|Year : 2020 | Volume
| Issue : 1 | Page : 11-15
Curative effect of aqueous extract of the bark of Boswellia dalzielii on flutamide-induced testicular toxicity in Wistar rat
Seno Akpobasaha1, Joseph O Ezugworie1, Nto J Nto1, Uloaku A Nto-Ezimah2
1 Department of Anatomy, Faculty of Basic Medical Sciences, College of Medicine, University of Nigeria Enugu Campus, Enugu, Nigeria
2 Department of Chemical Pathology, Faculty of Medical Sciences, College of Medicine, University of Nigeria Enugu Campus, Enugu, Nigeria
|Date of Submission||15-Aug-2019|
|Date of Decision||12-Jan-2020|
|Date of Acceptance||19-Feb-2020|
|Date of Web Publication||03-Apr-2020|
Dr. Nto J Nto
Department of Anatomy, Faculty of Basic Medical Sciences, College of Medicine, University of Nigeria Enugu Campus, Enugu.
Source of Support: None, Conflict of Interest: None
Background: Male reproductive dysfunction is a common health concern and can affect the quality of life. This study evaluated the curative effect of aqueous extract of the bark of Boswellia dalzielii on flutamide-induced testicular toxicity in Wistar rat. Materials and Methods: Sixteen male Wistar rats were randomly divided into four groups of four rats each, labeled Groups A–D. Group A was the negative control, Group B (positive control) received 10 mg/kg body weight of flutamide. Groups C and D were given 10 mg/kg body weight of flutamide per day for 7 days and discontinued, they were treated with 100 and 300 mg/kg body weight of aqueous extract of B. dalzielii, respectively, from days 8 to 21. All administrations were carried out orally. Results: There was a dose-dependent increase in body weight, raised serum testosterone levels, and increased cellular proliferation following administration of B. dalzielii on flutamide-induced testicular toxicity. Conclusion: The result of this study suggests that the aqueous extract of the bark of B. dalzielii has a therapeutic effect on flutamide-induced testicular toxicity.
Keywords: Aphrodisiac, boswellia dalzielii, flutamide, male reproductive dysfunction, testes
|How to cite this article:|
Akpobasaha S, Ezugworie JO, Nto NJ, Nto-Ezimah UA. Curative effect of aqueous extract of the bark of Boswellia dalzielii on flutamide-induced testicular toxicity in Wistar rat. Int J Med Health Dev 2020;25:11-5
|How to cite this URL:|
Akpobasaha S, Ezugworie JO, Nto NJ, Nto-Ezimah UA. Curative effect of aqueous extract of the bark of Boswellia dalzielii on flutamide-induced testicular toxicity in Wistar rat. Int J Med Health Dev [serial online] 2020 [cited 2020 Jul 11];25:11-5. Available from: http://www.ijmhdev.com/text.asp?2020/25/1/11/281892
| Background of the Study|| |
Male reproductive dysfunction is becoming a public health concern and can affect the quality of life. Available literature suggests an increase in the prevalence of male reproductive dysfunction.,, Rosen documented a 31% prevalence rate. Male infertility is on the rise and has been attributed to erectile dysfunction and testicular defect. Exposure to antiandrogen has been reported to induce testicular defects., The effects of antiandrogen are peculiar to the type of the antiandrogen.
Flutamide, a nonsteroidal antiandrogen, has been known to impair male reproductive function.,, It inhibits the uptake of androgen and the nuclear binding of androgen on target tissues. Elevated testosterone levels, loss of germ and Sertoli cells More Details, hypertrophy of interstitial cells, and low sperm count have been observed in experimental animals following exposure to flutamide.,,,
Plant materials have long been used in traditional medicine as an aphrodisiac and for the treatment of male reproductive dysfunction.,, The screening of plant materials used in phytotherapy for their medicinal properties is on the rise. One of such plants is Boswellia dalzielii, a tree of the species Boswellia, of the family Burseraceae. It is native to West Africa, the Savannah Region in particular.,
The bark-decoction of B. dalzielii is used in traditional medicine in Northern Nigeria for its aphrodisiac and fertility potentials. Our previous report also suggests that aqueous extract of the bark of B. dalzielii prophylaxes the toxic effect of flutamide on the testis., The biologically active compounds in the stem bark of B. dalzielii include flavonoids, tannins, saponins, alkaloids, anthracene, and cholesterol.,,, Antioxidant assays show presence of protocatechuic and gallic acids and also 4′-methoxy-(E)-resveratrol 3-O-rutinoside, incensole, and β-sitosterol., The aim of this study was to evaluate the curative effect of aqueous extract of the bark of B. dalzielii on flutamide-induced testicular toxicity in Wistar rats.
| Materials and Methods|| |
Collection of plant
The stem bark of B. dalzielii was harvested by a local phytotherapist in Kano State, Northwestern Nigeria. It was authenticated at the Department of Botany, University of Nigeria, Nsukka (voucher number UNH252).
Preparation of plant extract
The stem bark of B. dalzielii was dried at room temperature and ground to powder using a mortar and pestle. The powdered sample was sieved using a 2-mm mesh to remove residue. The dried powdered sample (100g) was dissolved in distilled water (200mL) and allowed to soak for 24h. The solution was filtered through a Whatman filter (125mm) paper no. 42. The filtrate was evaporated to dryness and stored in a refrigerator at 4°C.
Sixteen male Wistar rats (10 weeks old) were purchased from the animal house of the Department of Pharmacology and Toxicology, Faculty of Pharmaceutical Sciences, University of Nigeria, Nsukka. The experimental animals were housed in netted iron cages in group of four, fed with grower’s mash, and given water ad libitum. The animals were allowed to acclimatize for 2 weeks before the experiment under standard environmental conditions.
The experimental animals were randomly divided into four groups of four rats each. The period of administration was 21 days. Group A (negative control) received 5mL of normal saline only for 21 days. Group B (positive control) received 10 mg/kg body weight of flutamide only per day for the 21-day period. Group C (curative Group I) was given 10 mg/kg body weight of flutamide per day for 7 days and discontinued, they were then treated with 100 mg/kg body weight of aqueous extract of B. dalzielii from days 8 to 21. Group D (curative Group II) was given 10 mg/kg body weight of flutamide per day for 7 days and discontinued, they were treated with 300 mg/kg body weight of aqueous extract of B. dalzielii from days 8 to 21.
The weight of the animals was taken and recorded on days 1, 7, 14, and 21.
On day 22, the experimental animals were anesthetized with 25% urethane at a dose of 0.6mL/100g. The blood was collected for hormonal assay via left ventricular cardiac puncture. The testes were rapidly dissected for processing and light microscopic study. The weight of the testes was taken and recorded.
The collected blood was kept in non-heparin vacuum container and was span at 2500rpm in a bio-centrifuge (MSE, 0-5122A, East Sussex, UK) for 10 min. ECOBAS-6000 hormone-analyzing machine was used to analyze serum testosterone, leutinizing hormone (LH), and follicle-stimulating hormone (FSH) levels.
Light microscopic study
Modified Davidson’s fluid was used in fixing the testes for 24h., Tissue processing for microscopic examination was carried out following standard protocol. Three micrometer sectioned paraffin-embedded tissue was stained with hematoxylin and eosin.
Result was reported as mean ± standard deviation. Data were analyzed by using Statistical Package for the Social Sciences (SPSS) software program, version 23.0 Version 23.0., IBM Corp, Armonk, NY. Tests conducted include one-way analysis of variance (ANOVA) and Student t test. A value of P < 0.05 was considered statistically significant.
| Results|| |
A dose-dependent increase was observed in body weight in the curative groups (C and D). The increase in body weight [Table 1] was significantly higher (P < 0.05) in the curative groups (C and D) as compared to the controls (A and B).
Weight of testis
The weight of the testes was comparable in both the treatment and control groups [Table 2].
Serum testosterone levels [Table 3] were higher in the curative groups (C and D) as compared to the positive controls (Group A). The negative control (Group B) showed significantly high (P < 0.05) testosterone level as compared to all other groups. The serum FSH and LH levels were low in the curative groups as compared to the controls.
|Table 3: The levels of testosterone, FSH, and LH of the experimental models|
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Light microscopic study
An apparently normal testicular architecture and cellular composition was observed in the positive control [Figure 1]. Loss of germ and Sertoli cells, reduction of spermatozoa within the lumens of the seminiferous tubule, and hypertrophy of interstitial cells of Leydig were evident in the negative control [Figure 2]. The curative groups (C and D) showed increased cellular proliferation [Figure 3] and [Figure 4].
|Figure 1: An apparently normal testicular architecture and cellular composition observed in the negative control (arrows)|
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|Figure 2: Loss of germ and Sertoli cells, reduction of spermatozoa within the lumens of the seminiferous tubule, and hypertrophy of interstitial cells of Leydig as evident in the negative control (arrows)|
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|Figure 3: The low-dose curative group (C) showing increased cellular proliferation|
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|Figure 4: The high-dose curative group (D) showing increased cellular proliferation|
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| Discussion|| |
This study evaluated the curative effect of aqueous extract of the bark of B. dalzielii on flutamide-induced testicular toxicity in Wistar rats. Our results showed high serum testosterone level, loss of germ and Sertoli cells, reduced number of spermatozoa within the lumen of the seminiferous tubule, and hypertrophy of interstitial cells of Leydig following flutamide administration. This is in agreement with our previous report and the findings of a study by Oremosu et al. and Hassan et al. Flutamide administration may have altered Sertoli function and adversely affected the interstitial cells of Leydig and hence the disruption in hypothalamo-pituitary-testicular activities.,,
Administration of aqueous extract of the bark of B. dalzielii on flutamide-induced testicular reprotoxicity resulted in a dose-dependent increase in body weight and raised serum testosterone level. Testosterone is an anabolic steroid, which enhances metabolism, increases tissue and muscle mass, and consequently increases body weight.,, Testosterone exerts an androgenic effect by modifying deoxyribonucleic acid transcription and protein synthesis via its activities with androgen receptors in its unchanged form or as 5α-dihydrotestosterone. Testicular sections show increased cellular proliferation, size of seminiferous tubules, and interstitial cells of Leydig. Increased serum testosterone level has been implicated in morphological changes in testicular histoarchitecture.,,
The findings of this study suggest that aqueous extract of the bark of B. dalzielii exerted therapeutic effect on flutamide-induced testicular toxicity. This may be as a result of the biologically active compounds in the stem bark of B. dalzielii, which include flavonoids, tannins, saponins, alkaloids, anthracene, and cholesterol.,,, Flavonoid and saponin have been documented to have antioxidant activities., The antioxidant activities of flavonoids revive and fortify the reproductive system., Saponins have been reported to enhance spermatogenic stem cell and Leydig cell survival against reprotoxic agents.,
Protocatechuic acid has been isolated from the aqueous extract of the stem bark of B. dalzielii., It possess free radical-scavenging capacity and has therapeutic effect against oxidative damage. Oxidative stress is a major factor in the etiology of male infertility. Protocatechuic acid content of the stem bark of B. dalzielii may have counteracted oxidative damage in the testes from flutamide toxicity and consequently exerting a therapeutic effect.
| Conclusion|| |
The result of this study suggests that the aqueous extract of the bark of B. dalzielii has a therapeutic effect on flutamide-induced testicular toxicity.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Figure 1], [Figure 2], [Figure 3], [Figure 4]
[Table 1], [Table 2], [Table 3]