Carrascosa Jose P, Cotán David, Jurado Inmaculada, Oropesa-Ávila Manuel, Sánchez-Martín Pascual, Savaris Ricardo F, Tan Justin, Sánchez-Alcázar Jose A, Tan Seang L, Horcajadas José Antonio
1 SINAE, Sevilla, Spain.
2 Centro Andaluz de Biología del Desarrollo, Sevilla, Spain.
Reprod Sci. 2018 Jul;25(7):985-999. doi: 10.1177/1933719117732165. Epub 2017 Oct 5.
Intrauterine devices (IUDs) have been widely used to prevent pregnancies with great efficacy during decades. It has been demonstrated that IUD alters the endometrial gene expression, but there is no scientific data about how copper, a metal commonly used in these devices, by itself, is able to influence the processes of endometrial receptivity and apoptosis in decidualized human endometrial stromal cells. Five endometrial samples were obtained from fertile women and processed by a standard protocol to obtain human endometrial stromal cells for in vitro studies. Stromal cells were cultured in vitro and decidualized for 8 days. At day 6, copper was added to the treatment group or camptothecin as positive control for apoptosis until day 8. Five endometrial samples were used in each group. The aim of this study was to analyze the effect of copper in apoptosis and necrosis by flow cytometry, to visualize the apoptotic microtubule network during apoptosis by immunofluorescence, and finally to determine the gene expression profile of a panel of 192 genes related to endometrial receptivity and immune system by quantitative reverse transcription PCR (RT-qPCR). Copper, compared to the decidualized group, induced changes in the gene expression by an order of magnitude in 49 genes (42 up- and 9 downregulated). This alteration in the decidualization gene signature by copper includes 19 genes involved in the endometriosis pathology and others related to other gynecological disorders such as preeclampsia and infertility. Our results indicate that copper does not increase the apoptosis level induced by the decidualization treatment. However, copper alters the gene expression of some biomarkers of endometrial receptivity and immune response.
几十年来，宫内节育器（IUDs）已被广泛用于高效预防妊娠。已有研究表明，IUD会改变子宫内膜的基因表达，但对于这些装置中常用的金属铜本身如何影响人蜕膜化子宫内膜基质细胞的子宫内膜容受性和凋亡过程，尚无科学数据。从有生育能力的女性身上获取了5份子宫内膜样本，并按照标准方案进行处理，以获得用于体外研究的人子宫内膜基质细胞。将基质细胞在体外培养并蜕膜化8天。在第6天，将铜添加到治疗组，或将喜树碱作为凋亡的阳性对照，直至第8天。每组使用5份子宫内膜样本。本研究的目的是通过流式细胞术分析铜对凋亡和坏死的影响，通过免疫荧光观察凋亡过程中的凋亡微管网络，最后通过定量逆转录PCR（RT-qPCR）确定一组192个与子宫内膜容受性和免疫系统相关基因的基因表达谱。与蜕膜化组相比，铜诱导49个基因（42个上调和9个下调）的基因表达发生了一个数量级的变化。铜对蜕膜化基因特征的这种改变包括19个参与子宫内膜异位症病理过程的基因以及其他与子痫前期和不孕症等其他妇科疾病相关的基因。我们的结果表明，铜不会增加蜕膜化处理诱导的凋亡水平。然而，铜会改变一些子宫内膜容受性和免疫反应生物标志物的基因表达。
