化疗损伤性早发性卵巢功能不全小鼠模型的建立  

Establishment of chemotherapy-induced premature ovarian insufficiency model in mice

作　　者：刘娟[1] 谭宗建[1] 彭维晖[2] 雷薇[1] 刘人杰 黄盛文[3] Liu Juan;Tan Zongjian;Peng Weihui;Lei Wei;Liu Renjie;Huang Shengwen(Department of Reproductive Medicine,Guizhou Provincial People’s Hospital,Guizhou,Guiyang 550002,China;Department of Otolaryngology and Head-Neck,Guizhou Provincial People’s Hospital,Guizhou,Guiyang 550002,China;Prenatal Diagnosis Center,Guizhou Provincial People’s Hospital,Guizhou,Guiyang 550002,China)

机构地区：[1]贵州省人民医院生殖医学科,贵州贵阳550002 [2]贵州省人民医院耳鼻咽喉头颈外科,贵州贵阳550002 [3]贵州省人民医院产前诊断中心,贵州贵阳550002

出　　处：《发育医学电子杂志》2023年第5期346-351,共6页Journal of Developmental Medicine (Electronic Version)

基　　金：国家自然科学基金(82260299)。

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�Objective To establish a mouse model of chemotherapy-induced premature ovarian insufficiency(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 the last intraperitoneal injection,serum anti-Müllerian hormone(AMH),follicle stimulating hormone(FSH)and estradiol(E2)were detected by enzyme linked immunosorbent assay.Bilateral ovarian tissues were stained with hematoxylin and eosin to count follicles of different levels.The one-way analysis of variance and LSD-t test were used for statistical analysis.Result By the end of administration,two mice in the 10-day group died,while all mice in the other groups survived.Before intraperitoneal injection,there were no significant differences in body weight among all groups(P>0.05).When collecting specimens,the body weight of mice in 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 that in the control group[(20.28±0.61)g,all P<0.001)].After withdrawal of cisplatin,the estrous cycle in the 7-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

关 键 词：早发性卵巢功能不全 顺铂 化疗 动物模型 

分 类 号：R730.53[医药卫生—肿瘤] R-332[医药卫生—临床医学]