妊娠期环境雌激素双酚A暴露对子代成年雌鼠的生殖损伤作用

doi:10.11843/j.issn.0366-6964.2019.07.016

摘要/Abstract

摘要： 为探究母鼠妊娠期暴露双酚A（BPA）对子代成年雌性小鼠生殖损伤作用及DNA甲基转移酶（DNMT）基因表达量的影响，将120只妊娠母鼠随机分为6组，每组20只。各组分别在母鼠孕0.5~17.5 d内按0、2.5、5、10、20、40 mg·（kg·d）^-1剂量灌胃BPA。每组随机选取10只子代成年（56 d）雌鼠，麻醉后处死，收集血液、子宫和卵巢；统计子鼠成年时子宫和卵巢指数；HE染色观察子代雌鼠卵巢的组织形态学变化；放射性免疫法检测子代雌鼠血清中卵泡刺激素（FSH）、黄体生成素（LH）、雌二醇（E₂）和孕酮（P₄）水平；酶联免疫吸附法（ELISA）检测子代雌鼠卵巢内雌激素受体α（ERα）、雌激素受体β（ERβ）的含量；实时荧光定量PCR法检测子代雌鼠卵巢内ERα、孕酮受体（PgR）、DNMT1、DNMT3A和DNMT3B mRNA转录水平。结果显示，BPA显著降低子代雌鼠血清FSH、LH、E₂和P₄水平，显著提高子代雌鼠卵巢ERα和ERβ含量，显著降低子代雌鼠卵巢ERα、PgR和DNMT mRNA的转录水平（P<0.05或P<0.01）。BPA暴露子代小鼠成年时卵巢和子宫指数增大，HE染色观察卵巢实质发生萎缩，初级和次级卵泡数量增加。本研究表明，妊娠期暴露BPA可促进成年子鼠卵巢产生卵泡，显著降低生殖激素的含量及活性，显著增加卵巢激素受体的含量，抑制卵巢激素受体和DNMT的转录，对子代产生生殖损伤。

Abstract: This experiment was conducted to investigate the effect of bisphenol A (BPA) exposure during pregnancy on the genotoxicity of adult offspring females and gene expression levels of DNA methyltransferase (DNMT). One hundred and twenty pregnant mice were randomly divided into six groups, twenty mice in each group. The pregnant mice were administered with BPA at the doses of 0, 2.5, 5, 10, 20, 40 mg·(kg·d)^-1, from the pregnancy day 0.5 to day 17.5. Ten adult offspring females (56 d) randomly selected from each group were sacrificed after anesthesia and the blood, uterus and ovaries were collected. Then the uterine and ovarian index were calculated; the ovarian histomorphology were observed after hematoxylin-eosin staining (HE). The activity of follicle stimulating hormone (FSH), luteinizing hormone (LH) and the contents of estradiol (E₂), progesterone (P₄) in serum were detected by radioimmunoassay. The contents of ovarian estrogen receptor α (ERα) and estrogen receptor β (ERβ) were detected by ELISA. The mRNA relative transcription levels of ERα, progesterone receptor (PgR), DNMT1, DNMT3A, and DNMT3B in ovaries were detected by real-time quantitative PCR (RT-qPCR). The results showed that:1) Compared with the control group, BPA significantly reduced the serum levels of FSH, LH, E₂ and P₄; 2) BPA significantly increased the ovarian ERα and ERβ levels; 3) BPA significantly reduced the mRNA relative transcript levels of ovarian ERα, PgR and DNMT in the adult offspring females (P<0.05 or P<0.01); 4) The ovarian index were increased in the adult offspring mice exposed to BPA, but the ovarian parenchyma was atrophied and the number of primary and secondary follicles were increased. Our results reveal that BPA exposure to during pregnancy can promote the production of ovarian follicles in offspring females, significantly increase the contents of ovarian hormone receptor, inhibit the transcriptional activity of ovarian DNMT, and reduce the contents of reproductive hormone and activity, causing reproductive damage to adult offspring females.

中图分类号:

R994.6

魏媛媛, 崔宇擎, 韩超, 钟秀会, 包永占, 史万玉. 妊娠期环境雌激素双酚A暴露对子代成年雌鼠的生殖损伤作用[J]. 畜牧兽医学报, 2019, 50(7): 1466-1474.

WEI Yuanyuan, CUI Yuqing, HAN Chao, ZHONG Xiuhui, BAO Yongzhan, SHI Wanyu. Reproductive Injury Effects of Bisphenol A Exposure during Pregnancy on Offspring Adult Females[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(7): 1466-1474.

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