Institute of Endemic Diseases, Key Laboratory of Trace Elements and Endemic Diseases, National Health Commission of the People's Republic of China, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, 710061, People's Republic of China.
Department of Gynecology and Obstetrics, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, China.
Biol Trace Elem Res. 2021 Nov;199(11):4119-4128. doi: 10.1007/s12011-020-02524-x. Epub 2021 Jan 6.
Taking excessive sodium fluoride may cause female reproductive dysfunction, but underlying molecular mechanism is unclear. The ovarian granulosa cells are the key endocrine cells releasing reproductive hormones. The miRNAs in the granulosa cells play an important function in regulating reproduction. The aim of this study is to explore the role of miRNAs in granulosa cell apoptosis and autophagy, as well as estradiol (E) release in response to excessive sodium fluoride. The ovarian granulosa cells (KGN cells) were treated in vitro by different concentrations of sodium fluoride (NaF) for 24 h. The level of estradiol (E) in the incubation medium was measured by ELISA kits. The total RNA and protein were collected and purified from KGN cells. The expression of miRNAs was detected by the real-time PCR. The signal molecules involved in cell apoptosis and autophagy were detected by the real-time PCR and Western blotting. Six miRNAs in granulosa cells were significantly up- or downregulated by NaF and selected for real-time PCR analysis. The miR-378d was the most significantly upregulated one dose dependently by NaF. It was positively correlated to the extent of apoptosis but negatively correlated to the level of autophagy in KGN cells in response to NaF. In addition, miR-378d promoted E release in response to 1 and 2 mM NaF but reduced E release in response to 4 and 8 mM NaF treatments. It is concluded that expression of miR-378d in ovarian granulosa cells is negatively correlated to the autophagy and E release and positively correlated to cell apoptosis under the influence of NaF.
过量的氟化物可能导致女性生殖功能障碍，但潜在的分子机制尚不清楚。卵巢颗粒细胞是释放生殖激素的关键内分泌细胞。颗粒细胞中的 miRNAs 在调节生殖中起着重要作用。本研究旨在探讨 miRNAs 在颗粒细胞凋亡和自噬以及雌二醇（E）释放对过量氟化物反应中的作用。体外用不同浓度的氟化钠（NaF）处理卵巢颗粒细胞（KGN 细胞）24 小时。用 ELISA 试剂盒测量孵育培养基中雌二醇（E）的水平。从 KGN 细胞中收集和纯化总 RNA 和蛋白质。通过实时 PCR 检测 miRNAs 的表达。通过实时 PCR 和 Western blotting 检测参与细胞凋亡和自噬的信号分子。NaF 使 6 种颗粒细胞中的 miRNAs 显著上调或下调，并选择进行实时 PCR 分析。miR-378d 是受 NaF 影响最显著的一个剂量依赖性上调。它与 KGN 细胞中 NaF 诱导的凋亡程度呈正相关，但与自噬水平呈负相关。此外，miR-378d 促进了 1 和 2 mM NaF 处理时的 E 释放，但减少了 4 和 8 mM NaF 处理时的 E 释放。结论是，在 NaF 的影响下，卵巢颗粒细胞中 miR-378d 的表达与自噬和 E 释放呈负相关，与细胞凋亡呈正相关。
