富硒酵母对斑点叉尾蛔肝脏的毒性作用及蛋白质组学的影响

摘要

实验探讨了富硒酵母对斑点叉尾鮰肝脏的毒性作用及蛋白质组学的影响,结果表明,饲养56d后正常组和有机硒组采用原子荧光法检测硒含量均值为0.15和0.81mg/kg.H.E染色显示有机硒组肝细胞有明显的疏松化,部分发生脂肪变.通过二维凝胶电泳结合ImageMaster软件分析,鉴定出8个表达差异蛋白质点,其中3个在有机硒组表达上调,5个在有机硒组特异表达,经液相色谱串联质谱分析鉴定,这些蛋白质分别是伴侣蛋白TCP1一亚基8、甘油醛一磷酸脱氢酶,4SNc-Tudor蛋白、腺苷激酶、酮糖移转酶、丙氨酞-tRNA合成酶.富硒酵母饲料对斑点叉尾鮰肝具有明显的毒性效应,有机硒调控4SNc-Tudor蛋白,通过信号传导通道加强机体免疫.%To investigate the influence on channel catfish (lctalurus punctatus) liver toxicity and liver proteome change by the use of selenium-enriched yeast, Se-concentration in channel catfish liver was determined and the pathological change was monitored after selenium-enriched yeast feeding for 56 days. Proteins extracted from the liver of selenium-enriched yeast feeding channel catfishes and non-feeding controls were separated by two-dimensional gel electrophoresis (2-DE), differentially expressed proteins between the two groups were analyzed using ImageMaster software and identified by online reversed-phase nano-flow liquid chromatography coupled with ion trap mass spectrometry(LC-MS/MS). The average Se-concentration of liver form the non-feeding controls and the selenium-enriched yeast feeding ones were 0.15 mg/kg and 0.81 mg/kg. Significant difference was detected between these two groups (P < 0. 05). H. E staining of pathology from channel catfish liver showed that the selenium-enriched yeast group has obvious cell loose and cell fatty degeneration. 15 differetial protein were found by 2-DE separation and ImageMaster software analysis. Of them, 8 over-expressed proteins in selenium-enriched yeast were selected for MS analysis and identified successfully, including 3 u-regulated proteins and 5 uniquely expressed proteins in selenium-enriched yeast group compared with the controls. These differential proteins correspond to chaperonin TCP1 subunit 8 and glyceraldehyde-3-phosphate dehydrogenase,4SNc-Tudor domain protein, adenosine kinase a, transketolase and alanyl-tRNA synthetase-like et al. Obvious toxicity was detected in the channel catfish liver after feeding with selenium-enriched yeast. Organic selenium might strengthen immune through 4SNc-Tudor protein involved signal transmission pathway. Differential proteins identified in our work could be specific biomarkers related to toxicity of organic Se to channel catfish.