In Vitro, In Vivo, and In Silico Analysis of Pyraclostrobin and Cyprodinil and Their Mixture Reveal New Targets and Signaling Mechanisms

Pyraclostrobin
and cyprodinil are broad-spectrum fungicides that
are used in crops to control diseases. However, they are excessively
used and, as a result, end up in the environment and threaten human
health and ecosystems. Hence, knowledge of their mechanisms of action
is critical to revealing their environmental fate and negative effects
and regulating their use. In the present study, we conducted a comprehensive
study to show the adverse effects of pyraclostrobin, cyprodinil, and
their mixture using zebrafish larvae and different cell lines. Several
end points were investigated, including mortality, development, gene
expression, reporter assays, and molecular docking simulations. We
found that both compounds and their mixture caused developmental delays
and mortality in zebrafish, with a higher effect displayed by pyraclostrobin.
Both compounds altered the expression of genes involved in several
signaling pathways, including oxidative stress and mitochondrial function,
lipid and drug metabolisms, the cell cycle, DNA damage, apoptosis,
and inflammation. A noteworthy result of this study is that cyprodinil
and the mixture group acted as NFκB activators, while pyraclostrobin
demonstrated antagonist activity. The AHR activity was also upregulated
by cyprodinil and the mixture group; however, pyraclostrobin did not
show any effect. For the first time, we also demonstrated that pyraclostrobin
had androgen receptor antagonist activity.