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Effects of ambergris on appetite and serum endocrine hormonal levels in skinny sufferers

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The current study evaluated the effects of ambergris on some endocrine hormones, serum lipids, body weights and appetite. A total forty subjects were recruited to receive randomly 415 mg /day of either ambergris (Am; n=20) or placebo (PL; = 20) for 4 weeks. Blood samples were used for the assessments of serum lipids, testosterone, estradiol, growth hormone (GH), prolactin, insulin, thyroxin (T4), and cortisol. Data show significant increase in testosterone, estradiol, prolactin, insulin, cortisol, thyroxin (T4) levels and body weights after ambergris dosing only while growth hormone showed non-significant changes in both groups. A significant increase in total cholesterol (Tc), low density lipoprotein cholesterol (LDLc) and high density lipoprotein cholesterol (HDLc) while significant decrease in triglycerides (TGs) levels in ambergris group were observed. We conclude that ambergris ingestion resulted in; increase of both sexual desire and body weights due to its effect on some endocrine hormones.
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Research Article
EFFECTS OF AMBERGRIS ON APPETITE AND SERUM ENDOCRINE HORMONAL LEVELS IN
SKINNY SUFFERERS
MOHAMED-I KOTB-EL-SAYED1, ZAKARIA-Y AL-SHOAIBI2
1Biochemistry and Molecular Biology Department, Faculty of Pharmacy, Helwan University, Ain Helwan, Helwan P.O. Box 11790, Cairo,
EGYPT. 2Organic Chemistry Department, Faculty of Pharmacy, University of Sana’a, Madbah P.O. Box 19065, Sana’a, Yemen.
Email: mohamed.kotb71524@gmail.com
Received:14 May 2012, Revised and Accepted:25 June 2012
ABSTRACT
The current study evaluated the effects of ambergris on some endocrine hormones, serum lipids, body weights and appetite. A total forty subjects
were recruited to receive randomly 415 mg /day of either ambergris (Am; n=20) or placebo (PL; = 20) for 4 weeks. Blood samples were used for the
assessments of serum lipids, testosterone, estradiol, growth hormone (GH), prolactin, insulin, thyroxin (T 4), and cortisol. Data show significant
increase in testosterone, estradiol, prolactin, insulin, cortisol, thyroxin (T4) levels and body weights after ambergris dosing only while growth
hormone showed non-significant changes in both groups. A significant increase in total cholesterol (Tc), low density lipoprotein cholesterol (LDLc)
and high density lipoprotein cholesterol (HDLc) while significant decrease in triglycerides (TGs) levels in ambergris group were observed. We
conclude that ambergris ingestion resulted in; increase of both sexual desire and body weights due to its effect on some endocrine hormones.
Keywords: Ambergris, Insulin, Prolactin, Testosterone, Appetite
INTRODUCTION
Many factors in our food, beverages or ingestible of folklore use;
affecting body metabolic controllers leading to changes our appetite
and body weights. Ambergris (amber or anber) is an internal
pathological secretion of only 1% of all Sperm blue Whales (Physeter
macrocephalus L. = P. catodon) due seemingly to the irritation of the
stomach by the beaks of octopus and certain shellfish. It is
commercially available from certain companies e.g. Cadima Pathé
(France); La Via del Profumo (Italy), Bernard Perrin Cour tage
(France); Ambergris.co.nz (New Zealand) 1, 2, 3. The animal releases
amber spontaneously to found floating on the sea, cast on the
seacoast of warm countries, or it is collected from intestine of the
whale after its death 4. However, it is a fragrant substance of dark
brown to black but, on exposure to sunlight, air and sea water, it
gradually fades to a light gray and other chemical processes break
ambrein down into a mixture of derivative products 3, 5, 6.
Since there are many constituents have been isolated from
ambergris including up to 46 % of cholestanol type steroids. The
major constituent of ambergris the triterpenoid ambrein (1-ambra-
8, 13, 18 triene; C30H52O; 25-45 w/w); a conversion product of
cholesterol and is the major component, have musk-like odour so it
is used as perfume and its chemical structure was confirmed by
NMR. In addition, ambrein derivatives include; epicoprostanol (3a
hydroxy-5-b-cholestanol; a sterol; 30-40% w/w), coprostanone,
chlolestanone, cholesterol, epicholestanol porphyrine, copper and
fatty acids 4-10.
Moreover, many reports are available on the folklore importance of
crude ambergris, on account of some excellent medicinal properties
it possesses. It has been prized, not only for its use in perfumery, but
also for its alleged restorative and aphrodisiac properties 3, 5, 6. It has
been used by Ancient Egyptians for scenting cigarettes 6, 11 and in
Asia, besides being used as a drug, it also employed as a spice for
food and wines 12. In the eastern discipline of medicine, ambergris
containing compounds and pastes were thought to be excellent
curative for some nervous disorders, as well as replenish and
aphrodisiac 5, 9. A primary screening profile regarding the
cholinergic and/or adrenergic effects of some fractions of
ambergris has been reported earlier 13, and a few reports regarding
the effects of ambrein on the cardiovascular system 14, blood glucose
level 15, edema 16 and mode of action in antinociception 17 have
been published from the same laboratory. Similarly, Epicoprostanol
has been evaluated from the same laboratory for some hormonal
levels, anti-inflammatory and antipyretic activity, changes in plasma
biochemistry, protection against experimental models of insulitis
and smooth muscle responses have already been published 10, 18-23.
In Yemen and other Arabic countries, ambergris has been used in
folk medicine for gaining weight and as aphrodisiac. Its effect on
weight gaining may continue for several years after use of ambergris
especially if its users were at younger age or at age before puberty.
These folklore uses of ambergris attract many skinny individuals to
use it particularly here in Yemen. Therefore, the purpose of present
study is to evaluate the ambergris effects at its folklore dose on
levels of some endocrine hormones [testosterone, estradiol, GH,
prolactin, insulin, cortisol, and thyroxin (T4)], levels of serum lipids,
changes in menstruation, appetite, and body weight in Yemeni
skinny subjects.
MATERIALS AND METHODS
Equipments
Electric balance (Sartorius AG- Göttingen Germany BP310S),
Centrifuge (Hernie Z 400, Wehingen), ELIZA reader (Humareader
Human Company 2106/1682), UV/Visible Spectrophotometer
(Shimadzu), refrigerator, deep freeze, micropipette of different size,
and plastic syringe were used.
Ambergris source, description and dose
Source and description of ambergris
Ambergris samples were purchased from AL-Nasheri, local market
in Sana’a of Yemen Republic (They import it from Nisha
International Pte Ltd, Singapore). Description of ambergris: Samples
of ambergris were blackish to pale gray type, which is called Dokhni
ambergris as a traditional name. Ambergris gets its name from the
French “ambre gris” (gray amber) to distinguish it from the
fossilized resin, brown amber. Ambergris described according to
Nisha International Pte Ltd, as raw material of ambergris results
from a pathological condition of the sperm whale Physeter
macrocephalus L. syn P. catodon L. In the normal course of events,
calculus (sand and stones) or cachalot is regularly ejected from the
digestive tracts from adult sperm whales. Fresh material is almost
black turning to light gray as it matures. It contains 46% of
cholestanol type sterols including (+)-epi-coprosterine and the
triterpene alcohol (-)-ambreine (25-45%), which is odorless, but this
material is the precursor to other fragrant compounds formed by
auto-oxidation, sunlight, and seawater such as (-)-gamma-
cyclogeranyl chloride and (-)-gamma-bicyclohomofarnesal. The
identity of this material was verified by the Research Center of
College of Pharmacy, Sana’a University, Sana’a of Yemen and a
voucher specimen was kept on record (RC-23112010).
Asian Journal of Pharmaceutical and Clinical Research
Vol 5, Suppl 3, 2012 ISSN - 0974-2441
Vol. 4, Issue 3, 2011
ISSN - 0974-2441
Academic Sciences
M I Kotb El-Sayed and Z Y Al-Shoaibi.
Asian J Pharm Clin Res, Vol 5, Suppl 3, 2012, 138-142
139
Ambergris dose
A folklore dose of ambergris is approximately 11.6 grams/1 Kg of
honey bees or / 1500 mL of milk and taken as one teaspoonful after
12 hour of overnight fasting for 4 weeks. In the present study we
depend on the above folklore dose except using skim instead honey
bees or milk. The samples were grounded and weighed to give 415
mg in each single dose, which added freshly and daily to 50 ml of
skimmed milk, coded and preserved in the refrigerator not for more
than 12 hours before use.
Participants and protocol design
Inclusion criteria
In the present study a randomized double-blind placebo-controlled
trial design was used. Forty participants of average ages (21±1.3),
their gender was 20 male and 20 female. They were skinny and want
to gain weight. A written informed consent was obtained from all
participants in the study. The study followed guidelines of the
Declaration of Helsinki and Tokyo for humans. The study had
Medicine and Health Sciences College of Sana’a University Ethics and
Human Experimentation Committee approval.
Exclusion criteria
Subjects suffering from any acute, chronic, or parasitic diseases and
those with anorexia nervosa were excluded from the present study.
In addition, smokers, lactating- or pregnant-women, on steroid-,
insulin-, thyroxin-, bromocriptine-, or tamoxifen-therapy were
excluded from the present study.
Protocol design
Participants were assigned randomly to either therapeutic
(Ambergris; Am) or sub-therapeutic (placebo; PL) using a numbered
series of opaque sealed envelopes prepared in advance of the trial.
Participants in two groups were receiving 415 mg of either
ambergris (Am) or starch (PL) in 50 mL of skimmed milk/day after
12 hours of overnight fasting for 4 weeks. Participants instructions:
All subjects were instructed by the research nutritionist and medical
supervisor to keep a written record of their food intakes (3 days
including one weekend day), body weight (weekly), number of
sexual desire (weekly) and menstrual changes (at end of the month).
Blood sample withdrawal
All blood samples were collected after 12 hour of overnight fasting,
between 8:00 a.m. and 10:00 a.m. to minimize the effect of either
food or daily fluctuation. Technician staffs at biochemistry research
laboratory of medicine and health sciences collage of Sana’a
University were blind to both research protocol and coded samples.
Blood samples were collected before randomization to obtain base
line data and at 2 hour after last dose of ambergris or placebo at end
of 4 weeks. Serum was separated, preserved at 20 0C for
measurements of growth hormone (GH), prolactin, insulin, thyroxin
(T4), cortisol, testosterone (males), and estradiol (females) by ELISA
method (within 2 days of collection). In addition, serum levels of
triglycerides (TGs), total cholesterol (Tc), low density lipoprotein
cholesterol (LDLC), and high density lipoprotein cholesterol (HDLC)
were measured by colorimetric methods (within one week).
Biochemical estimation/assay
All hormones were measured by ELISA, growth hormone was
measured as described by Colao et al. 24, testosterone was measured
as described by Tietz 25, estradiol was measured as described by
Racliff et al. 26, cortisol was measured as described by Crapo 27,
thyroxin (T4) was measured as described by Schuurs, and Van
Weeman 28, insulin was measured as described by Frier et al. 29 and
prolactin was measured as described by Uotila et al. 30 (DRG
International, Inc., USA). Serum lipids were measured by
colorimetric methods; TGs were estimated using the phosphate
oxidase method as described by Trinder 31, total cholesterol was
estimated using the Chod pap method as described by Zoppi and
Fellini 32, LDLC was measured as described by Kerscher et al. 33, HDLC
was estimated using the dextran sulphate Mg (II) method as
described by Wieland and Siedel 34.
Statistical analysis
All results were expressed as mean ± S. E. M. Results were analyzed
using student-t-test to compare data at baseline with those at end
point of treatment within each group, while p-value < 0.05 was
considered statistically significant (using SPSS statistics software
program, version 15).
RESULTS
Data show significant increase in both sex hormones either in males
or females (testosterone and estradiol respectively; P < 0.001 and P
< 0.01 respectively) after ambergris dosing, while non-significant
change in placebo group. In addition, the measured pituitary
hormones (GH and prolactin) showed slightly different pattern, GH
show non-significant difference in both groups after dosing either
ambergris or placebo, while significant increase (P < 0.01) in
prolactin after ambergris dosing only. Moreover, insulin, cortisol and
T4 showed significant increase (P < 0.001, P < 0.01, and P < 0.01
respectively) after ambergris dosing, while non-significant change in
placebo group [data shown in Tables 1 and Fig 1].
Fig 1: Effects of ambergris (Am) or placebo (PL) dosing (415
mg/day for 4 weeks) on testosterone, estradiol [A], growth
hormone, prolactin [B], cortisol, thyroxine and insulin [C]
serum levels in its users (P < 0.001 vs. baseline data). T or
Testo = testosterone; E or Estra = estradiol; G or GH= growth
hormone; P or Prola = prolactin; Cor or Cort = cortisol; T4 =
thyroxine; Ins = insulin; B = before; and A = after.
M I Kotb El-Sayed and Z Y Al-Shoaibi.
Asian J Pharm Clin Res, Vol 5, Suppl 3, 2012, 138-142
140
Table 1: Effects of Ambergris or Placebo Oral Dosing on Serum Levels of Testosterone, Estradiol, GH, Prolactin, Insulin, Cortisol and
Thyroxin (T4) Hormones in its Users
Treatments
Testosterone
(ng/mL)
n=10 (male)
Estradiol (Pg/mL)
n=10 (Females)
GH
(ng/mL)
n=20
Insulin (µIU/mL)
n=20
Cortisol (ng/mL)
n=20
T4 (µg/dL)
n=20
Before
After
Before
After
Before
After
Before
After
Before
After
Before
After
Before
After
Placebo
(415 mg
starch/day
for 4
weeks)
7.07±0.
65
7.06±0.6
6
34.19±1.
28
34.16±1.2
2
1.53±0.
34
1.57±0.
34
13.3±1.
12
13.36±1.
09
19.73±1.
11
19.84±1.0
4
117.58±8
.37
118.37±7.6
10.45±0.
27
9.93±0.58
Ambergris
(415 mg
ambergris/
day for 4
weeks)
6.58±0.
54
8.55±0.6
5
32.62±1.
45
47.38±2.7
2
1.82±0.
44
1.83±0.
43
17.7±1.
16
28.7±3.1

16.82±1.
46
22.57±1.6
8
110.07±9
.08
165.27±8.3
9
10.98±0.
21
12.17±0.2
2
n = number of subjects (mean ± S.E.M); GH = Growth Hormone; T4 = Thyroxin;  = P < 0.001 vs. baseline data in each group.
In the same line, serum lipids data showed significant increase (P <
0.001) in Tc, LDLc and HDLc while significant decrease (P < 0.001) in
TGs levels after ambergris dosing while non-significant changes in
all serum lipids of placebo group [data shown in Table 2].
Table 2: Effects of Ambergris or Placebo Oral Dosing on Serum Lipids in its Users
Treatments
TGs (mg/dL)
n=20
Tc (mg/dL)
n=20
LDLC (mg/dL)
n=20
HDLC (mg/dL)
n=20
Before
After
Before
After
Before
After
Before
After
Placebo (415 mg
starch/day for 4 weeks)
103 ± 2.4
103.45±2.5
110.9±7.4
110.8±7.2
56.5±3.6
56.6±3.7
65.45±3.8
65.16±3.7
Ambergris (415 mg
ambergris/day for 4
weeks)
106.15±2.8
98.1±3.15
87.9±6.5
119.1.4±6.5
35.05±3.7
53.9±3.5
46±3.2
60.2±3.4
n = number of subjects (mean ± S.E.M); TGs = Triglycerides; Tc = Total cholesterol; LDLC = Low Density Lipoprotein Cholesterol; HDLC =
High Density Lipoprotein Cholesterol;  = P < 0.001 vs. baseline data in each group.
Data of body weights showed highly significant increase (P < 0.001)
at all interval points after ambergris dosing when compared with
base line data of body weights while non-significant changes in body
weights of placebo group at all interval points, except slightly
significant increase at 3rd and 4th weeks when compared with base
line data of body weights [data shown in Fig 2].
Fig 2: Effects of ambergris (Am) [A] or placebo (PL) [B] dosing
(415 mg/day for 4 weeks) on body weights means in its users (
P < 0.01,  P < 0.001 vs. baseline data). W = week.
Table 3 showed significant increase in percentage of sexual desire
and appetite after ambergris dosing than placebo. Moreover,
significant changes in menstruation (amenorrhea) after ambergris
dosing while none changes with placebo.
Table 3: Effects of ambergris or placebo oral dosing on sexual
desire, menstruation and appetite in its users
Treatments
Changes
Sexual desire
n = 20
Menstrual
disturbance
n = 10
Appetite
n = 20
Number
%
Number
%
Number
%
Placebo
(415 mg
starch/day
for 4
weeks)
No
change
0
%
No
Change
0
%
Increase
d in 2
individu
als
1
0
%
Ambergris
(415 mg
ambergris/
day for 4
weeks)
Increase
d in 13
individu
als
65
%
Observed
as
amenorrh
ea in 6
individua
ls
6
0
%
Increase
d in 18
individu
als
9
0
%
DISCUSSION
In Arabian society ambergris was named 'anbar’ and from this word
the European name ambergris was derived. They used ambergris to
treat heart and brain diseases, headaches, rheumatism, constipation,
common colds, and as aphrodisiac 10, 35, 36. However, the biochemistry
of ambergris in the scientific literature is still largely on a
speculative level and there is no published data about ambergris
effects on most of human endocrine hormones.
The results of the current study show increase of serum testosterone
levels in male subjects after ambergris ingestion. These results are in
M I Kotb El-Sayed and Z Y Al-Shoaibi.
Asian J Pharm Clin Res, Vol 5, Suppl 3, 2012, 138-142
141
agreement with study of Taha and Islam 19 whom states that
ambrein (as major constituent of ambergris) treatment was found to
elevate plasma testosterone levels. In addition, the present results
were in the same line with Taha & his college 37 whom reported that
administered 100 and 300 mg of ambrein/kg rat bodyweight,
increased sexual behavior in male rats via increased their number of
penile erections in the absence of females, as well as increased
intromissions and anogenital investigatory behavior in the presence
of females.
Since, there are many other constituents have been isolated from
ambergris, including ambrein non-volatile derivatives;
epicoprostanol, coprostanone, chlolestanone, cholesterol,
epicholestanol porphyrine, copper and fatty acids 7, 8. The elevation
of both testosterone and estradiol levels with ambergris ingestion
could not be explained by effect of ambrein only but also by effect of
non-volatile derivatives of it, which could be act as a precursor for
the synthesis of steroid hormones including testosterone, estradiol,
and cortisol hormones. This explanation depends on a similarity in
chemical structure (the structure of ambrein betrays its cholesterol
origin and structural similarity to steroid nucleus) and stay just
theoretical explanation and needs further future research.
Increased total thyroxin (T4) serum levels with ambergris ingestion
might be due to previously increased glucocorticoids which
stimulate protein breakdown in peripheral tissues, exposing
tyrosine to being involved in thyroid hormone synthesis 38 or due to
estrogen-induced increase in the serum concentration of thyroxin-
binding globulin 39.
Here, the decreased serum levels of TGs after ambergris ingestion
could be explained by formerly increased levels of cortisol, insulin
and thyroxin. Insulin and thyroxin activate lipoprotein lipase in
blood leading to the clearance of plasma from triglycerides while
cortisol induces lipolysis particularly in peripheral tissue leading to
decrease levels of TGs and increased levels of free fatty acids. The
increased free fatty acids lead to insulin resistance as reported by
Boden 40. Thereby the resulted increase in insulin level as reported
here after ambergris ingestion, might be due to the decreased
sensitivity of insulin receptors by elevated cortisol and thyroxin
levels. Studies in man have found that glucocorticoids can decrease
insulin receptor binding affinity without decreasing insulin receptor
numbers 41, decrease receptor number and affinity 42, 43. On the other
hand, central actions of glucocorticoids may enhance vagal
stimulation of insulin secretion 44.
These results and its explanation, in contrast to the results reported
by Taha 15, which states that ambrein reduce the blood glucose levels
of normal and moderately alloxan-diabetic rats but did not reduce
the blood glucose levels of severely-diabetic rats. This difference
might be due to using ambrein alone (not a crude ambergris) in
treating induced diabetes in rats.
As mentioned before, the structure of ambrein betrays its
cholesterol origin. However, increased levels of total and individuals
of cholesterol could attributed to that free fatty acid liberated from
lipolysis; favorably oxidized in the presence of higher level of total T4
into acetyl-CoA, which acts as building units for cholesterol
synthesis. In addition, ambergris containing ambrein derivatives act
as precursors of cholesterol synthesis 5, 6.
In the results of the present study, ambergris ingestion shows
significant increase in body weight of its users. However, many
reports and information from unpublished data mentioned that
ambergris used as an appetizer, to increase body weight in both
male and female. It was used usually with honey bees. In Arabic folk
medicine, it is used to increase appetite, sexual ability, and body
weight and these reports in the same line with the results of the
present study. Increased weights of subjects as an effect of
ambergris ingestion, may be due to the effects of elevated anabolic
hormones including; insulin, testosterone, and estradiol which acts
to increase protein synthesis and water retention respectively . This
hypothesis in some, in agreement with Tietz 45 who reported that
androgens and growth hormones increases protein synthesis and
serum protein levels. The non-significant increase in GH level in
serum of subjects, might due to a response to the increased levels of
cortisol, steroid hormones and increased glucose level according to
hypothesize insulin resistance.
These data supported by recorded changes in appetite, sexual desire
and menstrual changes after ambergris use. As a result of increased
levels of testosterone, estradiol, and prolactin with ambergris
ingestion; sexual desire increased and menstrual cycle disturbed or
ceased respectively. Since increased level of prolactin stimulate
corpus luteum to secrete progesterone and inhibits ovulation.
In addition, possibly increased levels of insulin and thyroxin result in
improvement of appetite. In contrast, glucocorticoids oppose other
actions of insulin, including its effect to reduce central appetite 46.
Moreover, sex hormones, i.e. estrogen, progesterone, and androgens,
play an important role in the regulation of appetite and energy
metabolism 47. Testosterone is known to stimulate appetite 47. In
contrast to estrogen, progesterone seems to stimulate appetite.
Several studies in women have demonstrated a distinct increase in
food intake in the premenstrual period of the menstrual cycle, when
progesterone levels are high 48, 49, 50.
We conclude that ambergris ingestion at its folklore dose resulted in
an increase of both sexual desire and body weights due to its effect
on some endocrine hormones. Thereby the ambergris-modified
testosterone levels support the folk use of this drug as aphrodisiac
or sexual stimulant. Moreover, ambergris increases body weight in
the region of protein synthesis without obesity, due to the effects of
increased anabolic hormones and increased lipolysis. We
recommend for further intensive research; to determine ambergris
effects on other endocrine hormones such as progesterone and
blood glucose; to raise awareness in the minds of its users where
amber considered as hormonal therapy. The ambergris might be
used as nutritional therapy (as an anabolic agent) for patients with
anorexia nervosa. However, the present study showed that
ambergris increases prolactin, cortisol and thyroxin, thereby
ambergris avoided to prescribe for infertile female, diabetic and
hyperthyroidism suffering patients.
Acknowledgments
This study was not supported by any financing or grants from the
university or the government and fully covered by the authors. We
are indebted to all the subjects whom participated in this study and
would like to thank technician staff (in biochemistry department of
Medicine and Health Sciences College of Sana’a University in Yemen)
and the nurses (in Al-Kuwait Hospital, in Sana’a of Yemen) for their
help during the experiments and for blood samples collection from
participants.
Conflict of interest: The authors declare that there are none conflict
of interest.
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... influence on some endocrine hormones. There was substantial rise in testosterone, estradiol, prolactin, insulin, cortisol, thyroxin (T4) levels and body weights after ambergis administration [9]. ...
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