摘要
用150ng·L^(-1)环境雌激素乙炔基雌二醇(17α-ethinylestradiol,EE2)、300μg·L^(-1)环境抗雄激素氟他胺(Flutamide,Flu)单独处理以及150ng·L-1 EE2与300μg·L^(-1) Flu联合处理对斑马鱼(Danio rerio)雄性成鱼进行了30d浸浴处理,并用组织学的方法检测了上述物质对斑马鱼精巢发育和精子发生的影响;同时,采用定量RT-PCR的方法研究了EE2和Flu影响斑马鱼精子发生的分子差异。组织学结果显示,EE2处理后斑马鱼精巢壁变厚,生殖细胞大量丢失;Flu处理后精子细胞数量下降,出现空腔,早期生殖细胞数量有增加的趋势;联合处理组斑马鱼精巢生殖细胞数量急剧减少,空腔化严重。定量RT-PCR结果显示,所有药物处理均能显著下调鱼类发育和繁殖相关转录因子基因(dmrt1、sf1)和生殖细胞标记基因(dmc1、nanos1)的表达(p<0.05);EE2单独处理、EE2与Flu联合处理还能显著下调鱼类类固醇合成酶基因p450-11β、cyp19a1a、cyp17α1和cyp17α2的表达(p<0.05),Flu单独处理只显著下调了p450-11β的表达水平(p<0.05),表现出一定的分子差异。研究提示EE2、Flu及两者联合处理均严重干扰斑马鱼精子发生过程,导致生殖细胞数量明显下降;上述药物对斑马鱼类固醇合成及配子发生相关基因表达的影响既有相似之处,也有明显不同。
In this study zebrafish （Danio rerio） males were exposed to 17a-ethinylestradiol （EE2, 150 ng·L^-1）, flutamide （Flu, 300μg·L^-1） and their combination （EE2, 150 ng ·L^-1＋ Flu, 300 μg·L^-1） for 30 days, and the influences of these drugs on spermat.ogenesis of zebrafish males were analyzed by histology. Meanwhile, we used quantitative RT-PCR （qRT-PCR）, to shed light on the possible differential mechanisms involved. Histological results showed thickened testicular epithelium and considerable loss of germ cells after EE2 treatment; decreased and increased number of spermatozoa and early stages of germ cell, respectively, after Flu treatment and sharply decreased number of all stages of germ cells in combination group. Moreover, qRT-PCR results showed dramatically reduced expression of transcription factors involved in steroidogensis regulation （dmrtl , s fl ） and germ cell marker genes （dmcl , nanosl ） in all exposed groups（p〈0.05）, qRT PCR results also showed down-regulated expression of gene responsible for 11-KT （11-keto testosterone） production （p450-11β） in all treated groups （p〈 0.05）, and reduced expressions of genes involved in estrogen （cyp19a1a）, androgen （cyp17a1） and progesterone （cyplfa2） productions after EE2 and combination treatment（p〈0.05）. However, the expressions of cyp19a1a, cyp17a1 and cyp17a2 remained unchanged after Flu treatment （p〈 0.05）. The study suggested that EE2, Flu and their combination seriously disrupted spermatogenesis in zebrafish, resulting in dra- matically decline of germ cell number; therefore, both similar and differential mechanisms were involved in disrupted spermatogene- sis of zebrafish treated with the drugs above, respectively.
出处
《重庆师范大学学报（自然科学版）》
CAS
CSCD
北大核心
2016年第4期15-21,共7页
Journal of Chongqing Normal University:Natural Science
基金
重庆师范大学校级项目(No.13XLZ08)
淡水鱼类资源与生殖发育教育部重点实验室开放课题基金(No.FFRD-2015-02)
关键词
乙炔基雌二醇
氟他胺
斑马鱼
精巢发育
精子发生
基因表达
17a-ethinylestradiol
flutamide
zebrafish
testicular development
spermatogenesis
gene expressions
