Arsenite: Toxicity and genotoxicity in a mouse Leydig cell line

Arsenic is a prevalent environmental contaminant recognized for its detrimental effects on several biological systems, particularly the male reproductive system. Although genotoxic effects linked to arsenite (As) exposure have been shown in various cell and tissue types, research concerning the male reproductive system is limited. In this study, the cytotoxic, apoptotic, and genotoxic effects of As exposure were evaluated using in vitro assays in TM3 Leydig cells, cells that play a critical role in testicular function. Leydig cells were exposed to increasing As concentrations (1, 2, 4, and 6 µM) for 24 h. Cell viability, oxidative stress (ROS production, SOD, CAT, GPx, and GSH levels), apoptosis (propidium iodide/Hoechst double fluorescent staining), and genotoxicity (micronucleus, comet, and chromosomal aberration tests) were analyzed. The administration of As markedly diminished cell viability in a concentration-dependent manner and inhibited antioxidant enzyme activity, resulting in a redox imbalance. An increase in the rate of apoptotic cells was noted in parallel to the As concentration. Genotoxicity analyses revealed an elevation in DNA damage, chromosomal aberrations, and micronucleus formation. In conclusion, As induces multifaceted toxic effects in TM3 Leydig cells by disrupting the oxidative balance, triggering apoptosis, and damaging genome integrity. This is the first in vitro study demonstrating genotoxicity of As in TM3 cells and offers an important contribution to the literature on male reproductive toxicity.