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Methotrexate induces germ cell apoptosis and impairs spermatogenesis in a rat

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Abstract

Purpose

The primary toxic effects of methotrexate (MTX) are myelosuppression and/or intestinal mucositis. The objective of the present study is to investigate the effect of MTX on germ cell apoptosis and spermatogenesis in a rat.

Methods

Male Sprague–Dawley rats were divided into three experimental groups: control rats treated with vehicle; MTX-2 rats treated with one dose (20 μg/kg) of MTX given IP and killed on the second day; and MTX rats treated with IP MTX (20 μg/kg) and killed on day 4. Johnsen’s criteria and the number of germinal cell layers in the testes were used to categorize the spermatogenesis. TUNEL assay was used to determine germ cell apoptosis. Western blotting was used to determine Bax and Bcl-2 protein levels. Statistical analysis was performed using the non-parametric Kruskal–Wallis ANOVA test, with p less than 0.05 considered statistically significant.

Results

On day 2, MTX-treated animals demonstrated minimal changes in the histological parameters of spermatogenesis, but germ cell apoptosis increased significantly (threefold increase, p = 0.002) compared to control rats. On day 4, MTX-treated rats demonstrated a trend toward a decrease in germ cell apoptosis, compared to day 2, and showed histological signs of impaired spermatogenesis (decreased number of germ cell layers and Johnsen’s criteria). A significant increase in cell apoptosis in MTX-treated rats was correlated with higher Bax/Bcl-2 protein levels.

Conclusions

MTX induced germ cell apoptosis and impaired spermatogenesis in rat testes.

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Acknowledgments

This work was supported by a research grant from Israeli Ministry of Health No. 3/6164 from 01/06/2010.

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Authors and Affiliations

  1. The Bruce Rappaport Faculty of Medicine, Laboratory of Intestinal Adaptation and Recovery, Technion-Israel Institute of Technology, Haifa, Israel

    Igor Sukhotnik, Omri Nativ, Alex Roitburt & Daniel Bejar

  2. Department of Pediatric Surgery, Bnai Zion Medical Center, 47 Golomb St, P.O. 4940, 31048, Haifa, Israel

    Igor Sukhotnik & Jorge G. Mogilner

  3. Department of Urology, Bnai Zion Medical Center, Haifa, Israel

    Ofer Nativ

  4. Section of Pediatric Surgery, C.S. Mott Children’s Hospital, University of Michigan Medical School, Ann Arbor, MI, USA

    Arnold G. Coran

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  1. Igor Sukhotnik
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  2. Omri Nativ
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  3. Alex Roitburt
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  4. Daniel Bejar
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  5. Arnold G. Coran
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  6. Jorge G. Mogilner
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  7. Ofer Nativ
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Correspondence to Igor Sukhotnik.

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Sukhotnik, I., Nativ, O., Roitburt, A. et al. Methotrexate induces germ cell apoptosis and impairs spermatogenesis in a rat. Pediatr Surg Int 29, 179–184 (2013). https://doi.org/10.1007/s00383-012-3197-0

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  • DOI: https://doi.org/10.1007/s00383-012-3197-0

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