Decrease in melatonin precedes follicle-stimulating hormone increase

during perimenopause
Olli Vakkuri, Aarre Kivel\l=a"\, Juhani Lepp\l=a"\luoto, Maija Valtonen and Antti Kauppila
Departments of Physiology and Obstetrics and Gynecology, University of Oulu, Kajaanintie, Oulu, Finland; Veterinary Research Station
University of Kuopio. Kuopio, Finland

Vakkuri O, Kivel\l=a"\A, Lepp\l=a"\luotoJ, Valtonen M, Kauppila A. Decrease in melatonin precedes follicle-

stimulating hormone increase during perimenopause. Eur J Endocrinol 196;135:18\p=n-\92. ISSN
0804\p=n-\4643
Melatonin, the hormone of the pineal gland, which in animal studies has been found to inhibit aging
processes, is secreted in smaller amounts towards senescence. Menopause, an aging process in women,
is known to be associated with typical changes in gonadotropin and sex steroid secretion. Our main
objective was to study the possible role of melatonin in the hormonal regulation of menopause. This
study focused on detailed changes in melatonin and follicle-stimulating hormone (FSH) secretion
cross-sectionally in pre- to postmenopausal females. Special attention was paid to females aged around
50 years, which is the mean menopausal age. Seventy-seven healthy female volunteers aged 30\p=n-\75
years were the subjects of this study. Melatonin was measured radioimmunologically from nocturnal
urine collected between 20.00 and 08.00h, and FSH and melatonin from blood samples taken at
09.00 h. Nocturnal urinary excretion of melatonin was found to decline significantly from premeno-
pause to postmenopause. The youngest premenopausal women (age group 30\p=n-\39 years) excreted the
highest amounts of melatonin (21.1 \m=+-\2.2pmol/h, mean\m=+-\sem,N 17). In the age group 40\p=n-\44
=

years the excretion declined by 41% (p < 0.05). The second significant decline (35%, p < 0.05) took
place between the age groups 50\p=n-\54 years and 55\p=n-\59 years. A declining trend as a function of age
was also seen in morning serum melatonin, Serum FSH rose sharply to high levels before the age of 50
(p < 0.01) and remained at a high level thereafter. Urinary melatonin correlated negatively with
serum FSH (r = \p=n-\0.32. p < 0.05). In conclusion, the inverse changes in melatonin and FSH secretion
during the perimenopausal years, with the sharpest decline in nocturnal excretion of melatonin far
before menopause, suggest that melatonin may be permissively linked to the initiation of menopause.
O Vakkuri. Department of Physiology. University of Oulu, Kajaanintie 52 A, 90220 Oulu, Finland

Menopause is a unique phenomenon in that female
reproductive competence goes away as a result of
cessation of ovarian function far before senescence. It
involves several hormonal changes, of which a decrease
in the synthesis of ovarian steroids and inhibin and an
increase in the synthesis and release of pituitary FSH
and LH are the most typical (1, 2). Although it has been
proposed that the sensitive ovarian follicles are
exhausted by menopause (3), it is not known what
factors determine the timing of menopause and what
mechanisms trigger the menopausal development.
Synthesis and secretion of melatonin, the pineal
hormone, takes place during the night, the dark time
of the day. Thus, it mediates rhythmic information of
night length to the organism (4). In addition to its
diurnal and seasonal significance, melatonin may be
involved essentially in the timing of puberty. Circulating
melatonin levels have been observed to decrease during
childhood, from under 5 years up to puberty, which
may be critical for the initiation of puberty (5-7).
Melatonin secretion has been reported to decrease also
after puberty as a function of age (6, 8-10). It is not
known whether this decrease in adults is gradual and
associated with normal aging or whether it is linked to
the cessation of female reproduction during menopause.
In previous studies mentioned above, data on meno¬
pausal melatonin changes are fragmentary. Fernandez
et al. (11) reported a decrease in morning serum
melatonin but, controversially, no changes in urinary
excretion of melatonin in some cohorts of menopausal
women as compared with fertile women. Therefore, we
were interested to study the pattern of melatonin
secretion throughout the perimenopausal years more
thoroughly.
Subjects and material
The subjects of this study were female volunteers aged
30-75 years who gave their informed consent. Most of
the women were healthy but some were treated for a
benign disease (due to uterine fibroma, genital prolapse
or urinary incontinence) not related to ovarian func¬
tion. The subjects were asked to give their menstrual
records in order to assess the reproductive state

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 35 90

30
S 150
O
o E
. 25
60
I
.2 20 -
(

S 15 C
«
T50H
30 =
c
'c 10 t**\**É ^
iÉ»
30 35 40 45 50 55 60 65 70 75
5
I FEMALE AGE
°-¿g»-,-8-88g'¿
. Fig. 2. Serum melatonin as a function of female age in a scattergram
50 55 60 65 70 75
30 35 40 45
presentation with linear regression analysis (y -0.13x + 18.2). All
=

FEMALE AGE serum samples were collected at 09.00 h.

25 r 75

at the time of use was 96%. Serum FSH was analyzed

" 20
with a commercial time-resolved FIA kit developed for
50 human FSH (Delfia. Wallac, Turku, Finland).
15 Statistical analyses were carried out using one-way
V-
_ analysis of variance. Correlation analyses and curve
10
fittings of the raw data were based on the FigP graphics
ç
-

25 — program.

5
2
Results
A scattergram with curve fittings of the results of the
30-39 40-44 45-49 50-54 55-59 >60 urine melatonin and serum FSH analyses is presented in
FEMALE AGE GROUP Fig. 1 A. Urinary excretion of nocturnal melatonin was
approximately 20pmol/h in premenopausal women
Fig. 1. Nocturnal melatonin (solid triangle) excretion into urine and of < 40 years but about 8 pmol/h in postmenopausal
FSH (open circle, sampled at 09.00 h) as a function of female
women of > 60 years. The 60% decrease was
serum

age. presented as a scatter-plot with curve fittings (A) and in six age < 0.01). The best curve fitting for this
groups (B. solid column for melatonin and open column for FSH: significant (p
means ±sem). *p < 0.05 for melatonin; ##p < 0.01 for FSH. decrease is cubic spline (Fig. IA).
The melatonin data of Fig. IA divided into six age
groups (Fig. IB) shows that melatonin excretion
(premenopausal postmenopausal women). The
and decreased by 41% (p < 0.05) from 21.1 ± 2.2 pmol/h
subjects received hormonal treatment for at least
no (mean ± sem, 17) in premenopausal women of
=

1 year before the study. The total number of subjects < 40 years to 12.5 ± 2.2 pmol/h (N 15) in women =

was 77. The data were pooled according to six age of 40-44 years. Thereafter, melatonin excretion
groups: 30-39 (N=17), 40-44 (N=15), 45-49 decreased to 11.2 ± 1.3 (N 13) in women of 50-54
=

(N 13), 50-54 (N 14), 55-59 (N 8) and over

= = = years (p > 0.05 compared with age groups 40-44 and
60 (N 10) years. = 45-49 years) and then by 35% (p < 0.05) to 7.3 ± 0.9
Melatonin secretion was followed by measuring the (N 8) pmol/h in women of 55-59 years.
=

nocturnal excretion of melatonin into urine. This was
collected between 20.00 and 08.00 h. Additionally,
blood samples were taken for serum FSH and melatonin
measurements at 09.00h. In fertile women, sampling
was carried out without any menstrual timing because
according to our (12) and other studies (13, 14)
melatonin does not significantly vary during the
menstrual cycle. The whole collection procedure was
carried out over a 2-month period (October-November).
Urine and serum melatonin were determined radio-
immunologically as described previously (15) using
[2-12,I]melatonin (16) as tracer. The purity of the tracer
Serum FSH level was generally < 10 IU/1 before
menopause and > 40 IU/1 after the age of 50 years.
These FSH observations could be curve-fitted according
to the exponential sigmoid model (an exponential
increase between two plateaus). More exactly, serum
FSH rose from 4.0 ±0.6 IU/1 (40-44 years) to
15.3 ±4.0 IU/1 (45-49 years) and then furthermore
to 41.8±7.8IU/1 (50-54 years); < 0.01 in both
cases (Fig. IB). Thus, the FSH rise started about 5 years
later than the decline in urinary melatonin. The changes
in serum FSH during perimenopause correlated
negatively to the corresponding changes in urinary

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excretion of melatonin (y = —5.38x + 37.9: r =body size in children, rather than to decreased secretion
—0.32,
< 0.05). of melatonin per day (6, 27), when taking into account
A scattergram of serum melatonin levels at 09.00 h also lack of pineal growth during childhood (28). In
(Fig. 2) shows that the morning level in the youngest adults the decrease is likely to be associated with a
age group was 53.3 ± 7.3 pmol/1, whereas the level in decreasing activity of the pineal gland, which is a process
the oldest age group was 33.5 ± 6.5 pmol/1. The associated with aging. The corresponding decline has
difference was not significant. The curve fitting used also been observed in GH and several other hormones
in Fig. 2 for serum melatonin data is based on linear (29, 30).
regression analysis (y —0.13x + 18.2). The negative
= Our results on an age-related decrease in urinary
correlation (r —0.12) between serum melatonin and melatonin are in general agreement with those on
=

age was not statistically significant (p > 0.05). serum melatonin by Waldhauser et al. (6). However, a
direct comparison is not possible because Waldhauser et
al. (6) had only four adult age groups with an age range
Discussion
of 15-20 years/group. They found decreased levels in
In the present study we observed a clearcut decrease nocturnal serum melatonin after menopause in older
(60%) in nocturnal melatonin excretion from fertile women, but not premenopausally. Due to the wide age
premenopausal ages of < 40 years to postmenopause of limits of the cohorts, closer inspection about the
> 60 years. This finding is in accordance with and association of melatonin to menopause was not
confirms the previous results on changes in circulating possible. It must be pointed out also that Waldhauser
melatonin as a function of human age. After the first et al. (6) measured nocturnal serum melatonin from a
controversial results in the early 1980s (17-23), an single blood sample, which might therefore not be as
age-related decline in melatonin secretion and a informative as whole nocturnal urine is.
negative correlation between melatonin secretion and We measured also morning serum melatonin in
age from childhood to senescence has been reported conjunction with FSH measurements. Morning mela¬
unambiguously in several studies (6, 8, 9, 24, 25). The tonin gives only limited information because, in
different findings reported in the early 1980s may be contrast to high nocturnal levels, morning levels (at
partly due to methodological reasons, to sampling only 09.00 h. 1-2 h after awakening) are fairly low and very
during daytime or to a limited age range monitored for close to low daytime levels (31). Additionally, we
melatonin secretion. performed the study in autumn when day length is
Our study focused especially on the rate and pattern getting shorter (in November, < 8 h in Oulu) and
of the decline in melatonin excretion from premeno¬ therefore there might have been differences in the
pause to postmenopause, i.e. throughout the mean diurnal rhythmic phase between the subjects. However,
menopausal age, which has been observed to be 51 a trend to lower serum melatonin levels was observed as
years in Finnish women (26). In our curve-fitting a function of age. It should be pointed out that
calculations the cubic spline was the best depictor ofthe Fernandez et al. (11) found a significant decrease in
age-related changes in urinary excretion of nocturnal morning serum melatonin, but only when the compar¬
melatonin. This indicates that the decline was not ison was carried out to the melatonin level in the
steady. Furthermore, when presented as age groups of follicular phase, not to that of the luteal phase. This
5 years we observed two steps in the decline: the first was surprising because, according to recent observa¬
one (41%) when moving from premenopausal women tions, circulating melatonin has no menstrual variation
(< 40 years) to women of 40-44 years, and the (12-14).
second one (35%) when moving from women of 50-54 The inverse change in melatonin as compared to FSH
years to women of 55-59 years. In contrast to our from premenopause to postmenopause is the most
results, Fernandez et al. (11) did not find any changes in interesting result of the present study. The first decrease
urinary excretion of melatonin during menopause, seen in nocturnal melatonin levels (5, 6) in the course of

possible because they had only three age groups
which did not cover the whole age range between
fertile and postmenopausal ages.
The rate, gradual or stepwise, of the melatonin
decrease with age is interesting and may have
physiological significance. According to Waldhauser
et al.(5, 6), the decline in melatonin levels is biphasic:
the first decline taking place from early childhood to
adolescence is steep (75-80%), whereas the second
one seen in adults is much smaller. In both cases the
decline was found to be steady without any clear
steps. Physiologically, the first decline has been
connected to intense growth and therefore to increasing
childhood and the corresponding increase in nocturnal
FSH levels may be closely associated with changes
occurring during growth and Anally with the triggering
of pubertal development. The slower and more gradual
decrease in adult people has been connected to aging.
The age-related decrease in melatonin secretion may
have some clinical implications. Decreased circulating
melatonin may increase the susceptibility of the
organism to oxidative damage, because recent studies
refer to a natural anti-aging hormone character of
melatonin, protecting the organism against the aging
processes of free radicals and thus also against cancer
(10, 32). Normal melatonin secretion also appears to be

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necessary for keepingup the circadian pacemaker
system (33). According to recent in vitro studies,
melatonin is involved in programmed cell death
(apoptosis) via interleukins (34). Additionally, in rats
the injection of synthetic melatonin has been shown to
increase their physical condition and life time (35).
However, due to pharmacological doses of melatonin
used, especially in the older studies, all results cannot be
regarded as physiologically relevant.
The relationships of melatonin, FSH (and LH) and sex
steroids take a new steady state menopausally. In
particular FSH increases sharply during menopause, as
seen also in the present study. This increase is a
consequence of a decreased secretion of estradiol and
other sex steroids, as well as inhibin (1, 2). A rise in FSH
is one of the best menopausal indicators. It has been
observed that administration of melatonin to postmeno¬
pausal women suppresses LH levels (36), which is also
in accordance with the inverse relationship between
melatonin and gonadotropins. Melatonin has been
found to be anti-estrogenic but stimulatory to proges¬
terone production (37). According to our results, the
clearest changes in melatonin secretion take place
before menopause, i.e. in women just over the age of 40
years. Thus, the changes in melatonin and FSH are
analogous to those in puberty, but not simultaneous.
Because the changes in melatonin precede those in
FSH, melatonin may have a permissive role in the
development of gonadal atrophy. Thus, melatonin may
be one factor in the cascadial timing mechanisms of the
menopause, although the exact mechanism (i.e. the
relationship between melatonin, hypophysis and ovary
with exhausting follicles) cannot be evaluated in this
study.
Inconclusion, our results on premenopausal decline
in urinary excretion of melatonin provide some
evidence for a connection between melatonin and
menopause. Based on inverse changes between mela¬
tonin and FSH, and also taking into account temporal
relationships, i.e. the early premenopausal decrease in
melatonin, we suggest that melatonin may trigger
analogously
menopausal development, as concluded
with respect to puberty.
Acknowledgments. This study was supported by the Fund of
Gynecological Research via the Oulu University Scholarship
Foundation.
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Received February 8th, 1996
Accepted May 20th, 1996
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