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目的观察PeroxiredoxinⅡ对小鼠植入前胚发育的影响,并探讨其可能的机制。方法收集昆明种小鼠的1细胞胚,采用微滴培养法连续培养48h,观察不同浓度PeroxiredoxinⅡ抗体对植入前胚发育的影响,以胚胎发育到各期的比例为判断发育效果的标准。同时,应用共聚焦扫描显微镜术结合特异性荧光探针2′,7′二氯氢化荧光素双乙酸酯,检测小鼠胚中氧自由基水平。结果抗体添加组和未添加组的1细胞胚经24h培养后96%以上发育成2细胞胚,但经48h培养,抗体添加组的胚发育明显受抑,多数停滞于2细胞期。激光共聚焦成像的结果表明,PeroxiredoxinⅡ抗体添加组的2细胞胚内荧光强度明显增强。结论PeroxiredoxinⅡ抗体可引起2细胞胚发育阻滞,这可能与胚内的氧自由基水平升高有关,提示植入前胚内PeroxiredoxinⅡ可减少或消除细胞内氧自由基,促进植入前胚的发育。
Objective To observe the effect of Peroxiredoxin Ⅱ on the development of preimplantation embryos in mice and to explore its possible mechanism. Methods The 1-cell embryos of Kunming mice were collected and cultured continuously with droplet culture method for 48 hours. The effects of different concentrations of Peroxiredoxin Ⅱ antibody on the development of preimplantation embryos were observed. The ratio of embryo development to each phase was the standard for judging the developmental effect. At the same time, confocal scanning microscopy combined with specific fluorescent probe 2 ’, 7’ dichlorofluorescein diacetate, detection of mouse embryonic oxygen free radicals. RESULTS: More than 96% of the 1-cell embryos were cultured in 2 h after embryo culture. However, after 48 h of culture, the embryo development of the antibody-added group was significantly inhibited, most of which stopped at the 2-cell stage. Laser confocal imaging results showed that Peroxiredoxin II antibody group significantly increased the fluorescence intensity of 2-cell embryos. Conclusion PeroxiredoxinⅡantibodies can cause the development of 2-cell embryos, which may be related to the increased level of oxygen free radicals in embryos, suggesting that pre-implantation of PeroxiredoxinⅡ can reduce or eliminate intracellular oxygen free radicals and promote the development of preimplantation embryos .