温补肾阳药对甲状腺切除的肾阳虚大鼠肝线粒体蛋白质组的影响  

Effects of warm and tonify kidney-yang herbs on liver mitochondria proteome of kidney-yang deficiency rats

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

机构地区：[1]浙江中医药大学生命科学学院,浙江杭州310053

出　　处：《中国中药杂志》2009年第10期1251-1256,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(30371710);浙江省中医药管理局重点项目(2006Z002)

摘　　要：目的:研究温补肾阳药对肾阳虚动物肝线粒体蛋白质组的影响。方法:12只SD大鼠,随机分为3组,每组4只。模型组和治疗组切除两侧甲状腺,正常组做相应的假手术,但不切除甲状腺。喂养7 d后模型组和治疗组大鼠出现拱背耸毛、大便稀溏等肾阳虚的症状。从第8天开始,治疗组每天喂饲温补肾阳药1次,给药剂量6.7 g.kg-1,其他2组喂饲等体积的生理盐水,连续6 d后,断头处死3组大鼠,分离提取其肝线粒体蛋白质,分别用荧光染料Cy3,Cy5标记,每个Cy3及Cy5标记的样品与等量Cy2标记的内标混合后在同一块凝胶中进行电泳分离,经不同光激发后扫描得到不同样品的蛋白质组图谱。经DeCyder 6.5软件进行差异蛋白质组分析,并对差异蛋白质进行质谱鉴定。结果:双向电泳后,平均每块胶检测到997个蛋白质点,其中所有图谱共有的点有687个,并从中筛选到22个差异蛋白,其中热休克蛋白60、过氧化氢酶、谷胱甘肽过氧化物、氨甲酰磷酸合成酶Ⅰ,ATP合成酶、乳铁传递蛋白、ATP酶质子通道蛋白、电子传递黄素蛋白和类钙离子激活中性蛋白酶12在肾阳虚状态下表达量均增加,而二氢硫辛酰胺支链酰基转移酶、脂酰辅酶A脱氢酶、鸟氨酸转氨酶和GTP结合调节蛋白在肾阳虚状态下表达量均降低。经温补肾阳药治疗后,热休克蛋白60、过氧化氢酶、谷胱甘肽过氧化物、ATP合成酶、乳铁传递蛋白、ATP酶质子通道蛋白、电子传递黄素蛋白二氢硫辛酰胺支链酰基转移酶、脂酰辅酶A脱氢酶和GTP结合调节蛋白表达量均增加,而氨甲酰磷酸合成酶Ⅰ和类钙离子激活中性蛋白酶12表达量减少。结论:温补肾阳药可能通过调节物质代谢、保护线粒体膜稳定性和维护细胞内信号传导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 thyroidectomized kidney-yang deficiency rats. Method： Twelve rats were divided into normal group, model group and treated group; each group had four rats. The rats of model group and treated group were excised the two side thyroid gland, and the rats of normal group were done the homologous operation, but didn＇t excise the thyroid gland. After seven days, the rats of model group and treated group appeared the symptoms of kidney-yang deficiency. From the eighth day, the rats of treated group were fed warm and tonify kidney-yang herbs 6.7 g · kg ^- 1 once daily, and the rats of other two groups were fed the equal normal. All rats of three groups were killed by decollation after six days of treatment, and liver mitoehondria proteins were separated. Each liver mitochondria protein sample was labeled with Cy3 or Cy5 randomly, and one Cy3-labeled sample and one Cy5-labeled sample were mixed on the same 2-D get along with a Cy2- labeled mixture of all samples as an internal standard and run on the same gel. The gels were scanned under different wavelength light after clectrophoresis. All images were analyzed by DeCyder 6.5 software, and the different proteins were identified by mass spectrum. Result： The expression of HSP60, catalase, glutathione peroxidase, carbamoylphosphate synthetase I , ATP synthase, lactotransferrin, H ^＋ -transporting two-sector ATPase, alpha-ETF and calpain 12 were increased in the thyroidectomized kidney-yang deficiency rats, while the expression of oxoisovalerate dehydrogenase, Acyl-CoA dehydrogenase, omithine aminotransferase, and GTP-binding regulatory protein were decreased. After the kidney-yang deficiency rats were treated with warm and tonify kidney-yang herbs, the ex- pression of HSP60, catalase , glutathione peroxidase, oxoisovalerate dehydrogenase , Acyl-CoA dehydrogenase, ATP synthase, lacto- transfcrrin, H^＋ -transporting two-sector ATPase, alpha-ETF and GTP-bi

关 键 词：蛋白质组 肾阳虚 温补肾阳药 线粒体 

分 类 号：R285.5[医药卫生—中药学]