15 male rats were grouped and administered crude ethanolic extracts of Garcinia cambogia seeds to test the effects on the histology of the testis and sperm counts. Group A served as the control while Groups B and C received 100 and 200 mg/kg body weight of extracts, respectively. The administration was done orally once a day, six days a week for six weeks. The routine histological preparation at the end of administration revealed increase in the interstitial spaces, degeneration of the Ledgid cells anddistortion in the arrangement of the cells of spermatogenic series. The sperm counts revealed a significant increase in the experimental groups when compared statistically with the control (

Adesanya, OA

Jimoh, OR

Josiah, SJ

Oluyemi, KA

Omotuyi, IO

Oyesola, TO

English

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African Journal of Biotechnology Vol. 6 (10), pp. 1236-1238, 16 May, 2007     Available online at http://www.academicjournals.org/AJB ISSN 1684–5315 © 2007 Academic Journals      Short Communication  Effects of crude ethanolic extract of Garcinia cambogia on the reproductive system of male wistar rats (Rattus novergicus)  Oluyemi Kayode A.1*, Jimoh Olusegun R.2, Adesanya Olamide A.1 Omotuyi Idowu O.3, Josiah Sunday J.3 and Oyesola Tolulope O.4.  1Department of Anatomy, School of Basic Medical Sciences, Igbinedion University, Okada. P.M.B 0006, Benin City. Nigeria. 2Department of Anatomy, College of Health Sciences, University of Ilorin, P.M.B. 1515, Ilorin, Kwara State, Nigeria. 3Department of Biochemistry, School of Basic Medical Sciences, Igbinedion University, Okada. P.M.B 0006, Benin City. Nigeria. 4Department of Physiology, School of Basic Medical Sciences, Igbinedion University, Okada. P.M.B 0006, Benin City. Nigeria.  Accepted 27 December, 2006  15 male rats were grouped and administered crude ethanolic extracts of Garcinia cambogia seeds to test the effects on the histology of the testis and sperm counts. Group A served as the control while Groups B and C received 100 and 200 mg/kg body weight of extracts, respectively. The administration was done orally once a day, six days a week for six weeks. The routine histological preparation at the end of administration revealed increase in the interstitial spaces, degeneration of the Ledgid cells and distortion in the arrangement of the cells of spermatogenic series. The sperm counts revealed a significant increase in the experimental groups when compared statistically with the control (p<0.05). It was 81.5 ± 13.62 × 106/ml and 70 ± 12.98 × 106/ml in groups Band C, respectively, as compared to 59.8 ± 2.14 × 106/ml in the Control group.  Key words: Garcinia cambogia, sperm count, histology, Leydig cells, interstitial spaces.   INTRODUCTION  With the shifting of attention from synthetic drugs to natural plant products, the use of plant extracts for disease treatment is now on the increase. Plants that were once considered of no value are now being investigated, evaluated and developed into drugs with little or no side effects. One of such plant is Garcinia cambogia (bitter kola), a plant found in the moist forest and grows as a medium size tree, up to 12 meters high in places like Nigeria, United State, South East Asia, India and Central Africa. The active constituent of G. kola is a dimeric flavonoid molecules fused  together- biflavonoid.   Biflavonoids  are     *Corresponding author. E-mail:Kayodedanatomist@yahoo.com. Tel: +2348059860788. potent antioxidants. Other constituents include xanthones and benzophenones (Ebong and Korubo-Owiye, 1996; Encyclopedia,  2002).  Several  works  done  on  G.  kola have confirmed its hypolipidermic (Koshy and Vijaya-lakshmi, 2001; Iwu, 1993; BBC News, 1999; Oluyemi et al, 2007), antifungal (Mackeen et al., 2002), antican-cerous (Ho et al., 2002; Pan et al., 2001), antihistaminic (Nakatani and Atsumi et al., 2000), antimicrobial (BBC News, 1999: Iwu et al., 1999), erythropoietic (Oluyemi et al, 2007), antiviral (Chen et al., 1996; Iwu et al., 1999) and antiulcerogenic effects (Mahendran et al., 2002). Though its effects on the reproductive system have been investigated by Akpantah et al. (2003) and Akinloye et al. (1999), their findings are however at variance.   The present study aims at furthering tests on the effects of G. kola on the reproductive system of male wis-tar rats and probably resolves the conflicting reports. Oluyemi et al.        1237    Table 1.  Sperm counts of wistar rats treated with crude ethanolic extract of G. cambogia.  Parameter Control A Group B Group C Dose Saline water 100 mg/kg body weight 200 mg/kg body weight Sperm density (million/ml) 59.8±0.96 70.0 ± 5.80Ψ 81.5 ± 6.08Ψ  n= 5 for all group.  Values are recorded as Mean ± SEM.  ΨValues are significantly different from control (P< 0.05).    MATERIALS AND METHODS  Animals  15 Adult male rats weighing between 120 – 135 g were obtained from the animal  house  of  the  Igbinedion  University,  Okada,  Edo  State were used for this experiment. The rats were kept in the animal control room, acclimatized for two weeks before the experiment commenced. The rats were fed on standard diet (Bendel Feed and Flour Mills Ltd), water was given ad libitum and maintained under standard conditions. The animal room was well ventilated with a temperature range of 25 - 27oC under day/night 12-12 h photoperiodicity. The rats were grouped into three groups (5 rats each) A (0 mg/kg), B (100 mg/kg), and C (200 mg/kg).   Plant materials  G. kola was obtained from a local market in Ile - Ife, Osun State and authenticated by the Botany Department, Igbinedion University. The outer coats were removed and the seed sun dried. The dried seeds were grounded into fine powder and the crude ethanolic extraction done using 70% alcohol. The solution was filtered after 24 h while the filtrate was concentrated to a semi solid form using the rotary evaporator, weighed and the solutions were prepared as100 and 200 mg/ml.    Experimental design  The administration of the extract was totally by gavage. Proper concentrations were administered by the use of metal oropharyn-geal canula and calibrated hypodermic syringe. The administration of G. kola extract was done once in a day, 6 days of the week and for the period of 6 weeks. The control group received no extract while groups B and C received 100 and 200 mg/kg body weight of the extracts respectively. The animals were sacrificed after the administration of extracts stopped.    Testis collection and semen analysis  The testes were dissected out via the inguinal region; the caudal epididymis were dissected free and suspended in the semen diluting fluid (sodium bicarbonate-formalin diluting fluid), containing 1% acetic acid, 4% NaHCo3  and 35% Formalin (Monica, 2000). The rest of the testis was fixed in 10% buffered formalin. The sperm count was done using haemocytometer counting chamber of the Neubauer type.    Routine histological preparation  The histology of the testes and epididymis were done by modifi-cation of method described by Akpantah et al. (2003). The organs were cut in slabs of about 0.5 cm thick transversely and fixed in 10% buffered formalin for a day after which it was transferred to 70% alcohol for dehydration. The tissues were passed through 90% and absolute alcohol and xylene for different durations before they were transferred into two changes of molten paraffin wax for 1 h each in an oven at 65 oC for infiltration. They were subsequently embedded and serial sections cut using rotary microtome at 6 microns. The tissues were fixed into albumenised slides and allowed to dry on hot plate for 2 min. The slides were dewaxed with xylene and passed through absolute alcohol (2 changes); 70% alcohol, 50% alcohol and then to water for 5 min. The slides were then stained with haematoxylin and eosin. The slides were mounted in Canada balsam. Photomicrographs were taken using X40 objectives.   Statistical analysis  All data from the sperm count were compare using a two-way ANOVA at p<0.05.   RESULTS AND DISCUSSION  Table 1 shows the effects of the crude ethanolic extract of G. cambogia on the sperm count of wistar rats. There were significant increases when compared with the control. This could be as a result of the presence of biflavonoid and xanthone in the plant. These compounds are potent antioxidants which are capable of increasing the production of testosterone, the key hormone involved in the production and maturation of spermatozoa in the seminiferous tubules of the testis (Ganong, 2003; Guyton and Hall, 1998). Akpantah et al. (2003) has found that G. cambogia increases the peripheral testosterone level. This result agrees with the work of Akinloye et al. (1999) who recorded an increase in the sperm count after feeding 35 mg/kg of G. cambogia to rabbits for 8 weeks.  Figures 1A – C show the effects of graded dosages of G. cambogia on the histology of the testes of the control and experimental rats. G. cambogia extract causes increase in the interstitial spaces, reduction in the Leidig cells population in the interstitial spaces, slight reduction in the seminiferous luminal spermatozoa concentration, contraction of the seminiferous tubules, and derangement of the cells of spermatogenic of series. These findings agreed with the work of Akinloye et al. (1999) but are at variance with the work of Akpantah et al. (2003) who recorded no histological difference in the testes of wistar rats. The reduction in sizes of the seminiferous tubules might be due to the contractile activities of the fibroblast of the seminiferous tubular wall. This  is  because  testos-  1238           Afr. J. Biotechnol.    A                                                                                C B    Figures 1A to C. Photomicrograph of the histology of the testis of the animals.    terone causes the contraction of the fibroblasts which aid the movement of the non-motile spermatozoa into the epididymal tubule where the final maturation into motile spermatozoa takes place (Peter et al., 1999). This might be the reason for the increase in the interstitial spaces. Figures 2A - C show the micrographs of the epididymis of the control and experimental rats. They show reduction in the sperm concentration in the tubules of the epididymis of both groups and increase in the interstitial spaces of the group which received 100 mg/kg body weight of extract. These agree with the work of Akpantah et al. (2003). The histological preparations of the testis and epididymis revealed toxic effects which were more pronounced in the group that received 200 mg/kg body weight of extract. This shows a dose dependent toxic effect. This is likely due to the presence of garcinone E in G. cambogia. This compound has been found to possess cytotoxic effects against liver cells (Ho et al., 2002).   REFERENCES  Akinloye AK., Igharna OO, Olaniyi MO, Alaka OO, Oke BO (1999). Preliminary investigations on the effects of Garcinia kola, (bitter kola). on the rabbit’s testis and epididymis. Trop. Vet. 18: 49-54.  Akpantah AO, Oremosu AA, Ajala MO, Noronha CC, Okanlawon AO (2003). the effect of crude extract of Garcinia Kola seed on the histology and hormonal milieu of male Sprague-Dawley rats’ reproductive organs. Niger. J. Health Biomed. Sci. 2(1): 40-46. BBC News. (1999). Garcinia cambogia. Thursday, August 5. Chen S-X, Min W, Boon-Nee L (1996). Active constituents against HIV-1 protease from Garcinia mangostana. Planta Med. 62 (4): 381-382 Ebong OO, Korubo-Owiye T (1996). Comparison of the effects of the seeds of Garcinia cola and Cola acuminata on gastric acid secretion in rats. West Afr. J. Pharmacol. Drug Res. 12: 51-54.         A                                                                                      B   C    Figures 2A to C. Photomicrograph of the histology of the epididymis of the animals.    Encyclopedia of Clinical Toxicology (2002). Garcinia kola. Pp 485-486. Ganong WF (2003) Review of Medical Physiology. Lange Twenty first edition. McGraw-Hill Companies, USA. 23: 429-430. Guyton AC, Hall JE (1998). Pocket companion to Textbook of Medical Physiology. 9th Edition, W.B. Saunders Company, India. 80: 691 Ho CK, Huang YL, Chen CC (2002). Garcinone E, a Xanthone derivative, has potent cytotoxic effects against Hepatocellular carcinoma cell lines. Planta Med. 68(11): 975-979. Iwu M (1993). Handbook of African Medicinal Plants. CRC press, BocaRatton, FL. Iwu MW, Duncan AR, Okunji CO (1999). New antimicrobials of plant origin. In: J Janick (ed.), Perspectives on new crops and new uses. ASHS Press, Alexandria, VA. pp. 457–462. Koshy AS, Vijayalakshmi NR (2001). Impact of certain flavonoids on lipid profiles. Protein action of Garcinia cambogia flavonoids. Photother. Res, Aug., 15 (5): 395-400. Mackeen MM, Ali AM, Lajis NH, Kawazu K, Kikuzaki H, Nakatani N (2002). Antifungal Garcinia acid esters from the fruits of Garcinia atroviridis. Z Naturforsch. © Mar-Apr., 57 (34): 291-295. Mahendran P, Vanisree AJ, Devi CSS (2000). The antiulcer activity of Garcinia cambogia extract against indomethacin-induced gastric ulcer in rats. Phytother Res. 16(1): 80-83. Monica C (2000). District Laboratory Practice in Tropical Countries, Part 2. Cambridge Low-Price Edition. Cambridge University Press. UK. p. 403. Pan MH, Chang WL, Lin-Shiau SY, Ho C-T, Lin JK (2001). Induction of apoptosis by garcinol and cucumin through cytochrome C release and activation of caspases in human leukaemia HL-60 cells. J. Agric. Food. Chem. 29(3): 1464-1474.  Peter LW, Lawrence HB, Martin MB (1999). Gray’s Anatomy: Anatomical Basis of Medicine and Surgery. Churchill Livingstone, Britain: 2(2): 76. Oluyemi KA, Omotuyi IO, Jimoh OR, Adesanya Olamide A, Saalu CL, Josiah SJ (2007). Erythropoietic and anti-obesity effects of Garcinia cambogia (bitter kola) in Wistar rats. Biotechnol. Appl. Biochem. 46: 69–72. (Printed in Great Britain).      

Effects of crude ethanolic extract of Garcinia cambogia on the reproductive system of male wistar rats (

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