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The effect of increasing doses of Methotrexate (Folate Antagonist) on sperm production in male Mouse

تأثير جرعات متزايدة من الميثوتركسات (معاكس حمض الفوليك) في إنتاج النطاف عند ذكور الفأر

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 Publication date 2005
  fields Biology
and research's language is العربية
 Created by Shamra Editor




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Increasing intramuscular doses of Methotrexate (25 mg / kg, 50 mg / kg, 100 mg / kg, 150 mg/ kg, 200 mg / kg, 250 mg / kg, 300 mg / kg) were tested on Swiss strain Mouse by intramuscular injection. The reproductive potency was evaluated by total sperm count and movement of the left testis and it's epididymis (15) days after injection. The weights of the latter of all tested animals were registered. The result indicate that Methotrexate, a Folate antagonist, is a negative factor for sperm production in a dose related manner. It was evident that the high doses (ex. Dose 200 mg / kg) lead to oligospermia. It was concluded that Methotrexate inhibits the reproductive potency during spermatogenesis and causes cytotoxicity in high doses. Further investigations are needed.

References used
Herbert, V: Development of Human Folate Deficiency. In: Picciano, M. F., Stokstool, E. L. R., Georgy, J. F., eds: Folic acid metabolism in health and disease (contemporary issue in Clinical Nutrition). Vol.13, New York: 1990, 13: 195 – 210
Lindenbaum, J., Allen, R. H: Clinical Spectrum and Diagnosis of Folate Deficiency in: Bailey, L. B., ed. Folate in health and disease. New York, NY: Marcel Dekker, inc, 1995: 43 – 73
Herbert, V: Biochemical and hematological lesions in Folic acid deficiency. Am. J. Clin. Nutr. 20: 562 – 569, 1967

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Intramuscular injection of increasing doses of Methotrexate Folateantagoniste (25, 50, 100, 150, 250, 300 mg/ kg) on Swiss strain males mice , resulted in decreasing Spermatogenesis at stage VII whereas higher doses had lethal effects. In fact do ses of 25 and 50 mg per kg were tolerable by mice and the cytotoxic tolerance level decreased at higher doses (100 and 150 mg/kg). Acute cytotoxic effect was noted at doses of 200 mg/kg and resulted in destruction of cell line. Doses of 250 and 300 mg/kg had lethal effect and the mice died 1 week after injection. In conclusion Methotrexate had cytotoxic effects on sperm cell lines. 100mg/kg doses decreased spermatogenesis while 200 mg/kg doses caused greater side effects and higher doses were lethal to the mice 1 week after injection.
Mice embryos were irradiated in utero by gamma rays (٠،٢،٤،٦ Gy) at ١٠،١٢،١٤،١٦،١٨ days of gestation. Histological study was carried out on the first premandibular molar after ٢،٤،٦،٨ days of irradiation, to investigate the effect of irradiation on different stages of molar development.
Increasing dose of Mithotrexate (25, 50, 100, 150, 200 mg/kg) were tested on Swiss strain mouse by one intramuscular injection, and their effects were studied (15) days later. The mean values of total spermatogonia and those of them undergoing mito tic division (in the metaphase) were evaluated in seminiferous tubular cross section, and the mitotic index (MI) of the spermatogonia was also calculated. Statistical analysis was performed by using IBM-Spss 19 program and for mean values comparison (one way ANOVA: post Hos Test, Multiple Comparison, and Tukey HSD) was used, while mitotic index was calculated by using Target Variable. Our results showed that Methotrexate injection didn’t lead to a significant decrease in spermatogonia mean values in the tested animals compared to the control, while it led to a dose dependent gradual decrease in the mean values of the spermatogonia undergoing mitotic division , and the greatest reduction was noticed in the two high doses: 150, 200, mg/kg (7.10±9.4, 3.65±2.15 respectively) compared to the control (37.40±2.26), and which led also to a severe reduction in the mitotic index (20.53%, 6.36% in the doses: 100, 200 mg/kg respectively) compared to the control (61.79%).
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