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COH 对子宫内膜间质细胞缝隙连接蛋白 43 表达的影响。

Effects of COH on the expression of connexin43 in endometrial stromal cells.

机构信息

People's Hospotal of Fu Yang, No. 63 Lu ci Street, Yingzhou District, Fu Yang, Anhui Province, China; Shanghai First Maternity and Infant Hospital, Tongji University, NO. 2699 Gaoke West Road, Pudong New Area, Shanghai, China.

Shanghai First Maternity and Infant Hospital, Tongji University, NO. 2699 Gaoke West Road, Pudong New Area, Shanghai, China.

出版信息

Taiwan J Obstet Gynecol. 2019 Sep;58(5):592-597. doi: 10.1016/j.tjog.2019.07.003.

Abstract

OBJECTIVE

The purpose of this study was to investigate the effect of controlled ovarian hyperstimulation (COH) on gap junction, and to induce the effect of an estrogen level overdose on gap junction in vitro by COH. Here, we mainly focus on connexion43 (Cx43), progesterone receptor (PR) and prolactin-related protein (PRP), and CyclinD3 genes expression, as well as the expression of Cx43 protein, were investigated.

MATERIALS AND METHODS

Mature BDF-1 mice were divided into different COH, and the mouse uterus was isolated, Paraffin sections evaluate the effect of COH on mouse uterine endometrial morphology. The other part was used for the extraction of mouse uterine endometrial stromal cells (ESC), some related gene changes are detected. Human ESC were isolated from human endometrium by primary culture, the estrogen concentrations 10 mol/L, 10 mol/L were added, the changes of Cx43 gene and related proteins were detected, too.

RESULTS

(1) HE staining showed that in the ovulatory endometrium of mice in the high super ovulation group, uterine glands in the stromal layer were significantly increased, the relative vascular tissues was less abundant. (2) In three groups of COH mice, the expression of Cx43, PR, and PRP genes in ESC was significantly different (P < 0.05). (3) In vitro ESC in the COH group showed significant differences in Cx43, PR, and CyclinD3 gene expression (P < 0.05), and showed an obvious dose effect. In addition, Western blot analysis showed that the Cx43 protein and Cx43 gene expression were similar.

CONCLUSIONS

(1) Animal experiments study showed that Cx43 gene expression in ESC was significantly decreased in hyper COH, in addition, the advance in gene expression was significantly earlier, suggesting decidualization appeared significantly earlier. (2) In vitro COH demonstrated when the estrogen concentration used was higher, the expression level of Cx43 gene and protein was lower. Combined with animal experiments, the endometrium decidualization was advanced in mice that were underwent hyper COH, which may reflect the endometrial receptivity.

摘要

目的

本研究旨在探讨控制性卵巢过度刺激(COH)对缝隙连接的影响,并通过 COH 诱导体外雌激素水平过量对缝隙连接的影响。在这里,我们主要关注连接蛋白 43(Cx43)、孕激素受体(PR)和泌乳素相关蛋白(PRP)以及细胞周期蛋白 D3 基因的表达,以及 Cx43 蛋白的表达。

材料和方法

将成熟的 BDF-1 小鼠分为不同的 COH 组,分离小鼠子宫,石蜡切片评估 COH 对小鼠子宫内膜形态的影响。另一部分用于提取小鼠子宫内膜基质细胞(ESC),检测相关基因的变化。通过原代培养从人子宫内膜中分离人 ESC,分别加入 10nmol/L 和 100nmol/L 的雌激素,检测 Cx43 基因和相关蛋白的变化。

结果

(1)HE 染色显示,在高超排卵组小鼠的排卵期子宫内膜中,基质层中的子宫腺明显增加,相对血管组织较少。(2)在三组 COH 小鼠中,ESC 中 Cx43、PR 和 PRP 基因的表达有显著差异(P<0.05)。(3)COH 组 ESC 中 Cx43、PR 和 CyclinD3 基因表达有显著差异(P<0.05),且呈明显的剂量效应。此外,Western blot 分析显示 Cx43 蛋白和 Cx43 基因表达相似。

结论

(1)动物实验研究表明,超 COH 中 ESC 中 Cx43 基因表达明显降低,此外,基因表达的提前程度明显更早,提示蜕膜化明显提前。(2)体外 COH 显示,当使用的雌激素浓度较高时,Cx43 基因和蛋白的表达水平较低。结合动物实验,超 COH 小鼠的子宫内膜蜕膜化提前,可能反映了子宫内膜的容受性。

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