天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (5): 460-464.doi: 10.11958/20201085

• 细胞与分子生物学 • 上一篇    下一篇

IKCa1在围着床期子宫内膜的表达及其作用

陈洪燕1，孙星宇2，李亚玲1，何金三3，刘玲2△   

  1. 1西南医科大学附属医院药学部（邮编646000），2生殖中心，3综合采购部
  • 收稿日期:2020-04-20 修回日期:2021-03-05 出版日期:2021-05-15 发布日期:2021-05-25
  • 通讯作者: 刘玲 E-mail:eye99@163.com
  • 基金资助:
    四川省科学技术计划项目（f150075），泸州市科技局项目［2016-S-67（17/23）］

The expression and role of IKCa1 in endometrium during peri-implantation

CHEN Hong-yan1, SUN Xing-yu2, LI Ya-ling1, HE Jin-san3, LIU Ling2△   

  1. 1 Department of Pharmacy, 2 Reproductive Medicine Center, 3 Department of General Purchasing, the Affiliated 
    Hospital of Southwest Medical University, Luzhou 646000, China
  • Received:2020-04-20 Revised:2021-03-05 Published:2021-05-15 Online:2021-05-25

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摘要： 摘要：目的　检测中电导钙激活性钾离子通道（IKCa1）在围着床期子宫内膜的表达及其在胚胎着床过程中的作用。方法　分别取分泌和增生早、中、晚期子宫内膜组织各5例。应用qPCR和免疫组化技术检测各期IKCa1 mRNA和蛋白的表达。体外培养人子宫内膜癌Ishikawa细胞，不同浓度（0、10、20、30 µmol/L）IKCa1阻滞剂TRAM-34干预后，qPCR检测细胞IKCa1表达，CCK-8法检测细胞增殖能力，Transwell实验检测细胞体外胚胎黏附能力，Western blot检测细胞上皮-间质转化（EMT）相关蛋白E盒结合蛋白1（Zeb1）和上皮型钙黏蛋白（E-cadherin）表达。结果　IKCa1在人子宫内膜中呈时序性表达，分泌期表达水平较增生期显著增高，以分泌中期最高。TRAM-34抑制IKCa1表达后，Ishikawa细胞增殖和胚胎黏附能力减弱；EMT标志蛋白E-cadherin表达显著上调，Zeb1表达显著下调（P＜0.05）。结论　IKCa1可能参与围着床期子宫内膜容受性的建立，并通过影响子宫内膜细胞增殖和EMT调控胚胎着床。

关键词: 瞬时电导钙激活钾通道, 子宫内膜, 胚胎植入, 上皮-间质转化, 钙黏着糖蛋白类, E盒结合锌指蛋白1

Abstract:

Abstract: Objective　To detect the expression of intermediate-conductance-Ca2+- activated K+ channels (IKCa1) in endometrium during peri-implantation and explore the role of IKCa1 in embryo implantation. Methods　The endometrial tissue samples of early, middle and late secretory stages and early, middle and late proliferative stages were taken respectively in 5 cases. Immunohistochemistry and quantitative PCR (qPCR) were used to detect the expression of IKCa1 mRNA and protein in endometrial tissues of different stages. Ishikawa cells were cultured in vitro and treated with different concentrations (0, 10, 20 and 30 μmol/L) of IKCa1 blocker TRAM-34. The expression of IKCa1 was detected by qPCR, cell proliferation was detected by CCK8, the cell adhesion was detected by Transwell and the expressions of EMT related proteins, E-box binding protein 1 (Zeb1) and E-cadherin were detected by Western blot assay. Results　The expression of IKCa1 was time-dependent in human endometrium. The expression level of IKCa1 was significantly higher in secretory phase than that in proliferative phase, especially in middle secretory phase. The expression of IKCa1 was inhibited by TRAM-34 in Ishikawa cells. The cell proliferation and embryo adhesion decreased, the expression of EMT marker protein E-cadherin was significantly up-regulated, and the expression of Zeb1 protein was significantly down regulated (P＜0.05). Conclusion　IKCa1 may be involved in the establishment of endometrial receptivity during peri-implantation, and may regulate embryo implantation by influencing endometrial cell proliferation and EMT.

Key words: intermediate-conductance calcium-activated potassium channels, endometrium, embryo implantation, epithelial-mesenchymal transition, cadherins, zinc finger E-box-binding homeobox 1