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目的:研究不同浓度的白芸豆提取物PHA(植物凝集素)对小鼠胚胎体外发育的影响。方法:实验一,采用添加不同浓度PHA的M16培养液培养小鼠2-细胞期胚胎,观察72 h后的发育情况,记录囊胚及各阶段胚胎的数目。实验二,用与实验一相同浓度的PHA预处理从1-细胞到囊胚不同发育阶段的胚胎24 h,然后移入不含PHA的培养液中继续培养,观察到囊胚或孵化囊胚的发育率。结果:含低质量浓度PHA的培养液有促进小鼠胚胎发育的作用,即50,100 mg·L-1的PHA可显著增加囊胚的数量。而含高质量浓度PHA(>1 000 mg·L-1)的培养液会使胚胎发育停滞在1-细胞到囊胚的不同时期,并随浓度增加表现为副凋亡或致死现象。结论:不同浓度PHA可对小鼠胚胎的体外发育具有不同的作用。1-细胞阶段的胚胎对PHA处理具有较高的敏感性。在作用时间上,24 h PHA预处理即可表现出对胚胎发育的促进或抑制作用。低浓度的PHA可促进胚胎发育,而高浓度的PHA则导致小鼠胚胎发育停滞,出现副凋亡或死亡。
Objective: To study the effects of PHA (phytohemagglutinin) extracted from white kidney beans on the in vitro development of mouse embryos. Methods: In experiment 1, mice were cultured in M16 medium supplemented with different concentrations of PHA to observe the development of embryos at 72 h. The numbers of blastocysts and embryos at various stages were recorded. In Experiment 2, embryos from 1-cell to different stages of blastocyst were pretreated with the same concentration of PHA as Experiment 1 for 24 h and then transferred to PHA-free medium for further culture to observe the development of blastocyst or hatched blastocysts rate. Results: The medium containing low concentration of PHA could promote embryonic development in mice. The PHA at 50 and 100 mg · L -1 significantly increased the number of blastocysts. However, medium with high concentration of PHA (> 1000 mg · L -1) could retard embryonic development in different stages of 1-cell to blastocyst and showed the phenomenon of accessory apoptosis or lethality as the concentration increased. Conclusion: Different concentrations of PHA may have different effects on in vitro development of mouse embryos. The 1-cell stage embryos are highly sensitive to PHA treatment. In the role of time, 24 h PHA pretreatment can show the promotion or inhibition of embryonic development. Low concentrations of PHA can promote embryonic development, while high concentrations of PHA lead to the development of mouse embryo stagnation, the occurrence of secondary apoptosis or death.