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机构地区:[1]武汉大学资源与环境科学学院
出 处:《农业环境科学学报》2005年第6期1065-1068,共4页Journal of Agro-Environment Science
基 金:国家自然科学基金资助(20177018);国家自然科学基金(20577036)
摘 要:用差速离心和核酸酶消化法从鲫鱼肝组织中提取和纯化线粒体,用不同浓度(0、10、20、30、40、50mg·L-1)的六氯苯对线粒体进行体外染毒(35℃,2h),并对线粒体蛋白质进行SDS-PAGE分析。结果显示,在一定浓度范围(0~50mg·L-1)内,随着六氯苯浓度的增加,线粒体表达的蛋白质条带呈现先增加后减少的现象,另外多肽B(Mr为63kD)的表达量明显减小甚至消失,表明六氯苯可能引起线粒体内遗传体系发生改变,导致线粒体蛋白质表达出现差异。这一结果为进一步研究六氯苯对线粒体的毒性作用机理具有一定意义,也为在亚细胞水平上研究污染物的毒性效应提供了一种可能的方式。It has been known that mitochondria from animal cells contain genetic materials uniquely besides cell nucleolus, which means intact and functional mitochondria plays an important role in organism. Hexachlorobenzene is one of the persistent organic pollutants (POPs). Therefore, developing a simple and effective procedure for determination of toxicological impact of the chemical is of significantly essential at subcellular level. The mitochondria were isolated from liver of Carassius auratus and purified through differential centrifugation and nuclease digestion. The isolated mitochondria were infected by different concentrations of hexachlorobenzene (0, 10, 20, 30, 40, 50 mg·L^-1) using outside body toxic infection (incubation 2 hours at 35℃), and the mitochondrial protein were analyzed by SDS-PAGE. The results showed that the amount of the expressing band of mitochondrial protein increased first (from 14 to 15 bands at 10, 20 mg·L^-1) and then decreased (from 14 to 13 bands at 50 mg·L^-1); while, the protein band B (polypeptide B, Mr: 66KDa) became thin gradually with the increasing of hexachlorobenzene concentrations, which indicated that hexachlorobenzene might change the mitochondrial genetic system potentially, resulted in differential expression of mitochondrial protein. These results had some significance for further studying the toxicity mechanism of hexachlorobenzene, and offered a potential method to study the toxicity of contaminant at subcellular level.
关 键 词:六氯苯 鲫鱼肝脏 线粒体蛋白质 表达差异 SDS-聚丙烯酰胺凝胶电泳
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