胰岛素样生长因子-I调节胚胎着床的体外研究

Investigation in vitro the role of IGF-I in role of IGF-I in regulating embryo implantation

目的 :观察胰岛素样生长因子 I (IGF I)对体外培养的滋养层细胞粘附到细胞外基质的影响 ,研究IGF I在胚胎着床过程中的调节作用。方法 :原代培养滋养层细胞 ,建立体外滋养层细胞粘附模型 ,IGF I按不同的时间和剂量处理细胞 ,用酶联免疫检测仪测定光密度值代表粘附细胞的相对数目。免疫细胞化学法检测粘着局部磷酸化粘着斑激酶 (pFAK)的表达。结果 :随IGF I浓度的增加 ,滋养层细胞的粘附数目明显增多 ,差异具有显著性意义 (P <0 0 5 ) ,10 0nM时粘附细胞数目最多 ,但与 10nM时相比 ,差异无显著性意义 (P >0 0 5 )。相同浓度的IGF I处理时 ,随时间的延长 ,粘附细胞数目也明显增多 ,差异具有显著性意义 (P <0 0 5 ) ,10nMIGF I培养 2h达最佳量效反应 (P <0 0 0 1)。加入抗整合素αvβ3 抗体能明显阻止其促粘附作用(P <0 0 0 1)。免疫组化染色显示 10nMIGF I处理后 ,粘着局部出现pFAK ,而对照组则无pFAK表达。结论 :IGF I能促进滋养层细胞粘附到纤维粘连蛋白 (FN) ,且这种粘附过程具有明显的时间和剂量依赖性 ,生理剂量的IGF I通过刺激整合素与配体结合 ,激活FAK ,进而激活整合素介导的信号传导通路 ,显著促进滋养层细胞粘附到细胞外基质。

Objective:To investigate the role of IGF-I in regulating Embryo Implantation, the effect of IGF-1 on the in vitro adhesion of trophoblast cells was observed. Methods:The in vitro trophoblast cell adhesion model was set up by primary culturing trophoblast cell from early gestation at artificial abortion. The trophoblast cells were incubated with different doses of IGF-I at different times. Examining optical density using enzyme-linked immunoassay appearance ～ assessed the amount of adhered cells. The expression of phosphorylated focal adhesion kinase was determined by immunocytochemistry. Results:The amount of adhered trophoblast cells significantly increased with the increasement of dose (P<0.05), The amount was the highest when incubated with 100 nmol/L IGF-I, but there was no difference compared with dose of 10 nmol/L (P>0.05). The amount of adhered trophoblast cells also significantly increased with the extension of time (P<0.05), the maximal response was obtained at 10 nmol/L IGF-I with 2 h incubation(P<0.001), but antibody against avB3 13 3 integrin significantly abolished the effects of IGF-I in these cells(P<0.001). Immunocytochemistry staining show pFAK was localized in the focal adhesions of the trophoblast cells treated with 10 nM IGF-I, whereas in the untreated trophoblast cells, no p-FAK was detectable. Conclusion:IGF-I stimulated trophoblast cells adhesion to fibronection in a time- and dose-dependent manner, IGF-1 significantly promoted adhesion of trophoblast cells to extracellular matrix by promoting integrins attach to their ligands, activating FAK and integrin signaling pathway.