Le Aiwen, Wang Zhong Hai, Dai Xiao Yun, Xiao Tian Hui, Zhuo Rong, Zhang Baozhen, Xiao Zhonglin, Fan Xiujun
Department of Obstetrics and Gynecology, Affiliated Shenzhen Nanshan People's Hospital of Guangdong Medical University, Shenzhen, Guangdong 518052, P.R. China.
Laboratory for Reproductive Health, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, Guangdong 518055, P.R. China.
Exp Ther Med. 2017 Dec;14(6):5949-5955. doi: 10.3892/etm.2017.5278. Epub 2017 Oct 10.
The aim of the present study was to investigate the effects of Icaritin on the proliferation and decidualization of endometrial stromal cells (ESCs). A total of 20 specimens of endometrium were collected during hysterectomy at the Gynecology Department of Shenzhen Nanshan People's Hospital (Shenzhen, China) between August 2014 and December 2015. The endometrium was digested with high concentrations of collagenase and DNase and filtered with meshes, and then the glandular epithelial and stromal cells were separated by the adhesion purification method. The purity of stromal cells was identified by vimetin and cytokeratin 7 immunostaining. The estradiol + progesterone (E2+P4) and/or cyclic adenosine monophosphate (cAMP) were added to induce an decidualization model, which was used to analyze the effect of Icaritin on the decidualization ability of the human ESCs. The decidualization markers of human ESCs, prolactin (PRL) and insulin-like growth factor-binding protein 1 (IGFBP-1), was analyzed by reverse-transcription quantitative polymerase chain reaction measurements of the mRNA levels, PRL immunostaining and ELISA analysis of the IGFBP-1 protein levels in the cells or cell culture supernatant separately. The results demonstrated that treatment with E2+P4 and/or cAMP for 96 h was able to induce decidualization in ESCs, and that the cells demonstrated polygon-shaped epithelioid changes. The cell nuclei revealed multinuclear changes, and the cells were also observed to be large and round in shape. The PRL expression and upregulated IGFBP-1 mRNA and protein levels in the E2+P4+cAMP treatment group indicated successful decidualization of the model. However, the addition of Icaritin inhibited the expression of PRL and IGFBP-1 mRNA, as well as IGFBP-1 protein in the induced ESCs compared with groups without Icaritin. These results suggest that Icaritin was able to inhibit the expression of decidualization-related genes in ESCs . However, the exact mechanisms require further investigation.
本研究旨在探讨淫羊藿素对子宫内膜基质细胞（ESCs）增殖和蜕膜化的影响。2014年8月至2015年12月期间，在深圳市南山人民医院（中国深圳）妇科进行子宫切除术时，共收集了20份子宫内膜标本。用高浓度的胶原酶和脱氧核糖核酸酶消化子宫内膜，并用滤网过滤，然后通过黏附纯化法分离腺上皮细胞和基质细胞。通过波形蛋白和细胞角蛋白7免疫染色鉴定基质细胞的纯度。添加雌二醇+孕酮（E2+P4）和/或环磷酸腺苷（cAMP）诱导蜕膜化模型，用于分析淫羊藿素对人ESCs蜕膜化能力的影响。分别通过逆转录定量聚合酶链反应测量细胞或细胞培养上清液中mRNA水平、PRL免疫染色和IGFBP-1蛋白水平的ELISA分析，对人ESCs的蜕膜化标志物催乳素（PRL）和胰岛素样生长因子结合蛋白1（IGFBP-1）进行分析。结果表明，用E2+P4和/或cAMP处理96小时能够诱导ESCs蜕膜化，细胞呈现多边形上皮样变化。细胞核显示多核变化，细胞也观察到体积大且呈圆形。E2+P4+cAMP处理组中PRL表达以及IGFBP-1 mRNA和蛋白水平上调表明模型成功蜕膜化。然而，与未添加淫羊藿素的组相比，添加淫羊藿素抑制了诱导的ESCs中PRL和IGFBP-1 mRNA以及IGFBP-1蛋白的表达。这些结果表明淫羊藿素能够抑制ESCs中蜕膜化相关基因的表达。然而，确切机制需要进一步研究。
