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目的:检测Survivin在围植入期小鼠子宫内膜组织中的表达,探讨其在胚胎植入过程中的作用。方法:选择成年雌性昆明小白鼠42只,阴道脱落细胞涂片法确定动情周期。将处于动情周期的雌鼠与性成熟雄鼠合笼后次晨检查阴道分泌物涂片,有精子者记为妊娠0 d。采用免疫组织化学和RT-PCR法分别检测妊娠0,3.5,4.5,5.5,6.5,7.5及9 d各组子宫内膜组织中Survivin蛋白及mRNA水平的变化。结果:①Survivin蛋白在妊娠0 d小鼠子宫内膜组织中表达较低,其阳性产物光密度(OD)值为0.058 1。之后逐渐增高,至妊娠5.5 d达峰值为0.140 7。之后逐渐下降,至妊娠9 d降至0.074。统计学检验结果提示:Survivin蛋白在胚胎植入期(4.5 d,5.5 d,6.5 d)表达较高,其OD值与胚胎植入前期(0 d,3.5 d)及胚胎植入后期(7.5 d,9 d)的差异均有显著性(P<0.05)。②Survivin mRNA的表达在妊娠0 d小鼠子宫内膜组织中较低,其与内参相比的相对表达量为0.330 4。之后逐渐增高,至妊娠6.5 d达峰值为1.310 7。之后逐渐下降,至妊娠9 d降至0.598。统计学检验结果提示:Survivin mR-NA在胚胎植入期表达较高,其OD值与胚胎植入前期及胚胎植入后期的差异均有显著性(P<0.05)。③Survivin蛋白及mRNA在胚胎植入后期小鼠子宫内膜组织中的表达略高于胚胎植入前期,但差异无显著性(P>0.05)。结论:Survivin在胚胎植入前期和后期均低表达,植入期高表达。这种变化提示Survivin是影响胚胎植入的重要因素之一。
OBJECTIVE: To detect the expression of Survivin in endometrial tissue during peri-implantation period and to explore the role of Survivin in embryo implantation. Methods: Forty-two adult female Kunming mice were selected and the estrous cycle was determined by vaginal smear method. The estrous cycle of female mice and sexual maturity males cage the next morning to check the vaginal discharge smear, sperm were recorded as pregnancy 0 d. Immunohistochemistry and RT-PCR were used to detect the changes of Survivin protein and mRNA levels in the endometrium of each group at 0, 3.5, 4.5, 5.5, 6.5, 7.5 and 9 d. Results: ①Survivin protein was low expressed in the endometrium of mice on day 0 of pregnancy, and the OD value of the positive product was 0.058 1. Then gradually increased to reach the peak value of 0.1407 on 5.5 d of pregnancy. After that, it decreased gradually to 0.074 after 9 d of pregnancy. The results of statistical analysis indicated that Survivin protein was highly expressed during embryo implantation (4.5 d, 5.5 d and 6.5 d), and its OD value was significantly higher than that of prevital implantation (0 d, 3.5 d) and embryo implantation (7.5 d , 9 d) were significantly different (P <0.05). The expression of Survivin mRNA was lower in the endometrium of mice on day 0 of gestation, and its relative expression relative to the internal reference was 0.330 4. Then gradually increased to peak at 6.5 d of pregnancy reached 1.310 7. After that, it gradually decreased to 0.598 by the 9th day of gestation. The results of statistical analysis indicated that mR-NA of Survivin was highly expressed during embryo implantation, and the OD value of Survivin mR-NA was significantly different from that of embryo implantation and later embryo implantation (P <0.05). ③ The expression of Survivin protein and mRNA in endometrium of embryo implantation stage was slightly higher than that of preimplantation stage, but the difference was not significant (P> 0.05). CONCLUSIONS: Survivin is low in both early and late stages of embryo implantation and is highly expressed during implantation. This change suggests that Survivin is one of the important factors affecting embryo implantation.