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目的研究肾阴虚、肾阳虚模型大鼠股骨皮质骨差异表达的蛋白质,从功能蛋白质组角度探讨肾阴虚、肾阳虚证的实质。方法将90只大鼠按随机数字表法分为正常组、肾阴虚组和肾阳虚组,每组30只,获取造模成功且表现稳定大鼠的股骨皮质骨,分别进行皮质骨总蛋白的提取制备、双向电泳(2-DE)、凝胶图谱的获取与分析、差异蛋白的质谱分析及相关功能的生物信息学检索。结果 3组大鼠股骨皮质骨总蛋白2-DE凝胶图谱可辨识蛋白点数目均为470个,与正常组比较,肾阴虚、肾阳虚组各有12个差异蛋白,其中有6个是肾阴虚、肾阳虚组共有的差异蛋白,4个表达趋势一致,分别为表达上调的载脂蛋白A-1、α烯醇化酶,表达下调的GDP解离抑制因子β、原肌球蛋白异构体2α-3链;有2个蛋白表达趋势相反,Ⅰ型胶原α-1链在肾阴虚组中表达上调,在肾阳虚组中表达下调;线粒体热休克蛋白60在肾阴虚组中表达下调,在肾阳虚组中表达上调;肾阴虚、肾阳虚组各有6个独自的差异蛋白,其中肾阴虚组有3个蛋白表达上调,3个蛋白表达下调;肾阳虚组6个蛋白表达均下调。结论大鼠股骨皮质骨中,4个共同且表达趋势一致的差异蛋白可以视为肾阴虚、肾阳虚证的共性功能蛋白质组;2个表达趋势相反及12个肾阴虚、肾阳虚组独自的差异蛋白可归为相应证型的差异功能蛋白质组。
Objective To study the differentially expressed proteins of the femur cortical bone in rats with deficiency of kidney yin and deficiency of kidney yang model and to explore the essence of deficiency of kidney yin deficiency and deficiency of kidney yang from the perspective of functional proteome. Methods 90 rats were randomly divided into normal group, kidney yin deficiency group and kidney yang deficiency group, 30 rats in each group. The cortical bone of the femur was obtained successfully and the rats were stable. Protein extraction and preparation, two-dimensional electrophoresis (2-DE), gel mapping acquisition and analysis, differential protein mass spectrometry and related functions of bioinformatics search. Results The number of distinguishable protein spots on the total protein 2-DE of the femur in the three groups were all 470. Compared with the normal group, there were 12 differential proteins in the kidney yin deficiency and kidney yang deficiency groups, of which 6 Is the common protein in kidney-yin deficiency and kidney-yang deficiency group. The four expressions are the same, which are up-regulated apoA-1, a-enolase, down-regulated GDP dissociation inhibitor β, Protein isoforms 2α-3 chain; two proteins showed the opposite trend. The type Ⅰ collagen α-1 chain was up-regulated in the kidney-yin deficiency group and down-regulated in the kidney-yang deficiency group. The mitochondrial heat shock protein 60 There were 6 independent proteins in kidney yin deficiency group and kidney yang deficiency group, of which 3 proteins were up-regulated and 3 proteins were down-regulated in kidney-yin deficiency group; Kidney Yang group 6 protein expression were down. CONCLUSIONS: Four common proteins in the femur and cortical bone of rats can be regarded as the common functional proteome of kidney-yin deficiency syndrome and kidney yang deficiency syndrome. The two expression trends are opposite and 12 kidney yin deficiency and kidney yang deficiency syndrome The group of individual differential proteins can be classified as differentially functional proteomes of the corresponding syndromes.