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目的:探讨转录因子C/EBP-β在人子宫内膜基质细胞体外蜕膜化过程中的表达变化及其作用。方法:采用8-Br-cAMP和MPA对人子宫内膜基质细胞体外蜕膜化处理,并通过瞬时转染的方法下调C/EBP-β的表达,分别收集蜕膜化处理不同时间及干扰前后的细胞及其培养上清液,采用Western blot和荧光适时定量RT-PCR分别在蛋白质水平和mRNA水平检测转录因子C/EBP-β在蜕膜化过程中的表达变化,采用化学发光法检测蜕膜化不同时间点培养液中泌乳素(PRL)的浓度及干扰前后PRL表达水平的变化。结果:免疫印迹结果显示人子宫内膜基质细胞在体外蜕膜化过程中C/EBP-β蛋白的表达随蜕膜化时间的延长逐渐升高(0.068、0.140、0.316、0.595),与对照组相比,差异有统计学意义(P<0.05);荧光适时定量RT-PCR也证实C/EBP-βmRNA的表达随蜕膜化时间的延长明显增加(0.020、0.054、0.094、0.486),与对照组相比差异有统计学意义(P<0.05)。化学发光法检测到在蜕膜化过程中细胞分泌的PRL水平随蜕膜化时间延长逐渐升高,差异有统计学意义(P<0.05),而转染C/EBP-βsiRNA后细胞分泌的PRL浓度则明显降低,差异有统计学意义(P<0.05)。结论:转录因子C/EBP-β在人子宫内膜基质细胞体外蜕膜化进程中上升性表达,在蜕膜化过程中可能发挥着重要的促进作用。
Objective: To investigate the change and its role of transcription factor C / EBP-β in the process of human endometrial stromal cells decidualization in vitro. Methods: Human endometrial stromal cells were treated with 8-Br-cAMP and MPA for decidualization in vitro. The expression of C / EBP-β was down-regulated by transient transfection. The expression of C / EBP-β in the process of decidualization was detected by Western blot and fluorescence quantitative RT-PCR at the protein level and mRNA level, respectively. The chemiluminescence The concentration of prolactin (PRL) and the expression of PRL before and after the intervention in different time points of membrane culture. Results: Western blotting showed that the expression of C / EBP-βprotein in human endometrial stromal cells was gradually increased with the decidualization time (0.068,0.140,0.316,0.595) in vitro. Compared with the control group (P <0.05). Fluorescence quantitative RT-PCR also confirmed that the expression of C / EBP-βmRNA increased significantly with the time of decidualization (0.020,0.054,0.094,0.486), compared with the control The difference was statistically significant (P <0.05). Chemiluminescence showed that the level of PRL secreted by cells during decidualization increased gradually with the time of decidualization, the difference was statistically significant (P <0.05), while the level of PRL secreted by cells after transfection with C / EBP-βsiRNA The concentration was significantly lower, the difference was statistically significant (P <0.05). CONCLUSION: The transcription factor C / EBP-β is up-regulated during the decidualization of human endometrial stromal cells in vitro and may play an important role in the process of decidualization.