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): 275-286 275 www.ijcpr.net 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 www.ijcpr.net 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 www.ijcpr.net 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 www.ijcpr.net 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 www.ijcpr.net 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 dissolvesin 100ml of confirming prostatic hyperplasia. The remaining distilled water subjects were then treated with the extract in Hence; 1mlwill 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 www.ijcpr.net 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) Intl. J Chem Pharm Res 2013;2(10): 275-286 281 www.ijcpr.net 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): 275-286 282 www.ijcpr.net 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). 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