干扰podoplanin的表达对人早孕期蜕膜基质细胞生物学功能影响的研究

发布时间：2018-03-29 21:13

本文选题：podoplanin　切入点：CLEC-2　出处：《苏州大学》2012年硕士论文

【摘要】：目的：探讨跨膜糖蛋白podoplanin在人早孕期母胎界面的表达及其对蜕膜基质细胞增殖及侵袭等生物学功能的影响。方法：收集正常早孕期蜕膜及绒毛组织，应用本实验室建立的胰酶消化、密度梯度离心法分离和培养蜕膜基质细胞(decidual stromal cells，DSCs)和滋养细胞（trophoblasts，，Tros）。采用免疫组织化学、免疫细胞化学以及流式细胞术分别检测podoplanin和及其受体C型凝集素样受体-2（C-type lectin-like receptor-2，CLEC-2）的表达水平和特点；并以siRNA干扰技术沉默蜕膜基质细胞podoplanin的表达，然后采用MTT比色法检测其活力、BrdU细胞增殖ELISA法检测其增殖能力、以流式细胞术分析其凋亡情况以及Transwell法分析podoplanin沉默后细胞侵袭能力的改变。结果：正常早孕蜕膜基质细胞和滋养细胞均表达podoplanin和CLEC-2，podoplanin在DSCs的表达水平显著高于Tros者，而CLEC-2在Tros和DSCs中表达水平均较低；采用siRNA干扰podoplanin在DSCs的表达后，在显著降低细胞活力并抑制细胞增殖和侵袭能力的同时，促进了该细胞的凋亡。结论：人早孕期母-胎界面均表达podoplanin及其受体CLEC-2，且podoplanin在蜕膜基质细胞的表达水平较高，提示podoplanin可能通过与内源性受体CLEC-2的相互作用参与调节DSCs的生物学功能并维持正常妊娠。Podoplanin可能在促进蜕膜基质细胞的增殖和侵袭、抑制其凋亡的信号通道中发挥着中介作用，这种作用有利于维持细胞自身的正常生物学功能和调节滋养细胞侵袭过程，从而最终有利于胎盘的形成和正常妊娠的维持。
[Abstract]:Objective: To investigate the expression of transmembrane glycoprotein Podoplanin in human fetal maternal interface of decidual stromal cell proliferation and invasion of biological function influence.
Methods: collect decidua and villi of early pregnancy, using trypsin digestion established in our laboratory, isolation and culture of decidual stromal cells by density gradient centrifugation (decidual stromal, cells, DSCs) and trophoblast cells (trophoblasts, Tros). By immunohistochemistry, immunocytochemistry and flow cytometry were used to detect Podoplanin and its receptor C type lectin like receptor -2 (C-type lectin-like receptor-2, CLEC-2) expression levels and characteristics; and the siRNA interference technology to silence Podoplanin expression of decidual stromal cells, then MTT assay was used to test the activity, detect the proliferation ability of BrdU cell proliferation by ELISA method, analysis of Podoplanin silencing cells invasion cell apoptosis Transwell method and flow cytometry.
Results: Podoplanin and CLEC-2 were expressed in normal decidual stromal cells and trophoblast cells, Podoplanin expression level in DSCs was significantly higher than that of Tros, but the level of CLEC-2 was lower in Tros and DSCs expression by siRNA interference in Podoplanin; the expression of DSCs, a significant reduction in cell viability and inhibit cell proliferation and invasion ability at the same time the cell apoptosis, promote.
Conclusion: Podoplanin and its receptor CLEC-2 were expressed in human first trimester maternal fetal interface, and Podoplanin in decidual stromal cells expressed higher levels, suggesting that Podoplanin may interact with endogenous CLEC-2 receptor involved in the biological function of the regulation of DSCs and.Podoplanin in normal pregnancy may promote the proliferation and invasion of decidual stromal cells, plays a mediating role the signal channel inhibits cell apoptosis, this effect is conducive to the normal biological function of keeping cell and regulation of trophoblast invasion process, which is conducive to the maintenance of the placenta formation and normal pregnancy.

【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

【参考文献】