液相蛋白芯片法测蜕膜细胞与滋养层细胞共培养体系中基质金属蛋白酶的表达

目的：探讨蜕膜基质细胞与绒毛外滋养层细胞共培养体系中细胞滋养层细胞侵入过程中基质金属蛋白酶表达水平的变化及在母胎界面间基质金属蛋白酶对细胞滋养层细胞侵袭行为的调控。 方法：实验于2005—09／12在南方医科大学南方医院生殖中心及病生实验室完成。①分别进行蜕膜基质细胞与绒毛外滋养层细胞的体外分离培养与纯化。②在Transwell—Col Insert细胞培养板内进行蜕膜细胞与滋养细胞体外共培养及直接混合培养。③利用Luminex xMAP对培养上清液中多种基质金属蛋白酶蛋白表达的含量变化进行分析。 结果：①与绒毛外滋养层细胞相比，基质金属蛋白酶1，3在共培养体系与混合培养体系中的表达依次升高，差异有显著性（P〈0．05），基质金属蛋白酶2，7，9在共培养体系与混合培养体系中的表达依次降低，差异有显著性（P〈0．05），且在共培养体系中以基质金属蛋白酶2，9表达尤为显著，基质金属蛋白酶8，12，13在各培养体系中均为低表达。②两种共培养体系中，除基质金属蛋白酶-13变化不显著外，其余基质金属蛋白酶均在Tranwell共培养体系下显著升高（P〈0．05）。④基质金属蛋白酶1，3，7，8两两之间正相关，基质金属蛋白酶2，7，9两两间正相关，相关关系均显著（P〈0．05），且关系较密切（r〉0.5548）。 结论：共培养条件下滋养层细胞侵入过程中基质金属蛋白酶量的变化及之间的相互关系与胚胎着床关系密切。

Expression of matrix metalloproteinase by Luminex xMAP in co-cultured system of decidual cells and cytotrophoblasts

AIM： To explore the variations of matrix metalloproteinases （MMPs） expressing during cytotrophoblast （CTB） invasion in co-cuhured system of decidual stromal cells （DSCs） and extravillous CTB, and study the regulation of MMPs on invasiveness of CTB in embryouterine interface.
METHODS： The experiment was carried out in Center of Reproductive Medicine and Laboratory of Pathophysiology in Nanfang Hospital of Southern Medical University from September to December 2005. （1） Extravillous CTB and DSCs were in vitro isolated, cultured and purified, separately. （2）DSCs and CTB were in vitro co-cultured and mixed cultured directly on Transwell-Col insert cell culture plate.（3）Luminex xMAP system was used to obtain the supernatants of culture media and detect the concentration of MMPs simultaneously.
RESULTS： （1）Compared with incubations of extravillous CTB, the expressions of MMP-2, -7 and -9 were significantly lower by turns （P 〈 0.05）, while the expressions of MMP-1 and -3 in order were significantly higher in co-culture system and mixed culture system （P 〈 0.05）. Especially MMP-2 and -9 expressed markedly in co-culture system, but MMP-8, -12 and -13 all lower expressed in two systems. （2）Nearly all the expressions of MMPs were dramatically increased in Transwell co-culture system except MMP-13 （P 〈 0.05）.（3）Positive correlativity was intensively associated between MMP-1, -3, -7, and -8, and between MMP-2, -7 and -9 （r 〉 0.554 8, P 〈 0.05）.
CONCLUSION： In the invasion of CTB of in vitro co-cultured system of DSCs and CTB, the variations of MMPs and the interaction between the two are intensively associated with the embryo implantation.