| This study was mainly approached the antioxidation effect of adding Baicalin(Bai), Ligustrazine(Lig)and Berberine(BR)in mCZB respectively on 2-cell mouse embryos in vitro culture ,and comparedthe effect of froze-thaw, pregnancy rate and litter rate after embryos transfer etc. in order to furtherinvestigate the value of these three Chinese Medicine effective constituents on improving themicro-environment of preimplantation embryos in vitro culture on mouse and enhancing the embryoquality and the developmental potential of embryos after embryos transfer , which can provide valuabledata and access to enhance the mammalian embryo culture medium composition and culture condition.The group added antibiotics to mCZB was regarded as the control, while the others supplemented with0.1μg/ml BR, 0.5μg/ml Lig and 4μg/ml Bai respectively as experiment groups in the first experiment tocompare the rates of hatched embryos, the contents of nitric oxide(NO) and malondialdehyde(MDA) in themicrodrop, and the cell amount of hatched embryos. As a result, the rates of hatched embryos from thethree groups of Bai ( 80.2% , 81.3% ), Lig (78.9%,77.6% )and BR (76.7% ,71.6%) was significantly higher(P<0.01) than control group (50.7%,47.8%) . The content of NO decreased at 72h as compared with that of24h in every experiment group, but it increased to the highest point at 120h. As a whole, raise trend wasevident for the content of MDA in every group at 72h and 120h, and the content of MDA in the control washigher than that in experiment groups at 120h except for Bai group. According to the cell amount ofhatched embryos, the difference between experiment groups(BR(87.2±8.6), Lig(83.9±7.7) ,Bai(81.9±6.2))and control group(77.4±5.6)were extremely obvious (P<0.01). The result showed that these three kinds ofChinese Medicine effective constituents could efficiently promote early embryos development and increasethe cell amount of hatched embryo, and the contents of NO and MDA were affected as well in vitro cultureon mouse.Based on the first experiment, the second one in which each group were conserved throughconventional freezing process from 2-cell embryo in vitro culture to the stage of morula and blastocyst wasoriented to compare the effects of froze-thaw in each group and the difference between the two methods.The results of morula and blastocyst showed that in the first procedure the development rate of blastocystcultured for 8～14 hours after froze-thaw in the Bai(65.8%)was superior(P<0.05)to that of the control(48.6%)and the BR(46.7%), and the Lig(81.1%)was significantly higher (P<0.01) than that ofthe control and the BR(46.7%). In the second procedure, the Ba(i80.6%)was significantly higher (P<0.01)than the control(55.2%)and the BR(47.6%)in vitro culture. Through analysis of the development rate ofblastocyst cultured for 6～8 hours in the first procedure, there was no significant difference(P>0.05)ineach group. In the second procedure, there was no significant difference(P>0.05)in each group, but theeffect of Chinese Medicine was superior(P<0.05)to that of control group(61.2%)in vitro culture. Theresults indicated there was no significant difference(P>0.05)in the different procedures. Bai and Lig cansignificantly enhance the development quality of embryos and the development rate of freezing embryos invitro culture.Based on Experiment two, froze-thaw embryos were cultured to the stage of blastula and thentransferred to uterus in experiment three. As a result, the pregnancy rate from the three groups of Bai(56.3% ),Lig (52.6% )and BR (55.0%) was higher than that of control group (46.2%),but there was notobvious difference between experiment groups and control group;the birth rate from the three groups ofBai (63.3% ), Lig(67.3% )and BR (60.5%) was higher than control group (52.8%),but there was notsignificant difference either.These results indicated the three Chinese Medicine effective constituents(Bai, Lig , BR)can distinctlyimprove the quality of 2-cell mouse embryo in vitro culture and be propitious to the developmentalpotential of embryos following embryos transfer after froze-thaw embryos.
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