The present work describes the screening and characterization of some common endocrine disrupting chemicals for their (anti)androgenic activities. Various chemicals (mostly pesticides and pharmaceuticals) were screened with the NIH3T3 cell line stably expressing human androgen receptor (hAR) and luciferase reporter gene for their ability to stimulate luciferase activity or inhibit the response that was evoked by 0.4nM testosterone. The most potent anti-androgenic compounds identified in our assay included chlorpyrifos, endosulfan and piperophos. Finally, the chemicals were analyzed for their effects on steriodogenesis in rat Leydig cells. Piperophos and chlorpyrifos showed a significant decrease in testosterone biosynthesis by Leydig cells. RT-PCR studies showed decrease in the expression of key steroidogenic enzymes: cytochrome P450scc, 3beta-HSD and 17beta-HSD and immunoblot analysis demonstrated a decrease in steroidogenic acute regulatory (StAR) protein expression by both these chemicals. Chlorpyrifos also showed a decrease in LH receptor stimulated cAMP production. In conclusion, we demonstrate that commonly used pesticides like chlorpyrifos and piperophos pose serious threat to male reproductive system by interfering at various levels of androgen biosynthesis.
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