兔胚胎-胎仔发育毒性试验中环磷酰胺给药方式的探讨

Administration mode of cyclophosphamide for the embryo-fetal developmental toxicity test in rabbits

目的探讨环磷酰胺不同给药途径、给药剂量对妊娠家兔胚胎及胎仔的影响,以确定诱导胎兔畸形最佳的给药方式。方法孕兔分为生理盐水对照C组、静脉注射Y1组(15 mg/kg)、皮下注射低剂量Y2组(20 mg/kg)和皮下注射高剂量Y3组(30 mg/kg),各组均在GD10~13(受孕当天为GD0)分别按相应途径给药。GD28解剖,检查黄体数、着床数、连胎子宫重、吸收胎率;取胎仔,检查胎仔性别、身长、尾长、活胎数、死胎数,并按要求进行外观畸形、内脏畸形和骨骼畸形的检查。结果环磷酰胺静脉注射组及皮下注射低剂量组孕兔胎仔均出现了畸形,两组外观畸形率分别为30.77%和95.65%,骨骼畸形率分别为7.69%和73.91%,内脏畸形率分别为20.51%和47.83%。皮下注射高剂量组吸收胎发生率为100%。结论孕兔在GD10~13皮下注射环磷酰胺20 mg/kg可用作家兔胚胎-胎仔毒性发育试验的阳性给药方法。该方法给药周期短,操作方便,吸收胎少,胎仔畸形率高,可为同行选择合适的兔胚胎-胎仔发育毒性阳性模型提供依据。

Objective To explore the effects of cyclophosphamide administered by different routes or in different doses on the embryo-fetal development in pregnant rabbits, and to determine the optimal mode of cyclophosphamide administration to induce fetal malformation. Methods Pregnant rabbits were divided into control group C （saline） , group YI （intravenous injection of 15 mg/kg cyclophosphamide,）, group Y2 （subcutaneous injection of low dose cyclophosphamide, 20 mg/kg） , and group Y3 （subcutaneous injection of high dose cyclophosphamide, 30 mg/kg）. Each rat was administrated according to the corresponding mode once daily on GDIO - 13. The day of conception was designated as GDO. The pregnant rabbits were sacrificed and dissected on GD28. Then, the number of corpora lutea and implantation, the weight of uterus with contained fetus, and fetal resorption rate were detected, the fetuses were removed and the fetal sex, body length, tail length, the number of live births and stillbirths were recorded, and the appearance of deformities, visceral deformities and skeletal malformations were detected. Results Pregnant rabbit fetuses in the cyclophosphamide intravenous injection group and subcutaneous injection of low dose cyclophosphamide group showed deformities. The appearance malformation rates in the two groups were 30. 77% and 95.65% , the skeletal deformity rates were 7.69% and 73.91% , and the visceral abnormality rates were 20. 51% and 47.83 % , respectively. The fetal resorption rate in the high dose cyclophosphamide subcutaneous injection group was 100%. Conclusions Subcutaneous injection of 20 mg/kg eyclophosphamide to pregnant rabbits at GD10 - 13 can be used as a positive administrationmethod for rabbit embryo-fetal developmental toxicity test. Thismethod has the advantages of short administration period, easy operation, few fetus resorption, and high rate of fetal malformation, thus, providing the evidence for selection of appropriate model of rabbit embryo-fetal developmental toxicity.