Khazaei Mozafar, Roshankhah Shiva, Ghorbani Rostam, Chobsaz Farzaneh
Fertility and Infertility Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Int J Fertil Steril. 2011 Oct;5(3):142-7. Epub 2011 Dec 22.
Sildenafil is a selective inhibitor of cyclic-guanosine monphosphat-specific phosphodiesterase type 5. It increases intracellular nitric oxide (NO) production in some cells. There are reports on its positive effect on uterine circulation, endometrial thickness, and infertility improvement. Endometrial epithelial cells (EEC) play an important role in embryo attachment and implantation. The present work investigates the effect of sildenafil on human EEC and their NO secretion in vitro.
In this experimental in vitro study, endometrial biopsies (n=10) were washed in a phosphate buffered solution (PBS) and digested with collagenase I (2 mg/ml in DMEM/ F12 medium) at 37°C for 90 minutes. Epithelial glands were collected by sequential filtration through nylon meshes (70 and 40 μm pores), respectively. Epithelial glands were then treated with trypsin to obtain individual cells. The cells were counted and divided into four groups: control and 1, 10, and 20 μM sildenafil concentrations. Cells were cultured for 15 days at 37ºC and 5% CO2; the media were changed every 3 days, and their supernatants were collected for the NO assay. NO was measured by standard Greiss methods. Data were analyzed by one way ANOVA.
There was no significant difference between groups in cell count and NO secretion, but the level of NO increased slightly in the experimental groups. The 10 μM dose showed the highest cell count. EEC morphology changed into long spindle cells in the case groups.
Sildenafil (1, 10, and 20 μM) showed a mild proliferative effect on human EEC numbers, but no significant change was seen in NO production.
西地那非是一种环磷酸鸟苷特异性5型磷酸二酯酶的选择性抑制剂。它能增加某些细胞内一氧化氮（NO）的生成。有报道称其对子宫循环、子宫内膜厚度及改善不孕有积极作用。子宫内膜上皮细胞（EEC）在胚胎着床和植入过程中起重要作用。本研究旨在探讨西地那非对人EEC及其体外NO分泌的影响。
在这项体外实验研究中，将10份子宫内膜活检组织在磷酸盐缓冲液（PBS）中冲洗，然后在37℃下用I型胶原酶（在DMEM/F12培养基中浓度为2mg/ml）消化90分钟。通过依次经尼龙网（孔径分别为70和40μm）过滤收集上皮腺体。然后用胰蛋白酶处理上皮腺体以获得单个细胞。对细胞进行计数并分为四组：对照组以及西地那非浓度为1、10和20μM的组。细胞在37℃、5%二氧化碳条件下培养15天；每3天更换一次培养基，并收集上清液用于NO检测。采用标准格里斯方法测定NO。数据采用单因素方差分析进行分析。
各组细胞计数和NO分泌之间无显著差异，但实验组中NO水平略有升高。10μM剂量组的细胞计数最高。病例组中EEC形态转变为长梭形细胞。
西地那非（1、10和20μM）对人EEC数量有轻微的增殖作用，但NO生成未见显著变化。
