菲对泥鳅(Misgurnus anguillicaudatus)肝性腺指数和卵黄蛋白原转录水平的影响

Effects of Phenanthrene on Hepatic Gonadal Index and Vitellogenin Gene Expression in Misgurnus anguillicaudatus

为探究菲(phenanthrene,PHE)对鱼类的内分泌干扰效应及生殖毒性损伤,将泥鳅(Misgurnus anguillicaudatus)暴露于菲浓度为1.33、1.77、2.36和3.13 mg·L^(-1)的水体中28 d,分别于7、14和28 d取样,采用称重法测定肝脏指数、性腺指数;qRT-PCR法检测卵黄蛋白原基因(vtg1、vtgc)在泥鳅肝脏和性腺的转录水平;酶联免疫吸附法测定卵黄蛋白原(vitellogenin,VTG)含量。结果显示:(1)PHE胁迫使雄性泥鳅肝脏指数升高、性腺指数下降;高浓度PHE胁迫引起雌性泥鳅肝脏指数上升,性腺指数升高;(2)PHE胁迫导致泥鳅vtg1和vtgc表达量显著升高,雄性泥鳅vtg1和vtgc转录水平与菲胁迫浓度之间呈现低浓度诱导、高浓度抑制的趋势;(3)PHE暴露使泥鳅肝脏、性腺VTG含量显著升高,且在PHE较高浓度组中尤为明显。这表明,一定浓度PHE暴露可以导致泥鳅肝脏和性腺发育异常;较低浓度PHE暴露可诱导肝脏和性腺vtg1、vtgc转录水平的表达,而高浓度PHE则抑制上述基因表达,PHE胁迫引起vtg1和vtgc表达水平存在差异;PHE可以显著诱导肝脏和性腺VTG的产生。因此,高浓度PHE干扰了泥鳅的性腺作用轴。

To study the effects of phenanthrene(PHE)on fish endocrine disruption and reproductive toxicity,loach(Misgurnus anguillicaudatus)was exposed to 1.33,1.77,2.36 and 3.13 mg·L^(-1)for 28 d.During the PHE treatment,the liver and gonad tissues of loach were collected on 7,14 and 28 d.The hepatosomatic index(HSI)and gonadosomatic index(GSI)were measured by weighing method,the expression of vitellogenin 1(vtg1)and vitellogenin c(vtgc)in the liver and gonad of M.anguillicaudatus was detected by qRT-PCR,and the content of vitellogenin(VTG)was determined by enzyme linked immunosorbent assay(ELISA).The results showed that:(1)PHE stress significantly increased HSI and decreased GSI of male,and significantly increased HSI and GSI of female.(2)PHE stress caused significant increase of vtg1 and vtgc expression in M.anguillicaudatus,and the expression of vtg1 and vtgc in male M.anguillicaudatus showed a trend of low concentration induction and high concentration inhibition.(3)PHE exposure significantly increased VTG content in liver and gonad of M.anguillicaudatus,especially in the group with higher concentration of PHE.It is suggested that certain concentration of PHE could induce abnormal development of liver and gonad of M.anguillicaudatus.Additionally,the lower concentration of PHE could induce expression of vtg1 and vtgc in liver and gonad,while the higher concentration of PHE could inhibit the expression of those two genes,and the expression of vtg1 and vtgc were different under PHE stress.Moreover,PHE could induce the production of VTG in liver and gonad of loach.Therefore,the gonadal axis of M.anguillicaudatus was disturbed by PHE contamination.