TLR4对母-胎界面滋养层细胞生物学行为的影响

The effect of TLR4 on biological behavior of trophoblast cells in maternal-fetal interface

目的:观察Toll样受体(TLR4)在着床前胚泡和子宫内膜的表达,进一步探讨TLR4对滋养层细胞生物学行为的影响。方法:采用免疫组织化学染色法对TLR4蛋白在着床前小鼠胚泡和子宫内膜的表达进行定位分析;利用小鼠胚泡与人蜕膜细胞共培养着床模型,观察抗TLR4抗体对胚泡滋养层细胞粘附和铺展的影响。结果:TLR4蛋白在小鼠胚泡的内细胞群和滋养层细胞的细胞质内呈阳性表达;不同浓度TLR4抗体组的粘附率与对照组相比,无统计学意义(P>0.05);100ng/ml、200ng/ml和400ng/ml组的铺展率分别为(64.3±4.0)%、(51.3±4.1)%、(46.2±4.2)%,均低于对照组的(83.5±3.3)%(P<0.05);扩展面积分别为(63.5±5.0)×10~3、(60.2±5.2)×10~3、(58.5±4.8)×10~3μm,均明显低于对照组(194.12±8.24)×10~3μm(P<0.01),并且其扩展面积以剂量依赖的方式受到抑制。结论:TLR4不影响胚泡滋养层细胞的粘附,可能与胚泡的侵入和滋养层细胞的进一步生长有关。

Objective:To observe the expression of TLR4 in blastocyst and endometrium befor implantation and to explore its effect on the biological behavior of trophoblast cells.Methods:The expressions of TLR4 protein in mouse blastocyst and endometrium before implantation were shown with immunohistochemistry,and the attachment and outgrowth of blastocyst trophoblast cells were observed by anti-TLR4 antibody with co-culture implantation model of mouse blastocyst and human decidual cells.Results:TLR4 protein was positively expressed in the cytoplasm of the inner cell mass and trophoblast cells of mouse blastocyst.The ratio of attachmen was no significant difference between different concentration of TLR4 antibody group 78.52±2.47、82.3±2.6、77.3±3.0 and control group 87.4±2.3(P>0.05).The ratio of outgrowth in 100 ng/ml group 64.3±4.0 was lower than that of control group 83.5±3.3(P<0.05).The ratio of outgrowth in 200 ng/ml group 51.3±4.1 and 400 ng/ml group 46.2±4.2 was significantly lower than that of control group(P<0.01).The spreading areas of different concentration groups 63.5±5.0、60.2±5.2、58.5±4.8 was significantly lower than that of control group 194.1±8.2(P<0.01),and its expanded area was inhibited in a dose-dependent manner.Conclusion:TLR4 does not affect the attachment of blastocyst trophoblast cells,which may be related to the invasion and the further development of embryonic trophoblast cells.