母鼠孕期和哺乳期连续染毒双酚A对子代雄鼠睾丸发育的影响

doi:10.11843/j.issn.0366-6964.2019.05.010

摘要/Abstract

摘要：

旨在探究双酚A（BPA）对子代雄鼠睾丸发育的影响。本研究将8周龄体重18~22 g的SPF级母鼠随机分为7组，每组4个重复，每个重复5只，A组每天给予普通蒸馏水，B组每天饮水给予0.05 mg·kg^-1 BPA，C组每天饮水给予0.5 mg·kg^-1 BPA组，D组每天饮水给予5 mg·kg^-1 BPA，E组每天饮水给予10 mg·kg^-1 BPA，F组每天饮水给予20 mg·kg^-1 BPA，G组每天饮水给予50 mg·kg^-1 BPA。F0代母鼠自怀孕起直至F1代子鼠断奶止饮水染毒BPA，F1代雄鼠于断奶（21日龄）处死。ELISA结果显示，母鼠染毒BPA剂量5 mg·kg^-1以上时，子代血清和睾丸组织BPA含量显著增加（P<0.05）。睾丸器官指数测定结果证明，染毒BPA剂量大于等于20 mg·kg^-1可导致子代雄鼠睾丸指数显著增大（P<0.05）。H&E染色显示，母鼠染毒BPA剂量大于等于10 mg·kg^-1可导致睾丸生精小管萎缩，小管间隙变大。彗星试验结果证明，染毒BPA大于等于5 mg·kg^-1可使子代睾丸细胞核DNA损伤显著上升（P<0.05）。免疫组化结果显示，母鼠染毒BPA剂量大于等于20 mg·kg^-1时子代睾丸雄激素受体（AR）表达量显著减少（P<0.05）。转录组测序结果显示，母鼠染毒50 mg·kg^-1BPA后子代雄鼠睾丸剪切体代谢通路U5 snRNA亚基编码基因Snrnp40上调，剪切体通用蛋白组件编码基因Hnrnpu下调，导致mRNA转录后修饰第一步受阻，荧光定量PCR结果与转录组测序结果一致，证实了转录组结果。结果表明，母鼠暴露于低剂量BPA可以引起子代睾丸发育异常，其分子机制可能与剪切体进行mRNA转录后修饰第一步反应受阻有关。

Abstract:

This study was conducted to investigate the effects of maternal exposure to BPA on testicular development of male offspring mice. Eight-week-old SPF female mice weighing 18-22 g were randomly divided into 7 groups (group A, B, C, D, E, F and G) with 4 replicates in each group, 5 per replicate. Group A was given normal distilled water daily, and group B was given 0.05 mg·kg^-1 BPA daily, group C was given 0.5 mg·kg^-1 BPA daily, group D was given 5 mg·kg^-1 BPA daily, group E was given 10 mg·kg^-1 BPA daily, group F was given 20 mg·kg^-1 BPA daily, group G was given 50 mg·kg^-1 BPA daily. F0 female mice were exposed to BPA from pregnancy day 1 to the lactation day 21. F1 male mice were sacrificed for sampling at weaning (21 d). The results showed that the BPA content in the serum and testis of the male offspring mice significantly increased when the dose of BPA was more than 5 mg·kg^-1 (P<0.05). The dose of BPA greater than or equal to 20 mg·kg^-1 resulted in a significant increase in testicular index in male offspring mice (P<0.05). H&E staining showed that the testicular tubules shrunk and the small tube gap became larger when the dose of BPA was greater than or equal to 10 mg·kg^-1. The results of comet assay showed that the damage of testicular DNA significantly increased in the progeny testis DNA when the BPA exposure dose was greater than or equal to 5 mg·kg^-1 (P<0.05). The results of immunohistochemistry showed that the expression of androgen receptor (AR) in the testes of the offspring significantly decreased when the dose of BPA was greater than or equal to 20 mg·kg^-1 (P<0.05). Transcriptome sequencing showed that the expression of Snrnp40 encoding U5 snRNA subunit up-regulated in spliceosome pathway, and the expression of Hnrnpu encoding splicing universal protein component down-regulated, which led to the first steps of mRNA post-transcriptional modification were blocked in male offspring mice when the dose of BPA was 50 mg·kg^-1. The results of the real-time PCR were consistent with those of the transcriptome sequencing. The result indicate that female mice exposing to low doses of BPA can cause testicular dysplasia of male offspring mice, and its molecular mechanism may be related to the hindered reaction of the first step of mRNA post-transcriptional modification.

中图分类号:

S852.21

张石磊, 包佳鹭, 史万玉, 钟秀会. 母鼠孕期和哺乳期连续染毒双酚A对子代雄鼠睾丸发育的影响[J]. 畜牧兽医学报, 2019, 50(5): 993-1005.

ZHANG Shilei, BAO Jialu, SHI Wanyu, ZHONG Xiuhui. Effects of Maternal Exposure to Bisphenol A during Pregnancy and Lactation on the Development of Testis in Male Offspring Mice[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(5): 993-1005.

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