吡虫啉对大鼠卵巢颗粒细胞孕酮合成的影响  

Effects of imidacloprid on progesterone synthesis in rat granulosa cells

作　　者：高二二 骆海燕 淡清华 任启语 高小博 陆彩玲 GAO Er-er;LUO Hai-yan;DAN Qing-hua;REN Qi-yu;GAO Xiao-bo;LU Cai-ling(Department of Genetics,National Research Institute for Family Planning,Beijing 100081;Graduate School of Peking Union Medical College,Beijing 100730)

机构地区：[1]国家卫生健康委科学技术研究所遗传优生中心,北京100081 [2]北京协和医学院研究生院,北京100730

出　　处：《生殖医学杂志》2024年第5期641-646,共6页Journal of Reproductive Medicine

基　　金：国家自然科学基金项目(82070327);中央级公益性科研院所基本科研业务项目(2023GJZD01)。

摘　　要：目的探究新烟碱类杀虫剂吡虫啉(IMI)对大鼠卵巢颗粒细胞孕酮合成的影响。方法使用不同浓度(0、100、200、500、1000μmol/L)的IMI分别处理大鼠卵巢颗粒细胞48 h,收集细胞及细胞培养液上清,电化学发光法检测孕酮和雌二醇(E_(2))水平;Western Blot法检测IMI处理后颗粒细胞孕酮合成关键酶甾体激素合成急性调节蛋白(StAR)的相对表达量;使用PI3K/AKT通路抑制剂LY294002与1000μmol/L IMI共处理颗粒细胞48 h后,收集细胞培养液上清检测孕酮水平。结果IMI处理大鼠卵巢颗粒细胞48 h,与对照组相比,随着IMI浓度的增加,细胞培养上清液中孕酮水平呈剂量依赖性增加(P<0.05),E_(2)水平无显著变化(P>0.05);Western Blot结果显示,与对照组相比,不同浓度的IMI处理颗粒细胞48 h后,StAR蛋白的相对表达量均显著升高(P<0.01)。PI3K/AKT通路抑制剂LY294002与1000μmol/L的IMI共处理颗粒细胞48 h后,与对照组相比,LY294002显著降低颗粒细胞基础孕酮的合成水平(P<0.01);与IMI共处理能显著抑制IMI刺激孕酮合成的效果(P<0.001)。结论IMI影响大鼠卵巢颗粒细胞孕酮的合成,其机制可能与PI3K/AKT信号通路的激活有关。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(0,100,200,500,1000μmol/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 1000μmol/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 StAR 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 1000μmol/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.

关 键 词：吡虫啉 颗粒细胞 孕酮 PI3K/AKT 

分 类 号：R994.6[医药卫生—毒理学]