosterone assay and 8.8 and 13.6%, respectively in the LH assay. serum
Quantitative analyses of urinary steroids The method for the quantitative determination of urinary steroid fractions has been described in previous reports (Matsumoto et al. 1966; Seki Se Matsumoto 1967). One hundred ml of each urine sample was first treated with limpet /J-glucuronidase. After extraction of the steroids with chloroform, steroid sulphate was hydrolyzed by the method of Burstein Se Lieberman (1958). Each of the steroid hydrolysates thus obtained was washed with 1 NaOH, H.,0 and separated by elution chromatography on Amberlite IRC-50 using methanol:ethanol:water (3:9:8 v/v) as eluent. The steroids extracted following hydrolysis with /i-glucuronidase were separated into cortisol + cortisone + tetrahydrocortisol (3«,1 l/?,17a,21-tetrahydroxy-5/i-pregnan-20-one) + tetra¬
hydrocortisone (3ci,l7a,21-trihydroxy-5/?-pregnane-ll,20-dione), allotetrahydrocortisol
(3a,ll/?,17a,21-tetrahydroxy-5a-pregnan-20-one), ll/j-hydroxyaetiocholanolone (3«,ll/idihydroxy-5/?-androstan-17-one), 11-ketoaetiocholanolone (3cc-hydroxy-5/i-androstane11.1
7-dione), 11/J-hydroxyandrosterone (3a,ll/J-dihydroxy-5«-androstan-17-one),
aetio-
cholanolone
(3a-hydroxy-5/?-androstan-17-one), 5/?-pregnane-3a,17a,20a-triol, androsterone and 5/?-pregnane-3a,20a-dioL The other hydrolysate was separated into dehydro¬ epiandrosterone and 5-pregnene-3/i,20a-diol. Steroid fractions thus obtained were quantitatively
determined by Porter-Silber reagent, Zimmermann reagent, acid and Oertel-Eik-Nes reagent (Oertel Se Eik-Nes 1959), respectively.
sulphuric
RESULTS surgery and fiberoptic bronchoscopy under general or local anaesthesia on serum levels of LH and testosterone The pre-, during- and post-anaesthesia levels of serum LH and testosterone in male patients are shown in Tables 1 and 2. Fig. 1 shows the average per¬ centage changes from the pre-anaesthesia levels in the same patients. In sur¬ gical patients, the level of serum LH increased significantly from that of
Effects of major
SERUM LH
0
05
2-3
1
Fig
12
4
DAYS
HOURS
L
major surgery and fiberoptic bronchoscopy under general or local anaesthesia on serum levels of LH and testosterone in males. The results are expressed as the average percentage change from the pre-anaesthesia level. Conditions of experiments are shown in Tables 1 and 2. Differences from "Pre-anaesthesia" (P): I < 0.05, 2 < 0.01. Effects of
the control (pre-anaesthesia) 30 min and 1 h after the beginning of general anaesthesia. In spite of continuing surgical stress and general anaesthesia, the LH concentration then decreased toward the control level 2-3 h after the beginning of anaesthesia. A significant decrease in the level of serum LH was noted on the 2nd post-operative day, but the level increased slightly on the 7th day following surgery under general anaesthesia, thought the increase was not statistically significant. Although the duration of fiberoptic broncho¬ scopy under general anaesthesia ranged from 40-60 min instead of 2-4 h in surgery, the change in the serum levels of LH during and following the bron¬ choscopy under general anaesthesia was similar to that found in surgery (Table 1, Fig. 1). However, the rate of change was lower in the bronchoscopy than in surgery. On the other hand, the level of LH exhibited no significant change in the bronchoscopy under local anaesthesia (Table 1, Fig. 1).
Table i. Effects of major surgery and fiberoptic bronchoscopy under general or local anaesthesia on serum level of LH (Mean + se ug/100 ml) in males.
Before anaesthesia
Surgery
Bronchoscopy
Bronchoscopy
under general anaesthesia
under general anaesthesia
under local anaesthesia
4.5 ± 0.5
(If
.7 ± 0.6
(16)
4.3 ± 0.8
(10)
(10) ( 6)
After onset of anaesthesia 30 min 1 h
9.5 ±
0.8*;;-(12)
8.6 ±
0.8**(15)
4.5 ± 0.8
10.0 ±
1.0**(18)
9.4 ±
1.1**(15)
4.2 ± 0.4
5.0 ± 0.9
2-3 h
Post-anaesthesia 2nd day 4th day 7th day
2.9 ±
(10)
0.4*s-(16) (17) 0.8 (17)
6.1 ± 0.8
4.5 ±
(12)
0.4**(15)
4.6 ± 0.7 6.0 ±
5.1 ± 1.1 5.0 ± 0.8
7.0 ± 1.2
( 9)
4.6 + 0.6
(10) ( 9) ( 9)
Differences from "Before anaesthesia" (P): ** < 0.01 (i-test for paired observations). The duration of major surgery and fiberoptic bronchoscopy under general and local anaesthesia was 2-4 h, 40-60 min and 30 min, respectively. ( ) indicates number of patients at each point.
levels of testosterone progressively decreased during the opera¬ value significantly smaller than the control level was found 2-3 h after the beginning of anaesthesia. The lowest concentration was obtained on the 2nd post-operative day when a significant decrease in the level of LH was noted. Significantly decreased levels of serum testosterone were maintained until the 7th post-operative day when the level of serum LH showed a slight increase. The levels of serum testosterone showed no significant decrease in fiberoptic bronchoscopy under general and local anaesthesia, except for that obtained on the 2nd day following bronchoscopy under general anaesthesia, when the serum LH was found to be slightly but significantly decreased The
serum
tions, and
a
(Table 2, Fig. 1). on serum level of testosterone in pre- and male patients post-operative Since the results shown in Tables 1 and 2 and Fig. 1 suggested some decrease of testicular sensitivity to gonadotrophins under major surgical stress, the ex¬ periment shown in Table 3 was performed. However, the rate of increase in
Effect of HCG
Table 2. Effects of major surgery and fiberoptic bronchoscopy under general or local anaesthesia + se «g/100 on serum level of testosterone (Mean ml) in males.
Surgery
under general anaesthesia
Bronchoscopy
Bronchoscopy
under general anaesthesia
under local anaesthesia
0.77 ± 0.04
(18)
0.57 ± 0.04
(16)
0.73 ± 0.04
(10)
30 min
0.83 ± 0.06
(12)
0.62 ± 0.04
0.72 ± 0.05 (18) 0.47 ± 0.09**(10)
0.58 ± 0.04
(15) (15) (12)
0.67 ± 0.05
1 h
(10) ( 6)
0.22 ± 0.29 ± 0.51 ±
0.47 ±
0.04*(15)
0.68 ± 0.07
( 9)
0.67 ± 0.05 0.74 ± 0.05
Before anaesthesia After onset of anaesthesia
2-3 h
Post-anaesthesia 2nd day 4th day 7 th day
0.03**(16) 0.03**(17) 0.05**(17)
Differences from "Before anaesthesia"
observations). ( ) indicates number
Effect of HCG
of
patients
on serum
0.63 ± 0.04
at
*
(P):
each
0.53 ± 0.04
< 0.05,
**
< 0.01
0.67 ± 0.05
(i-test
for
Table 3. level of testosterone in pre- and
post-operative
Mean ±
After operation 6th day2 6th
day3
Differences from "Before
observations). 1
2 3
se
males.
injection of HCG'
3 h
operation
paired
point.
Alter
Before
(10) ( 9) ( 9)
6 h
(«g/100 ml)
10
0.68 ± 0.12
0.76 ± 0.11
0.91 ± 0.10*
10
0.32 ± 0.04
0.45 ± 0.04**
10
0.49 ± 0.06
0.57
0.46 ± 0.04* 0.65 ± 0.08*
injection" (P):
*
< 0.05,
**
± 0.06*
< 0.01
(¿-test
for
paired
Two thousand units of HCG was injected intramuscularly at 9.00 a. m. Control serum samples were obtained immediately before the HCG injection. Post-operative and pre-operative samples were obtained from the same patients. Post-operative samples were obtained from different groups of patients.
testosterone level similar to that in
post-operative patients was pre-operative patients, though the pre- and post-injection values were significantly lower in the post-operative patients than in the pre¬ operative patients, respectively.
following
Effects of surgical stress
HCG injection in
urinary steroid excretion and serum levels of patients under adrenal suppression Since the results shown in Tables 1-3 and those obtained by Tcholakian Sc Eik-Nes (1971) in dogs seemed to suggest a shift in the androgen biosynthetic pathway of the testis following surgical stress, urinary steroids were measured under adrenal suppression with dexamethasone in 6 male patients before and 6 days after pulmonary lobectomy. Metabolites of cortisol decreased markedly testosterone
fiftieth after dexamethasone administration. Under adrenal suppression, urinary excretion of aetiocholanolone plus androsterone de¬ creased significantly from the pre-operative level on the 6th post-operative to
less than
on
and LH in male
one
the
Effects of
Table 4. stress on urinary steroid excretion and serum levels of testosterone and LH in 6 males under adrenal suppression with dexamethasone.
surgical
No
suppression
Before op.
urinary
steroids
Metabolites of cortisol2
11-Oxy-17-ketosteroids 11-Deoxy-17-ketosteroids 5-Pregnene-3/?,20a-diol 5/LPregnane-3a,20a-diol 5/?-Pregnane-3a,17a,20a-triol Serum testosterone and LH Testosterone LH
Adrenal
Before op.1 Mean ±
se
1
2
After
op.1
(mg/24 h)
3.18 ± 0.75 0.89 ± 0.24 1.92 ± 0.46
0.06 ± 0.02
0.06 ± 0.03
0.14 ± 0.05
0.06
1.24 ± 0.20
0.52 ± 0.08s
< 0.10 < 0.20 0.98 ± 0.20
