温补肾阳药对激素依赖性肾阳虚动物肝线粒体蛋白质组影响的研究  

Effects of Warm and Tonify Kidney-Yang Herbs on Liver Mitochondria Proteome of Kidney-Yang Deficiency Rats

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

机构地区：[1]浙江中医药大学生命科学系,杭州310053

出　　处：《中国药学杂志》2007年第3期169-175,共7页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金资助项目(30371710)

摘　　要：目的利用双向凝胶内差异显示电泳技术,分析温补肾阳药对激素依赖性肾阳虚动物肝线粒体蛋白质组影响。方法正常大鼠、肾阳虚大鼠及温补肾阳药治疗大鼠肝线粒体蛋白质样品分别用荧光染料Cy2,Cy3,Cy5标记,经双向电泳后用不同波长光激发扫描后得到不同样品的蛋白质组图谱,经DeCyder软件分析,以肾阳虚动物与正常动物肝线粒体蛋白质相差1·2倍以上的蛋白作为差异蛋白,共获得16个差异蛋白质,经质谱测定和与蛋白质文库比对,其中11个蛋白质得到鉴定。结果肾阳虚动物由于肾上腺皮质功能受损,引起热休克蛋白60和70含量增加,可能起到调整处于应激状态的细胞存活、保护细胞免受损伤的作用;肾阳虚动物糖代谢和脂代谢功能降低,而蛋白质代谢和核酸代谢功能增强,表现为二氢硫辛酰胺脱氢酶和脂酰辅酶A脱氢酶活性降低,而肌氨酸脱氢酶、氨甲酰磷酸合成酶和亚硫酸氧化酶活性增加;肾阳虚动物由于三羧酸循环和脂肪酸β-氧化功能障碍,引起ATP合酶、醛脱氢酶和NADH脱氢酶活性代偿性增加,试图通过增加ATP的产量以缓解能量代谢的不足;但肾阳虚动物由于鸟氨酸氨基转移酶活性降低,导致储存能量的肌酸合成降低,使产生的ATP大部分通过热能形式消耗掉,从而导致肾阳虚的临床虚寒症状。应用温补肾阳药治疗后使热休克蛋白60更趋增加,可能加强对细胞的保护作用;温补肾阳药增加二氢硫辛酰胺脱氢酶的活性,可通过改善糖代谢和促进三羧酸循环功能使机体的能量代谢增强;温补肾阳药还可以通过增加ATP合酶和醛脱氢酶的含量,使ATP的产量增加,缓解能量代谢功能障碍。结论温补肾阳药可以通过调整线粒体内多种与能量代谢相关蛋白质表达而起到治疗肾阳虚的作用。OBJECTIVE To study the effects of warm and tonify kidney-yang herbs on the liver mitochondria proteome of the kidney-yang deficiency rats by two-dimensional difference gel electrophoresis （2-D DIGE） . METHODS Every liver mitochondria protein sample from the normal rats, the kidney-yang deficiency rats and the kidney-yang deficiency rats treated with warm and tonify kidney-yang herbs was labeled with Cy2, Cy3 or Cy5 respectivley. Each Cy3-labeled 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, using DIGE technology with internal standard. MALDI-TOF MS was used to provide sensitive and accurate mass spectral data for 16 unique proteins that were changed in abundance more than 1.2 folder up or down between normal and kidney-yang deficiency rats, and 11 proteins were successfully identified. RESULTS In the kidney-yang deficiency rats, the expression of heat shock protein 60 and 70 was increased, which contributed to protect the cell from damage. The metabolism of carbohydrate and lipids was reduced due to the decrease of the 2-oxoisovalerate dehydrogenase and Acyl-CoA dehyclrogenase. The metabolism of protein and nucleic acid was enhanced because of the increase of the sareosine dehydrogenase, carbamoyl- phosphate synthetase Ⅰ andsulfite oxidase. For the inhibition of tricarboxylic acid cycle and β-oxidation of fatty acid, the expression of the ATP synthase, mitochondrial aldehyde dehydrogenase and NADH dehyclrogenase was increased compensationally and the energy metabolism was alleviated by increasing the output of ATP. The scarcity of ornithine aminotransferase in the kidney-yang deficiency rats resulted in deficient creatine phosphate which was need for depositing energy and most of ATP was consumed as heat energy. Therefore, further deficiency of energy led to deficiency-cold symptom. After the rats suffering kidney-yang deficiency were treated with warm and tonify kidney-yang herbs, the expression of heat shock protein 6

关 键 词：蛋白质组 肾阳虚 温补肾阳药 线粒体 能量代谢 

分 类 号：R284[医药卫生—中药学]