Endocrine Abstracts

Introduction: Atrazine is one of the most widely used pesticides worldwide and a common contaminant in human drinking water. Atrazine is also a potent endocrine disruptor capable of causing a variety of negative health outcomes, including decreased testosterone production and infertility. Although harmful effects of atrazine on male reproductive health have been reported, little is known about its molecular mechanisms. Within the seminiferous tubules, Sertoli cells are responsible for ensuring a suitable environment for spermatogenesis, providing mechanical and nutritional support to germ cells. However, the cytotoxic and metabolic effects of atrazine on Sertoli cells remain unknown.

Aim of this study: In this work, we aimed to elucidate the effects of the endocrine disruptor atrazine on the nutritional support of spermatogenesis by studying its effect on the metabolic profile and mitochondrial function of Sertoli cells.

Materials and methods: Mouse Sertoli cells (TM4 cell line, n=10) were exposed to biologically relevant concentrations of atrazine (in µg/l: 0.3, 3, 30, 300 and 3000). After 24 h, cytotoxicity was accessed. The mitochondrial activity and total ROS production were accessed by JC-1 dye and CM-H₂DCFDA probe, respectively. FRAP assay was used to measure the antioxidant potential of culture media. Lactate dehydrogenase (LDH) protein levels were analyzed by Western Blot and the glycolytic function was evaluated by Seahorse XF Glycolysis Stress Test Kit.

Results: Despite no cytotoxicity was observed, our results show a decreased metabolic activity when cells were exposed to 300 µg/l and 3000 µg/l of atrazine for 24 h. Additionally, a dose-dependent decrease in the expression of LDH was observed after exposure. Although the mitochondrial function of Sertoli cells was not affected by atrazine, we observed a tendency for increased endogenous ROS production in the highest atrazine concentrations (300 µg/l and 3000 µg/l) associated with a decreasing trend in the antioxidant potential of the culture media in the same concentrations, suggesting a pro-oxidant status.

Conclusions: Our data suggest that atrazine interferes with the glycolytic metabolism of mouse Sertoli cells, by reducing oxidoreductase activity and LDH expression. Since lactate is the preferred energetic source for developing germ cells, the nutritional support of spermatogenesis may be compromised. In addition, our results suggest that atrazine may induce a prooxidant status, leading to increased ROS production and oxidative stress. Overall, the endocrine disruptor atrazine induces a pro-oxidant state and modulates Sertoli cells metabolic support of spermatogenesis which can compromise it and lead to male infertility.