摘    要:目的：从蛋白组学整体水平研究儿童哮喘稳定期不同中医证候的血清差异蛋白，为规范儿童哮喘治疗的证候辨证和分型标准，为阐明证候的生物学本质奠定科学基础。方法：采用同位素标记相对和绝对定量标记结合二维液相色谱串联质谱蛋白组学技术的方法，来筛选鉴定肺脾气虚证，肺肾阴虚证和脾肾阳虚证儿童哮喘患者血清蛋白中差异表达的蛋白，并通过生物信息学进行分析。结果：共鉴定总血清蛋白260种，从中筛选出与其他组别均表达差异的脾肾阳虚证儿童哮喘差异蛋白质共51种（取变化倍数<0.8或>1.2）;差异蛋白主要参与20种生物过程，实现8种分子功能，并定位在17种细胞结构。结论：获得脾肾阳虚组儿童哮喘患者血清中的特征性蛋白51种，这些特征性蛋白可以作为鉴别小儿哮喘脾肾阳虚证的候选蛋白。

Abstract

OBJECTIVE: To study serum differential proteins of different TCM syndromes in the stable serum albumin group in children, to standardize the syndrome differentiation and classification criteria for childhood asthma treatment, and to lay a scientific foundation for clarifying the biological nature of syndromes. METHODS: Isotopically labeled relative and absolute quantitative markers were used in combination with two-dimensional liquid chromatography tandem mass spectrometry to screen and identify differentially expressed proteins in serum proteins of children with asthma due to deficiency in lung and spleen qi, lung-kidney yin deficiency and spleen-kidney yang deficiency. , and analysis through bioinformatics. RESULTS: A total of 260 proteins were identified, and 51 kinds of proteins with different expressions between the spleen-kidney yang deficiency syndrome and other groups were screened out (change in magnification <0.8 or >1.2); The differential proteins were mainly related with 20 biological processes, 8 molecular functions and 17 cell structures. CONCLUSION: 51 differentially expressed proteins in the serum of children with asthma in the spleen-kidney Yang deficiency group. These differential proteins can be used as candidate target proteins for differentiating TCM syndromes in children with asthma.

关键词：儿童哮喘; 蛋白组学; 脾肾阳虚证；中医证候   

Keywords: Childhood asthma; proteomics; Spleen and Kidney Yang Deficiency; TCM syndromes

目    录

1 引言 4

2 材料与方法 5

2.1 实验样本 5

2.2 实验试剂 5

2.3 主要实验仪器 5

2.4 实验方法 5

2.4.1 血清差异蛋白的鉴定及定量 5

2.4.2 iTRAQ的标记及检测 6

2.4.3 毛细管高效液相质谱检测及鉴定 7

2.4.4 质谱数据统计分析 7

2.4.5 生物信息学分析 7

3 实验结果与分析 7

3.1 脾肾阳虚组特征蛋白 8

3.2 差异蛋白GO分析 定量蛋白组学技术筛选脾肾阳虚证儿童哮喘特征性蛋白:http://www.chuibin.com/yixue/lunwen_205895.html