六味地黄汤对甲亢型肾阴虚大鼠肝线粒体蛋白质组的影响

目的 应用双向电泳技术和质谱技术研究六味地黄汤对甲亢型肾阴虚大鼠肝线粒体蛋向质组的影响.方法 双向电泳分离甲状腺素造成的肾阴虚模型和六味地黄汤治疗大鼠肝线粒体蛋白质,银染后扫描得到蛋白质组图谱,用Image Master2D6.0软件进行差异蛋白质组分析,筛选差异在2倍以上的蛋白质进行质谱鉴定和生物信息学分析.结果 大鼠Major urinary protein precursor、Peroxiredoxin-6、内质网蛋白29前体、大鼠转甲状腺素蛋白D链表达升高,78 kDa葡萄糖调节蛋白前体、谷胱甘肽硫转移酶Mu 1、热休克蛋白60、肽基脯氨酰顺反式异构酶、核转录因子-κB 2、H~+-转运ATP合酶、BNIP1 protein表达均降低.结论 六味地黄汤滋阴补肾作用机理可能与下调线粒体内H~+-转运ATP合酶表达量从而减缓能量代谢、抑制NF-κB过量表达和活化以调节炎性细胞因子紊乱状态和凋亡抑制状态、上调PDX6表达对抗氧自由基损伤以及增强性激素分泌水平相关.

Effects of Liuwei Dihuang Decoction on hepatic mitochondria proteome of kidney-yin deficiency rat induced by hyperthyrea

Objective Two-dimensional electrophoresis and mass spectrum were used to study the effects of Liuwei Dihuang Decoction on hepatic mitochondria proteome of kidney-yin deficiency rat induced by hyperthyrea. Methods The protein of liver mitochondria from kidney-yin deficiency rats and the kidney-yin deficiency rats treated with Liuwei Dihuang Decoction were isolated by two-dimensional electrophoresis, then stained with silver staining method. The gels were respectively imaged by UMAX ImageScanner and spot-features of proteome difference were analyzed by ImageMaster2D6.0 software. MALDI-TOF MS was used to identify the different protein with abundant changes more than 2 folder between normal rats and kidney-yin deficiency rats. Results In the treated group it was found that major urinary protein precursor, peroxiredoxin-6, endoplasmic reticulum protein 29 precursor, chain D rat transthyretin were increased, while 78 kDa glucos eregurated protein precursor, glutathione S-transferase Mu 1, 60 kDa heat shock protein, peptidyl-prolyl cis-trans isomerase, H~+ -transporting ATP synthase, nuclear factor kappa B 2, BNIP1 protein were decreased. Conclusion The mechanism of nourishing kidney-yin action of Liuwei Dihuang Decoction lies in its relieving energy metabolism, regulating inflammatory factorderangement and apoptosis inhibition, antagonizing free radical damage, enhancing gonadal hormone level by the down regulation of NF-κB, H~+-transporting ATP synthase, and the up-regulation of PDX6.