Acta Obstet Gynecol Scand 55: 387-394, 1976

HISTAMINE METABOLISM AND FEMALE SEX HORMONES IN WOMEN F. Jonassen, G. Granerus and H. Wetterqvist From the Department of Obstetrics and Gynecology (Head: Prof. N . Wiqvist). the Department of Clinical Physiology I . (Head: Prof. A . Carlsten), and the Department of Clinical Chemistry (Head: Prof. S . Lindstedt), Sahlgren's Hospital. Universitv of Gothenburg. Sweden

Abstract. Oral combined contraceptives did not seem to alter histamine metabolism in females. During treatment with gonadotrophic hormones in four amenorrhoeic patients there was a tendency towards increasing excretion of methylhistamine (MeHi) followed by a sudden decrease corresponding to changes in the urinary estrogen. The excretion of methylimidazoleacetic acid (MeImAA) seemed to parallel that of MeHi. The findings support the hypothesis that an endogenous surge of estrogen may influence histamine turnover in women. Women of postmenopausal age have about the same histamine metabolism as younger menstruating women. Estrogen medication relieved symptoms of hot flushes or sweats but did not seem to affect the histamine turnover.

INTRODUCTION Histamine metabolism seems to be influenced by the normal menstrual cycle (11). The women investigated showed individual variations in the excretion of both histamine and its metabolites. There was no significant difference in the excretion during the pre- and post-ovulatory phase. However, the urinary excretion of methylimidazoleacetic acid (MeImAA) showed a tendency to increase at the time of ovulation. Also there seemed to be a significant correlation between the excretion of estrogen and that of methylhistamine (MeHi). In adult women Green et al. (7) reported that estrogens might influence the urinary excretion of histamine and MeHi. The cases, however, were few in number and the analyses were not performed under standardized dietary conditions. The present investigation represents an extension of our previous studies on the influence of female sex hormones on histamine metabolism and included the effect of oral contraceptives, ovulation

stimulation with gonadotrophic hormones and estrogen substitution at postmenopausal age. MATERIAL AND METHODS Sixteen healthy women were treated with different oral contraceptives over 17 tablet periods and a total of 77 urine samples were collected. In 10 of these women urine were also collected during their normal menstrual cycle (39 samples). The excretion of histamine (Hi) and methylhistamine (MeHi) was measured. A second group comprised four women with secondary amenorrhoea. Two of the subjects were studied during one period of stimulation with gonadotrophic hormones, one was studied during two periods and the fourth during four periods of stimulation. In this group the excretion of histamine and metabolites was studied as well as that of total estrogen and luteinizing hormone (LH). The progesterone level in plasma was measured to decide whether ovulation had taken place. A third group included 20 postmenopausal women. Ten of them had subjective symptoms such as hot flushes and/or sweats. The other ten had no subjective symptoms at all (Table I). In this group of women urine collection was performed twice on two consecutive days, and this was repeated approximately four weeks later. Histamine and MeHi in the urine were determined. At one of these days serum was taken for analysis of LH and follicle-stimulating hormone (FSH) and estrogen excretion was determined in the urine. The 10 women with symptoms were studied in the same way before and during treatment with estrogen (ethinylestradiol 20 p g per 24 hours) four weeks later. Urine was collected in 24-hour portions. The urinary samples were immediately placed in a refrigerator at +4T. When samples for hormone analyses had been withdrawn the urine was mixed with hydrochloric acid ( 1 M). All collections of urine were performed under standardized dietary conditions (4). Histamine (Hi) in the urine was measured by a bioassay technique according to Wetterqvist &White (19) and the values given are expressed as pg histamine base per 24 Actu Obstet Gynecol ScundS.5 (1976)

388

F. Jonassen et al.

Table I. Pertinent data on 20 postmenopa~sulwomen in group 4 A-K had no symptoms and treatment, L-U had symptoms of hot flushes and/or sweats and were treated with estrogen. +, Hot flushes or sweats; + +. both symptoms present; (+), almost free from flushes andfor sweats; 0, no symptoms

Subject

A B C D E F G H I K L M N 0 P

Q R

S T U

Age (y.) 50 51 51 56 57 58 58 60 60 65 48 51 52 54 55 56 57 57 58 58

Parity

Weight (kg)

Smoking cigarettes/day

Time since menopause (Y.)

60

< 10

59

0 0

3 2 3 2 2 1 6 12 16

64 66

0

1 3

70 65

< 10

3

64 64

0 0

< 10 0

61 65 60 62 59 66 65 60 61 60 64

0 2

0

10 2 1

< 10

3

0

3 1

0

< 10

0 0 0 0 0 0

64

hours, corrected for recovery. The identity of histamine was checked according to Reuse (15). Methylhistamine (MeHi) in the urine was determined as described by Fram & Green (3). White (20) and Granerus et al. (6). The values are given a s pg methylhistamine base per 24 hours, corrected for recovery. I-methyl-4-imidazoleacetic acid (MeImAA) in the urine was determined as described by Granerus & Magnusson ( 5 ) with slight modifications (4, 1 l), corrected for recovery and expressed as mg per 24 hours.

3 5 1 3 4

Symptoms

Symptoms after treatment

++ + ++ ++ ++ + ++ + ++ +

Excretion of luteinizing hormone (LH) in urine was measured according to a technique described by Wide & Porath (22), Wide (21) and Wide et al. (23). The values are expressed in I.U. per 24 hours. LH in serum was determined according to the same technique and expressed in ng per ml. The follicle-stimulating hormone (FSH) in serum was determined according to Wide et al. (23) and is expressed in ng per ml. The total estrogen excretion was determined in urine

Table 11. Urinary excretion of histamine ( H i )and methylhistamine (MeHi)in pg buse per 24 horrrs in seven cycles from six women taking oral combined contraceptives (group I ) Subject

Compound

Samples

Mean +S.E.

Range

Tablet taken ~~

A

B C C D

E F

Hi MeHi Hi MeHi Hi MeHi Hi MeHi Hi MeHi Hi MeHi Hi MeHi

5 5

46+ I 246+29

42- 50 167-323

Follinyl

5 5

14k 2 233 +25

10-20 158-295

Conlunett

5 5

16+ I 230+35 IS+ 2 220+3 1

12- 19 180-334

Conlunett

15- 20

Conlunett

23k 2 211+17

10- 38 128-346

Piloval

16t 2 209+32 36+ 8 166k25

lo- 21 I 19-30 I 21- 65

Anovlar mite

5 5

12

12 5 5 5 5

Acra Obstet Gynecol Scand 55 (1976)

151-302

110-231

Lyndiol mite

Histamine me~abolismand female sex hormones

389

120100.

80 LH IU/24hrs

4020.

80 601

Hi ~ g / 2 4 hrs loo

MeImAA rnq/24hrs

n

nMflnnI-9

4.

.

2-

4

8

I2

I6

I

20

24

28

0"

I

M

Fig. I . Urinary excretion of histamine (Hi), methylhistamine (MeHi), methylimidazoleacetic acid (MeImAA), estrogen (Estr) and luteinizing hormone (LH) in a woman

(age 26, one pregnancy, non-smoking) during one menstrual cycle before (left) and (right) three months later during a period o n oral contraceptives (Follinyl).

according to the method of Brown (1) and Ittrich (8) and the values are given in pg per 24 hours. Progesterone analyses in plasma were performed as described by Johansson (lo), modified by Ellingboe et al. (2). The analyses of estrogens and progesterone were assayed at the Department of Clinical Chemistry, Sahlgren's Hospital.

fS.E.). Linear regressions were calculated according to the method of least squares.

Drugs

In the first group of women: Conlunett=norethisterone 1 mg + mestranol 0.1 mg; Follinyl=norgestrel 0.5 mg + ethinylestradiolO.05 mg; Lyndiol mite=lynestrenol2.5 mg + mestranol 0.075 mg; Anovlar mite=norethisterone acetate 3 mg + ethinylestradiol 0.05 mg; Piloval=quingestanol acetate 0.5 mg + ethinylestradiol 0.05 mg; Astra 2028=norethisterone 1 mg + ethinylestradiol 0.05 mg. In the second group of women: Humegon=human menopausal gonadotrophin (HMG); Pregnyl= human chorionic gonadotrophin (HCG). In the second group of women: Linoral=ethinylestradi01 0.01 mg. Statistical methods

Conventional statistical methods were used for the calculation of means and standard error of means (mean

RESULTS AND COMMENTS Six women on oral contraceptives excreted histamine and methylhistamine in the urine (Table 11) in quantities within the previously published limits of the normal menstrual cycle (4, 11). In nine other women where the subjects served as their own controls there were no differences in the mean urinary excretion of histamine and MeHi before and during treatment with oral contraceptives. (Hi before treatment 3 7 f 5 pg/24 hours, during treatment 3 2 f 4 pg/24 hours, MeHi before 192k 15 pg/24 hours, during treatment 195f 13 pg/24 hours.) Fig. 1 illustrates the excretion during one.menstrua1 cycle and a period with oral contraceptives on one healthy female (the sixteenth). N o significant changes in urinary histamine or MeHi were observed. At one single day (the fourteenth) the Acra Obster Gynecol Scund 55 (1976)

390

F. Jonassen et al. LH in serum ng/ml 4j

r,

200 1 Estr p g / 2 4 h r s loo

1

2001

400

Fig. 2 . Urinary excretion of histamine (Hi) and methylhistamine (MeHi) in a woman (subject F in Table 11, smoking) during a period on oral contraceptives (Lyndiol mite; left). Twelve months later one menstrual cycle was examined when the woman had been without medication for some months (right). During this cycle the estrogen excretion (Estr) in the urine and luteinizing hormone (LH) in serum were also determined.

1

1

MeHi p g / 2 4 hrs 2oo

20

10

20

10

excretion of MeImAA rose substantially. This was preceded by a peak in histamine on the eleventh day and peak in MeHi on the twelfth day (right). Fig. 2 shows on the other hand one woman (subject F in Table 11) first studied when taking oral contraceptives (Lyndiol mite). She then stopped the medication and the investigation was resumed 12 months later. In this particular case there was a higher urinary excretion of both histamine and MeHi during the menstrual cycle. Figs. 3-6 show the cases in group 2 . Four women with secondary amenorrhoea were treated with human menopausal and human chorionic gonadotrophin during altogether eight periods. The doses injected are presented together with the excretion values of hormones and histamine and metabolites in the figures. The excretion levels of plasma progesterone verified ovulation on all occasions. Three of the patients seemed to react in the same

HCG I

HMG

manner. They showed a tendency to increased urinary output of MeHi in parallel with increased estrogen excretion (Figs. 3 and 4). After the maximal estrogen response had been reached there was a decrease in the MeHi values. As shown in Fig. 3, subject excreted increasing amounts of both MeHi and MeImAA in parallel with an increased urinary estrogen excretion. A subsequent decrease was observed immediately after the period of stimulation. However, in a second course of gonadotrophin stimulation of subject 3 no significant elevation of MeHi output in connection with the estrogen peak could be observed (Fig. 4 right and Fig. 5). The fourth patient (Fig. 6) reacted with increasing urinary excretion of histamine but not of MeHi or MeImAA. In a second attempt to stimulate ovulation there was a weaker hormonal response and a slight if any increase in urinary histamine. Fig. 7 and Table I11 show the results of measure-

HCG

c

HMG

0 -

2001

MeImAA m g / 2 4 hrs

30 Days

3

I

200,

,

,

,

5

10

15

, hkrn~",",,,

20 Days

Acta Obstet Gynecol Scand 55 (1976)

1,1 !$, 5

10

Fig. 3. Urinary excretion of histamine (Hi), methylhistamine (MeHi), methylimidazoleacetic acid (MeImAA) and estrogen (Estr) in subject 1 (left) and subject 2 (right) with secondary amenorrhoea (group 2). Subject 1 was treated with human menopausal gonadotrophin ( H M G t 7 5 , 150 or 225 I.U. daily-as indicated at the top of the figures. Subject 2 was treated with doses of 150,225 or 300 I.U. daily. At the point marked by the arrow both women were injected with 9000 I.U. of human chorionic gonadotrophin (HCG).

, 15

2 0 Days

-

HCG i HMG

HCG i

I

2001

Hi pg/24hrs

Histamine metcibolism cind female sex hormones

Ioo]n 400 1

1

-

n RTC?

Inn

n

n

n n

n

1

MeHi

pq/24 hrs

Fig. 4. Urinary excretion of histamine (Hi), methylhistamine (MeHi), estrogen (Estr) and luteinizing hormone (LH) in subject 3 with secondary amenorrhoea (group 2). She was treated during the first period (left) with 150 I.U. of human menopausal gonadotrophin (HMG) daily and during the second period (right) with doses of 150 or 225 I.U. daily as indicated at the top of the figures. At the points marked by the arrows she was injected with 9000 I.U. of human chorionic gonadotrophin (HCG).

ments in the 20 postmenopausal women in group 3. The mean excretion values for histamine and MeHi in the group were within the normal limits previously published for fertile women (4, 11) and no differences were found between the subjects with and those without symptoms. The 10 women with symptoms were then given estrogen orally each day for at least four weeks while the other 10 received no treatment. There was a slight positive correlation between the excretion values for estrogen and for urinary histamine in the symptom free untreated group (r=0.58) (Fig. 7, left). Estrogen treatment did not alter the urinary excretion of histamine and MeHi. Of the hormones analysed only FSH showed a decrease and this observation, together with the clinical improvement, indicated that the dose given was adequate.

DISCUSSION The current study illustrates the difficulties involved in trying to understand the r6le of female sex hormones in histamine metabolism in man. Firstly, it is obvious that there are large individual variations in excretion values of histamine and MeHi, even in women without abnormal changes in the

391

internal secretion of sex hormones. It is known that the methodological error is on average ? l o % for the analyses of histamine and MeHi (Wetterqvist, unpublished). Secondly, there is an upper limit for the ability of a subject to cope with an investigation of this type. More samples from more women would have been desirable but this was not possible for practical reasons. Thirdly, the processes underlying the turnover of female sex hormones in the body are not yet fully understood. We have not included studies of releasing factors in our schedule nor did we analyse adrenal secretions. The possibility also remains that the histamine turnover may affect the secretion of female sex hormones via a local or general feedback mechanism. Fourthly, the principles for the regulation of the total histamine turnover in man are largely unknown. The importance of the composition of the food ingested for histamine metabolism is undisputable (4), but the functional state of the gastrointestinal tract (17) as well as physiological changes in the kidneys may also be of some importance. Green et al. (7) found increased values for histamine and/or MeHi in the urine of two amenorrhoeic patients in whom an increase in the endogenous estrogen levels had been induced by administration

-'

HMG

HCG

200,

-'

HCG

3

Estr pg / 2 4 hrs loo

Hi p g / 2 4 hrs

'"1

r?

1

-nn

Fig. 5 . Urinary excretion of histamine (Hi), methylhistamine (MeHi), estrogen (Estr) and luteinizing hormone (LH) in subject 3 with secondary amenorrhoea (group 2) during two further periods of hormone stimulation. During these periods she was treated with 150 or 225 I.U. of human menopausal gonadotrophin (HMG) daily as indicated at the top of the figures. At the points marked by the arrows she was injected with 9000 I.U. of human chorionic gonadotrophin (HCG). Acta Obstet Gvnecol ScandS5 (1976)

392

F. Jonassen et NI.

HMG

~- -

HCG

HCG

4001

cr-1' n

1

200,

1

400 1

1

MeHi pg124hrr 2oo

Fig. 6. Urinary excretion of histamine (Hi), methylhistamine (MeHi), estrogen (Estr) and luteinizing hormone (LH) in subject 4 with secondary amenorrhoea (group 2) during two periods of hormone stimulation. She was treated with 75, 150 or 225 I.U. of human menopausal gonadotrophin (HMG) daily as indicated at the top of the figures. At the points marked by the arrows she was injected with 9000 I.U. of human chorionic gonadotrophin

(HCG).

of gonadotrophic hormones. The changes in MeHi values were not impressive whereas the histamine tended t o increase. However, there did not seem to be any absolute correlation between high estrogen values and a high urinary histamine output, nor did Green et al. collect urine under standardized dietary conditions. Previous results from this department showed a probable correlation between the endogenous estrogen levels and histamine turnover in that histamine, MeHi and MeImAA excretion increased at midcycle ( 1 1). The present study seems to support this observation. Stimulation of amenorrhoeic women with gonadotrophin inducing a marked increase in the endogenous estrogen levels resulted in a slight augmentation of histamine and/or metabolite output in 4 of 8 trials. Oral contraceptives (estrogen or gestagen dominated) did not influence histamine turnover significantly. A similar lack of effect was found in the guinea-pig (14) whereas a combination of estrogen and progesterone increased histamine output in the mouse (13) and decreased the excretion in the rat (12). These findings illustrate marked differences in Acta Obstet Gynecol Scand 55 (1976)

histamine turnover between different species. It should, however. be born in mind that the oral contraceptives represented a long-lasting medication whereas the animals were subjected to an acute hormonal stimulation by single injections. The mechanism behind the steroid induced effect upon histamine turnover in the human female is difficult to explain. It has been shown that estrogens effect on histamine depletion in the uterine tissue (9. 16). It seems, however, not probable that only local alterations within the reproductive organs may be reflected in the urinary output of histamine since mast cells are presented in nearly all tissues throughout the human body. Climacteric symptoms are mainly characterized by the occurrence of hot flushes and sweats. The pathogenesis of this syndrome is not wellunderstood. It has been suggested that the vascular symptoms should be the result of an instability between the hypothalamus and the autonomic nervous system brought about by a decline in estrogen. Hot flushes in the same region of the body can be induced by-'intravenous injection of histamine and it could be speculated that this substance might be involved in the pathogenetic events. The present results show that there is no difference in the excretion of histamine and MeHi between postmenopausal women with and without climacteric symptoms. Nor is there any difference in this respect between postmenopausal and fertile

5 -

Erlr

5 -

Ngl24hra

Ertr pg /24 hrs

Fig. 7. Urinary excretion of histamine (Hi), methylhistamine (MeHi) and estrogen (Estr) in 10 women of menopausal age without any symptoms or treatment (left) and in 10 other women of the same age but with symptoms of hot flushes and/or sweats (right). X denotes values obtained without any treatment. Circles denote values obtained when the women were treated with ethinylestradiol (20 pg daily). (Left upper part: y=2.6&+6.52).

Histamine metabolism and female sex hormones

393

Table 111. Urinar-y excretion of histamine (Hi) and inethylhistamine ( M e H i ) in pg brise per 24 hours in 20 menopausal women (group 3) The first 10 (A-K) had no symptoms and treatment. The others (L-U) had symptoms of hot flushes and/or sweats and were treated with ethinylestradiol at the time of the second urine collection (20 pg daily) Subject

Compound

Samples

Treatment

Mean 2S.E.

Range

A-K

Hi MeHi Hi MeHi Hi MeHi Hi MeHi

20 20 20 20

No No No No No No Yes Yes

282 3 197k16 27f 2 202k19 22If- 3 192f I I 23k 4 195flS

lo- 49 92-354 10- 46 76364 7- 59 109-289 7- 88 1 10-247

L-u

19 19 20 20

women ( I I). It is hard to believe that the probable linear distribution of the histamine excretion values in the symptom-free women as compared to the lack of linearity in patients with symptoms has a n y meaningful significance. It has been observed that patients undergoing cryotherapy for premalignant cervical lesions get temporary hot flushes during the thawing of the cervical tissue. Measurement of urinary histamine and M e H i showed a n increased

excretion for a t least four hours following the treatment (18). It seems probable that postmenopausal hot flushes are not elicited b y a systemic release of histamine into the circulation. ACKNOWLEDGEMENTS We wish to thank the Department of Clinical Chemistry, Sahlgren’s Hospital, for valuable help with hormone analyses. This study was supported by grants from the Medical Faculty, University of Goteborg, and from the Goteborg Medical Society.

REFERENCES 1. Brown, J. E.: A chemical method for the determination of oestriol, oestron and oestradiol in human urine. J Biochem60: 185, 1955. 2. Ellingboe, J., Nystrom, E. & Sjovall, J.: Liquid gel chromatography of steroids on hydrophobic sephadex derivatives. Acta Endocrinol61, Suppl. 138: 20, 1969. 3. Fram, D. H. & Green, J. P.: The presence and measurement of methylhistamine in urine. J Biol Chem 240: 2036, 1965. 4. Granerus, G.: Urinary excretion of histamine, methylhistamine and methylimidazoleacetic acids in man under standardized dietary conditions. Scand J Clin Lab Invest 22, Suppl. 104: 59, 1968. 5. Granerus, G. & Magnusson, R.: A method for semiquantitative determination of 1-methyl-4-immidazole-

acetic acid in human urine. Scand J Clin Lab Invest 17: 483, 1965.

6. Granerus, G., Wetterqvist, H. & White, T.: Histamine metabolism in healthy subjects before and during treatment with aminoguanidine. Scand J Clin Lab Invest 22, Suppl. 104: 39, 1968. 7. Green, J. P., Fram, D. H. & Kase, N.: Methylhistamine and histamine in the urine of women during the elaboration of estrogen. Nature 204: 1165, 1964. 8. Ittrich, G.: Untersuchungen iiber die Extraction des roten Kober-Farbstoffs durch organische Losungsmittel zur Ostrogenbestimmung in Harn. Acta Endocrinol35: 34, 1960. 9. Iversen, 0. H.: Mast cells in the myometrium of the human cervix uteri and changes caused by androgenic and estrogenic hormones. Acta Pathol Microbiol Scand49: 337, 1960. 10. Johansson, E. D. B.: Progesterone levels in peripheral plasma during the luteal phase of the normal menstrual cycle measured by a rapid competitive piotein binding technique. Acta Endocrinol (Kbh) 61: 592, 1969. 11. Jonassen, F., Granerus, G . & Wetterqvist, H.: Histamine metabolism during the menstrual cycle. Acta Obstet Gynecol Scand 55: 297, 1976. 12. Jonassen, F. & Wetterqvist, H.: Effects of female sex hormones on histamine metabolism in rats. To be published. 13. Jonassen, F. & Wetterqvist, H.: Effects offemale sex hormones on histamine metabolism in mice. To be published. 14. Jonassen, F. & Wetterqvist, H.: Effects of female sex hormones on histamine metabolism in guinea-pigs. To be published. 15. Reuse, J. J.: Comparisons of various histamine antagonists. Br J Pharmacol3: 174, 1948. 16. Szego, C. M.: RBIe of histamine in mediation of hormone action. Fed Proc24: 1343, 1965. 17. Tham, R.: Liberation of histamine in man. Gas chromatography of ring methylated imidazoleacetic acids in urine. Scand J Clin Lab Invest 18: 603, 1966. 18. Tronstad, S. E., Bergstrom, H., Granerus, G., Jonassen, F. & Wetterqvist, H.: Flush och okad urinActu Obstct Gynecol Scund 55 (1976)

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19. 20. 21. 22.

F. Jonassen et al. utsondring av histamin och methylhistamin vid kryobehandling av cervix uteri. Medicinska Riksstamman, p. 327, 1973. Wetterqvist, H. &White, T.: Bioassay of histamine in human urine. Scand J Clin Lab Invest 25: 325, 1970. White, T.: Histamine and methylhistamine in cat brain and other tissues. Br J Pharmacol26: 494, 1966. Wide, L.: Radioimmunoassays employing imrnunosorbents. Acta Endocrinol 63, Suppl. 142: 207, 1970. Wide, L. & Porath, J.: Radioimmunoassays of proteins with the use of Sephadex-coupled antibodies. Biophys Acta 130: 257, 1966.

Acta Obster Gynecol Scand 55 (1976)

23. Wide, L., Nillius, S. J., Gemzell, C. & Roos, P.: Radioimmunosorbent assay of follicle-stimulating hormone in serum and urine from men and women. Acta Endocrinol73, Suppl. 174: 7, 1973. Submitted for publicution March 17, 1975

Fredrik Jonassen Department of Obstetrics and Gynecology Sahlgren’s Hospital 413 45 Gothenburg Sweden