联苯胺作用下的鱼肝脏线粒体蛋白质表达差异研究

Differences of Expression for Mitochondrion Proteins in Fish Liver Under Exposure of Bianiline

用具有三致作用且广泛存在于环境中的联苯胺对鱼肝脏线粒体进行了体外染毒,并进行了线粒体蛋白质的SDS-PAGE分析。结果表明,在一定范围内(0～50μg·mL-1),随着联苯胺剂量的增加,线粒体蛋白质表达条带数逐渐减少,且随染毒剂量的逐渐增大同时,分子量较小的差异表达条带数呈现逐渐消失的趋势。这一结果对进一步研究化学致癌物对线粒体的损伤机理有重要意义。

It has been known that mitochondrion proteins from animal cells contain genetic materials besides cell nucleolus, which means intact and functional mitochondrion plays an important role in existence of life. Bianiline, as many precursors of artificial chemicals and being regarded as a risky pollutant on environment, is entering our environment through a variety of approaches, therefore, development of a simple and effective procedure for determination of toxicological impact of the chemical is of significant essence at sub- cell level. In the present study, we tested the differences of expression for mitochondrion proteins in fish liver under exposure of bianiline with the technique of SDS- PAGE. The mitochondrion proteins in fish liver were firstly separated from the fish liver by homogenization and centrifugations in the presence of Dnase. Then the purified mitochondrion proteins were treated with different concentrations of 0, 10, 20,30, and 50 μ g· mL- 1 for bianiline with an incubation period of 2 hours at 35 ℃ . To display the distribution of expressive strips of the mitochondrion proteins, the technique of SDS- PAGE was utilized accordingly. It was found that as the increase of the concentrations of bianiline, the number of the displayed strips firstly enhanced, from 12 to 14 strips for 0 and 10 μ g· mL- 1, respectively, and then decreased by 4 strips for the treatment at 20 μ g· mL- 1. It has also been discovered that the low molecules of the proteins exhibited more sensitive response than the high molecules under the exposure of bianiline, suggesting that as the chemical structures containing genes for the proteins be more stable than the low molecules, therefore, they are more tolerant to xenobiotics than the low molecules. As the importance of the mitochondrion proteins, it may be concluded that the procedure developed in our lab may be a new way in evaluation of risk of chemicals on environment.