摘要 【摘要】 目的 探讨腹腔内注射顺铂建立化疗损伤性早发性卵巢功能不全（premature ovarian insufficiency，POI）小鼠模型的方法。 方法 采用7~8 周龄C57BL/6J 雌性小鼠40 只，随机分为4 组，每组10 只。实验组腹腔注射顺铂2.5 mg/（kg·d），分为顺铂4 d 组、7 d 组和10 d 组；对照组腹腔注射生理盐水连续10 d。停药后每天进行阴道脱落细胞涂片，监测动情周期。停药后14 d，采用酶联免疫吸附试验测定血清抗米勒管激素（anti-Müllerian hormone，AMH）、卵泡刺激素（follicle stimulating hormone， FSH）及雌二醇（estradiol，E2）。取双侧卵巢，苏木精- 伊红染色计数各级卵泡。统计学方法采用单因素 方差分析和独立样本t 检验。 结果 至给药结束时，顺铂10 d 组小鼠死亡2 只，其余组小鼠未出现死亡。 腹腔注射前，各组间小鼠体质量差异无统计学意义（P 值均>0.05）；取材时，顺铂4 d 组、7 d 组及10 d 组 小鼠的体质量［（17.43±0.53）、（14.03±0.79）、（12.10±0.47）g］均低于对照组［（20.28±0.61）g］，差异有统计学意义（P 值均<0.001）。停药后，顺铂7 d 组和10 d 组动情周期［（8.30±1.34）、（9.63±1.06）d］均长于对照组［（4.90±0.74） d］，差异有统计学意义（P值均<0.001）。停药14 d时血清性激素水平：与对照组比较，顺铂4 d 组、7 d 组及10 d 组的AMH 水平［（13.31±0.68）、（7.04±0.59）、（5.62±0.86）μg/L］及E2 水平［（64.76±2.54）、（40.78±1.66）、（33.95±2.49）ng/L］均下降（P 值均<0.05）；顺铂7 d 组及10 d 组的FSH水平［（36.49±2.81）、（41.92±2.28）μg/L］均升高（P 值均<0.001）。停药14 d 时卵泡计数：与对照组比较，顺铂4 d 组的窦卵泡数量［（8.1±1.2）个］ 减少（P<0.05）；顺铂7 d 组及10 d 组的原始卵泡［（25.4±2.7）、 （25.0±3.2）个］、初级卵泡［（23.2±2.5）、（22.5±3.7）个］、次级卵泡［（10.4±1.8）、（10.1±1.3）个］及窦卵泡［（7.7±1.8）、（7.6±1.4）个］均下降（P<0.05），闭锁卵泡［（9.2±1.8）、（9.5±1.8）个］明显增多（P<0.01）。 结论 小鼠连续7 d 腹腔内注射顺铂2.5 mg/（kg·d），可以有效建立化疗损伤性POI 小鼠模型，造模方法简便，且能较好地模拟POI 患者的卵巢状态及激素水平。
Abstract: 【Abstract】 Objective To establish a mouse model of chemotherapy-induced premature ovarianinsufficiency (POI) by intraperitoneal injection with cisplatin. Method Forty female C57BL/6J mice aged 7-8 weeks were randomly divided into four groups, with ten mice in each group．The mice in experimental groups were intraperitoneally injected with cisplatin 2.5 mg/(kg·d), and divided into 4-day, 7-day, and 10-day groups. The mice in the control group were intraperitoneally injected with normal saline for ten days. After the medicine withdrawal, vaginal smears were collected daily to monitor estrous cycles. 14 days after thelast intraperitoneal injection, serum anti-Müllerian hormone (AMH), follicle stimulating hormone (FSH) andestradiol (E2) were detected by enzyme linked immunosorbent assay. Bilateral ovarian tissues were stainedwith hematoxylin and eosin to count follicles of different levels. The one-way analysis of variance and LSD-ttest were used for statistical analysis. Result By the end of administration, two mice in the 10-day groupdied, while all mice in the other groups survived. Before intraperitoneal injection, there were no significantdifferences in body weight among all groups (P>0.05). When collecting specimens, the body weight of micein 4-day, 7-day and 10-day groups [(17.43±0.53), (14.03±0.79) and (12.10±0.47) g] were lower than thatin the control group [(20.28±0.61) g, all P<0.001)]. After withdrawal of cisplatin, the estrous cycle in the7-day group and 10-day group [(8.30±1.34) and (9.63±1.06) d] was longer than that in the control group[(4.90±0.74) d, all P<0.001)]. 14 days after the end of cisplatin administration, AMH levels [(13.31±0.68),(7.04±0.59) and (5.62±0.86) μg/L] and E2 levels [(64.76±2.54), (40.78±1.66) and (33.95±2.49) ng/L]in 4-day, 7-day and 10-day group were lower than those in the control group (all P<0.05), while FSH level[(36.49±2.81) and (41.92±2.28) μg/L] in 7-day and 10-day group were higher than that in the control group(all P<0.001). 14 days after the end of cisplatin administration, the number of antra follicles in 4-day group(8.1±1.2) was lower than that in the control group (P<0.05); the number of primordial follicles in 7-day and10-day group (25.4±2.7 and 25.0±3.2), primary follicles (23.2±2.5 and 22.5±3.7), secondary follicles(10.4±1.8 and 10.1±1.3) and antra follicles (7.7±1.8 and 7.6±1.4) were lower than those in the controlgroup (all P<0.05); and the number of atretic follicles in 7-day and 10-day group (9.2±1.8 and 9.5±1.8)were higher than those in the control group (P<0.01). Conclusion A 7-day continuously intraperitonealinjection with cisplatin 2.5 mg/(kg·d) could effectively establish the chemotherapy-induced POI mousemodel. The method was simple and could simulate the ovarian status and hormone levels of POI patients.
2023, Vol. 11 
