Micromorphometric and Stereological Effects of Ethanolic Extracts of Garcinia cambogia seeds on the Testes and Epididymides of Adult Wistar Rats
A Adesanya Olamide, A Oluyemi Kayode, A Ofusori David, O Omotuyi Idowu, U Okwuonu Christina, O Ukwenya Victor, A Adesanya Rotimi
epididymis, garcinia cambogia, morphometry, sperm concentration, testis
A Adesanya Olamide, A Oluyemi Kayode, A Ofusori David, O Omotuyi Idowu, U Okwuonu Christina, O Ukwenya Victor, A Adesanya Rotimi. Micromorphometric and Stereological Effects of Ethanolic Extracts of Garcinia cambogia seeds on the Testes and Epididymides of Adult Wistar Rats. The Internet Journal of Alternative Medicine. 2006 Volume 5 Number 1.
The objective of this study was to evaluate testicular and epididymal alterations resulting from the administration of ethanolic extract of Garcinia cambogia by morphometric methods. Fifteen (15) rats weighing between 120-135g were used for the study. These were divided into control and experimental groups of 5 rats each. They were given ethanolic extract at doses of 0.00mg/kg B.W (control), 100mg/kg B.W (Group B), 200mg/kg B.W (Group C) respectively by gastric lavage for 6 weeks. Five slides chosen at random from the testicular slides of control and experimental were evaluated and analyzed. There was a reduction in the germinative cell thickness of the seminiferous tubules in the treated group compare with control. There was a significant increase in the sperm counts but reduction in motility in the treated groups in a dose dependent manner compare with control (P<0.05). The volume density ratio of lumen was increased in the treated groups which receive the higher dose of extract compared with the control.
The presence of biflavonoids and xanthone in G. kola seeds have been confirmed15. These compounds are potent antioxidants3,7,16. Administration of G. kola seed extracts caused an increase in testosterone production in Sprague-Dawley rats17,18 due to the anti-oxidant properties of its constituents.
The use of quantitative methods of light and electron microscope levels has increased remarkably over the last few years. These methods collectively called “morphometry” or “stereology” allow the number, volume, surface area and lineal extent of structures in three dimensions to be assessed from two dimensional images of light and electron microscopy19,20.
The aims of this study were to investigate the effects of methanolic extract of G. Kola seeds on testes and epididymides using morphometric and stereologic methods.
Materials And Methods
Fifteen (15) adult male wistar rats weighing between 120-135g 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 standard diet (Rat's pellet, Bendel feeds and flour mills Limited, Edo state, Nigeria. Water was given
The plant material,
The administration of the extract was totally done by gavage using metallic oropharyngeal canula and calibrated hypodermic syringe. The administration of
The testes and the epididymides were dissected free, weighed in a torsion balance and fixed in 10% buffered formalin. Routine histological slides preparations were done using Erlich's haematoxylin and eosin method as described by Oluyemi et al7. The stained sections were subjected to micro-morphometric analysis according to Huttunen et al21
The sperm count was analyzed using Neubaeur's counting chamber.
The values are recorded as mean ± standard deviation. The statistical significance of difference in the mean and standard deviation (p<0.05) was analyzed by two-way ANOVA comparison of each of the test groups and the control.
Morphometric analysis of Garcinia kola treated Testis: The diameter of seminiferous tubules showed an increment in the treated group as compared with the control (Table 1). This observation was pronounced in group C which received 200mg/kg B.W of the extract (Table 1). The germinative cell thickness and germinative cell thickness/ radius, showed a gradual reduction in the treated group as compared with the control. This observation is dose dependent.
Semen Analysis: The sperm concentration presents a dose dependent significant increment (P<0.05) as shown in Table 2 in contrast to dose dependent significant decrement observed in sperm motility.
Volume Density of Seminiferous Tubules: The volume density of seminiferous tubules was observed to present a non significant increment in the epithelium and lumen of the treated group as shown in Table 3. a dose dependent reduction was observed in the treated group (Table 3)
Volume Density of Epididymis: Table 4 presents the volume density of the epididymis. It shows a significantly higher volume of epithelium in a dose dependent manner when compared with the control. The luminal volume of Group B is not significantly different from that of the control while the volume of the Group C is significantly lower than that of the Group A and Group B. The connective tissue volume shows a dose dependent increase in both treatment groups compared to the control.
The morphometric analysis of the seminiferous tubules (Tables 1) of the present work justifies the increment in sperm counts as found in the epididymal lumen (Table 2). The increase in the diameter of the seminiferous tubules and the decrease in the germinative cell thickness could be as a result of rapid spermiogenesis resulting in the depopulation of the peripherally placed spermatids and increase in luminal sperm concentration. Spermatids develop into mature spermatozoa through the process of spermiogenesis22. Rapid spermiogenesis is facilitated by antioxidants and increase in the peripheral testosterone level. These two factors are properties of G.kola's biflavonoids and xanthones7,23. The decrease in the motility of the spermatozoa could be due to the rapidity of development. The spermatozoa may need a moderate but progressive development for them to have excellent motility. This may also be due to the presence of some toxic component like benzophenone in the ethanolic extracts of G. kola. Cotterih et al24, has reported the antioxidant potency of Kolaviron- an antioxidant extract of G.kola. Other antioxidant like carotenoid has been found to protect spermatogenesis in animals exposed to toxicant25. Anti-oxidants such as carotenoids are well known as highly efficient scavengers of singlet oxygen and other excited species (oxidants). Apigenin-based flavonoids represent 60% of the total flavonoids present in the diethyl-ether fraction of G. kola26. Other studies in man, have shown that lycopene helps men with idiopathic infertility, with an improvement in male fertility especially sperm characteristics25.
However, the diameter of the ST is largest in the group receiving the 200mg/kg B.W of ethanolic extracts of G. kola. The diameter of the group receiving 200mg/kg B.W of ethanolic extracts of G. kola was significantly higher than that of 100mg/kg B.W extracts and the control. The volume density of the epithelium and lumen of the seminiferous tubules (Table 3) as assessed by the Weibel counting grids (point counting method) increase non-significantly as compared with the control (P>0.05). The volume density of C.T of the seminiferous tubules shows a dose dependent increase when compared with the control (Table 3). In relation to a report by Oluyemi et al7 ,there is a need for the development of more connective tissues to support the contractile mechanism involved in the migration of immotile spermatozoa, formed in the seminiferous tubule, into the epididymis where they acquire motility. This mechanical contraction is brought about by the fibroblasts in the connective tissue walls of the seminiferous tubules. Young and Heath27 affirmed that the contractile activity of fibroblasts increases in response to the testosterone level in the testis. This androgen (testosterone) is produced in the testis by the interstitial cells of Leydig in response to follicle stimulating hormone (FSH) produced by the anterior pituitary27. Akpantah et al23 found out that G. Kola extract increases the peripheral testosterone levels in wistar rats treated with 100mg/Kg B.W of the extract. Increased spermatogenic activity in seminiferous tubules associated with the administration of G. kola extract is hence due to the ability of the antioxidant compounds in the latter to increase peripheral testosterone levels7,23.
The significant decrease in the volume density of the lumen of the epididymis (Table 4) is most likely due to the significant increase in the epithelium (pseudostratified) of the epididymis with the corresponding increase in the volume density of the connective tissue of the epididymal wall. This is a very strong indication that G.Kola extract influences the epididymis in a positive direction. The ciliated epithelium is also important in the movement of the spermatozoa from the proximal to the distal portion of the tube prior to ejaculation which under the influence of sympathetic innervations. “The proximal portion of the muscular wall of epididymis exhibits slows rhythmic contractility which gently moves spermatozoa towards the ductus deferens. Distally the smooth muscle is richly innervated by the sympathetic nervous system which produces intense contractions of the lower part of the epididymis during ejaculation27.
In conclusion, this work further confirmed the spermatogenic and tissue enhancing effects of G. Kola extract in male Wistar rats.
The Authors hereby acknowledge with deep regret the wife of the Corresponding Author Late Mrs Olufunke R. Adesanya for her contribution in the typing of this manuscript shortly before her demise. May your gentle soul rest in the bosom of our Lord. Amen.