Toxicological Research

Korean Society of Toxicology & Korea Environmental Mutagen Society (한국독성학회)
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Potential role of inducible nitric oxide synthase (iNOS) activity in testicular dysfunction following co-administration of alcohol and combination antiretroviral therapy (cART) in diabetic rats: an immunohistochemistry study

  • Elna Owembabazi (School of Anatomical Sciences, University of the Witwatersrand) ;
  • Pilani Nkomozepi (Department of Human Anatomy and Physiology, University of Johannesburg) ;
  • Ejikeme F. Mbajiorgu (School of Anatomical Sciences, University of the Witwatersrand)
  • Received : 2023.03.23
  • Accepted : 2023.06.18
  • Published : 2024.01.15
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Abstract

Diabetes, alcohol abuse, and combination antiretroviral therapy (cART) use have been reported to cause multi-organ complications via induction of oxidative stress and inflammation. Moreover, these are the most common factors implicated in male reproductive dysfunctions. This study evaluated testicular oxidative stress, inflammation, apoptosis, and germ cell proliferation in diabetic rats receiving alcohol or cART and their combination. Thirty adult male Sprague Dawley rats were divided into five groups, each consisting of six rats; control, diabetic only (DM), diabetic treated with alcohol (DM+A), diabetic treated with cART (DM+cART), and diabetic treated with both alcohol and cART (DM+A+cART). After 90 days of treatment, the rats were terminated, and the testes were extracted and processed for immunohistochemistry analysis for oxidative stress, inflammatory cytokines, apoptosis, and cell proliferation marker. In comparison to the control, oxidative stress markers, inducible nitric oxide synthase (iNOS), malondialdehyde (MDA), and 8-hydroxydeoxyguanosine (8-OHDG) increased significantly in all treated groups. Expression of testicular proinflammatory cytokines, interleukin-1β, and tumor necrosis factor-α was upregulated in all treated groups, but interleukin-6 was upregulated in DM, DM +cART, and DM+A+cART treated groups and was downregulated in the DM+A treated group. All treated animal groups showed an upregulation of apoptotic marker (caspase 3) and a downregulation of proliferation marker (Ki-67). However, Ki-67 staining intensity significantly increased in treated animals compared to the control. These findings suggest that diabetes, alcohol abuse, cART use, and their combination via iNOS activity upregulation can induce inflammation and oxidative stress in testicular tissue, stimulating germ cell apoptosis and proliferation inhibition leading to failure of spermatogenesis.

Keywords

Acknowledgement

We appreciate the collaborative efforts of our colleagues Jaclyn Asouzu Johnson, Idemudia Eguavoen, and Vaughan Perry and extend special appreciation to Hasiena Ali for her laboratory assistance.

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