Regucalcin counteracts tert-butyl hydroperoxide and cadmium-induced oxidative stress in rat testis

J Appl Toxicol. 2017 Feb;37(2):159-166. doi: 10.1002/jat.3333. Epub 2016 Apr 25.

Abstract

Regucalcin (RGN) is a calcium (Ca2+ )-binding protein with multiple physiological roles and has also been linked to the suppression of oxidative stress. It is widely known that oxidative stress adversely affects spermatogenesis, disrupting the development of germ cells, and interfering with sperm function. The present study aims to analyze the role of RGN modulating testicular oxidative stress. To address this issue, seminiferous tubules (SeT) from transgenic rats overexpressing RGN (Tg-RGN) and wild-type (WT) were cultured ex vivo for 24 h in the presence/absence of pro-oxidant stimuli, tert-butyl hydroperoxide (TBHP, 250 and 500 μM) and cadmium chloride (Cd, 10 and 20 μM). Noteworthy, SeT from Tg-RGN animals displayed a significantly higher antioxidant capacity and diminished levels of thiobarbituric acid reactive substances relatively to their WT counterparts, both in control and experimental conditions. Regarding the antioxidant defense systems, a significant increase in the activity of glutathione-S-transferase was found in the SeT of Tg-RGN whereas no differences were observed in superoxide dismutase activity throughout experimental conditions. The activity of apoptosis executioner caspase-3 was significantly increased in the SeT of WT rats treated with 250 μM of TBHP or 10 μM of Cd, an effect not seen in Tg-RGN animals. These results showed that the SeT of Tg-RGN animals displayed lower levels of oxidative stress and increased antioxidant defenses, exhibiting protection against oxidative damage and apoptosis. Moreover, the present findings support the antioxidant role of RGN in spermatogenesis, which may be an important issue of further research in the context of male infertility. Copyright © 2016 John Wiley & Sons, Ltd.

Keywords: TBARS; cadmium; glutathione-S-transferase; oxidative stress; regucalcin; reproductive toxicity; seminiferous tubules; spermatogenesis; superoxide dismutase; tert-butyl hydroperoxide.

MeSH terms

  • Animals
  • Antioxidants / metabolism
  • Cadmium / toxicity*
  • Calcium-Binding Proteins / genetics
  • Calcium-Binding Proteins / physiology*
  • Carboxylic Ester Hydrolases
  • In Vitro Techniques
  • Intracellular Signaling Peptides and Proteins / genetics
  • Intracellular Signaling Peptides and Proteins / physiology*
  • Male
  • Oxidative Stress / drug effects*
  • Rats, Sprague-Dawley
  • Rats, Transgenic
  • Seminiferous Tubules / drug effects
  • Seminiferous Tubules / metabolism
  • Testis / drug effects*
  • Testis / metabolism
  • tert-Butylhydroperoxide / toxicity*

Substances

  • Antioxidants
  • Calcium-Binding Proteins
  • Intracellular Signaling Peptides and Proteins
  • Cadmium
  • tert-Butylhydroperoxide
  • Carboxylic Ester Hydrolases
  • Rgn protein, rat