Effect of ethanol extract of Zapoteca portoricensis stem on testosteroneinduced benign prostate hyperplasia (BPH) in adult male albino rats

2018 Australian Journal of Basic and Applied Sciences  
Citation:Joshua, et al., Effect of ethanol extract of Zapotecaportoricensis stem on testosterone-induced benign prostate hyperplasia (BPH) in adult male albino rats. of men greater than 50 year of age have symptoms arising from BPH and 20-30% of men reaching 80year of age require surgical intervention for the management of BPH. Despite the high impact of BPH on public health, however, the pathogenesis of BPH is still largely unresolved. Although ageing and genetic predisposition represent the
more » ... ion represent the central mechanism implicated, recent novel finding also highlighted the key-role of hormonal alterations, metabolic syndromes and inflammation (Parsons and Kashefi, 2008; Alberto et al., 2009) Symptoms of BPH are classified as storage or voiding. Storage symptoms include urinary frequency, urgency (compelling need to void that cannot be deferred), urgency incontinence and voiding at night (nocturia) which may contribute to insomnia. Voiding symptoms include urinary hesitancy (difficulty initiating the stream), straining to void, week or intermittent stream (start and stops), and incomplete bladder emptying. These storage and voiding symptoms are evaluated using the international prostate symptoms score (IPSS) questionnaire, designed to assess the severity of BPH (Black et al., 2006) . Over the years, physician advocated the use of lifestyle, voiding position, medications and surgery, for management and control of prostatic disorders especially BPH. The failure of lifestyle and voiding position in management of BPH has made the use of medication an alternative choice for initial therapy as seen in the Europe and in the USA. However there are common side effects associated with medications which include postural hypotension, h eadache, nasal congestion, weakness, decreased libido, and erectile dysfunction. Surgery, another alternative method of BPH treatment, has been discovered to be associated with dryness of ejaculation leading to sterility (Santillo and Lowe, 2006) . Herbal remedy has become a commonly sought treatment for BPH due to numerous side effects associated with other methods. For instance, saw palmetto extract from Serenoarepens and others are approved in European countries and they are available in USA. Other documented herbal medicines for BPH include beta-sitosterol from Hypeo sisrooperi (African star grass),Urtica dioica (stinging nettle) root andTelfairi aeccidentalis Hook F. (fluted pumpkin) seeds (Barry et al., 2011; Ejike and Ezeanyika, 2011) . However, there is absence of demonstrated scientific information on the treatment of BPH with stem extracts of Zapoteca portoricencis as used by the traditional medicine practitioners in Eastern Nigeria. Hence, the success of this research will help in advancing the search for BPH treatment with bearable or no side effect. MATERIALS AND METHODS Plant Materials The stems of Zapoteca portoricensis were collected from Orba, Nsukka, Enugu State of Nigeria. The plant materials were authenticated by Mr. Alfred Ozioko, a taxonomist at the Centre for Ethenomedicine and Drugs Development, a subsidiary of Bioresources Development and Conservation Program (BCDP), Nsukka, Enugu State. Chemicals and Assay kits All the chemicals used in this study were of analytical grade and were used as such without further purification. Fresh distilled water was used throughout the experimental period. Assay kits used in all the analysis in this study were products of Bioscience, Cusabio, Magiwel and Randox laboratories. Laboratory animals Adult male albino rats (230-400 g) obtained from the animal house of Biochemistry Department, University of Nigeria, was kept under standard environmental condition of 12/12 hours light/dark cycle. They were housed in polypropylene cages (5 animals per cage), and were maintained on mouse chow (Livestock Feeds Nigeria Ltd), provided with water ad libitum. They were allowed to acclimatize for 7 days to the laboratory conditions before the experiment. The use and care of the animals, and the experimental protocol were in strict compliance with the Institute of Laboratory Animals Research (ILAR) guidelines on the use and care of animals in experimental studies and approved by the local ethics committee of our institution. Preparation of ethanolic stem extract of Zapoteca portoricensis Fresh stems of Zapoteca portoricensis were collected, cleaned and air dried before being subjected to size reduction to a coarse powder with electric grinder. The stem powder (5kg) was dissolved in 10L of 99.7% ethanol for 72hours to achieve maximum extraction. The mixture was agitated using a magnetic stirrer and filtered with Whatman No.1 filter paper. The filtrate was evaporated to dryness until constant weight of the crude extract was obtained. The concentrated crude extract (86.5g) with a percentage yield of 1.73 was stored in an air tight bottle and kept in a refrigerator at 4 °C till used. Qualitative phytochemical analysis Preliminary phytochemical screening was performed to identify the presence of bioactive compounds in crude ethanolic stem extract of the Zapoteca portoricensis used in this study. The phytochemicals (such as flavonoids, glycosides, tannins, alkaloids, saponins, steroids and terpinoids) were tested for using the method of Sofowora (2008); modified by Tiwari et al. (2011) . Acute toxicity and lethality (LD50) test The oral acute toxicity of the ethanol extract was determined according to the method described by Lorke (1983). Determination of Doses The ethanol extract of Zapoteca portoricensis was subjected to acute toxicity studies to determine the dose for the in vivo studies according to the Organization for Economic Cooperation and Development guidelines (Deora et al., 2010) . In all cases, a 3000-mg/kg bwt oral dose of the test extract was found to be tolerable, as no mortality was observed during the study. On the basis of this study, the doses of 100 and 200 mg/kg bwt for the extract were selected. Preparation of 2 % v/v Tween 80 solution The 2 % v/v Tween 80 [polyoxyethylene (20) sorbitan monooleate] solution used in dissolving the extract and the drug (finasteride) was prepared by adding 2ml of the Tween 80 in a 100ml measuring cylinder containing 50 ml of distilled water. This was shaken thoroughly and distilled water added to make up the 100 ml mark. Preparation of testosterone propionate, finasteride and extract solutions Testosterone propionate solution used in this study was prepared by dissolving 5g of testosterone propionate in 1.25L of olive oil at a stock concentration of 4 mg/ml. Finasteride solution was prepared by dissolving 1g of finasteride in 0.1L of 2 % v/v Tween 80 solution at a stock concentration of 10 mg/ml. The ethanol stem extract was prepared by dissolving 10g of the crude extract in 0.1L of 2 % v/v Tween 80 solution at a stock concentration of 100 mg/ml. Experimental design for benign prostatic hyperplasia induction and treatment Animal grouping and BPH induction A total of 25 adult male albino rats (weighing 230-390g) were selected for this study. They were randomly divided into five groups (1, 2, 3, 4 and 5) of 5 animals each and each group housed in its own cage. Animals in Groups 2, 3, 4 and 5 were induced with BPH by exogenous administration of 10 mg/kg body weight testosterone propionate dissolved in olive oil which served as the vehicle. The administrations which were once a day by subcutaneous injection as outlined by Nandecha et al. (2010) , Ejike and Ezeanyika (2011) and Surendra et al. (2011) lasted for 14 days before commencement of treatment. The normal control group (group 1) received subcutaneous injection of olive oil in place of the hormone for the same duration. Animal grouping and BPH treatment At the end of 14 days induction, the animals in Groups 1 (normal controls) and 2 (positive controls) were given oral doses of 2% v/v Tween 80 solution. The animals in Group 3 were given oral dose of 10 mg/kg body weight finasteride, while those in groups 4 and 5 received oral doses of 100 mg/kg and 200 mg/kg body Citation:Joshua, et al., Effect of ethanol extract of Zapotecaportoricensis stem on testosterone-induced benign prostate hyperplasia (BPH) in adult male albino rats. weight extracts respectively. The oral administration was done once per day by the use of gavages for 21 days. The animals were weighed prior to the commencement of the experiment and subsequently every week till the end of the experiment as outlined by Nandecha et al.,(2010) . Collection of sera and tissue samples for analysis After 21 days of treatment, the rats were fasted for 12 hours, anesthetized by a brief exposure to trichloromethane vapour, and bled exhaustively by ocular puncture. Blood samples were collected, allowed to clot and centrifuged at 2000 × g for 10 min. The sera were carefully separated and used for biochemical analyses. Each rat was promptly dissected and the prostate carefully excised, freed of external fascias, washed in cold normal saline, blotted with filter paper and weighed on a sensitive balance to determine the mean prostate weight and the mean prostate-body weight ratio. rats. Group 1 = Normal control Group 2 = Positive control (Testosterone-induced BPH non-treated) Group 3 = Standard control (BPH + finasteride-treated) Group 4 = BPH + extract treated (100mg/kg body weight) Group 5 = BPH + extract treated (200mg/kg body weight) rats. Group 1 = Normal control Group 2 = Positive control (Testosterone-induced BPH non-treated) Group 3 = Standard control (BPH + finasteride-treated) Group 4 = BPH + extract treated (100mg/kg body weight) Group 5 = BPH + extract treated (200mg/kg body weight) Group 1 = Normal control Group 2 = Positive control (Testosterone-induced BPH non-treated) Group 3 = Standard control (BPH + finasteride-treated) Group 4 = BPH + extract treated (100mg/kg body weight) Group 5 = BPH + extract treated (200mg/kg body weight)
doi:10.22587/ajbas.2018.12.12.2 fatcat:kxsfgkayqveh7hwlv6sea5vkwy