网络首发:2016-01-20,
纸质出版:2016
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miR-29a促进小鼠子宫内膜的蜕膜化[J]. 中山大学学报(医学科学版), 2016,37(1).
Mir -29a Promotes Endometrial Decidualization in Mice[J]. Journal of Sun Yat-sen University (Medical Sciences), 2016, 37(1).
摘 要: 【目的】 探讨微小RNA29a(miR-29a)在小鼠子宫内膜基质细胞(ESC)蜕膜化中的作用。【方法】 收集孕第1~9天(D1 ~ D9)的小鼠子宫,通过实时定量PCR(qRT-PCR)的方法检测小鼠子宫组织中miR-29a的表达;构建去除卵巢及其甾体激素的处理模型,观察甾体激素对miR-29a表达的影响。采用雌二醇(E2)和孕酮(P4)对分离培养的小鼠子宫内膜基质细胞进行蜕膜化处理,并通过瞬时转染的方法下调miR-29a的表达,分别收集蜕膜化处理不同时间及干扰前后的细胞,采用qRT-PCR检测催乳素相关蛋白(dPRP)mRNA的表达,Western blot法检测细胞周期蛋白D3(cyclin D3)、孕酮受体(PR)蛋白的表达。 【结果】 在小鼠妊娠第1~5 天,子宫组织中的miR-29a 表达量较低,随着蜕膜化的进程(妊娠第6~9天),miR-29a的表达量均显著性增加(与妊娠D1相比);在去除卵巢及其甾体激素的处理模型中,一旦给予雌孕激素刺激,miR-29a的表达量显著增加;在小鼠子宫内膜基质细胞体外诱导蜕膜化过程中,qRT-PCR结果显示dPRP mRNA的表达以及miR-29a的表达随着蜕膜化时间的延长逐渐升高(与24 h相比),Western blot法显示蜕膜化标志分子cyclin D3、PR的表达也随着蜕膜化时间的延长逐渐升高;转染下调miR-29a后,蜕膜化标志分子及miR-29a的表达也相应下降。【结论】 妊娠早期miR-29a表达于母胎界面,且受子宫内膜蜕膜化的影响,说明miR-29a具有促进子宫内膜蜕膜化的作用。
Abstract: 【Objective】 To explore the effect of micro RNA -29a (miR -29a) in mouse endometrial stromal cells (ESC) during the decidua process. 【Methods】 To collect uterus of mouse, which were pregnant for 1-9 d, detecting the miR -29a expression in uterine tissue of mice by real -time quantitative PCR (qRT -PCR). The effects of steroid hormones on the expression of miR -29a by constructing a mouse model of ovariectomized and processing of sex steroid hormones were observed. Use of estradiol (E2) and progesterone (P4) on decidual mouse endometrial stromal cells in vitro processing, and reduced miR -29a expression with the method of transient transfection. Cells in different time of decidualization and interference were collected respectively. The level of prolactin related proteins (dPRP) mRNA was detected by qRT -PCR, and the expression of decidualization marker molecule cyclin D3 and progesterone receptor (PR) was detected by Western blot. 【Results】 The expression of miR -29a was relatively low in the uterus of pregnant mice after 1-5 d. With the progress of deciduas (pregnant after 6-9 d), the level of miR -29a significantly increased (compared with that of pregnant mice after 1d). In the processing model of ovary removal and steroid hormones treatment, the expression of miR -29a increased significantly after given estrogen and progesterone stimulatio; qRT -PCR and Western blot showed that the level of dPRP mRNA, miR -29a, cyclin D3, PR increased with prolonging of decidualization time; the expression of decidual marker molecules and miR -29a also decreased after downregulation of miR -29a by transfection. 【Conclusion】 miR -29a expresses on the maternal fetal interface in early pregnancy, this is affected by decidua. So indicates that miR -29a can promote the progress of endometrial decidualization.
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