Investigating penthiopyrad's potential toxicity on male reproductive health via in vitro models and computational network toxicology and docking studies

Penthiopyrad, a novel chiral member of succinate dehydrogenase inhibitors (SDHI), exhibits a broad-spectrum fungicidal activity in agriculture. However, information on effects of penthiopyrad on male reproductive health is limited. Therefore, the potential toxic effects and mechanisms of penthiopyrad were investigated in the present study using an in vitro model of Leydig (TM3) cells. The treatment concentrations were 5–25 μM because the median inhibitory concentration (IC₅₀) value of penthiopyrad was found to be 58 μM. According to the findings, it induced reactive oxygen species (ROS) production and DNA damage (≥6,48 folds) through loss of mitochondrial membrane potential (MMP). Apoptosis was induced but not autophagy and necrosis. Aiming to gain deeper insight into the mechanisms underlying the in vitro toxicological profile reported for penthiopyrad, a series of chemoinformatic studies -including a Network Toxicology (NT) approach and molecular docking analysis-were integrated. As a result, the correlation between 'male infertility' and 'mitochondrial dysfunction' -highlighted as key terms based on in vitro assays outcomes and penthiopyrad's known profile-was examined, revealing mitochondrial respiratory complexes I–V as potential key targets. Penthiopyrad demonstrated the ability to form favourable interactions with key amino acid residues that may play a critical role in impairing mitochondrial function by interfering with electron transport and ATP synthesis. The computational findings reinforce the reported in vitro assays, as oxidative stress, loss of MMP, and apoptosis as significant contributors to male reproductive toxicity as a non-target organism.