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目的:以小鼠为模型,评估卵细胞胞质内单精子注射(ICSI)技术的安全性。方法:模拟临床ICSI技术,通过孤雌激活、免疫荧光、胚胎移植、早期植入点检测、中期顶臀径检测等实验方法综合评估该技术。结果:ICSI导致小鼠植入前胚胎发育能力显著下降,尤其是8细胞之后的胚胎发育率下降极显著(P<0.01),而且在ICSI胚胎的雄原核上检测到异常的H3K9双甲基化荧光。进一步检测移植后的ICSI胚胎发育情况,结果显示,与对照(正常受精)相比,E5.5 d早期植入率无显著差异(P=0.6),但是E9.5 d的“正常胚胎”百分率却显著降低(P<0.01),即使在这些“正常的ICSI胚胎”中也出现了高比例的发育滞后现象。结论:ICSI有可能通过对雄原核H3K9双甲基化的影响进而影响胚胎生长发育。
OBJECTIVE: To evaluate the safety of intracytoplasmic sperm injection (ICSI) technique in mouse as a model. Methods: To simulate clinical ICSI technique, this technique was comprehensively evaluated by experimental methods such as parthenogenetic activation, immunofluorescence, embryo transfer, early implantation point detection and mid-peak hip-hop detection. RESULTS: ICSI resulted in a significant decrease in embryo development before implantation, especially after 8-cell embryo development (P <0.01), and abnormal H3K9 dimethylation was detected on the pronucleus of ICSI embryos Fluorescence. Further examination of ICSI embryo development after transplantation revealed no significant difference in early implantation rates at E5.5d compared to control (normal fertilization) (P = 0.6), but at E9.5d, “normal embryo ”The percentages were significantly lower (P <0.01), with a high percentage of developmental lag even in these“ normal ICSI embryos. ” Conclusion: It is possible that ICSI may affect the growth and development of embryos through the influence on the methylation of H3K9 in pronucleus.