两种造模方法导致大鼠卵巢早衰模型的比较研究

Comparative study between two methods for premature ovarian failure model establishment in rats

目的探讨两种不同方案制造卵巢早衰（POF）大鼠动物模型的差异，以寻找合适的动物模型制备方法，为POF的治疗提供理论依据。方法选择9—10周龄雌性Wistar大鼠60只为研究对象，随机分为实验造模A组、实验造模B组和对照组各20只；实验造模A组采用环磷酰胺给药方案，首次给予负荷剂量50mg／kg，以8mg·kg^-1·d^-1的剂量连续腹腔注射14d；实验造模B组给药方案采用一次性腹腔注射环磷酰胺120mg／kg联合皮内注射白消安12mg／kg。结果实验造模A、B组大鼠均可见总卵泡数、次级和窦状卵泡数明显减少，闭锁卵泡增多、卵巢间质出现纤维化，卵巢功能损伤；实验造模A组动情周期改变发生在注射药物7—10d左右，实验造模B组动情周期改变发生在结束注射后第2天。实验前后实验造模A组的大鼠体质量减少明显，相对质量小于对照组（P〈0．05）；实验造模B组大鼠体质量减少不明显（P〉0．05）；实验造模A、B组大鼠卵巢质量均小于对照组（P〈0．05）。实验造模A、B组卵泡刺激素（FSH）含量均高于对照组，雌二醇（E2）均低于对照组（P〈0．05）；但实验造模A、B组之间差异无统计学意义（P〉0．05）。实验造模A组死亡率高于实验造模B组（P〈0．05）。结论一次性腹腔注射环磷酰胺120mg／kg联合皮内注射白消安12mg／kg造模法成模时间短、简便易行、成功率高、死亡率低，是建立化疗所致POF动物模型的一种较好选择。

Objective To compare the difference between two plans for premature ovarian failure （POF） model establishment in rats, in order to figure out suitable method for animal model establishment and provide theoretical basis for POF treatment. Methods A total of 60 rats aged from nine to ten weeks were included in this research. THe rats were randomly assigned into experimental group A for model establishment, experimental group B for model establishment, and control group, 20 rats in each group. In experimental group A for model establishment, cyclophosphamide （CTX） was administrated via intraperitoneal injection： the load dose at the first time was 50 mg/kg, followed by 8 mg· kg^-1· d^-1 for 14 days continually. In experirrrental group B for model establishment, dosage regimen was CTX intraperitoneal injection at one time with the dose of 120 mg/kg combined with intracutaneous injection of busulfan with the dose of 12 mg/kg. Results The two experimental groups showed obviously decrease in total number of follicles, subprime and sinus follicles, increase in atresia follicles, and ovarian interstitial fibrosis, ovarian function damage; in experimental group A for model establishment, change in estrous cycle occurred at 7-10 days after injection, while in experimental group B for model establishment, estrous cycle change occurred after 2 days of injection. In experimental group A for model establishment, body weight of rats decreased after treatment, and relative quality was less than that in control group （ P〈0. 05 ） ; in experimental group B for model establishment, decrease in rats body weight was not obvious （ P〉0. 05 ） ; in the two experimental groups, ovary quality of rats was less than that in control group （ P〈0. 05 ） . The levels of follicle-stimulating hormone （FSH） in the two experimental groups were higher than those in control group, and estradiol （ E2 ） levels in the two experimental groups were significantly lower than those in control group （P〈0. 05 ） ; however, there was no significant difference between the two groups. Mortality in group A for model establishment was higher than that in group B （P〈0. 05） . Conclusion One time intraperitoneal injection of CTX with the dose of 120 mg/kg combined with intraeutaneous injection of busulfan with the dose of 12 mg/kg can establish animal model in relatively short time, and it is easy to perform with high successful rate and low mortality. And it is a better selection for animal model establishment for chemical therapy induced POF.