Effects of imidacloprid on progesterone synthesis in rat granulosa cells
Objective:To explore the effect of imidacloprid(IMI)on progesterone synthesis in rat ovarian granulosa cells.Methods:Rat granulosa cells were treated with different doses of IMI(0,100,200,500,1 000 μmol/L)for 48 h,and the levels of progesterone and estradiol in granulosa cells and their culture supernatant were measured by electrochemiluminescence immunoassay.The expression of key enzyme related with progesterone synthesis,namely steroidogenic acute regulatory protein(StAR)in primary cultures of rat granulosa cells was detected by Western Blot.The PI3K/AKT pathway inhibitor LY294002 was co-treated with 1 000 μmol/L IMI in rat granulosa cells for 48 h,and cell culture supernatant was collected to detect progesterone levels.Results:When rat granulosa cells were treated with IMI for 48 h,the progesterone level in the culture supernatant of granulosa cells showed concentration-dependent increase in all groups except the control group(P＜0.05),but the estradiol levels did not significantly change with the increase of IMI dose(P＞0.05).Western Blot analysis showed that the protein expression of St AR was significantly increased after the treatment with different doses of IMI for 48 h compared with the control group(P＜0.01).The PI3K/AKT pathway inhibitor LY294002 co-treated with 1 000 µmol/L IMI in rat granulosa cells for 48 h significantly reduced basal progesterone levels in granulosa cells compared with the control group(P＜0.01).Additionally,LY294002 co-treated with IMI significantly inhibited IMI-stimulated progesterone synthesis(P＜0.001).Conclusions:IMI affects the progesterone synthesis in primary cultures of rat granulosa cells,and the mechanism may be related to the activation of the PI3K/AKT signaling pathway.
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维普
