论文部分内容阅读
目的:探讨Bax-小干扰RNA(small interfering RNA,siRNA)和Bak-siRNA对小鼠胚胎发育及凋亡的效应。方法:通过对体外培养小鼠早期胚胎激光打孔,完成Bax-siRNA(Bax-siRNA组)、Bak-siRNA(Bak-siRNA组)和Bax-Bak-siRNA(Bax-Bak-siRNA联合组)的转染,并以阴性siRNA和纯水为阴性对照组和空白对照组,对确定转染效果的胚胎观察形态变化、生长发育状况及囊胚形成率,应用Hoechst 33342和碘化丙啶的方法分析各转染组小鼠早期胚胎细胞凋亡和增殖程度以及凋亡颗粒率。结果:鼠胚激光打孔后可成功转染Bax-siRNA、Bak-siRNA和Bax-Bak-siRNA。Bak-siRNA组和Bax-Bak-siRNA联合组鼠胚的囊胚形成率显著高于阴性对照组(P<0.01),而且凋亡颗粒率显著低于阴性对照组(P<0.01)。Bax-siRNA组的囊胚形成率和凋亡颗粒率与阴性对照组比较均无统计学差异(P>0.05)。结论:Bak-siRNA和Bax-Bak-siRNA可以改善小鼠胚胎的囊胚形成率,减少胚胎细胞的凋亡。
Objective: To investigate the effect of small interfering RNA (siRNA) and Bak-siRNA on mouse embryonic development and apoptosis. Methods: Bax-siRNA (Bax-siRNA group), Bak-siRNA group (Bak-siRNA group) and Bax-Bak-siRNA group (Bax-Bak-siRNA group) were completed by laser drilling of early embryos in vitro. Transfection, negative control and pure water as negative control group and blank control group, to determine the morphological changes of transfection embryos, growth and development and blastocyst formation rate, the application of Hoechst 33342 and propidium iodide method analysis The apoptosis and proliferation of early embryos and the rate of apoptotic granules in each transfection group were observed. Results: Bax-siRNA, Bak-siRNA and Bax-Bak-siRNA were transfected successfully after laser drilling. The blastocyst formation rate of Bak-siRNA group and Bax-Bak-siRNA group was significantly higher than that of negative control group (P <0.01), and the apoptotic granulocyte rate was significantly lower than that of negative control group (P <0.01). The blastocyst formation rate and apoptotic granulocyte rate in Bax-siRNA group were not significantly different from those in negative control group (P> 0.05). Conclusion: Bak-siRNA and Bax-Bak-siRNA can improve the blastocyst formation rate of mouse embryos and reduce the apoptosis of embryonic cells.