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目的研究原代培养蜕膜细胞在孕酮和米非司酮作用后EP3-Ⅲ的表达变化。方法蜕膜组织收集自2013年5-12月就诊于辽宁省计划生育科研院门诊手术室要求人工流产的妊娠妇女12例。采用复合酶对蜕膜组织进行消化,全组份蜕膜细胞自然传代法纯化,人泌乳素(PRL)免疫组化法鉴定原代培养蜕膜细胞。RT-PCR方法检测EP3-Ⅲ在人蜕膜组织和原代培养细胞中的表达情况。采用实时定量PCR方法检测孕酮和米非司酮作用后EP3-Ⅲ在原代蜕膜细胞中的表达变化。结果分离培养的蜕膜基质细胞呈长梭形和不规则星形,95%细胞胞浆PRL阳性。RT-PCR检测到EP3-Ⅲ在人蜕膜组织和原代培养细胞中均有表达。孕酮作用后,EP3-Ⅲ在原代蜕膜细胞中表达明显升高,而米非司酮作用后EP3-Ⅲ表达明显降低,差异有统计学意义(P<0.05)。结论 EP3-Ⅲ在人早孕蜕膜中的表达变化对妊娠的维持有重要作用。
Objective To study the changes of EP3-Ⅲ expression in progenitor cultured decidual cells after progesterone and mifepristone treatment. Methods Decidual tissues were collected from 12 pregnant women who asked for artificial abortion in the outpatient operating room of Family Planning Research Institute of Liaoning Province from May to December in 2013. The decidual tissue was digested by complex enzyme, all the decidual cells were purified by natural passage, PRL immunohistochemistry was used to identify primary cultured decidual cells. The expression of EP3-Ⅲ in human decidual tissue and primary cultured cells was detected by RT-PCR. Real-time quantitative PCR was used to detect the expression of EP3-Ⅲ in primary decidual cells after progesterone and mifepristone treatment. Results The decidual stromal cells cultured in vitro were long spindle and irregular star, with 95% cytoplasmic PRL positive. The expression of EP3-Ⅲ in human decidual tissue and primary cultured cells was detected by RT-PCR. After progesterone treatment, the expression of EP3-Ⅲ in primary decidual cells was significantly increased, while the expression of EP3-Ⅲ was significantly decreased after mifepristone treatment (P <0.05). Conclusion The expression of EP3-Ⅲ in human decidual cells during pregnancy may play an important role in the maintenance of pregnancy.