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Table of Contents
ORIGINAL ARTICLES
Year : 2020  |  Volume : 25  |  Issue : 1  |  Page : 11-15

Curative effect of aqueous extract of the bark of Boswellia dalzielii on flutamide-induced testicular toxicity in Wistar rat


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 Submission15-Aug-2019
Date of Decision12-Jan-2020
Date of Acceptance19-Feb-2020
Date of Web Publication03-Apr-2020

Correspondence Address:
Dr. Nto J Nto
Department of Anatomy, Faculty of Basic Medical Sciences, College of Medicine, University of Nigeria Enugu Campus, Enugu.
Nigeria
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ijmh.IJMH_22_19

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  Abstract 

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 Top


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.[1],[2],[3] Rosen[4] documented a 31% prevalence rate. Male infertility is on the rise and has been attributed to erectile dysfunction and testicular defect.[4] Exposure to antiandrogen has been reported to induce testicular defects.[5],[6] The effects of antiandrogen are peculiar to the type of the antiandrogen.

Flutamide, a nonsteroidal antiandrogen, has been known to impair male reproductive function.[5],[6],[7] It inhibits the uptake of androgen and the nuclear binding of androgen on target tissues.[8] 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.[5],[6],[7],[9]

Plant materials have long been used in traditional medicine as an aphrodisiac and for the treatment of male reproductive dysfunction.[6],[7],[9] The screening of plant materials used in phytotherapy for their medicinal properties is on the rise.[6] 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.[6],[9]

The bark-decoction of B. dalzielii is used in traditional medicine in Northern Nigeria for its aphrodisiac and fertility potentials.[9] Our previous report also suggests that aqueous extract of the bark of B. dalzielii prophylaxes the toxic effect of flutamide on the testis.[5],[6] The biologically active compounds in the stem bark of B. dalzielii include flavonoids, tannins, saponins, alkaloids, anthracene, and cholesterol.[6],[9],[10],[11] Antioxidant assays show presence of protocatechuic and gallic acids and also 4′-methoxy-(E)-resveratrol 3-O-rutinoside, incensole, and β-sitosterol.[6],[12] 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 Top


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.[11]

Experimental animals

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.

Experimental design

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.

Body weight

The weight of the animals was taken and recorded on days 1, 7, 14, and 21.

Sample collection

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.

Hormonal assay

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.[13]

Light microscopic study

Modified Davidson’s fluid was used in fixing the testes for 24h.[6],[9] Tissue processing for microscopic examination was carried out following standard protocol.[5] Three micrometer sectioned paraffin-embedded tissue was stained with hematoxylin and eosin.

Statistical analysis

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 Top


Body weight

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).
Table 1: Body weight of experimental models

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Weight of testis

The weight of the testes was comparable in both the treatment and control groups [Table 2].
Table 2: Weight of the testis of experimental models

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Hormonal assay

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 Top


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[6] and the findings of a study by Oremosu et al.[5] and Hassan et al.[7] Flutamide administration may have altered Sertoli function and adversely affected the interstitial cells of Leydig and hence the disruption in hypothalamo-pituitary-testicular activities.[5],[6],[7]

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.[7],[14],[15] 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.[16] 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.[5],[6],[7]

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.[6],[9],[10],[11] Flavonoid and saponin have been documented to have antioxidant activities.[10],[11] The antioxidant activities of flavonoids revive and fortify the reproductive system.[10],[11] Saponins have been reported to enhance spermatogenic stem cell and Leydig cell survival against reprotoxic agents.[17],[18]

Protocatechuic acid has been isolated from the aqueous extract of the stem bark of B. dalzielii.[12],[19] It possess free radical-scavenging capacity and has therapeutic effect against oxidative damage.[12] Oxidative stress is a major factor in the etiology of male infertility.[20] 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 Top


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

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

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Nayernia K, Nolte J, Michelmann HW, Lee JH, Rathsack K, Drusenheimer N. In vitro differentiated embryonic stem cells give rise to males gametes. Dev Cell 2006;11:125-32.  Back to cited text no. 1
    
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World Health Organization.Laboratory manual for examination of human semen. 5th ed. WHO Press, World Health Organization, Geneva, Switzerland.2010; pp 4-197.  Back to cited text no. 2
    
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NIH consensus conference on impotence. JAMA 1993;270:83-90.  Back to cited text no. 3
    
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Rosen RC. Prevalence and risk factors of sexual dysfunction in men and women. Curr Psychiatry Rep 2000;2:189-95.  Back to cited text no. 4
    
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Oremosu AA, Arowosaye VO, Akang EN, Bassey RB. Effect of Cissus populneal and Panax ginseng on flutamide-induced testicular defect in pre-pubertal male rats. Br J Med Med Res 2013;3:173-81.  Back to cited text no. 5
    
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Akpobasaha S, Ezugworie JO, Nto NJ. Protective effect of aqueous extract of the bark of Boswellia dalzielii on flutamide-induced testicular reprotoxicity in Wistar rat.Acta Sci Med Sci 2019;3:114-18.  Back to cited text no. 6
    
7.
Hassan LA, Anyanwu GE, Nto NJ, Abireh IE, Akunna GG. Protective effect of aqueous extract of Cyperus esculentuson the flutamide induced testicular defect of male Wistar rats. SJAMS 2018;6:2391-5.  Back to cited text no. 7
    
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Elks J, Ganellin CR. The dictionary of drugs: Chemical data: chemical data, structures and bibliographies. Pennsylvania, PA: Springer; 2014. pp. 573.  Back to cited text no. 8
    
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Ezugworie JO, Akpobasaha S, Nto NJ, Finbarrs-Bello E, Agu AU, Esom EA. The histological and hormonal effect of aqueous extract of Boswellia dalzielii on the testis of male Wistar rat. Int J Pure App Biosci 2018;6:31-6.  Back to cited text no. 9
    
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Adoum AO. Determination of toxicity effects of some savannah plants using brine shrimp test (BST). Int J P App Sci 2008;2:1-5.  Back to cited text no. 10
    
11.
Abdulazeez M, Hassan HS, Usman MA. Preliminary phytochemical and antispasmodic studies of the stem bark of Boswellia dalzielii. Niger J Pharm Sci 2009;8:1-6.  Back to cited text no. 11
    
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Liu CL, Wang JM, Chu CY, Cheng MT, Tseng TH. In vivo protective effect of protocatechuic acid on tert-butyl hydroperoxide-induced rat hepatotoxicity. Food Chem Toxicol 2002;40:635-41.  Back to cited text no. 12
    
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Atlas MR, Alfred E, Lawrence C. Laboratory manual experimental microbiology. St. Louis, MO: Mosby; 1995.  Back to cited text no. 13
    
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Powers M. Performance-enhancing drugs. In: Houglum J, Harrelson GL. editors. Principles of pharmacology for athletic trainers. 2nd ed. SLACK Incorporated, Thorofare, NJ, United States. 2015;345.  Back to cited text no. 14
    
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Atuadu VO, Anyanwu GE, Finbarrs-Bello E. Effect of ethanolic rind extract of Citrullus lanatus on the Hpg axis and sexual behavior of male Wistar rats. Int J Pure App Biosci 2018;6:1-7.  Back to cited text no. 15
    
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Ferrando AA, Sheffield-Moore M, Paddon-Jones D, Wolfe RR, Urban RJ. Differential anabolic effects of testosterone and amino acid feeding in older men. J Clin Endocrinol Metab 2003;88:358-62.  Back to cited text no. 16
    
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Ang HH, Lee KL, Kiyosh M. Sexual arousal in sexually sluggish old male rats after oral administration of Eurycoma longifolia. J Basic Clin Physio Pharmacol 1997;15:3-40.  Back to cited text no. 17
    
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Minyoung JI, Naojiro M, Masayasu Y, Hiroshi I. Effect of protopanaxatriol saponin on spermatogenic stem cell survival in bulsulfan-treated male mice. Reprod Med Biol 2007;6:99-108.  Back to cited text no. 18
    
19.
Alemika TE, Onawunmi GO, Olugbade TO. Antibacterial phenolics from Boswellia dalzielii. Niger J Nat Prod Med 2006;10:108-10.  Back to cited text no. 19
    
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Aitken RJ, Roman SD. Antioxidant systems and oxidative stress in the testes. Oxid Med Cell Longev 2008;1:15-24.  Back to cited text no. 20
    


    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4]
 
 
    Tables

  [Table 1], [Table 2], [Table 3]



 

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