Original Article Biological Sciences

Transcription

Original Article Biological Sciences
Original Article
Biological Sciences
International Journal of Chemical And Pharmaceutical Research
ISSN 2319-1716
Volume 2, Issue 10, October 2013.
© Glorigin Lifesciences Private Limited.
EFFECT OF ONION, GARLIC AND ARIDAN AQUEOUS EXTRACT ON EXPERIMENTALLY
ENLARGED PROSTATE GLAND IN ADULT WISTAR RATS
Odumosu F.M1*, Olokodana .O1, Huthman I.O2, Akinyemi R.A3, Adesanya O.A1, Adefule A.K1
Abstract
This work focused on determining the effect caused by the aqueous extract of onion, garlic and aridan on
experimentally enlarged prostate gland in adult wistar rats. Twenty male adult Wistar rats with body
weights ranging between 130-220g were randomly divided into four groups of five rats each. Group I:
Normal control (received 0.5ml of normal saline); Group II: Hormone treated control; were given
continuous dosage of 300µg and 80µg of testosterone and estradiol respectively subcutaneously in the
inguinal region on alternate days for four weeks. Groups III & IV were given continuous dosage of
300µg/ml and 80µg/ml of testosterone and estradiol respectively on alternate days for three weeks, after
which group III animals were given 4ml/mg body weight of extract(high dose) orally, while group IV
received 2ml/mg body weight of extract (low dose). Immediately after induction, some animals randomly
selected were sacrificed for the gross and histological inspection of prostate gland. These procedures were
also carried out after four weeks of treatment with the extract, with the prostate taken and processed
routinely for paraffin embedding and stained with H&E. Treatment with extracts caused a significant
decrease in the size of the enlarged prostate in a dose related manner (P≤0.05) when compared with the
hormone treated control group which were not treated (with the extract) after the period of induction. This
shows that the herbal drug administration (for a month) reduced the size of the prostate significantly
(P≤0.05) both at high and low dosages respectively.
Author Affiliations: 1 Department of Anatomy, Olabisi Onabanjo University (Ikenne), Ago-Iwoye, Ogun
State, Nigeria; 2Department of Community Health, Sultan Abdur-Rahman School of Health Technology,
Gwadabawa, Sokoto State, Nigeria; 3Department of Anatomy, University of Ibadan, Ibadan, Oyo State,
Nigeria.
Keywords: onion, garlic, aridan, prostate gland, aqueous extract, benign prostatic hyperplasia, Wistar rats.
* Corresponding Author: Odumosu F.M, Dept. of Anatomy, Faculty of Basic Medical Sciences, Olabisi
Onabanjo University (Ikenne), Ago-Iwoye Ogun-State, Nigeria, Phone: +234-7030604589, Email:
[email protected]
Intl. J Chem Pharm Res 2013;2(10): 275-286
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Odumosu F.M, et al.,
Experimentally enlarged prostate gland in adult Wistar rats
―zones‖ was first proposed by McNeal in 1968.
1. INTRODUCTION
The prostate is a walnut-sized gland that
McNeal found that the relatively homogeneous
forms part of the male reproductive system. The
cut surface of an adult prostate in no way
gland is made of two lobes, or regions, enclosed
resembled ―lobes‖ and thus, led to the
[1]
by an outer layer of tissue
. It is the only
description of ―zones‖
[10]
He has described four
exocrine organ located in the midline in humans
different regions of the prostate, the largest of
and similar animals. It lies immediately inferior
the regions being the anterior fibro- muscular
to the bladder, posterior to the pubic symphysis,
and
and anterior to the rectum
[2]
non-glandular
region.
The
remaining
. It is a compound
glandular prostate is subdivided into three zones:
tubuloalveolar exocrine gland of the male
peripheral (PZ), central (CZ) and the traditional
reproductive system in most mammals.[3][4]. The
zones (TZ) in relation to the urethra. The
prostate in men, like the breast in a woman, is a
peripheral zone is located posterior –laterally,
gland that produces and secretes fluid and
the central zone is located at the base, and the 2
controls the flow of fluid. Cells lining the
lobes of the transitional zone locate along the
prostate gland make some of the semen that
proximal urethra. The lymph vessels of the
comes out of the penis at the time of sexual
prostate drain into internal iliac lymph nodes [11].
climax
[5]
. The mean weight of the "normal"
The function of the prostate is to secrete a
prostate in adult males is about 11 grams,
slightly alkaline fluid, milky or white in
usually ranging between 7 and 16 grams.[6]
appearance that usually constitutes 50–75% of
The
mainly
the volume of the semen along with spermatozoa
pseudostratified, comprising tall columnar cells
and seminal vesicle fluid. [12] The prostatic fluid
and basal cells which are supported by a
is expelled in the first ejaculate fractions,
fibroelastic
together with most of the spermatozoa. In
orientated
secretory
epithelium
stroma
smooth
is
containing
muscle
randomly
The
comparison with the few spermatozoa expelled
epithelium is highly variable and areas of low
together with mainly seminal vesicular fluid,
cuboidal or squamous epithelium are also
those expelled in prostatic fluid have better
present, with transitional epithelium in the distal
motility, longer survival and better protection of
regions of the longer ducts.
not
have
a
capsule
[7]
bundles.
The prostate does
rather,
integral
sperm is transmitted from the ductus deferens
It measures
into the male urethra via the ejaculatory ducts,
about 3 cm (about 1.2 inches) across.. Generally,
which lie within the prostate gland. Orgasm in
the prostate can be divided in two ways: by
men is achieved solely through stimulation of
fibromuscular band surrounds it
zone, or by lobe.
[9].
an
the genetic material. During male ejaculation,
[8]
The zone classification is
more often used in pathology. The idea of
Intl. J Chem Pharm Res 2013;2(10): 275-286
the prostate gland, such as prostate massage or
receptive anal intercourse. [13][14][15].
276
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Odumosu F.M, et al.,
Experimentally enlarged prostate gland in adult Wistar rats
The prostatic part of the urethra develops
relation between these conditions. Original
from the pelvic part of the urogenital sinus
hypotheses of an association between BPH and
(endodermal origin). Endodermal outgrowths
prostate
arise from the prostatic part of the urethra and
documenting the presence of both conditions at
grow into the surrounding mesenchyme. The
autopsy
glandular
prostate
prostate cancer frequently coexist; more than
differentiates from these endodermal cells, and
20% of men with prostate cancer also have BPH,
the associated mesenchyme differentiates into
and cancer is found incidentally in a significant
the dense stroma and the smooth muscle of the
percentage (10%–20%) of surgically removed
prostate.
[16]
epithelium
of
the
The prostate glands represent the
BPH
cancer
were
based
on
studies
[20]
. In more recent studies, BPH and
[21]
specimens
.
There
are
several
modified wall of the proximal portion of the
similarities between the 2 conditions: the
male urethra and arises by the 9th week of
prevalence of both BPH and prostate cancer
embryonic life in the development of the
increases in parallel with age; both are androgen
reproductive system.
dependent
There are three major prostate disorders;
[22]
; both respond to androgen-
deprivation treatments
[23]
; BPH and prostate
Prostatitis, Benign Prostatic Hyperplasia BPH
cancer share similar genetic alterations; and an
and
expanding body of evidence supports an
Prostate
cancer.
Prostatitis
is
an
inflammatory condition of the prostate. There
important
role
for
inflammation
in
both
[24]
are four different forms, each with different
conditions
causes and outcomes. Two relatively uncommon
are strong arguments that BPH and prostate
forms are: acute Prostatitis and chronic bacterial
cancer
prostatis are treated with antibiotics. Chronic
histology and anatomic location of these 2
non-bacterial Prostatitis comprissing 95% of
conditions support the generally accepted belief
prostatitis diagnosis is treated by large varities of
that BPH does not affect prostate cancer risk [25].
modalties including alpha blocker, phytotherapy,
BPH is characterized by hyperplasia of primarily
physical
anti
stromal and, to a lesser extent, epithelial cells,
histamine and anxiolytics, nerve modulator,
whereas prostate cancer involves complex
surgery
teraphy,
[17]
pscychotheraphy,
and more
[18].
More resently, a
. Despite these similarities, there
are
unrelated.
histopathologic
changes
Differences
in
the
in
the
glandular
[26]
combination of trigger point and psychological
epithelium
therapy has proved effective for category III
transition zone of the prostate, while only 24%
[19].
prostatic
of prostate cancers occur in the transition zone,
hyperplasia (BPH) and prostate cancer are
two-thirds in the peripheral zone, and the rest in
common urologic conditions in older men, and
the central zone [27].
Prostatitis
aswell
Benign
. Nearly all BPH arises in the
there is a large body of evidence supporting a
Intl. J Chem Pharm Res 2013;2(10): 275-286
277
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Odumosu F.M, et al.,
Symptoms
Experimentally enlarged prostate gland in adult Wistar rats
of
BPH
can
increase
the
used as a food source
[34]
. Most onion cultivars
detection rate of prostate cancer by increasing
are about 89% water, 4% sugar, 1% protein, 2%
the number and extent of urologic examinations
fibre and 0.1% fat. They contain vitamin C,
[26]
, and treatment of BPH via transuretheral
vitamin B6, folic acid and numerous other
resection of the prostate can result in the
nutrients in small amounts. They are low in fats
detection of incidental prostate cancer
[21]
. In
and in sodium, and with an energy value of
addition, BPH can increase prostate-specific
166kJ (40 kcal) per 100 g (3.5 oz) serving, they
antigen (PSA) levels, thereby increasing the
can contribute their flavour to savoury dishes
.
without raising caloric content appreciably.[35]
BPH also causes an enlargement of the prostate
Onions contain chemical compounds such as
gland, which could lead to increased biopsies
phenolics and flavonoids that basic research
because
shows to have potential anti-inflammatory, anti-
likelihood of detecting latent prostate cancer
of
an
examination (DRE)
abnormal
digital
[28]
rectal
[29]
, although increased
prostate size could also make it more difficult to
detect cancer
[30]
. Detection bias may be even
more of a concern in contemporary studies
cholesterol,
anticancer
and
antioxidant
[36]
properties
. These include quercetin and its
glycosides
quercetin
3,4'-diglucoside
and
quercetin-4'-glucoside. [37]
because of the widespread use of PSA screening
Garlic (Allium sativa), is a species in the
and the availability of pharmacologic treatments
onion genus [38], native to central Asia,[39] and
for BPH.
has long been a staple in the Mediterranean
Onion (Allium cepa) is the most widely
region, as well as a frequent seasoning in Asia,
cultivated species of the genus Allium.[31][32] . It
Africa, and Europe; and has since been used for
is a biennial plant but is usually grown as an
both culinary and medicinal purposes.[40] It is a
annual. Modern varieties typically grow to a
bulbous plant, growing up to 1.2 m (4 ft) in
height of 15 to 45 cm (6 to 18 in). The leaves are
height. It produces hermaphrodite flowers and
blueish-green and grow alternately in a flattened,
pollination occurs by bees and other insects. It
fan-shaped swathe. They are fleshy, hollow and
species have two other subspecies
[42]
[41]
or
cylindrical, with one flattened side. The base of
varieties.
the leaf is flattened, usually white sheath that
world for its pungent flavor as a seasoning or
grows out of a basal disc. From the underside of
condiment
the disc, a bundle of fibrous roots extends for a
research studies in humans, have suggested
short way into the soil. As the onion matures,
possible cardiovascular benefits of garlic.[44] The
food reserves begin to accumulate in the leaf
known vasodilative effect of garlic is possibly
bases and the bulb of the onion swells.[33] . Bulbs
caused
from the onion family are thought to have been
polysulfides to hydrogen sulfide in red blood
Intl. J Chem Pharm Res 2013;2(10): 275-286
278
. Garlic is widely used around the
[43]
by
. Animal studies, and some early
catabolism
of
garlic-derived
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Odumosu F.M, et al.,
Experimentally enlarged prostate gland in adult Wistar rats
cells (RBCs), a reaction that is dependent on
consist of a fleshy pulp with small, brownish-
reduced thiols in or on the RBC membrane. It
black seeds. The dry fruit has a pleasant aroma
has been found to reduce platelet aggregation
[53]
and hyperlipidemia. [45] Garlic is also alleged to
spice in Southern and Eastern Nigeria
help regulate blood sugar levels. Regular and
fruit is used to prepare soup for mothers from
prolonged use of therapeutic amounts of aged
the first day of birth to prevent post partum
garlic extracts lower blood homocysteine levels
contraction.
and
management
has
been
shown
to
prevent
some
. It is therefore, used as a popular seasoning
Its
fruits
of
are
used
[54]
. The
for
convulsions,
the
leprosy,
complications of diabetes mellitus.[46][47] When
inflammation, rheumatism, flatulence, jaundice
crushed, Allium sativum yields allicin, an
and fevers
antibiotic
[48]
and
antifungal
compound
[55]
. Its leaves are essential for the
treatment of epilepsy and present strong
[56]
(phytoncide). Fresh or crushed garlic also
molluscicidal activity
affords the sulfur-containing compounds alliin,
extract has also been shown to possess
ajoene, diallyl polysulfides, vinyldithiins, S-
hypoglycaemic properties
allylcysteine, and enzymes, B vitamins, proteins,
has also been proven to be used for the treatment
minerals, saponins, flavonoids, and Maillard
of gastrointestinal related clinical problems [58].
reaction
2. MATERIALS AND METHODS
containing
products,
which
compounds.
are
not
sulfur-
Furthermore,
a
. The aqueous fruit
[57]
. The root extract
2.1. Management
phytoalexin (allixin) was found, a nonsulfur
Twenty adult male wistar rats weighing
compound with a γ-pyrone skeleton structure
between 130g - 220g were obtained from
with antioxidant effects, antimicrobial effects,[49]
Nigerian Institute of Medical Research, Yaba.
antitumor promoting effects, inhibition of
Lagos. The rats were kept at the animal house
aflatoxin B2 DNA binding,
[50]
and neurotrophic
provided by the Department of Anatomy,
effects. The composition of the bulbs is
Olabisi Onabanjo University and divided into
approximately 84.09% water, 13.38% organic
four groups of 5 animals each after a period of
matter, and 1.53% inorganic matter, while the
one week acclimatization. They were fed with
leaves are 87.14% water, 11.27% organic
standard commercial rat pellet. Food, water and
matter, and 1.59% inorganic matter. [51]
air were given ad libitum. The animal room was
Aridan (Tetrapleura tetraptera) is a species
of flowering plant in the pea family native to
Western Africa.
[52]
It belongs to the mimosaceae
well ventilated with a temperature range of 2527
ºc
under
photoperiodicity.
day/night
The
12-12
hormonal
hour
induction
family. It is referred locally to as Aridan-Yoruba
(testosterone and oestradiol; 300µg/ml and
and oshosho in Ibo. It is generally found in the
80µg/ml respectively in alternate days for four
lowland forest of tropical Africa. The fruit
weeks)
Intl. J Chem Pharm Res 2013;2(10): 275-286
279
commenced
after
the
period
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of
Odumosu F.M, et al.,
Experimentally enlarged prostate gland in adult Wistar rats
acclimatization. After the period of induction,
residue = 46g; weight of onion, garlic and aridan
some of the animals were sacrificed to
dissolved in the filtrate = 14g
appreciate the gross and histological features
Since 14g of extract dissolvesin 100ml of
confirming prostatic hyperplasia. The remaining
distilled water
subjects were then treated with the extract in
Hence; 1mlwill dissolve= 14g ÷ 100ml.
high and low dosages (for four weeks in
Therefore; concentration of extract obtained =
alternate days) based on their body weight, and
0.14g/ml.
the prostate glands further processed.
2.4. Histology
Animals
2.2. Drug Administration
were
sacrificed
by
cervical
The hormones were diluted in corn oil into
dislocation. The thoracic vertebrate was opened
300µg/ml and 80µg/ml of testosterone and
under aseptic condition; the same procedure was
estradiol
were
performed throughout with prostates removed
administered exogenously via the inguinal
for histological section. The prostates were fixed
region for four weeks, for thrice a week in
in 10% formal saline, after complete fixation the
alternate days and the control group received an
blocks were embedded in paraffin and sections
equivalent volume of 0.5ml of normal saline.
cut at 5 micron which was then stained with H
The extracts were given at 4.0mg/ml and
and E and mounted in Canada balsam.
respectively.
Hormones
2.0mg/ml of high and low dosages
[59]
. The
Microscopic examination of the sections was
Testosterone was manufactured by Green Field
then carried out under a light microscope.
Pharm. (JIANG SU) Co., Ltd, China while
3. RESULTS
Estradiol was manufactured by Medipharm
Table 1: Prostate weight per 100g body
(Pvt.) Ltd., 108-Kotlakhpat Industrial Estate,
weight of the groups
Lahore.
2.3. Method of Extraction
Equal quantities of; onion bulb (Allium
cepa), garlic (Allium sativa) and aridan fruit,
haven removed the seed (Tetrapleura tetraptera)
were obtained at a local market in Ibadan, Oyo
State. 20g of each plant samples were taken (60g
in all); dried, grinded to powdered form and
soaked in 100ml of distilled water. After a
period of 48 hours, the solution was filtered;
volume of filtrate obtained = 54ml, weight of
Intl. J Chem Pharm Res 2013;2(10): 275-286
GROUPS
N
MEAN ± S.D
NORMAL CONTROL
5
0.05 ± 0.02
(NC)
HORMONE
5
0.11 ± 0.01
TREATED (HT)
LOW DOSE
5
0.08 ± 0.01α,β
EXTRACT (LDE)
HIGH DOSE
5
0.05 ± 0.01α,γ
EXTRACT (HDE)
The means of the groups were compared using
T-test at P < 0.05
α – No significant difference between NC and
LDE, HDE
β – There is a significant difference between HT
and LDE
γ - There is a significant difference between HT
and HDE
280
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Odumosu F.M, et al.,
Experimentally enlarged prostate gland in adult Wistar rats
Fig. 1: Graphical Representation of Table 1
Fig 3: Hormone Treated Group showing
Prostate with little (or no) prostatic secretion
and concretions, compressed excretory duct
and smooth muscle bundle and enlarged
glandular epithelium.
Fig 2: Normal Control Group showing
Prostate with normal Histoarchitecture
Prostate Stained with H & E (Mag.x1800)
Fig 4: Prostate Treated with Extract in Low
Dose shows gradual reversal to normal
Histoarchitecture
Prostate Stained with H & E (Mag.x1800)
PC- Prostatic Concretion, ED- Excretory Duct,
PS-
Prostatic
Secretion,
GE-
Glandular
Epithelium, SMB- Smooth Muscle Bundle
Prostate Stained with H & E (Mag.x1800)
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Odumosu F.M, et al.,
Experimentally enlarged prostate gland in adult Wistar rats
Fig 5: Prostate Treated with Extract in High Dose shows complete reversal to normal
Histoarchitecture
Prostate stained with H & E (Mag.x1800)
PC- Prostatic Concretion, ED- Excretory Duct, PS- Prostatic Secretion, GE- Glandular Epithelium,
SMB- Smooth Muscle Bundle.
4. DISCUSSION
With the values provided in Table 1, as
initial state). This could be attributed to the antiinflammatory properties of the extract
well as the pattern of data distribution in Fig. 1;
as
one would appreciate the anabolic effect haven
testosterone.
caused by the combined administration of the
hormones
[60][61]
well
as the
withdrawal
of
[36][49][55]
testicular
In addition; Fig. 2 shows the normal
. However, after the periods of
histoachitecture of the prostate gland, with the
high and low dosages administration; one would
prostatic concretions, excretory duct, prostatic
observe a clear reversal in the enlarged states of
secretion, glandular epithelium and smooth
the prostates (almost bringing them back to their
muscle bundles observed in their normal state.
Intl. J Chem Pharm Res 2013;2(10): 275-286
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Odumosu F.M, et al.,
Experimentally enlarged prostate gland in adult Wistar rats
Fig. 3 displays the anabolic effect haven caused
4. Tsukise, A.; Yamada, K. (1984). "Complex
by the hormones; resulting in hypertrophy of the
carbohydrates in the secretory epithelium of
epithelial cells, constriction of the excretory
the goat prostate". The Histochemical
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Journal
concretions and then compressed smooth muscle
doi:10.1007/BF01003614. PMID 6698810.
bundles.
After
the
period
of
extract
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5. Andrology Australia.
administration, Fig. 3 and Fig. 4 confirms the
6. Leissner KH, Tisell LE (1979). "The weight
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Nephrol.
CONCLUSION
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Further research upon the aqueous
extract of Allium cepa, Allium sativa and
13
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137–42.
PMID 90380.
7. "Prostate
Tetrapleura tetraptera might be a promising
Gland
Development".
ana.ed.ac.uk. Retrieved 2011-08-03.
treatment for individuals with Benign Prostatic
8. Raychaudhuri, B., Chaill, D.: Pelvic Fascia
Hyperplasia, as this work confirms its role in
in Urology, Ann R Coll Surg Engn: vol90:8,
reversing hypertrophied prostate glands to its
pg633-637, 2008
initial state in adult male wistar rats.
9. Instant Anatomy- Abdomen-vessels-veinsprostatic plexus. 2007-11-23
ACKNOWLEDGMENTS
We appreciate the supportive role of all
10. Mayers Robert P.: Structure of the Adult
the staff of the Department of Anatomy, Olabisi
Prostate from a Clinician's Stand Point,-
Onabanjo University (Ikenne) Ago- Iwoye,
Clinical Anatomy. Vol3 :(2000), page 214-
Ogun
215, June 2010.
State,
Nigeria
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their
valuable
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11. Aare Mehik; Epidemology and Diagnostical
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