前脑啡肽在子宫内膜间质细胞蜕膜化中的作用

Role of proenkephalin in endometrial stromal cell decidualization

摘要:

目的 探讨前脑啡肽（PENK）在人子宫内膜间质细胞（HESCs）蜕膜化前后的表达情况，以及分析PENK对蜕膜化发生的影响。方法 ①采用8-溴腺苷-3'，5'-环单磷酸和醋酸甲羟孕酮进行体外蜕膜化诱导，并通过F-actin染色、实时荧光聚合酶链反应（qRT-PCR）和Western blotting验证蜕膜化诱导的情况。②使用GEO数据库、qRT-PCR和Western blotting验证PENK在HESCs蜕膜化诱导前后的表达水平。③使用不同siRNA敲减HESCs中的PENK，并通过qRT-PCR和Western blotting检测敲减前后蜕膜标志物的表达，以此来研究PENK对蜕膜化的影响。④使用GEO数据库分析PENK在反复种植失败患者子宫内膜中的表达情况。结果 从形态学、转录水平和蛋白水平上证明了HESCs体外蜕膜化的成功诱导。蜕膜化诱导组PRL、IGFBP1 mRNA和IGFBP1蛋白相对表达量较对照组1高（P <0.05）。PENK在HESCs体外蜕膜化诱导过程中表达增加（P <0.05）。相比HESCs，PENK敲减的HESCs蜕膜化受到抑制，表现为蜕膜标志物PRL、IGFBP1和FOXO1表达减少（P <0.05）。GEO数据库提示与对照组子宫内膜相比，PENK在反复种植失败患者子宫内膜中表达可能下调（P <0.05）。结论 PENK是子宫内膜蜕膜化发生的关键基因。蜕膜化期间PENK表达增加，促使子宫内膜容受性建立。PENK表达减少可能与蜕膜化的异常，以及胚胎反复种植失败有关。

Abstract:

Objective To explore the expression of proenkephalin (PENK) in human endometrial stromal cells (HESCs) before and after decidualization and to analyze the effect of PENK on decidualization.Methods HESCs were subjected to in vitro decidualization using 8-bromoadenosine 3′,5′-cyclic monophosphate and medroxyprogesterone acetate. F-actin staining, qRT-PCR and Western blotting were used to verify the efficiency of decidualization induction. GEO database analysis, qRT-PCR and Western blotting were performed to validate the expression level of PENK in HESCs with or without decidualization stimulation. Different siRNAs were used to knock down PENK expression in HESCs, and the expression levels of decidualization markers before and after knockdown were measured by qRT-PCR and Western blotting to investigate the effect of PENK on decidualization. GEO database analysis was conducted to evaluate the PENK expression in the endometrium of patients with recurrent implantation failure.Results Successful induction of in vitro decidualization in HESCs was demonstrated at the morphological, transcriptional and protein levels. The mRNA levels of PRL and IGFBP1 as well as the protein level of IGFBP1 were higher in the decidualization-induced group than those in the control group 1 (P < 0.05). The expression level of PENK increased with the stimulation of decidualization in HESCs (P < 0.05). Compared to HESCs, HESCs with PENK knockdown resulted in significant inhibition of decidualization, as evidenced by decreased expression of decidualization markers PRL, IGFBP1 and FOXO1 (P < 0.05). The analysis of data from GEO databases suggested that PENK expression may be downregulated in the endometrium of patients with recurrent implantation failure compared to the control group (P < 0.05).Conclusions This study demonstrated that PENK is a key gene in endometrial decidualization. Increased PENK levels during decidualization contributes to the establishment of endometrial receptivity. Decreased expression of PENK may be associated with abnormal decidualization and the occurrence of recurrent implantation failure.