温补肾阳药对肾上腺切除的肾阳虚大鼠肝线粒体蛋白质组的影响  

The Effects of Warm and Tonify Kidney-Yang Herbs on the Liver Mitochondria Proteome of the adrenalectomized Kidney-Yang deficiency Rats

作　　者：卢德赵[1] 沃兴德[2] 唐利华[2] 李毅[2] 施孟如[2] 沃立科[2] 杨贞[2] 

机构地区：[1]北京中医药大学基础医学院,北京100029 [2]浙江中医药大学生命科学学院,浙江杭州310053

出　　处：《中华中医药学刊》2008年第4期744-747,共4页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(30371710);浙江省中医药管理局资助项目(2004C019);浙江省卫生科技计划项目(2004B137)

摘　　要：目的:应用凝胶内差异显示电泳技术和质谱技术研究温补肾阳药对肾上腺切除的肾阳虚大鼠肝线粒体蛋白质组的影响。方法:通过切除两侧肾上腺造成肾阳虚模型大鼠和温补肾阳药治疗大鼠肝线粒体蛋白质经荧光染料Cy3、Cy5标记,与等量Cy2标记的内标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。利用DeCyder软件进行差异蛋白质组分析,筛选出18个差异蛋白质进行质谱鉴定,其中15个蛋白质得到鉴定。结果:经温补肾阳药治疗后,肾阳虚动物氨甲酰磷酸合成酶、过氧化氢酶、二氢硫辛酸脱氢酶、热休克蛋白60、亚硫酸氧化酶、ATP合成酶、醛脱氢酶、丙酮酸脱氢酶、3-巯基丙酮酸转硫酶、腺苷酸激酶同工酶5、谷胱甘肽过氧化物酶表达均增高。结论:温补肾阳药可能通过加快糖、脂类、蛋白质和核酸分解代谢,加快ATP合成以及清除自由基和过氧化物,维持线粒体膜稳定性,从而起到治疗肾阳虚的作用。Objective ： Two- dimensional difference gel electrophoresis and mass spectrum was used to study the effects of Warm and Tonify Kidney - Yang Herbs on the Liver Mitochondria Proteome of the adrenalectomized Kidney - Yang deficiency Rats. Methods ： Every liver mitochondria protein sample from the kidney -yang deficiency rats and the kidney - yang deficiency rats treated with warm and tonify kidney - yang herbs was labeled with Cy3 or Cy5 randomly, Each Cy3 - labeled sample and Cy5 - labeled sample were mixed on the same 2 - D gel along with a Cy2 - labeled mixture of all samples as an internal standard and run on the same gel. The gels were scan under different wavelength light after electrophoresis. All images were analyzed by DeCyder software. 18 unique proteins were selected, and in which 15 protein were successfully identified by mass spectrum. Result ： After the kidney - yang deficiency rats were treated with warm and tonify kidney - yang herbs, the expression of Carbamoyl phosphate synthetase I , Catalase, Dihydrolipoyl dehydrogenase,heat shock protein 60, Sulfite oxidase, ATP synthase, Aldehyde dehydrogenase, pyruvate dehydrogenase, 3 - mereaptopyruvate sulfurtransferase, Adenylate kinase isoenzyme 5 and glutathione peroxidase were increased. Conclusion ： The warm and tonify kidney -yang herbs perform its therapeutical effect by accelerating the metabolism of carbohydrate,lipids ,nucleic acid and protein,and removing the free radicle and perhydride to improve the stability of mitochondria membrane.

关 键 词：肾阳虚 线粒体 蛋白质组 温补肾阳药 

分 类 号：R2-031[医药卫生—中西医结合] Q493.8[医药卫生—中医学]