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激素型肾阳虚动物肝线粒体蛋白质组与能量代谢相关性
引用本文:卢德赵,沃兴德,施孟如,李毅,唐利华.激素型肾阳虚动物肝线粒体蛋白质组与能量代谢相关性[J].中国生物化学与分子生物学报,2005,21(6):807-814.
作者姓名:卢德赵  沃兴德  施孟如  李毅  唐利华
作者单位:浙江中医学院生命科学系,杭州,310053
基金项目:国家自然科学基金(No.30371710)资助项目~~
摘    要:应用凝胶内差异显示电泳技术研究肾阳虚大鼠肝线粒体蛋白质组,并从肝线粒体蛋白质组角度阐述肾阳虚与能量代谢的关系.8个分别来自于肾阳虚大鼠和正常大鼠的肝线粒体蛋白质样品(各4个)分别用荧光染料Cy3、Cy5标记,以及8个样品等量混合物用Cy2标记作为内标,每一Cy3、Cy5标记样品与Cy2标记的内标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软件结合内标分析,以肾阳虚组动物与正常组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,实验共获得16个差异蛋白质,经质谱测定和与蛋白质文库比对,鉴定11个蛋白质.其中,肾阳虚动物热休克蛋白60和70、肌氨酸脱氢酶、氨甲酰磷酸合成酶、亚硫酸盐氧化酶、ATP合酶、醛脱氢酶和NADH脱氢酶表达量增加,而丙酮酸脱氢酶、α酮戊二酸脱氢酶、脂酰辅酶A脱氢酶和鸟氨酸氨基转移酶表达量降低.实验表明,肾阳虚动物能量代谢相关酶的变化与肾阳虚的临床虚寒症状有关.

关 键 词:蛋白质组  肾阳虚  肝线粒体  能量代谢  
收稿时间:2005-12-20
修稿时间:2005年1月25日

Relativity of Liver Mitochondria Proteome and Energy Metabolism in Shenyang Deficience Animals Induced by Hormone
LU De-Zhao,WO Xing-De,SHI Meng-Ru,LI Yi,TANG Li-Hua.Relativity of Liver Mitochondria Proteome and Energy Metabolism in Shenyang Deficience Animals Induced by Hormone[J].Chinese Journal of Biochemistry and Molecular Biology,2005,21(6):807-814.
Authors:LU De-Zhao  WO Xing-De  SHI Meng-Ru  LI Yi  TANG Li-Hua
Affiliation:(Department of Life Sciences, Zhejiang Traditional Chinese Medical College, Hangzhou 310053, China
Abstract:Two-dimensional differential gel electrophoresis (2-D DIGE) was used to study the difference of liver mitochondria proteome between the shenyang deficience rats and normal rats. The relation between shenyang dificience and energy metabolism were observed. Eight samples (four samples from normal rats and four from shenyang dificience rats) were labeled with Cy3 or Cy5. Each Cy3-labeled sample and Cy5-labeled sample were mixed on the same 2-D gel along with a Cy2-labeled mixture of all eight samples as an internal standard. The gels were imaged by Typhon 9400 using different emission filter respectively and spot-features were analyzed by DeCyder software in each paired normal/disease comparison. Using DIGE technology with internal standard, statistically significant quantitative comparisons of each protein abundance change could be made across multiple samples simultaneously without interference of gel-to-gel variation. 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 shenyang dificience rats, and 11 protein were successfully identified. In these proteins, the expression of heat shock protein60/70,sarcosine dehydrogenase,carbamoylphosphate synthetaseⅠ,ATP synthase,sulfite oxidase,mitochondrial aldehyde dehydrogenase precursor and NADH dehydrogenase increased, while the expression of ornithine amino-transferase,2-oxoisovalerate dehydrogenase,acyl-CoA dehydrogenase decreased in shenyang dificience rats. These findings indicate that the change of the enzymes relation to energy metabolism in shenyang dificience animals involved in the deficiency-cold symptom in the clinic.
Keywords:proteome  shenyang deficience  liver mitochondria  energy metabolism
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