胚泡表面Le~Y寡糖对小鼠胚泡纤维粘连蛋白、层粘连蛋白表达及分泌的影响

目的　体外观察 L e Y 寡糖对着床前小鼠胚泡细胞外基质 ( ECM)主要成分纤维粘连蛋白 ( FN)、层粘连蛋白 ( LN)的表达和分泌的影响。　方法　采用特异性单克隆抗体 AH6阻断胚泡表面 L e Y寡糖 ,运用半定量逆转录 -聚合酶链反应 ( RT-PCR)及免疫印迹法检测 L e Y寡糖对着床前小鼠胚泡 FN及 LN的表达和分泌的影响。　结果　在体外培养中 ,经 AH6封闭胚泡表面 L e Y 寡糖抗原后仅 1 .5h,胚泡 FN及 L N的分泌有明显升高 ( P<0 .0 1 ) ,并且持续 6 h以上 ,而胚泡 FN及 L N的基因转录表达变化不明显 ( P>0 .0 5)。　结论　胚泡表面的 L e Y寡糖抗原对着床前胚泡的 FN及 L N的分泌具有促进作用

Effects of Lewis Y oligosaccharide on secretion and gene expression of fibronectin and laminin in murine blastocyst

Objective: To investigate the effects of oligosaccharide Le Y on the expression and secretion of fibronectin (FN) and laminin (LN) in cultured murine blastocysts. Methods: The expression and secretion of fibronectin (FN) and laminin (LN) in cultured murine blastocysts were measured by immuno-dot blot and RT-PCR after blocking the oligosaccharide on blastocyst surface with specific monoclonal antibody AH6. The cultured blastocysts were treated with antibody AH6 for 3 hrs, then the antibody was removed and culture was continued. The expression and secretion of FN and LN were analyzed at 1.5, 3 and 6 hrs. Results: The results showed that, when oligosaccharide Le Y blocked by AH6 antibody, the secretion of FN and LN in blastocysts were graduall increased from 1.5 to 6 hrs (P<0.01); but gene expression of FN and LN had no significant change during the period of observation (P>0.05). Conclusion: The results indicate that Le Y may play a role in regulating the secretions of FN and LN of murine blastocyst at pre-implantation stage.