Effects Of Fenvalerate On Testosterone Concentration And Circadian Expression Of Related Genes In TM3 Cells

Objective: To explore the effects of fenvalerate on testosterone concentration and circadian expression of related genes in TM3 cells, to provide basic data for mechanistic studies.Method:（1）The mouse Leydig cell lines（TM3） was used to measure testosterone（T） levels by ELISA, the cell viability by CCK8,and the cell cycle by flow cytometry.（2） The quantitative PCR was used to detect gene expression.（3） Twenty four hours after fenvalerate treatment, cells were collected at different time points of a day（0:00, 4:00, 8:00,12:00, 16:00, 20:00, 24:00）, to detect circadian expression of genes by fluorescence quantitative PCR.（4） [Ca²⁺]i concentration was measured by flow cytometry.Results:（1） The doubling growth time of the mouse Leydig cell line（TM3） was 22.53 ± 2.84 h. Fenvalerate treatment significantly reduced testosterone concentration by 37% of the control.（2） In Leydig cells, the Bmal1 and StAR gene showed circadian expression, with peak and trough at 16:00 and 4:00 for Bmall, and 12:00 and 4:00 for StAR. Fenvalerate treatment up-regulated REV-erbα gene and down-regulated Bmal1, RORa and StAR gene.（3） Fenvalerate treatment resulted in an increase in [Ca²⁺]i concentration by 9.67%.Conclusion:1. Fenvalerate treatment reduced testosterone levels and changed circadian expressions of clock genes.2. The circadian effect of fenvalerate on testosterone levels might be due to an increase in [Ca²⁺]i concentration.