Assessment of Reproductive Toxicity of Hydroethanolic Root Extracts of Caesalpinia benthamiana, Sphenocentrum jollyanum, and Paullinia pinnata

Abstract

Purpose: Male sexual dysfunction negatively affects an individual's quality of life and thus its of prime public concern, hence the need to boost reproductive abilities in such individuals. This study assessed the effect of hydroethanolic root extracts of Caesalpinia benthamiana (CBRE), Sphenocentrum jollyanum (SJRE), and Paullinia pinnata (PPRE), commonly used as aphrodisiacs in Ghana, using male Sprague-Dawley rats.

Methods: Plasma testosterone, follicle-stimulating hormone, and luteinizing hormone were assayed in grouped rats treated orally with 1 mL/kg normal saline, 50 mg/kg monosodium glutamate (MSG), and 100, 300, or 1000 mg/kg CBRE, SJRE, and PPRE, respectively, for 60 days. Epididymis and testis weights were determined. Semen was assessed on spermatozoa count, motility, and morphology. Malonyladehyde formation in lipid-peroxidation assay and histological examinations were performed to assess pathological changes in testes. Testicular testosterone was also assayed.

Results: While MSG, CBRE, SJRE, and PPRE treatments did not result in significant reduction (p>0.05) in plasma testosterone, there was significant reduction (p≤0.05 -0.0001) in plasma luteinizing hormone, and follicle-stimulating hormone. The combined mean wet weights of epididymides and testes of all treated groups did not vary significantly (p>0.05) from the control. There was significant reduction (p≤0.0001) in sperm motility and count, with significant morphological changes (p≤0.05-0.001), ie, bent necks, tails, and midpieces, and multiple anomalies in the spermatozoa in extract and MSG-treated groups. There was also significant (p≤0.0001) reduction in testicular testosterone among all treatment groups.

Conclusion: Hydroethanolic CBRE, SJRE, and PPRE were found to have detrimental effects on reproductive function with prolonged usage and thus may not be safe to use in healthy males who intend to reproduce.

Keywords: herbal aphrodisiacs; monosodium glutamate; plasma follicle-stimulating hormone; plasma luteinizing hormone; sperm anomalies; sperm motility; testicular testosterone.

Figure 1

Plasma testosterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) concentrations in rats treated with normal saline (NS), monosodium glutamate (MSG), and hydroethanolic root-bark extracts of Caesalpinia benthamiana (CBRE), Sphenocentrum jollyanum (SJRE), and Paullinia pinnata (PPRE) for 60 days. Data presented as group means ± SEM, n=7. Significant differences between treatment and control: ns, p>0.05; *p≤0.05; **p≤0.01; ***p≤0.001; ****p≤0.0001 (one-way ANOVA followed by Dunnett’s multiple-comparison test).

Figure 2

Sperm counts of rats treated with normal saline (NS), monosodium glutamate (MSG), and various doses of hydroethanolic root-bark extracts of Caesalpinia benthamiana (CBRE), Sphenocentrum jollyanum (SJRE), and Paullinia pinnata (PPRE) for 60 days. Data presented as means ± SEM, n=7. Significant differences between treatment and control: *p≤0.05; ***p≤0.001; ****p≤0.0001 (one-way ANOVA followed by Dunnett’s multiple-comparison test).

Figure 3

Percentage sperm motility of rats treated with normal saline (NS), monosodium glutamate (MSG), and various doses of hydroethanolic root-bark extracts of Caesalpinia benthamiana (CBRE), Sphenocentrum jollyanum (SJRE), and Paullinia pinnata (PPRE) for 60 days. Data presented as means ± SEM, n=7. Significant differences between treatment and control: *p≤0.05; ***p≤0.001; ****p≤0.0001 (one-way ANOVA followed by Dunnett’s multiple-comparison test).

Figure 4

Sperm morphology at the end of 60 days of treatment with normal saline (NS), monosodium glutamate (MSG), and various concentrations of the hydroethanolic root-bark extracts of Caesalpinia benthamiana (CBRE), Sphenocentrum jollyanum (SJRE), and Paullinia pinnata (PPRE). Values are means ± SEM, n=7. ns, p>0.05; *p≤0.05; **p≤0.01, ***p≤0.001; ****p≤0.0001 (one-way ANOVA followed by Dunnett’s multiple-comparison test).

Figure 5

Malondialdehyde concentration indicative of lipid peroxidation of testes after treatment with normal saline (NS), monosodium glutamate (MSG), and various concentrations of hydroethanolic root-bark extracts of Caesalpinia benthamiana (CBRE), Sphenocentrum jollyanum (SJRE), and Paullinia pinnata (PPRE) for 60 days. Data presented as group means ± SEM, n=7. Significant differences between treatments and control: ns, p>0.05; *p≤0.05, ***p≤0.001 (one-way ANOVA using Dunnett’s multiple-comparison test).

Figure 6

Testicular testosterone concentration inf rats treated with normal saline (NS), monosodium glutamate (MSG), and various concentrations of hydroethanolic root-bark extracts of Caesalpinia benthamiana (CBRE), Sphenocentrum jollyanum (SJRE), and Paullinia pinnata (PPRE) for 60 days. Data presented as means ± SEM, n=7. Significant differences between treatments and control: ***p≤0.001; ****p≤0.0001 (one-way ANOVA followed by Dunnett’s multiple-comparison test).

Figure 7

Representative photomicrographs of histological assessment of testes of Sprague-Dawley rats after 60 days of treatment. (A) Normal saline–treated rats showing well-layered seminiferous tubules with different stages of spermatogenic cells; (B) 50 mg/kg monosodium glutamate (MSG)-treated rats showing severe atrophy in some seminiferous tubules and reduction in interstitial Leydig cells; (C) 1,000 mg/kg SJRE–treated rats showing mild atrophy of seminiferous tubules; (D) 1,000 mg/kg CBRE–treated rats showing severe atrophy in some seminiferous tubules; (E) 1,000 mg/kg PPRE–treated rats showing different degrees of atrophy (transverse section, 200×, H&E stain).

Figure 8

Representative photomicrographs of histological assessment of the epididymides of Sprague-Dawley rats after 60 days of treatment. (A) Normal saline–treated rats showing well-layered seminiferous tubules with different stages of spermatogenic cells; (B) MSG-treated rats showing necrosis in connective tissue; (C) 1,000 mg/kg SJRE–treated rats showing disorganization of cells and epididymitis; (D) 1,000 mg/kg CBRE–treated rats showing greatly convoluted tubules, but with very scanty spermatozoa; (E) 1,000 mg/kg PPRE–treated rats showing greatly convoluted tubules, but with scanty spermatozoa (transverse section, 200×, H&E stain).