| About 30-70% of embryos are abortive in the process of mammal embryonic development, most occur in early stage. Lots of developmental abnormalities of cells occur in embryogenesis in vitro. Cell apoptosis is an important way to remove the developmental abnormalities of cells. Research shows the clearance of the developmental abnormal cells in early embryos also realize by apoptosis. Apoptosis is more universal in preimplantation embryos, even in normal embryos. Apoptosis is a strictly controlled process of multiple genes, and apoptotic occurrence is the direct result of the expression of some apoptosis gene. In order to improve the survival ability of embryos in vitro, to explore by changing conditions of culture to add the apoptosis inhibitor to inhibit apoptosis of embryonic cells.It is found that apoptosis of embryos can be observed from morphological change. It is caused by changes of cytoskeleton, which involve the function of Rho associated kinase (ROCK). Y-27632 is widely used as specific inhibitor to Rho associated kinase. In recent years, it was found that signal conductions pathways in Rho associated kinase plays an important role in embryogenesis. It has been reported that Y-27632 has an effect on embryos development, and none has shown any consistent benefit but some statistics and data on the developmental rates of embryos, while it doesn't go further into the details of the molecular mechanism in embryo by expatiating the role of Y-27632. The change of apoptosis gene expression in early embryogenesis is of great significance to mechanism.This experiment is adopted morphological observation, TUNEL detection, real-time fluorescence quota PCR the method, respectively. This paper introduces specific inhibitor to Rho associated kinase Y-27632 cultured in vitro, which affect cell apoptosis of mouse early embryos and apoptosis gene, such as Bcl-2 family(Bax,Bcl-2). By analysis of the experimental results, Y-27632 may regulate expression of Bcl-2 family, and then change the quality of mouse embryos.Objective to observe the morphology, 10μM concentrations Y-27632 is the most suitable for mouse early embryogenesis. Comparing with control group, after 1 and 2 days two cell embryonic cells culture, morulas and blastocyst development rate have no significant difference(P>0.05). By embryonic cells account after 2 and 3 days culture, respectively, further investigation shows that The rate from the experiment group were significantly higher than the control group(P<0.05)two days later; but not significant difference(P>0.05) three days later. Experimental results of transplant recipients on both experimental group and control group show that the rate of birth has no significant difference (P>0.05). The results show that Y-27632 has some influence on the increase of early embryogenesis in number.By using TUNEL apoptosis histology staining in kit, both experimental group and control group showed no signs of apoptosis after 2 days culture, while after 3 days, The results of staining assay indicated that apparent apoptosis had taken place in experimental group. Experimental results indicate that embryos cultured short-term in Y-27632 are not apoptosis, while long-term culture lead to apoptosis.It is found that Y-27632 evidently affects Bax and Bcl-2 expression in the process of mouse early embryogenesis using real-time fluorescence quota PCR. After three days two cell embryonic cells cultured in 10μM concentrations Y-27632, then the expression of Bax was increased, while Bcl-2 reduced, it shows that long-term embryonic culture lead to cell apoptosis.
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