重组人钙调素对小鼠胚胎发育及凋亡的影响

Effect of recombinant human calmodulin on development and apoptosis of mouse preimplantation embryo in vitro

目的探讨外源重组人钙调素(rhCAM)对小鼠早期胚胎体外发育及凋亡的影响。方法小鼠超促排卵,收集2-细胞胚胎置于含不同浓度rhCAM的IVF-30培养液中,观察并记录胚胎形态,TUNEL法检测囊胚细胞数及囊胚细胞凋亡情况。结果20μg/ml rhCAM组的胚胎体外培养48、72 h的囊胚形成率和孵化囊胚率均高于对照组,该组体外培养72 h囊胚细胞凋亡指数显著低于对照组。200μg/ml rhCAM组的胚胎囊胚形成率低于对照组,凋亡指数显著高于对照组。结论一定浓度的rhCAM可促进胚胎体外发育并可抑制胚胎细胞的凋亡。

Objective： To evaluate the effect of recombinant human calmodulin （rhCAM） on the development and apoptosis of mouse preimplantation embryos in vitro. Methods.. Kunming female mice were super-ovulated with PMSG followed by hCG 48 hours later and mated. Forty hours after hCG treatment, two-cell mouse embryos were collected and cultured in the IVF- 30 media supplemented with rhCAM at different concentrations （0 μg/ml, 0.2 μg/ml, 2.0 μg/ml, 20 μg/ml and 200 μg/ml）. Embryo development was recorded every 24 hours. TUNEL method was used to detect the cell number and apoptosis in each blastocyst. Results. The rates of blastocyst formation and hatching in the group of 20 μg/ml rhCAM were markedly higher than those in the control group （0μg/ml rhCAM. ）. A lower apoptotic index in the blastocysts cultured for 72 hours in the group of 20 μg/ml rhCAM was also observed when compared with that in the control group. However, at the concentration of 200μg/ml rhCAM, the percentage of blastocyst formation and hatching was significantly lower than that in the control group, and a higher apoptotic index was observed in the blastocysts cultured for 72 hours in the presence of 200 μg/ml rhCAM. Conclusion： rhCAM at certain concentration can promote the development of mouse preimplantation embryo in vitro and suppress apoptosis of embryos.