控制性超促排卵大鼠模型的建立

Establishment of Controlled Ovarian Hyperstimulation Rat Model

目的选择理想的符合临床实际的控制性超促排卵(COH)大鼠模型,为后续的实验提供良好的实验基础。方法将75只大鼠随机平均分为正常组(N),模型10组(M10),模型20组(M20),模型40组(M40)和模型100组(M100),每组各15只。各模型组分别于动情期后一日的下午5:00给予腹腔注射孕马血清促性腺激素(PMSG)10单位、20单位、40单位和100单位,并于48 h后,再分别给予腹腔注射人绒毛膜促性腺激素(HCG)10单位、20单位、40单位和100单位,注射完后立即合笼,以次日行阴道涂片见大量精子记为怀孕1 d。在第8天,行剖宫探查,记录大鼠的妊娠情况和着床的胚泡数。另取25只大鼠,按上述方法随机分对照组和不同剂量干预组,每组5只,各组大鼠在妊娠第5天取材,处理后观察子宫内膜胞饮突和LIF蛋白的表达。结果各组大鼠交配率无统计学差异(P>0.05)。与N组相比,M10组大鼠怀孕率无明显降低(P>0.05),M20、M40和M100组大鼠怀孕率明显降低(P<0.01);和N组相比,M10组大鼠胚泡着床位数无明显增加(P>0.05),M20、M40和M100组大鼠胚泡着床数明显增高(P<0.05)。和N组相比,M10大鼠子宫内膜LIF蛋白表达量无明显差异(P>0.05);M20、M40和M100组大鼠子宫内膜LIF蛋白表达量明显降低(P<0.05);M40和M100组大鼠子宫内膜LIF蛋白表达量比M20组更少,但差异无统计学意义(P>0.05)。N组大鼠在妊娠第5天子宫内膜腺上皮细胞表面胞饮突丰富;M20大鼠子宫内膜腺上皮细胞表面几乎未见明显胞饮突表达。结论M20组大鼠妊娠率较正常组明显降低,且平均胚泡数较高,胞饮突提前出现,LIF表达较正常组降低,是较为符合临床实际的理想模型。

Objective To select an ideal controlled ovarian hyperstimulation(COH)rat model that is in line with clinical practice,and provide a good experimental basis for the follow-up experiment.Methods Seventy-five rats were randomly divided into normal group(N),model 10 group(M10),model 20 group(M20),model 40 group(M40)and model 100 group(M100),with 15 rats in each group.Each model group was given intraperitoneal injection of 10 IU,20 IU,40 IU and 100 IU of pregnant horse serum gonadotropin(PMSG)at 5:00 pm on the day after the estrus period.After 48 hours,the rats were given intraperitoneal injection of 10 IU,20 IU,40 IU and 100 IU of human chorionic gonadotropin(HCG),and immediately caged.The large number of sperm found on the vaginal smear the next day was recorded as the first day of pregnancy.On the next day,vaginal smear of seeing a large number of sperms was treat as the first day of pregnancy(D1).On the D8 days,record the pregnancy rate and the number of implanted blastocysts.Another 25 rats were randomly divided into 5 groups as above.The rats in each group were mated and collected the samples on the D5 days.The expression of the pinopodes and the LIF protein expression were observed.Result There was no significant difference in the mating rate between the five groups(P>0.05).Compared with N group,the pregnancy rate of M10 group was not significantly lower(P>0.05),while the pregnancy rate of M20,M40 and M100 groups were significantly lower than that of N group(P<0.01).The number of blastocysts of M20,M40 and M100 groups was significantly increased than that of N group(P<0.05).Compared with N group,the endometrial expression of LIF protein in Day 5 of M10 group was not significantly lower(P>0.05);the LIF protein in M20,M40 and M100 groups was significantly lower(P<0.05);the LIF protein in M40 group and M100 group was less than that in M20 group,but the difference was not statistically significant(P>0.05).The result of electron microscopy showed that there were abundant pinopodes in endometrial glandular epithelial cells of N group in Day 5,while the pinopodes was almost not expressed in the M20 group.Conclusion The pregnancy rate of rats in the M20 group was significantly lower than that in the normal group,and the average number of blastocysts was higher,the pinocytosis appeared earlier,and the expression of LIF was lower than that in the normal group,which was an ideal model in line with clinical practice.