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目的探讨骨桥蛋白(OPN)对小鼠胚泡黏附、扩展的影响及机制。方法采用纤维蛋白铺板微滴培养法培养胚胎,观察并统计重组小鼠OPN(rmOPN)、OPN抗体及精氨酸-甘氨酸-天冬氨酸多肽(RGD)对胚泡的脱带、黏附和扩展的影响;采用24孔板培养胚胎,酶联免疫吸附测定(ELISA)法检测胚泡培养液基质金属蛋白酶2和9(MMP-2-、9)的浓度。结果不同浓度的OPN组间胚泡的脱带率、黏附率与对照组相比无显著性差异(P>0.05);但1.0 mg/L和10.0 mg/L OPN组可促进小鼠胚泡提前扩展,72h囊胚扩展率与对照组相比差异有显著性,10.0mg/L组72h的扩展率显著高于OPN 0.1 mg/L组(P<0.05)。OPN抗体和RGD均显著抑制胚泡的脱带、黏附和扩展,与对照组相比,差异有显著性,且随着浓度的增加,抑制作用越明显。OPN促进胚泡分泌MMP-2及MMP-9,且存在时间和剂量依赖性。结论OPN在体外可通过促进小鼠胚泡的扩展和分泌MMP-2-、9来调节小鼠早期胚泡着床。
Objective To investigate the effect and mechanism of osteopontin (OPN) on adhesion and expansion of mouse blastocysts. Methods Fetal embryos were cultured with fibrin plating dextran. The expression of OPN (rmOPN), OPN antibody and arginine - glycine - aspartic acid polypeptide (RGD) The embryos were cultured in 24-well plates and the levels of matrix metalloproteinase 2 and 9 (MMP-2, 9) in blastocyst medium were detected by enzyme linked immunosorbent assay (ELISA). Results There was no significant difference in blastocyst debonding rate and adhesion between different concentrations of OPN (P> 0.05). However, OPN of 1.0 mg / L and 10.0 mg / L could promote early blastocyst The expansion rate of blastocyst at 72h was significantly different from that of control group. The expansion rate at 72h in 10.0mg / L group was significantly higher than that of OPN 0.1mg / L group (P <0.05). Both OPN antibody and RGD significantly inhibited the removal, adhesion and expansion of blastocyst. Compared with the control group, the OPN antibody and RGD showed significant difference, and the inhibition was more obvious with the increase of concentration. OPN promotes the secretion of MMP-2 and MMP-9 by blastocysts in a time- and dose-dependent manner. Conclusion OPN can regulate the implantation of early blastocysts in mice by promoting the expansion of mouse blastocysts and secrete MMP-2, 9 in vitro.