This web only provides the extract of this article. If you want to read the figures and tables, please reference the PDF full text on Blackwell Synergy. Thank you.
- Complementary Medicine -
Effects of Boesenbergia rotunda (L.) Mansf. on sexual behaviour
of male rats
Paiwan Sudwan1,2, Kanokporn
Saenphet1, Salika Aritajat1, Narit
Sitasuwan1
1Department of Biology, Faculty of Science,
2Department of Anatomy, Faculty of Medicine, Chiang Mai University,
Chiang Mai 50200, Thailand
Abstract
Aim: To study the effects of Boesenbergia rotunda
(Krachai) on sexual behaviour in male albino rats.
Methods: Thirty-two male Wistar rats were equally divided into four groups: experimental groups were gavaged with the
ethanolic extract of the rhizome of B.
rotunda at doses of 60, 120 and 240 mg/kg and a control group received distilled
water, for 60 days. Sexual behaviour, reproductive organs, diameter of seminiferous tubule, epididymal sperm density,
and androgenic hormones were evaluated.
Results: Within 30-min observation, there was no significant difference of
courtship behaviour, mount frequency (MF), intromission frequency (IF), mount latency (ML), intromission latency
(IL), copulatory efficiency or intercopulatory interval in male rats. In three 10-min intervals over a 30-min period,
courtship behaviour and MF during the first 10-min were significantly higher than those in the second and third
10-min observation in all groups, whereas IF had no significant difference. All doses of
B. rotunda extract significantly increased the relative testicular weight and the diameter of the seminiferous tubules. The dose of 60 mg/kg also
significantly increased the relative weight of the seminal vesicle. Nevertheless, the sperm density, serum testosterone
and androstenedione levels were not affected by the
B. rotunda extract. Conclusion: B.
rotunda does not affect sexual behaviour nor serum androgenic
levels. (Asian J Androl 2007 Nov; 9: 849_855)
Keywords: Boesenbergia rotunda; sexual behaviour; testosterone; androstenedione; reproductive organ; sperm density
Correspondence to: Dr Paiwan Sudwan, Department of Anatomy, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200,
Thailand.
Tel: +66-53-945-312 ext. 208, +66-081-366-1526
Fax: +66-53-945-304
E-mail: pasudwan@mail.med.cmu.ac.th
Received 2006-06-22 Accepted 2006-11-08
DOI: 10.1111/j.1745-7262.2007.00255.x
1 Introduction
Boesenbergia rotunda (L.) Mansf., called "Krachai" in Thai, is a herb of the Zingiberaceae family. The fresh
rhizomes are commonly used in Southeast and South Asia as a food ingredient. In Thailand, they are also used as a
folk medicine for health-promotion, antiflatulence, stomach discomfort, diuresis, leucorrhea, treatment of oral
diseases and antidysentery [1] without acute toxi-city [2]. The rhizomes of
B. rotunda were found to contain 1,5-Cineol,
Boesenbergin A, dl-Pinostrobin Corphor, flavonoid, Chromene [1], panduratin C,
panduratin A, hydroxypanduratin A, helichrysetin,
2',4',6'-trihydroxyhydrochalcone and uvangoletin [3]. In addition to the purposes of primary health
care, the rhizomes have been reported as having aphrodisiac properties
[4_7]. It is sometimes referred to as "Thai ginseng" [4] and has long been used among Thai men for sexual enhancement by using it as an ingredient of traditional
remedies for impotency [5] and a sexual enhancing herb [4_8]. Nevertheless, there is no animal model to confirm or
support these properties.
In the present study, we investigated the effect of
ethanolic extract from the rhizomes of B. rotunda
on sexual behaviour, reproductive organs, diameter of
seminiferous tubule, sperm density and sex hormones in
male albino rats to evaluate its properties.
2 Materials and methods
2.1 Animals
Thirty-two male and 20 female Wistar rats (Rattus
norvegicus), weighing 200_240 g and 6 weeks of age
were purchased from the National Laboratory Animal
Centre, Salaya, Nakhorn Pathom, Thailand. The animals were housed in groups (two rats per cage) under
standard conditions (inverted 12 h : 12 h light : dark cycle)
at 25 ± 2ºC and with free access to pelleted food and
water at the Animal Facility Unit, Department of Biology,
the Faculty of Science, Chiang Mai University. They
were accustomed to the housing conditions for at least 1
week before studying. All experimental procedures were
in accordance with institutional regulations for Animal
Care and Use (No. RE 001/06), Department of Biology,
Faculty of Science, Chiang Mai University.
2.2 Plants and extract preparation
The rhizomes of B. rotunda were collected from
Chiang Mai Province, Thailand and identified by Dr Charun
Maknoi, the Botany Section, Queen Sirikit Botanic Garden, Ministry of Natural Resources and Environment,
Mae Rim, Chiang Mai, Thailand (voucher specimen No.
06-051718). They were sliced, dried in an oven at
60ºC, ground, extracted with 50% ethanol in a soxhlet
apparatus and evaporated by rotary evaporation. This
extract was stored at 4ºC till use. All doses of
B. rotunda extract were diluted in distilled water at a low dosage
(60 mg/kg BW), at the amount equivalent to the
quantity of daily traditional use in humans (120 mg/kg BW)
and at a high dose (240 mg/kg BW). The amount of 120 mg/kg
BW was used by referring to Hem-hongsa and Chaophya Abhaibhubejhr Hospital (Muang,
Prachinburi, Thailand) [1, 9].
2.3 Experimental design
The animals were divided into three groups (n =
8) of male rats that were given the B.
rotunda extract at doses of 60, 120 and 240 mg/kg BW by oral
administration for 60 days and a control group
(n = 8), which received distilled water at 1 mL/day. Sexual behaviours
were tested twice in each of the 32 male rats between
days 26_30 and between days 56_60 of the B.
rotunda application by introducing a receptive female with estrous
phase into the observation cage. At day 61, the animals
were anesthetized and blood was collected for hormone
assay by cardiac puncture from the left ventricle. The
testis, cauda epididymis, seminal vesicle and prostate gland
were removed, trimmed of fat, and weighed. After weights
were recorded, the cauda epididymis was cut into small
pieces, homogenized in 10 mL of 0.9% NaCl and the sperm number was estimated in duplicate using a
hemocytometer (Improved Neubauer, Boeco, Germany).
2.4 Measurement of seminiferous tubule diameter
The left testis from each animal was fixed in Bouin's
solution for paraffin work and sectioned at 6 μm, stained
with hematoxylin and eosin (H&E) and periodic
acid-Schiff (PAS), and examined with a light microscope.
Approximately 20 (per testis) cross-sections of
seminiferous tubules with an apparent lumen were sampled. The
diameter of round tubules or the short axis of elliptical
tubules was measured using an ocular micrometer coupled to a 40 × objective lens.
2.5 Sexual behaviour testing procedure
The totals of 32 male rats were individually tested
for sexual behaviour in a rectangular glass observatory
cage and were given a 10-min accommodation for
adaptation period. Thereafter, an estrous female was
introduced and sexual behaviour was recorded along 30 min.
Video recordings were made throughout the whole period for following behavioural parameters: courtship
behaviour, sniffing, rubbing against or moving under
or over the female's torsal [10]; mount frequency (MF),
the number of mounts; intromission frequency (IF),
the number of intromissions; mount latency (ML), time
from introduction of the female to the first mount with
pelvic thrusting; intromission latency (IL), time from
introduction of the female to the first mount with
pelvic thrusting and vaginal penetration (intromission) [11].
Courtship behaviour and latencies were recorded in
seconds.
The following parameters were calculated: %
courtship behaviour, % mount, % intromission, copulatory
efficiency (number of intromissions/number of mounts)
and intercopulatory interval (average time between
intromissions) [12].
2.6 Hormone tests
Serum was prepared for the electrochemiluminescence immunoassay (ECLIA) and the
radioimmunoassay (RIA). The testosterone levels were determined by
using Elecsys Testosterone reagent kit (Roche
Diagnostics GmbH, Indianapolis, IN, USA) for the Elecsys 2010
immunoassay analyzer and androstenedione was determined by using a Coat-A-Count Direct Androstenedione
kit (Diagnostic Products Corporation, Los Angeles, CA,
USA) for Gamma counter with the cooperation of the
Reproductive Medicine Laboratory, Department of Obstetrics and Gynecology, Faculty of Medicine, Chiang
Mai University.
2.7 Data analysis
Data were expressed in mean ± SD. The mean
values of sexual behaviour for both doses and durations were
statistically analyzed by two-way analysis of variance
(ANOVA) layout completely randomized design, followed
by one-way ANOVA and subsequently least significant
difference (LSD) multiple comparisons test. Mean
values of other parameters were statistically analyzed by
one-way ANOVA followed by LSD. The percentages were analyzed using Chi-squared test. The SPSS 6.0
and 10.0 (SPSS Inc., Chicago, IL, USA) was employed
for all statistical analyses. Significance was inferred when
P ¡Ü 0.05.
3 Results
3.1 Sexual behaviour
There were no significant changes in the whole
30-min period of % courtship behaviour, % mounting, %
intromission, the time of courtship behaviour, IF, ML, IL,
copulatory efficiency and intercopulatory interval in male
rats (Tables 1, 2). However, the study of three
10-min intervals over a 30-min period revealed that the control
and treated groups of rats had significantly higher
courtship behaviour and MF during the first 10-min
observation than in the second and third 10-min observation
(Tables 3, 4), whereas both treated and control groups
had no significant difference in three 10-min periods of
IF during 30 min and they showed consistent frequency
along 30 min (Table 5).
3.2 Organ weights
All groups of male rats receiving B.
rotunda extract had significantly higher relative testis weight than the
control group. B. rotunda extract at the dose of 60
mg/kg BW also significantly induced the relative weights of the
seminal vesicles of the male rats (Figure 1). There were
no significant differences in the caudal epididymis or
prostate gland weights when compared with the control group.
3.3 Diameter of seminiferous tubule
For all groups of male rats receiving B.
rotunda extract, the diameters of seminiferous tubule were larger
than that the control group (Figure 2).
3.4 Sperm density
All doses of the ethanolic extract of B. rotunda
did not significantly affect the sperm density when
compared with the control (Figure 3).
3.5 Hormone levels
Serum testosterone and androstenedione levels were
not affected by B. rotunda extract (Figure 4).
4 Discussion
This study used different doses and durations of
B. rotunda extract to determine their effects on sexual
behaviour in male rats. The results showed clearly that
B. rotunda extract in all doses and durations did not
affect sexual behaviour when compared with the control
groups. Consequently, this research did not support the
belief that B. rotunda could enhance sexual behaviour.
Nevertheless, B. rotunda extract caused an increase
in the testicular weight and seminiferous tubular
diameter of all treated groups and in the seminal vesicle weight of
the lowest treated group. These weight gains may signify
a selective effect of B. rotunda as reported in
Lepidium meyenii [13]. Because B.
rotunda extract did not change the weights of the other sexual organs, sperm density,
serum testosterone or androstenedione levels after
being treated for 60 days, it is difficult to conclude that
ethanolic extract from B. rotunda could enhance sexual
activity. This plant may have an opportunity to make
itself fully apparent in other conditions as reported in
the experiments with other plant species in various doses
and times [14, 15], in various solvent extractions or
with methods of extraction [16], or in sexual condition
of male rats [11, 16_17]. Likewise, Watcho et
al. [14, 15] found that the rats treated with
Mondia whitei for 55 days and 1, 2, 4 and 6 h after treatments showed no
changes in testosterone concentration, but an
application of M. whitei for 8 days induced an increase in the
testicular weight, testicular testosterone level and sperm
density without affecting the accessory gland weights
[15]. Another study in aphrodisiac herbs, Tribulus
terrestris extract did not affect the sexual behaviours or
intracavernous pressure in normal rats, but it improved
those parameters in the castrated rats [17]. The study
of T. terrestris was followed in humans and showed a
lack of effect on testosterone and androstenedione levels
in young men receiving the extract at 1, 3, 8, 20 and 24
days from the beginning of the supplementation [18].
Thus, B. rotunda will be needed to demonstrate the
course and dose response relationship, animal conditions
or ultrastructural changes of testis.
In conclusion, the ethanolic extract of B.
rotunda, does not affect sexual behaviour or serum androgen
levels, but it enhances seminiferous tubule, testis and
seminal vesicle.
Acknowledgment
Thanks to the Scholarship of Commission on Higher
Education, Ministry of Education, Faculty of Medicine and
the research fund of Faculty of Science, Chiang Mai
University, Thailand for financial support. We also would
like to thank Mr Supap Saenphet, Mr Suluck Wutteeraphon,
Ms Warunya Ittipunkul and Ms Siriporn Nunta, for their
kind cooperation and technical assistance and to Ms Lucy
Coombs for proofreading the manuscript.
References
1 Hemhongsa P. Thai Herbal Health. 1st ed. Bangkok: The
WVO Office of Printing Mill; 1998. p46_7.
2 Chomchalow N, Bansiddhi J, MacBaine C. Amazing Thai
Medicinal Plants. Bangkok: Horticultural Research Institute
(HRI), Department of Agriculture and Horticultural Science
Society of Thailand (HSST). 2003. p10_1.
3 Cheenpracha S, Karalai C, Ponglimanont C, Subhadhirasakul
S, Tewtrakul S. Anti-HIV-1 protease activity of compounds
from Boesenbergia pandurata. Bioorg Med Chem 2006; 14:
1710_4.
4 Association of Traditional Medicine School. Traditional
Medicine. Bangkok: Traditional Medicine Association Printing;
1981. p5_6.
5 Pongpamorn PC. Textbook of Traditional Medicine. Bangkok:
Umnauysarn; 1982. p170_1.
6 Theingburanathum W, Dictionary: Diseases and Thai Herbs.
Bangkok: Uksornpittaya; 1995. p13.
7 Wutythamawech W. Encyclopedia of Thai Herbs I. Bangkok:
Phet 69 Printing; 2000. p40.
8 Deewiset K. Thai Pharmacy. Bangkok: The WVO Office of
Printing Mill; 1999. p36.
9 Chaophya abhaibhubejhr Hospital. Boesenbergia
rotunda (KraChay): Herbal Tea. Prachinburi: Herbal Product R&D
Project; 2005.
10 Sachs BD, Meisel RL. The physiology of male sexual behavior.
In: Knobil E, Neill JD, editors. The Physiology of
Repro-duction. Vol. 2. New York: Raven Press; 1988. p1393_403.
11 Carro-Juáreza M, Cervantesa E, Cervantes-Méndeza M,
Rodríguez-Manzo G. Aphrodisiac properties of
Montanoa tomentosa aqueous crude extract in male rats. Pharmacol
Biochem Behav 2004; 78: 129_34.
12 Ratnasooriya WD, Dharmasiri MG. Effects of
Terminalia catappa seeds on sexual behaviour and fertility of male rats.
Asian J Androl 2000; 2: 213_9.
13 Gonzales GF, Ruiz A, Gonzales C, Villegas L, Cordova A.
Effect of Lepidium meyenii (maca) roots on spermatogenesis
of male rats. Asian J Androl 2001; 3: 231_3.
14 Watcho P, Kamtchouing P, Sokeng S, Moundipa PF, Tantchou
J, Essame JL, et al. Reversible antispermatogenic and
antifertility activities of Mondia whitei
L. in male albino rat. Phytother Res 2001; 15: 26_9.
15 Watcho P, Kamtchouing P, Sokeng SD, Moundipa PF, Tantchou
J, Essame JL, et al. Androgenic effect of
Mondia whitei roots in male rats. Asian J Androl 2004; 6:
269_72.
16 Ang HH, Ngai TH. Aphrodisiac evaluation in non-copulator
male rats after chronic administration of Eurycoma
longifolia Jack. Fundam Clin pharmacol 2001; 15:
265_8.
17 Gauthaman K, Adaikan PG, Prasad RNV. Aphrodisiac
properties of Tribulus terrestris extract (protodioscin) in normal
and castrated rats. Life Sci 2002; 71: 1385_96.
18 Neychev VK, Mitev VI. The aphrodisiac herb
Tribulus terrestris does not influence the androgen production in young
men. J Ethnopharmacol 2005; 101: 319_23.
|