Laboratory of Reproductive Biology, School of Public Health, Joint International Research Laboratory of Reproduction & Development, Chongqing Medical University, Chongqing 400016, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2023 Feb 25;52(1):1-12. doi: 10.3724/zdxbyxb-2022-0669.
To explore the effect of exposure to di (2-ethyl) hexyl phthalate (DEHP) in early pregnancy on endometrial decidualization in mice and its relation with lncRNA -.
Early pregnancy mice were exposed to DEHP (1000 mg·kg·d) to construct the model. The uterus was collected on day 6 of pregnancy to detect its effect on decidualization by HE staining and immunofluorescence. A decidualization induction model of mouse endometrial stromal cells exposed to DEHP (0.1, 0.5, 2.5, 12.5, 62.5 μmol/L) was constructed. The changes of cell morphology were observed by light microscopy and phalloidin staining, and the expression of decidual reaction related molecular markers were detected by immunofluorescence, realtime RT-PCR and Western blotting. The expression of - in decidua tissue and cells was detected by realtime RT-PCR. Cellular localization of - was determined by lncLocator database and RNA FISH. AnnoLnc2 database was used to predict miRNAs bound to -.
The number of embryo implantation sites, uterine weight and uterine area were significantly lower in the DEHP exposed group than those in the control group, and the expression of the decidual reaction related molecular markers matrix metalloprotein 9 and homeobox A10 in the DEHP exposure group were also significantly lower than those in the control group (all <0.05). With the increase of DEHP concentration, the expression of in decidua cells was gradually decreased. 2.5 μmol/L DEHP exposed stromal cells failed to be fully decidualized andphalloidin staining showed abnormal cytoskeleton morphology. The expression levels of homeobox A10, bone morphogenetic protein 2 and proliferating cell nuclear antigen in the DEHP exposure group were significantly lower than those in the control group (all <0.05). The expression of - in DEHP exposed decidua tissue and cells was significantly reduced (both <0.05). - is mainly localized in the cytoplasm and - might bind to 45 miRNAs, among them, miR-138-5p, miR-155-5p, miR-183-5p and miR-223-3p were associated with endometrial decidualization.
DEHP exposure in early pregnancy may impair endometrial decidualization, and the damage may be associated with the down-regulation of -.
探讨孕期早期二（2-乙基）己基邻苯二甲酸酯（DEHP）暴露对小鼠子宫内膜蜕膜化的影响及其与长链非编码 RNA - 的关系。
采用 DEHP（1000mg·kg·d）暴露孕早期小鼠构建模型。取妊娠第 6 天的子宫，通过 HE 染色和免疫荧光检测其对蜕膜化的影响。构建 DEHP（0.1、0.5、2.5、12.5、62.5μmol/L）暴露的小鼠子宫内膜基质细胞蜕膜化诱导模型。通过光镜和鬼笔环肽染色观察细胞形态变化，通过免疫荧光、实时 RT-PCR 和 Western blot 检测蜕膜反应相关分子标志物的表达。通过实时 RT-PCR 检测蜕膜组织和细胞中 - 的表达。通过 lncLocator 数据库和 RNA FISH 确定 - 的细胞内定位。使用 AnnoLnc2 数据库预测与 - 结合的 miRNAs。
DEHP 暴露组胚胎着床部位数、子宫重量和子宫面积明显低于对照组，DEHP 暴露组基质金属蛋白酶 9 和同源盒 A10 等蜕膜反应相关分子标志物的表达也明显低于对照组（均<0.05）。随着 DEHP 浓度的增加，蜕膜细胞中 - 的表达逐渐降低。2.5μmol/L DEHP 暴露的基质细胞未能完全蜕膜化，鬼笔环肽染色显示细胞骨架形态异常。DEHP 暴露组的同源盒 A10、骨形态发生蛋白 2 和增殖细胞核抗原的表达水平明显低于对照组（均<0.05）。DEHP 暴露的蜕膜组织和细胞中 - 的表达明显降低（均<0.05）。- 主要定位于细胞质，- 可能与 45 个 miRNAs 结合，其中 miR-138-5p、miR-155-5p、miR-183-5p 和 miR-223-3p 与子宫内膜蜕膜化有关。
孕期早期 DEHP 暴露可能损害子宫内膜蜕膜化，这种损伤可能与 - 的下调有关。
