摘要
目的 建立孕激素功能不足导致的自然流产小鼠模型,为自然流产发生机制以及孕期补充孕激素临床规范的研究提供理想的动物模型.方法 成年雌性小鼠随机分成米非司酮（RU486）高剂量组（50 μg/100μL丙二醇）、中剂量组（25 μg/100 μL丙二醇）、低剂量组（10 μg/10 μL丙二醇）和溶剂对照组（100 μL丙二醇/只）,于妊娠8d经颈背部皮下注射RU486或溶剂后,分别于妊娠10d和分娩后观察妊娠状况和窝产仔数;于妊娠10d取各组小鼠外周血和子宫组织,对各组小鼠的血清雌二醇（E2）和孕酮（P4）水平进行检测;应用基因芯片技术,对实验组与对照组间的子宫组织差异表达基因进行筛查分析,并通过Western blot检测,对其中3个差异表达基因进行了验证.结果 高、中和低剂量组流产率分别为100％、46.0％和13.9％,且中剂量组妊娠鼠子宫内均为停育被吸收的胚胎,而低剂量组的仔鼠出生体质量显著小于对照组（P＜0.05）;与对照组相比,中剂量组妊娠鼠血清E2水平升高（P＜0.05）、P4水平降低（P＜0.001）,而低剂量组妊娠鼠血清中E2水平降低（P＜0.05）、P4水平升高（P＜0.05）.经基因芯片检测和Western blot验证表明,与对照组相比,细胞周期蛋白依赖性激酶4（CDK4）基因在中剂量组子宫组织中的表达量显著降低（P＜0.05）,促细胞凋亡因子（Bax）基因的表达量也显著性降低（P＜0.05）,而细胞周期蛋白D2（CCND2）基因的表达量则显著升高（P＜0.05）.结论 适当剂量的RU486可诱导胚胎着床后妊娠鼠出现妊娠丢失或新生仔鼠体质量降低的表型,可能成为一种新的可用于研究人妊娠期孕酮功能不足所致的流产或宫内生长受限等病理妊娠发生机制的动物模型.
Objective To establish spontaneous abortion mouse models of progesterone insufficiency induced by mifepristone （RU486） to provide an ideal animal model for the mechanism of spontaneous abortion and clinical practice of supplying progesterone in pregnancy period. Methods Adult female mice were randomly divided into a RU486 high-dose group （50 μg/100 gL propylene glycol/only）, middledose group （25 μg/100 gL propylene glycol/only）, low-dose group （10 μg/100 μL propylene glycol/only） and solvent control group （100 μL propylene glycol/only）. A subcutaneous injection of RU486 or solvent was given on pregnancy day 8, and the phenotype of pregnant mice and neonatal mice were observed on pregnancy day 10（d10） and after their delivery. The peripheral blood and uterus tissues of the mice in each group were collected on d10 to detect the levels of their serum estradiol （E2） and progesterone（P4）. The uterine tissue differentially expressed genes between the experimental and control groups were screened and analyzed by Gene Chip Technology, and three of them were validated by Western blot. Results The abortion proportions were 100%, 46.0% and 13.9% respectively in the high-dose group, middle-dose group and lowdose group, and some embryos stopped developing and were absorbed in the uterus were observed in the middle-dose group, whereas the newborn mouse birth weights in the low-dose group were significantly lower than those in the control group （P〈0.05）. Compared with those of the control group, the E2 levels of the pregnant mice serum of the middle-dose group were increased （P〈0.05） and P4 levels were decreased （P〈0.001）, while the E2 levels of the pregnant mice serum of the low-dose group were decreased （P〈0.05）, and P4 levels were increased （P〈0.05）. The Gene Chip and Western blot analysis and verification discovered that the expression of CDK4 in the uterine tissues was significantly reduced in the middle-dose group （P〈0.05） compared with the control group, the expression of Bax was also significantly decreased （P〈0.05）, whereas the expression of CCND2 significantly increased and was statistically significant （P〈0.05）. Conclusion The appropriate dose of RU486 able to induce the pregnancy loss of pregnant mice after embryo implantation or neonatal weight loss. It might become a new animal model to be used for study of pathological mechanisms of the abortion or intrauterine growth restriction induced by progesterone deficiency in women pregnancy period.
出处
《实验动物与比较医学》
CAS
2016年第5期327-333,共7页
Laboratory Animal and Comparative Medicine
