不同胚泡移植的液体量对小鼠胚胎分布和妊娠的影响及其作用机制

The effect of blastocyst transfer liquid volume on mouse embryonic distribution and pregnancy prognosis

目的探讨不同胚泡移植的液体量对小鼠胚胎分布和妊娠的影响及其作用机制。方法通过体外胚胎移植技术,观察不同剂量(3、5、10μL)的胚泡移植液体量对小鼠胚胎分布和妊娠的影响;通过实时定量PCR和免疫印迹技术,观察不同剂量的胚胎移植液体量对着床位点白血病抑制因子(Lif)和HoxA10表达量的影响。结果 (1)胚泡移植的液体量为3μL和5μL时,胚胎着床率分别为(83.4±10.9)%和(74.8±10.7)%,两者差异无统计学意义(P>0.05),且胚胎分布均匀,胎儿发育良好;当胚泡移植的液体量为10μL时,胚胎着床率为(35.0±17.5)%,与3μL和5μL组比较,其着床率明显下降(均P<0.01),且出现胚胎分布不均匀,有些胎儿出现流产现象。(2)与3μL组相比较,10μL组着床位点的Lif mRNA和HoxA10蛋白表达量明显下降(均P<0.01),而5μL组着床位点的Lif mRNA和HoxA10蛋白表达量无明显变化(均P>0.05)。结论体外小鼠胚泡移植术时,采用胚泡移植的液体量为3μL时,有利于胚胎的均匀分布和胎儿的生长发育。

Objective To investigate the effect of blastocyst transfer liquid volume on mouse embryonic distribution and pregnancy prognosis. Methods Through real - time quantitative PCR and immunoblotting techniques, the effect of different blastocyst transfer liquid volumes on the expression of leukemia inhibitory factor （Lif） and HoxA10 was observed. Results With 3 μL and 5 μL blastocyst transfer liquid, the embryo implantation rates were （ 83.4 ± 10. 9） % and （74. 8± 10. 7）%, respectively; with well- spaced embryonic distribution and well developed embryos; which was significantly reduced with 10 μL blastocyst transfer liquid [ （35.0 ± 17.5） %, P 〈 0. 01 ±, with aberrantly spaced embryos and miscarried embryos. With 10 μL blastocyst transfer liquid, the Lif mRNA and HoxA10 proteins were significantly down - regulated compared with 3 μL group （ P 〈 0. 01 ）. However, no significant difference was observed between 5 and 3 μL groups （P 〉 0. 05）. Conclusion When mouse blastocysts are transferred in vitro, 3 μL blastocyst transfer liquid benefits a uniform embryo distribution and development.