温补肾阳药对丙基硫氧嘧啶引起肾阳虚大鼠肝线粒体蛋白质组的影响  

Effects of Warming and Tonifyin Kidney-Yang Drugs on Liver Mitochondria Proteome of Rats with Kidney-Yang Deficiency

作　　者：卢德赵[1] 沃兴德[1] 李毅[1] 唐利华[1] 沃立科[1] 施孟如[1] 杨贞[1] 

机构地区：[1]浙江中医药大学生命科学学院

出　　处：《中华中医药杂志》2007年第2期102-107,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金资助项目(No.30371710);浙江省中医药管理局资助项目(No.2004C019);浙江省卫生科技计划项目(No.2004B137)

摘　　要：目的:应用凝胶内差异显示电泳技术研究温补肾阳药对药物性肾阳虚动物肝线粒体蛋白质组的影响。方法:正常大鼠、通过喂饲丙基硫氧嘧啶造成肾阳虚模型大鼠和温补肾阳药治疗大鼠的肝线粒体蛋白质经荧光染料Cy2、Cy3、Cy5标记,等量混合后在同一胶中进行电泳分离,经不同光激发后扫描得到不同样品的蛋白质组图谱。经DeCyder软件进行差异蛋白质组分析,筛选出16个差异蛋白质。经质谱测定及数据库比对,其中10个蛋白质得到鉴定。结果:肾阳虚时氨甲酰磷酸合成酶1、二氢硫辛酰胺支链酰基转移酶、泛醇-细胞色素C还原酶核心蛋白I、异戊酰辅酶A脱氢酶、热休克蛋白60、琥珀酰CoA合成酶、羟氨基辅酶A脱氢酶/酮脂酰辅酶乙酰辅酶A脱氢酶α亚基、鸟氨酸转氨甲酰酶、谷胱甘肽过氧化物酶表达量均增加,而鸟氨酸氨基转移酶表达量降低。应用温补肾阳药治疗后,上述蛋白质的表达量均有不同程度的纠正。结论:喂饲温补肾阳药的肾阳虚模型大鼠糖、脂类和蛋白质代谢均得以改善,可能与温补肾阳药维护线粒体膜稳定性有关。Objective： To study the effects of warming and tonifying kidney-Yang drugs on the liver mitoehondria proteome of rats with kidney-yang deficiency using two-dimensional difference gel eleetrophoresis （2-D DIGE）. Methods： Every liver mitoehondria protein sample from the normal rats, the kidney-yang deficiency rats and the kidney-yang deficiency rats treated with warm and tonify kidney-yang drugs were labeled with Cy3 or Cy5 randomly, Each Cy3-1abeled sample and Cy5- labeled sample were mixed on the same 2-D gel along with a Cy2-1abeled mixture of all samples as an internal standard and run on the same gel. All images were analyzed by DeCyder software DIGE technology with internal standard. 16 unique proteins were selected, and in which 10 protein were successfully identified. Results. In the kidney-yang deficiency rats, the expressions of earbamoyl-phosphate synthetase 1, dihydrolipoamide branched chain transaeylase , ubiquinol-eyteehrome e reductase, isovaleryl eoenzyme A debydrngenase, heat shock protein 60, sueeinyl-CoA synthetase, hydroxyamino-eoenzyme A dehydrogenase/ketoaeyl eoenzyme acetyl eoenzyme A dehydrngenase asubunit, ornithine transearbamylase and glutathione peroxidase all increased. While the expression of ornithine aminotransferase decreased. After the treatment of warming and tonifying kidney-yang drugs, the expressions of above-mentioned proteins all had rectification in different degrees. Conclusion： The metabolism of earbobydrate, lipids and protein had been rectified when the kidney-yang deficiency rats were fed with warming and tonifying kidney-yang drugs, which was likely attributed to the improvement of stability of mitoehondria membrane.

关 键 词：蛋白质组 肾阳虚 温补肾阳药 线粒体 

分 类 号：R285[医药卫生—中药学]