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Abdelmohsen1997 PDF
Abdelmohsen1997 PDF
3, 1997
This work aimed to study the changes in sex hormones and lipid profile in adult female albino
rats subjected to treatment with nicotine (N), immobilization stress (S), or their combinations
(N+S). These treatments were applied either for one day (T1) or daily for 10 days (T10), after
which rats in the estrus stage were used for the determination of plasma corticosterone (CS),
serum sex hormones as progesterone (P), estrogen (E), FSH, LH and serum lipid profile
including total cholesterol (TC), HDL-C, LDL-C, triacylglycerol (TG) and non esterified fatty
acids (NEFA).
It was clear that either N or S raised plasma CS and serum P levels in both the treatment
regimens and that N+S induced a higher level of these hormones compared to each treatment
alone. Serum E level was only elevated during T10 regimen only. An increase in serum LH
level was only observed after a single exposure to either N or S, however their combination
abolished the stimulatory effect induced by each treatment alone. Serum FSH was not altered
by exposure to either N or S alone in both regimens, but in the T10 regimen their combination
significantly lowered FSH level.
Regarding the effect on serum lipid profile, serum TC was increased in all T10 regimen
groups. LDL-C was increased by N+S treatment in both regimens, however no change in
HDL-C level was observed in all groups. Serum NEFA was increased in all the treated groups
during T10 regimen, while in the T1 regimen NEFA level was only elevated by the combination
N+S. Serum TG was insignificantly altered in all the treated groups. The observed changes in
the lipid pattern were attributed to the alterations occurred in CS and female sex hormones that
caused by N, S or their combinations. 1997 The Italian Pharmacological Society
KEY WORDS: nicotine, immobilization stress, corticosterone, progesterone, estrogen, FSH, LH, cholesterol,
lipoproteins, non esterified fatty acids.
and high serum non esterified fatty acids (NEFA) day, rats in the estrus stage were stressed for 1 h
were observed after N injection in animals [11] and before killing (N10+S1). Control untreated groups
after smoking in humans [12]. It was shown that long- receiving saline were run for each treatment regimen.
term consumption of oral N to monkeys induced an
atherogenic lipoprotein profile by enhancing lipolytic Methods of analysis
conversion of very low density lipoprotein (VLDL) to In all experiments, animals were killed by decapi-
LDL. However, other reports showed that N had no tation. The time of decapitation was kept between
effect on lipoproteins level and can be therapeutically 12–2 p.m. to minimize the variations in the measured
used in the antismoking programs [13]. hormonal level during the day. Blood samples were
Since it is generally believed that the perception of collected in heparinized and non-heparinized tubes.
emotional stress and smoking of nicotine are closely The separated serum samples were used for the esti-
associated, and since a possible site for their effects mation of progesterone (P) by RIA method using pro-
on reproduction and lipid profile is via changes in the gesterone (I125) coated tube kit obtained from Orion
hormonal pattern, the present study was performed to Diagnostics, Finland, while serum levels of estradiol
investigate the effect of the combination of N with (E), LH and FSH were determined by RIA method
stress on the level of plasma corticosterone, female using double antibody kits obtained from Diagnostic
sex hormones as well as lipid profile in female rats. Products Corporation (DPC), USA. Furthermore,
serum samples were used for the estimation of total
cholesterol (TC) [16], HDL-C [17], triacylglyercol
MATERIALS AND METHODS (TG) [18] and NEFA [19]. Serum LDL-C was calcu-
lated from the equation developed by Friedewald et
Animals al. [20]. Separated plasma was used for the estimation
Adult female albino rats (Wistar strain) weighing of corticosterone (CS) level [21].
150–200 g were used in this study. The rats were Statistical analysis of data was carried out using
exposed to a constant light-dark cycle to ensure reg- Student t-test.
ular vaginal smear patterns. Experiments were perfor-
med on rats chosen in the estrus stage, as determined
by the examination of their vaginal smears, since it RESULTS
was observed that the induction of immobilization
stress in rats caused the predominance of this stage. Effect on plasma corticosterone level
Data in Fig. 1 showed that plasma CS was increased
Treatments by 272% and 217% of the control value in response to
Nicotine (N) free base (Hopkin and Williams, Ltd S1 and N1 treatments respectively, while in the case of
England) diluted with saline was injected intraperi- S10 and N10 treatments the increase was found to be
toneally (i.p.) in a dose of 0.5 mg kg−1 body weight in 161% and 191% respectively. The increase in plasma
a single or repeated injections. This dose was reported CS induced by the combination of N1+S1 treatment
to produce a plasma N level in rats nearly equivalent was not significantly different from that produced by
to that resulted from the absorbance of N of the smoke each treatment alone, whereas the combinations
of 20 cigarettes [14]. This dose was also shown to N10+S10 and N10+S1 induced a higher level of the
increase plasma corticosterone and catecolamines in plasma hormone amounted to 353% and 332% of the
rats [7]. control value respectively.
Immobilization stress (S) was applied according to
the method described by Tache’ et al. [15]. Effect on serum sex hormones level
Either S1, N1 treatments or their combination caused
Design of experimental work a marked increase in serum P level (Fig. 2) reaching
Two experimental regimens were performed. A 220%, 310% and 345% of the control value respect-
single treatment regimen (T1) in which rats, chosen in ively. The increase in serum P by these treatments in
the estrus stage, were either subjected to S for one T10 regimen was less if compared to the corresponding
hour (S1), injected with a single dose of N (N1) or treatments in T1 regimen. The combination (N10+S1)
stressed for one hour during which N was injected 20 highly raised P level by 303% of control and this is
minutes before killing (N1+S1). The other treatment significantly higher than that induced by N1 treatment
regimen lasted for 10 days (T10). In this regimen, a alone. No significant changes in serum E (Fig. 3) were
group of rats was exposed to S for 5 h daily and observed in groups of T1 regimen, while an increase
injected i.p. with saline (S10), while N was injected amounting to 164%, 178% and 189% of the control
into two other groups either alone (N10) or in combi- was induced by S10, N10 treatments and their combi-
nation with stress (N10+S10), and the animals in the nation respectively. The increase in serum E caused
estrus stage were then killed 20 minutes after the last by the treatment (N10+S1) was not significantly higher
injection of saline or N. Another group of animals than that caused by N10 treatment alone. Serum LH
were injected daily with N for 10 days and in the last (Fig. 4) was slightly elevated by S1 and N1 treatments
Pharmacological Research, Vol. 35, No. 3, 1997 183
*
†
300 *
400 ‡
*
‡
Percentage of control
Percentage of control
*
300 *
* 200 *
* *
*
200
*
100
100
0 0
T1 T10 N10 + S1 T1 T10 N10 + S1
Fig. 1. Effect of nicotine (N), immobilization stress (S) and Fig. 3. Effect of nicotine (N), immobilization stress (S) and
their combinations on plasma corticosterone level for one their combinations on serum estradiol level for one (T1) or
(T1) or ten (T10) days. (Values are expressed as percentage of ten (T10) days. (Values are expressed as percentage of control
control±SEM) (n=8). h, (S); , (N); , (N+S). Control value ±SEM) (n=8). h, (S); , (N); , (N+S). Control value for
for plasma corticosterone in rats is 18.4±1.56 µg dl−1. serum estradiol in rats is 9.37±1.30 pg ml−1. *Significant
*Significant difference from control group at P<0.05. difference from control group at P<0.05.
†Significant difference of N+S from the corresponding
stressed group at P<0.05. ‡Significant difference of N+S
from the corresponding nicotine-treated group at P<0.05.
200
Percentage of control
*
400 *
†
* * *
‡ †
Percentage of control
300 100
* *
†
*
200
*
100 0
T1 T10 N10 + S1
Table I
Effect of nicotine (N), immobilization stress (S) and their combinations on serum total cholesterol (TC),
HDL-C, LDL-C, triacylglycerol (TG) and non esterified fatty acids (NEFA) for one (T1) or ten (T10) days.
(Values shown are means±SE for 6–8 animals in each group)
Group TC HDL-C LDL-C TG NEFA
(mg dl−1) (mg dl−1) (mg dl−1) (mg dl −1) (mM l −1)
Control T1 68.6±6.90 41.3±2.33 19.1±3.90 48.7±5.60 1.00±0.14
Control T10 60.0±3.00 33.8±2.90 14.3±2.20 58.7±5.50 0.94±0.11
to the effect of AVP which controls ACTH secretion exposure to a stressful stimulus results in an activation
[30]. So when another acute or chronic stimulus was of central opoid neuronal function with the release of
applied, a higher plasma CS response was observed. β-endorphin that exert an inhibitory influence on the
Furthermore, N and stress are postulated to act upon hypothalamo-pituitary-gonadal axis and directly
HPA axis via functionally separate pathways and that inhibits GnRH neurons [44].
there is little, if any cross adaptation [31]. Our results revealed that serum FSH was not affec-
Another finding of this study was that S in T1 and ted by exposure to either S or N alone in both treat-
T10 regimens produced a significant increase in serum ment regimens. Previous reports had described that
P level. The exact source of P is likely to be derived the FSH response to either stress or nicotine treat-
from the adrenal glands, as preclinical studies have ments was less pronounced than the LH response [45].
shown that stress increased P in ovarectomized female Generally, a relatively strong neuronal stimulus is
animals, as well as in castrated male animals [31]. required to produce a change in FSH level. The
Moreover, adrenalectomy has shown to abolish stress- potency of GnRH in inducing FSH secretion is only
induced P release [32]. The increase in ACTH during one fifth that of its LH releasing activity [46]. On the
stress is responsible for the increase in P production other hand, the groups (N 10+S10) and (N10+S1) showed
from the adrenals [33]. In addition, the increase in cir- a significant decrease in serum FSH. This was attri-
culating prolactin levels during stress might also con- buted to the elevation of P level in these groups which
tribute to the increase in P level since prolactin cause an inhibition in FSH release. This negative
potentiated the action of ACTH on the adrenals and feedback effect of P is mainly exerted at the hypothal-
this action was more pronounced on P production amic level where it decreases GnRH pulse frequency
[34]. N administration resulted in increasing serum P by inducing the release of β-endorphin [47].
level in both the treatment regimens. This might also It was clear that serum TC was significantly
be due to increased ACTH during N treatment. The increased in all T10 regimen groups. An increase in
changes observed in P level during T 10 regimen in LDL-C was only induced by the combination of N
response to S, N or their combinations are similar to with S while no change of HDL-C level was noticed.
those of CS. This might indicate a common regulation It seems likely that the increase in E and P levels
through ACTH, which exerts its major action on the observed in T10 regimen in response to N or S might
processes which are involved in the transformation of be responsible for the changes in serum lipid pattern.
cholesterol to pregnenolone [35]. It was reported [48] that estradiol causes an increase
From our results, it was shown that serum E level in total, LDL- and HDL-cholesterol when given to
was only increased during T10 regimen. This increase female rats and that the simultaneous P administration
is suggested to be mainly of adrenal origin, since this partly reversed the enhancing effect of estradiol on
response was reported to disappear in adrenalecto- HDL-C fraction. Estradiol alone or combined with
mized rats [36]. Moreover, from our data no corre- progesterone increased the synthesis of several apop-
lation was found between serum E and LH in different roteins in female rats [49]. Although both apo-B and
groups. It was reported that ACTH increased E level VLDL where shown to be increased by estrogen
in rats adrenals [37] as well as in human serum [38]. administration to postmenopausal women [50], how-
Since ACTH stimulates CS and P production in the ever studies in baboons suggested that increased LDL
biosynthetic reactions from cholesterol to pregneno- apoB in P treated animals resulted from higher con-
lone, it seems reasonable that ACTH would stimulate version of VLDL to LDL [51]. In addition, while
adrenal estrogens at the same site. estrogen induced-increase in HDL appears to be
An increase in serum LH levels was only observed attributable to increased apoA1 synthesis, the ability
after a single exposure to either S or N. It was of P to reverse such effect may be due to that P accel-
reported that acute exposure to S [39] or a single N erate apoA1 catabolism [52]. In addition, Bryant et al.
treatment [40] activates the various hypothalamic [53] showed that chronic S, through activation of cen-
norepinephrine nerve-terminal systems, exerting a tral endogenous opoid systems, causes impaired pro-
stimulatory effect on GnRH and subsequent LH tein synthesis which may result in a reduced or defec-
release [41]. This effect which is mediated via α-adre- tive production of LDL-receptors or recognition of
nergic neurons, appears to depend on the level of the apoprotein B of LDL.
circulating sex steroids [42]. When N was combined Serum NEFA was elevated, in the T1 regimen, only
with S in the T1 regimen, the stimulatory effect on by the combination [N+S]. It was shown that either
serum LH level which was induced by each treatment stress or nicotine [54] treatment stimulates the release
alone was abolished. Fuxe et al. [43] showed that the of epinephrine and norepinephrine in the circulation
combination of acute stress with acute nicotine within a few minutes and their combination might
counteracted the action of one another on the hypo- enhance their lipolytic effect. On the other hand,
thalamic norepinephrine nerve terminals. It was also increased NEFA level observed in all groups during
noticeable that serum LH level returned to the control T10 regimen could be a result of the delayed lipolytic
value after repeated exposure to either S, N or their effect of the elevated corticosterone observed in these
combinations. It has been reported that chronic groups. Glucocorticoids promote lipolysis via syn-
186 Pharmacological Research, Vol. 35, No. 3, 1997
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