Proton Pump Dysfunction and Upregulation of Caspase-3 Activity via Oxidative-Sensitive Signaling Mediate Rohypnol-Induced Testicular Toxicity

Abstract

Rohypnol is a benzodiazepine that is used for its anxiolytic, anticonvulsant, muscle relaxant, and hypnotic properties, but commonly abused. It has been reported to induce sexual dysfunction; however, its effect on testicular integrity and sperm quality is yet to be documented. This study evaluated the impact of Rohypnol on testicular integrity, circulatory testosterone, spermatogenesis, and sperm quality. The likely role of oxidative stress-dependent pathway as well as testicular proton pumps and caspase-3 activity in Rohypnol-induced modulation of male reproductive function was explored. In this study, adult male rats were randomized into six groups of six rats each: the control received 1 ml of distilled water as vehicle, while the low- and high-dose Rohypnol-treated groups received 2 mg/kg and 4 mg/kg of Rohypnol for four weeks. The control-, low-, and high-dose recovery groups received 2 mg/kg and 4 mg/kg of Rohypnol for four weeks, respectively, which was followed by a 4-week drug-free recovery period. Rohypnol treatments led to impaired steroidogenesis and spermatogenesis and low sperm quality. This was accompanied by a rise in the markers of testicular injury, oxidative stress, inflammation, and apoptosis. Also, the activities of testicular enzymatic antioxidants and proton pumps were significantly reduced. Although the impacts of low-dose Rohypnol treatment were partially reversible, those of the high-dose Rohypnol treatment were not. Histopathological findings on the testicular tissues were in tandem with the biochemical alterations. Thus, it is safe to infer that Rohypnol induces testicular damage by the suppression of Na+/K+-ATPase and Ca2+-ATPase and the activation of caspase-3 through an oxidative-sensitive signaling pathway.