Matsumoto Kenji, Yamauchi Nobuhiko, Watanabe Ryo, Oozono Shinji, Kubota Kaiyu, Nishimura Kyohei, Wood Chris, Soh Tomoki, Kizaki Kei-Ichirou, Hattori Masa-Aki
Department of Animal and Marine Bioresource Sciences, Graduate School Kyushu University, 6-10-1 Hakozaki, Fukuoka 812-8581, Japan.
Cell Tissue Res. 2009 Mar;335(3):575-83. doi: 10.1007/s00441-008-0734-1. Epub 2008 Dec 17.
The induction of the decidualization of endometrial stromal cells is possible in an in vitro cell culture system. However, thus far, methods differ according to species or cell type, and a more stable or universal system has not yet been developed. The purpose of the present study has been to establish an in vitro decidualization system in primary cultured rat endometrial stromal cells (RES). The RES were treated with medroxyprogesterone acetate and dibutyryl-cyclic adenosine monophosphate (MPA treatment), estradiol and progesterone, or arachidonic acid. After 24 h of treatment, cells responded to all of the stimulations by expressing desmin mRNA. However, decidual/trophoblast prolactin-related protein (dPRP) mRNA was only expressed in the MPA-treated cells. Desmin and dPRP mRNA were not expressed after MPA treatment of the RES derived from immature rat uteri. However, mRNA from both desmin and dPRP were expressed in RES derived from gonadotrophin-injected immature rats. The expression of matrix metalloproteinase-2 (MMP-2) and MMP-9 mRNA did not change after the decidual treatment of RES examined by real-time polymerase chain reaction. However, the results of gelatin zymography showed that the active forms of MMP-2 and MMP-9 significantly increased after in vitro decidualization (P < 0.05). We conclude that MPA treatment is the most effective method for stimulating decidualization in RES. Use of this system has revealed that sexual maturation and gonadotrophins are important for RES with regard to decidualization. Furthermore, the activity of MMP-2 and MMP-9 might increase during decidualization without a corresponding increase of the expression of these genes.
在体外细胞培养系统中诱导子宫内膜基质细胞蜕膜化是可行的。然而,到目前为止,方法因物种或细胞类型而异,尚未开发出更稳定或通用的系统。本研究的目的是在原代培养的大鼠子宫内膜基质细胞(RES)中建立体外蜕膜化系统。RES用醋酸甲羟孕酮和二丁酰环磷酸腺苷(MPA处理)、雌二醇和孕酮或花生四烯酸处理。处理24小时后,细胞通过表达结蛋白mRNA对所有刺激作出反应。然而,蜕膜/滋养层催乳素相关蛋白(dPRP)mRNA仅在MPA处理的细胞中表达。来自未成熟大鼠子宫的RES经MPA处理后,结蛋白和dPRP mRNA未表达。然而,来自注射促性腺激素的未成熟大鼠的RES中结蛋白和dPRP的mRNA均有表达。通过实时聚合酶链反应检测,RES蜕膜处理后基质金属蛋白酶-2(MMP-2)和MMP-9 mRNA的表达没有变化。然而,明胶酶谱分析结果显示,体外蜕膜化后MMP-2和MMP-9的活性形式显著增加(P<0.05)。我们得出结论,MPA处理是刺激RES蜕膜化的最有效方法。使用该系统表明,性成熟和促性腺激素对RES的蜕膜化很重要。此外,在蜕膜化过程中,MMP-2和MMP-9的活性可能增加,而这些基因的表达没有相应增加。