摘要
目的:应用双向电泳技术和质谱技术研究甲亢型肾阴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阴虚与能量代谢的关系。方法:双向电泳分离正常大鼠和甲状腺素造成的肾阴虚模型大鼠肝线粒体蛋白质,用银染方法染色,经扫描得到蛋白质组图谱,用ImageMaster 2D 6.0软件进行差异蛋白质组分析,筛选差异在2倍以上的蛋白质作为差异蛋白,进行质谱鉴定和生物信息学分析。结果:肾阴虚大鼠氨甲酰磷酸合酶Ⅰ、转甲状腺素蛋白D链表达量降低,短链-3-羟酰辅酶A脱氢酶、线粒体分裂蛋白1、谷胱甘肽硫转移酶Mu1、核有丝分裂器蛋白1、凋亡抑制蛋白3、热休克蛋白60、还原型烟酰胺腺嘌呤二核苷酸脱氢酶1β亚单位、核转录因子-kB2、BNIP1 protein、鸟氨酸转氨酶表达量均增加。结论:肾阴虚大鼠能量代谢活跃,线粒体内物质氧化分解作用加强,柠檬酸循环和氧化磷酸化加速。肾阴虚大鼠存在以NF-kB过量表达为主的免疫炎性细胞因子紊乱和凋亡抑制状态。这些发现提示肾阴虚证的发病机理和能量代谢亢进、炎性细胞因子紊乱、凋亡抑制状态有相关性。
Objective ： Two - dimensional electrophoresis and mass spectrum were used to study the difference of hepatic mitochondrial proteome between the Kidney - yin deficiency rats induced by hyperthyrea and the normal rats, and the relation between the kidney -yin deficiency and energy metabolism was observed. Methods ：The protein of liver mitochondria from normal rats and kidney - yin deficiency rats were isolated by two - dimensional electrophoresis, then stained with silver staining method. The gels were respectively imaged by UMAX Image Scanner and spot - features of proteome difference were analyzed by ImageMaster 2D 6.0 software. MALDI - TOF MS was used to identified the different protein which abundance changed more than 2 folder up or down between normal and kidney - yin deficiency rats. Results ： In the kidney - yin deficiency rats induced by hyperthyrea, the expression of Carbamoyl - phosphate synthase I , Chain D Rat Transthyretin decreased, while the expression of Hydroxyacyl eoenzyme A dehydrogenase （Short chain 3 hydroxya） , Mitochondrial fission 1 protein, Glutathione S transferase Mu 1,60 kDa heat shock protein, NADH dehydrogenase 1 beta subunit, Nuclear mitotic apparatus protein 1, Inhibitor of apoptosis protein 3, Nuclear factor kappa B2, BNIP 1 protein, Ornithine arninotransferase increased. Conclusion ： Energymetabolism was active in kidney - yin deficiency rats＇ mitochondria where the degradation and oxidization of nutritive material were being strengthened, Simultaneously, citric acid cycle and oxidative phosphorylation were being accelerated. There were inflammatory factor derangement and apoptosis inhibition resuiting from the over expressing of NF - kB in kidney - yin deficiency rats. These results give a indication that energy metabolism accentuation, inflammatory factor derangement, apoptosis inhibition, are correlated with the pathogenesis of kidney - yin deficiency.
出处
《中华中医药学刊》
CAS
2009年第12期2469-2473,共5页
Chinese Archives of Traditional Chinese Medicine
基金
浙江省中医药管理局重点项目(2006Z002)
关键词
肾阴虚
线粒体
蛋白质组
能量代谢
NF-KB
kidney-yin deficiency
mitochondria
proteome
energy metabolism
NF-kB
