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目的:应用相对和绝对定量同位素标记(iTRAQ)技术研究脾气虚大鼠回肠蛋白质组的表达,寻找有意义的差异蛋白,为探索脾气虚证的现代科学本质提供新的实验方法与依据。方法:12只健康雄性SD大鼠随机分为正常组和脾气虚组,脾气虚组采用饮食不节加力竭游泳方法造模,15d后分离提取回肠总蛋白,以iTRAQ试剂标记后进行质谱检测,并对差异表达的蛋白进行生物学分析。结果:与正常组比较,脾气虚组回肠271个上调蛋白,303个下调蛋白;Pathway显著性富集分析确定下调差异蛋白参与的主要生化代谢途径和信号转导途径有细胞外基质受体交互作用、黏着斑等19条,上调差异蛋白参与包括PPAR信号通路、甾类激素生物合成在内的通路24条。选择细胞外基质受体交互作用为兴趣通路,发现与小肠屏障功能密切相关的层粘连蛋白、胶原蛋白以及整合蛋白等19个蛋白发生下调。结论:脾气虚证可能与回肠细胞外基质受体交互作用通路改变及回肠黏膜屏障功能受损相关。
OBJECTIVE: To study the expression of ileal proteome in rats with spleen-qi deficiency syndrome by using relative and absolute quantitative isotope labeling (iTRAQ) technology and to search for meaningful differential proteins, providing a new experimental method and basis for exploring the modern science essence of spleen-qi deficiency syndrome. Methods: Twelve healthy male Sprague-Dawley rats were randomly divided into normal group and spleen-qi deficiency group. The deficiency of spleen-qi deficiency group was induced by diets without eating and exhausting. After 15 days, the total protein of ileum was extracted and labeled with iTRAQ reagent. , And biological analysis of differentially expressed proteins. RESULTS: Compared with the normal group, 271 up-regulated proteins and 303 down-regulated proteins were found in the ileum of spleen-Qi deficiency group. Pathway significant enrichment analysis identified the major biochemical metabolic pathways and signal transduction pathways involved in the down-regulation of differential proteins with extracellular matrix receptor interaction 19 adhesion spots and upregulation of differential proteins involved in 24 pathways including PPAR signaling pathway and steroid hormone biosynthesis. The interaction of extracellular matrix receptors was selected as the pathway of interest, and 19 proteins, including laminin, collagen and integrin, were found to be down-regulated in the intestinal barrier. Conclusion: The deficiency of spleen Qi may be related to the alteration of the pathway of interaction of extracellular matrix with ileum and the impaired barrier function of ileal mucosa.