Anti-Fertility Effects of Fractions from Carica Papaya (Pawpaw) Linn
Total Page:16
File Type:pdf, Size:1020Kb
Arabian Journal of Chemistry (2014) xxx, xxx–xxx King Saud University Arabian Journal of Chemistry www.ksu.edu.sa www.sciencedirect.com ORIGINAL ARTICLE Anti-fertility effects of fractions from Carica papaya (Pawpaw) Linn. methanol root extract in male Wistar rats Chinaka O. Nwaehujor a,*, Julius O. Ode b, Mercy R. Ekwere a, Rita I. Udegbunam c a Department of Biochemistry, Faculty of Basic Medical Sciences, University of Calabar, P.M.B. 1115 Calabar, Nigeria b Department of Veterinary Pharmacology and Toxicology, Faculty of Veterinary Medicine, University of Abuja, Nigeria c Department of Veterinary Surgery, Faculty of Veterinary Medicine, University of Nigeria, Nsukka, Nigeria Received 22 October 2012; accepted 8 October 2014 KEYWORDS Abstract The methanol root extracts of Carica papaya (Pawpaw) are used in eastern Nigeria for Antifertility; the treatment of malaria, hepatitis and jaundice. The aim of this study was to investigate the effects Carica papaya root; of the fractions isolated from C. papaya methanol root extract on fertility in male Wistar rats using CPFE 1; sperm counts, percentage defective sperm cells (morphology), biochemical and hormonal assays as Male Wistar rats; biomarkers. The roots of C. papaya were extracted using 80% methanol for 72 h. Oral acute toxicity Birth control; study was done with the crude extract for 24 h. The extract was fractionated by column chromatog- Sperm count raphy using petroleum ether, chloroform, ethyl acetate and methanol. The petroleum ether fraction was further fractionated on preparative TLC using ethyl acetate–methanol solvent systems to iso- late CPFE 1, CPFE 2 and CPFM 1. The 3 fractions (75 mg/kg) were used to treat male Wistar rats orally for 60 days. Animals were euthanized and testes collected, homogenized and used for sperm count and motility. Plasma and serum were used to assay biochemical parameters including aspar- tate aminotransferase (AST), blood urea nitrogen (BUN), total bilirubin (TB), alkaline phosphatase (ALP), alanine aminotransferase (ALT), gamma glutamyl transferase (GGT), triglycerides, hormones (LH and FSH). Histopathological study of the testes, kidney, heart and liver were con- ducted. Acute toxicity result showed that C. papaya root extract produced no mortalities at the dose of 2000 mg/kg but induced CNS-related symptoms as well as diuresis. The fractions significantly (P < 0.01) produced decreases in sperm counts and increased the percentage of defective sperm cells. There were significant (P < 0.05) increases in aspartate aminotransferase (AST) and blood urea nitrogen (BUN). Histopathological studies showed mild kidney and cardiac hyperaemia, slight * Corresponding author. Tel.: +234 8035450300. E-mail address: [email protected] (C.O. Nwaehujor). Peer review under responsibility of King Saud University. Production and hosting by Elsevier http://dx.doi.org/10.1016/j.arabjc.2014.10.018 1878-5352 ª 2014 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/). Please cite this article in press as: Nwaehujor, C.O. et al., Anti-fertility effects of fractions from Carica papaya (Pawpaw) Linn. methanol root extract in male Wistar rats. Arabian Journal of Chemistry (2014), http://dx.doi.org/10.1016/j.arabjc.2014.10.018 2 C.O. Nwaehujor et al. hepatic degeneration and severe necrosis of the germinal epithelium of the testes. This study calls for some level of caution in the use of these roots and its extracts/fractions in traditional medicine for treating diseases. On the other hand, it could be a good source of drug for birth control. ª 2014 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/). 1. Introduction In reproduction, various extracts of C. papaya have been shown to have antifertility activity in male (Chinoy and Nigeria, within a few years will be a leading country with the Padman, 1996) and female rats (Chinoy et al., 1997). It is also highest population growth in Africa. Since the population is reported to be an abortifacient and a lactogenic (Burkill, tremendously on the increase, economists have forecast that 1985). Aqueous extract of C. papaya leaf caused reduction in this may drastically affect the economic growth. Family plan- mean values of andrological parameters as a result of lesion ning has been promoted through several methods of contra- of the seminiferous tubule epithelium (Oyekunle and Omope, ception, although these contraceptives have various side 2010). The present study therefore is to determine the effects effects produced by their steroid content (Bingel and Benoit, of fractions from the methanol root extract of C. papaya on 1973). Therefore, there is a need for drugs which are effective some reproductive and biochemical parameters in male Wistar with lesser side effects. rats. Carica papaya (Pawpaw) is the sole species in the genus Carica of the plant family Caricaceae. Originally from South- 2. Materials and methods ern Mexico, Central America and Northern South America, the papaya is now cultivated in most countries with tropical 2.1. Preparation of the extract and fractionation climate like Nigeria (Oyekunle and Omope, 2010). C. papaya is known as okwulu bekee by the Igbo, Ibepe by the Yoruba Fresh C. papaya (Linn.) roots were obtained from Nsukka and kawuse by the Hausa tribes of Nigeria. It is a fast growing urban region in November 2012 and authenticated by Mr. semi-woody tree of about 6 m tall, stem normally not A. Ozioko, a taxonomist with the Biodiversity Development branched, and becoming hollow with age; male and female Centre Program (BDCP), Nsukka Enugu State, Nigeria. The flowers usually grow on separate plants, reaching maturity roots (2.5 kg) were chopped into pieces and air dried on a lab- within a year and living for 5–6 years only. It is widely culti- oratory bench. This was pulverized into coarse powder. 1000 g vated throughout West Africa around habitations in the forest of the dried pulverized roots was extracted by cold percolation zone (Burkill, 1985). in 80% methanol:water for 72 h with intermittent shaking Different parts of C. papaya plant are attributed with dif- every 2 h. This was filtered with Whatman No. 4 filter paper ferent medicinal values. For example, in African traditional and dried using a rotary evaporator (Bu¨ chi, Switzerland) at medicine, the boiled green leaves are combined with leaves 40 °C. The extract was loaded into a 4 cm·50 cm glass column of Azadirachta indica, Cymbopogon citratus, Psidium guajava pre-loaded with silica gel (Silica Gel 60–120 mesh, 60A, and stem bark of Alstonia boonei boiled together and the Oxford, India) and fractionated using petroleum ether, chloro- hot infusion is drunk as one wine glass full thrice daily in form:ethyl acetate (8:2) and ethyl acetate:methanol (3:7) as elu- the treatment of malaria (Gill, 1992). Its fresh leaves are also ents. Following partial purification, the petroleum ether efficacious in the treatment of gonorrhea, syphilis and amoe- fraction, which showed significant effects on sperm parameters bic dysentery (Gill, 1992). The milky juice of the unripe fruit was further subjected to ethyl acetate and methanol sub-frac- is a powerful abortifacient, anti-helminthic for roundworms, tionation using preparative thin layer chromatography stomach disorders and enlargement of liver and spleen (TLC). Three compounds were obtained from the sub-fraction (Gill, 1992). The seeds are also effective as a vermifuge and viz. CPFE 1 and 2, and CPFM 1. These were used for anti-fer- in the treatment of hypertension, diabetes mellitus and tility assay described below. hypercholesterolemia (Gill, 1992). Results from studies on biological activities of C. papaya parts, extracts and isolated 2.2. Animals compounds showed that the latex and root extracts inhibited Candida albicans while extracts of pulp and seeds showed Adult male Wistar rats weighing between 170 and 210 g were bacteriostatic properties against Staphylococcus aureus, used for the study. They were kept in the animal house of Escherichia coli, Salmonella typhi, Bacillus subtilis, and the Faculty of Veterinary Medicine, University of Nigeria, Entamoeba histolytica, in vitro (Emeruwa, 1982). Its root Nsukka under a well-ventilated environment. The animals aqueous extract has been shown to have a purgative effect were allowed free access to feed and water ad libitum. The (Akah et al., 1997). use of the animals conformed with internationally accepted C. papaya latex has been shown to have activity against C. principles for laboratory animal use and care as documented albicans (Giordani et al., 1996), Heligmosomoides polygyrus in the European Community guidelines, Council Directive, (Satrija et al., 1995), Ascaris suum and Ascaridia galli (Satrija 1986 (86/609/EEC), revised in Directive 2010/63/EU. The et al., 1994). Aqueous extract of C. papaya roots have shown experimental protocols were approved by the Ethics Commit- potential activity in the management of dengue fever (Nisar tee for Animal Experimentation, University of Nigeria, et al., 2011), antitumor and immune-modulatory activities Nsukka and in accordance with Nigerian Federal Government (Otsuki et al., 2010), hepatitis and jaundice in children. legislation on Animal care. All animals, including the control Please cite this article in press as: Nwaehujor, C.O. et al., Anti-fertility effects of fractions from Carica papaya (Pawpaw) Linn. methanol root extract in male Wistar rats. Arabian Journal of Chemistry (2014), http://dx.doi.org/10.1016/j.arabjc.2014.10.018 Anti-fertility effects of fractions from Carica papaya (Pawpaw) Linn. methanol root extract 3 group, were humanely euthanized before the onset of severe fields in each sample. The mean of the 3 estimations was used clinical conditions.