异烟肼致肝脏毒性的蛋白质组学分析

Proteomics analysis of isoniazid toxicity in rat liver

目的:应用蛋白质组学技术研究异烟肼致大鼠肝脏损伤时肝脏蛋白质表达谱的变化,为从分子水平上寻找毒性损伤标志物以及阐明异烟肼毒性机制奠定基础。方法:将Wistar大鼠随机分为生理盐水正常对照组和异烟肼组(400 mg.kg-1),分别连续灌胃14 d后处死大鼠,提取肝脏总蛋白,双向凝胶电泳分离蛋白质组成分,采用考马斯亮蓝R-250染色,经ImageMaster2D Platinum5.0软件分析比较两组图谱,并对差异蛋白斑点进行基质辅助激光解吸/电离飞行时间质谱(MALD-TOF-MS)分析鉴定。结果:发现11个差异有统计学意义的蛋白质点,鉴定出8个蛋白,其中,异烟肼处理组比对照组表达升高的蛋白质有鸟氨酸转氨酶、葡萄糖调节蛋白、乙醛脱氢酶和3α-羟甾类脱氢酶,比对照组表达降低的蛋白质有抗氧化酶B166、谷胱苷肽转硫酶、醛铜还原酶和碳酸酐酶。结论:异烟肼可能造成肝脏抗氧化系统和氧化应激异常,这对阐明异烟肼的肝损伤机制具有十分重要的作用。

Objective： In order to lay the foundation of finding the markers of toxic injury and clarifing the mechanism of isoniazid（INH） toxicity from the molecular level,proteomics technology was applied to study the variation of liver protein expression profile between normal rat and liver injured rat caused by INH.Methods： Male Wistar rats（n=18） were orally administrated with 0 and 400 mg·kg-1 INH for 14 days,respectively.The total liver proteins in INH and control groups were extracted and separated by two-dimensional polyacrylamide gel electrophoresis（2-DE） technology.Then the gels were stained by Coomassie Brilliant Blue R-250,and analyzed using Image Master 2D Platinum analysis software（version 5.0）.The differential expression of proteins was analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（MALDI-TOF-MS）.Results： Eleven protein spots with significant difference were found and eight of which were identified successfully.In these identified proteins,the expression levels of omithine aminotransferase,glucose regulated protein,aldehyde dehydrogenase and 3-alpha-hydroxysteroid dehydrogenase in the isoniazid treatment group went upward compared with the control group,while the expression levels of antioxidant enzyme B166,glutathione S-transferase,aldose reductase and carbonic anhydrase in the isoniazid treatment group went downward compared with the control group.Conclusion： INH may lead to liver disorders through anti-oxidative system and oxidation stress,which is important for further elucidating the mechanisms of liver injury caused by INH.