连续重复给予含左炔诺孕酮紧急避孕药对雌性大鼠生育力及其子代健康的影响  

Effects of consecutively repeated application of emergency contraceptive pills containing levonorgestrel on female fertility and the health of F1 offspring

作　　者：胡颖怡 倪其承 钟瑞华 杨文捷 李国停 周洁芸 郭湘洁 谢淑武 周佳铃 朱焰 Hu Yingyi;Ni Qicheng;Zhong Ruihua;Yang Wenjie;Li Guoting;Zhou Jieyun;Guo Xiangjie;Xie Shuwu;Zhou Jialing;Zhu Yan(Pharmacy School,Fudan University,Shanghai 200032,China;NHC Key Lab of Reproduction Regulation,Shanghai-MOST Key Laboratory of Health and Disease Genomics,Shanghai Institute for Biomedical and Pharmaceutical Technologies,Shanghai 200232,China;Shanghai Lab.Animal Research Center,Shanghai 201203,China;Shanghai University of Medicine&Health Sciences,Shanghai 201318,China)

机构地区：[1]复旦大学药学院,上海200032 [2]上海市生物医药技术研究院国家卫生健康委生育调节药械重点实验室上海生殖健康药具工程技术研究中心,上海200032 [3]上海市实验动物研究中心,上海201203 [4]上海健康医学院药学院,上海201318

出　　处：《中华生殖与避孕杂志》2024年第6期562-573,共12页Chinese Journal of Reproduction and Contraception

基　　金：上海市自然科学基金(20ZR1448200)。

摘　　要：目的探讨连续重复给予含左炔诺孕酮(levonorgestrel,LNG)紧急避孕药(emergency contraception pills,ECPs)对雌性大鼠生育力及其F1代仔鼠健康的影响。方法成年SPF级雌性大鼠于每个动情周期连续给予3次LNG-ECPs,分别给予3个(P-3)、6个(P-6)和12个(P-12)动情周期。雌性大鼠每个给药时程下,采用Excel产生随机数按体质量分层随机分为2组,分别为LNG-ECPs组和溶媒对照组,分别灌胃给予0.12 mg/kg LNG-ECPs和等体积溶媒。各组于末次给药后4 h分别取1/2动物(12~18只)进行解剖(6~9只)和交配(6~9只);剩余1/2动物(12~18只)于停药恢复3个动情周期后再分别进行解剖(6~9只)和交配(6~9只),计算脏器系数。采用酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测雌性大鼠血清中卵泡刺激素(follicle-stimulating hormone,FSH)、黄体生成素(lutenizing hormone,LH)、雌激素、孕激素、睾酮、抗苗勒管激素(anti-Müllerian hormone,AMH)以及游离甲状腺激素3(free thyroid hormone,fT3)的水平。雌性大鼠卵巢组织切片苏木精-伊红(hematoxylin and eosin,HE)染色后进行卵泡计数。记录LNG-ECPs给药结束和停药恢复期雌性大鼠妊娠率及窝仔数,测定F1代仔鼠生长指数以及主动和被动运动能力等指标。取P-12大鼠卵巢组织进行转录组学测序,建立LNG-ECPs致卵巢差异表达基因谱,分析LNG-ECPs致卵巢损伤的差异基因,并进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析。结果①连续给予3个和6个动情周期后,LNG-ECPs组雌鼠的血清激素水平和生育力与溶媒对照组相比,差异均无统计学意义(均P>0.05);F1代仔鼠生长指数和行为学结果与溶媒对照组仔鼠相比差异均无统计学意义(均P>0.05)。②连续给予12个动情周期后,与溶媒对照组相比,LNG-ECPs组大鼠血清中FSH[(0.21±0.17)U/L]、LH[(0.27±0.08)U/L]和孕激素[(0.68±0.23)μg/L]水平显著�Objective To explore the effects of consecutively repeated application of emergency contraception pills(ECPs)containing levonorgestrel(LNG)on the female fertility and the health outcomes of F1 generation rats.Methods Female SPF rats were intragastric administered with LNG-ECPs consecutively for 3(P-3),6(P-6)and 12(P-12)estrous cycles(three times in each estrous cycle),respectively.Under each administration schedule,rats were randomly divided into 2 groups according to body weight stratification using random numbers generated in Excel,i.e.LNG-ECPs group and solvent control group,administered with 0.12 mg/kg LNG-ECPs and corresponding volumes of 0.5%CMC-Na,respectively.Four hours after the last dosing,half of the animals(12-18)in each group were allotted randomly for dissection(6-9)and mating(6-9),respectively.The remaining half(12-18)were recovered for 3 estrous cycles,and then were randomly allocated for dissection(6-9)and mating(6-9).Organ coefficients were calculated.Serum levels of follicle-stimulating hormone(FSH),luteinizing hormone(LH),estradiol,progesterone,testosterone,anti-Müllerian hormone(AMH)and free thyroid hormone 3(fT3)were examined by enzyme linked immunosorbent assay(ELISA).Ovarian tissues were sectioned and stained with hematoxylin and eosin(HE)for follicle counting.In addition,the pregnancy rate and litter size of the female rats were recorded,and the growth indexes and behavioral parameters of the cubs were measured.Moreover,RNA sequencing(RNA-seq)of the ovarian tissues was performed to establish the differential expression gene profile of ovarian injury induced by LNG-ECPs.Then gene ontology(GO)function and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment were analyzed.Results 1)After consecutive administration for 3 and 6 estrous cycles,LNG-ECPs showed no significant impact on the serum hormone levels and female fertility(all P>0.05),and the growth indexes and behavioral parameters of the F1 generation(all P>0.05).2)After consecutive administration for 12 estrous cycles,the s

关 键 词：左炔诺孕酮 生育力 行为学 卵巢损伤 转录组学 

分 类 号：R965[医药卫生—药理学]