骨桥蛋白对小鼠胚泡黏附和扩展的影响及其机制  

Effect of osteopontin on mouse blastocyst adhesion and outgrowth and its mechanism

作　　者：卢瑾文[1] 杨菁[1] 李风和[1] 徐望明[1] 龙文[1] 徐兰萍[1] 苏红[1] 谢青贞[1] 

机构地区：[1]武汉大学人民医院生殖医学中心,武汉430060

出　　处：《解剖学报》2009年第5期794-797,共4页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30670233)

摘　　要：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 of osteopontin on mouse blastocyst attachment and outgrowth in vitro and its mechanism. Methods Mouse blastocysts were cultured in microdrops, and the culture plates were precoated with fibronectin. Blastocysts were cultured in vitro in Quinn＇ s medium containing various concentrations of recombinant mouse osteopontin, osteopontin antibody and RGD peptide. The percent of hatched blastocysts, blastocysts with adhesion and outgrowth were calculated at 24 hours,48 hours and 72 hours after culture. In another experiment, blastocysts were cultured in 24-well culture plate, the concentrations of matrix metalloproteinases （MMP）-2, -9 of culture medium were detected by ELISA. Results There were no significant difference in percent of hatching and attachment between control and OPN treated groups, however, the rates of outgrowth in OPN treated groups at concentrations of either 1.0mg/L or 10.0mg/L were higher than those in control group after 72 hours culturing（ P 〈 0.05 and P 〈 0.01, respectively）, and the blastocysts started to outgrowth in advance after 48 hours culturing, whereas mouse osteopontin antibody and RGD reduced the incidence of blastocyst hatching, attachment and outgrowth significantly in a dose-dependent manner. Osteopontin promoted the blastocysts secreting MMP-2, -9 in a close-dependent and time-dependent manner. Conclusion Osteopontin is involved in regulating embryo early implantation process in vitro by promoting blastocyst outgrowth and secreting MMP-2, -9.

关 键 词：骨桥蛋白 胚胎着床 黏附 扩展 基质金属蛋白酶 酶联免疫吸附测定法 小鼠 

分 类 号：R711.6[医药卫生—妇产科学]