"唾液-脾肾蛋白质关联"新假说的物质基础及脾肾虚病证结合新型无创性分子诊断模型研究

In recent years, Studies on saliva detection which are noninvasive and convenient have already aroused great interest. Saliva is considered as biological liquid of great scientific and clinical potential. And the advanced technologies of proteomics have provided a powerful methodology for the breakthrough of saliva detection. Based on the related theory of traditional Chinese medicine(TCM)- - "saliva is the liquid metaplasiaed by the Spleen and Kidney" , and combined with modern research achivements, this project presents a new hypothesis of "Saliva protein associated with the Spleen and Kidney", and uses new proteomics research technologies of SELDI-TOF-MS and iTRAQ, Selects patients with spleen deficiency or kidney deficiency syndrome according to the generally accepted standard of deficiency syndrome from five kinds of common diseases such as Chronic ulcerative colitis, Chronic gastritis, gastric cancer, chronic nephritis and type 2 diabetes mellitus, groups according to the rules " the same disease with different TCM syndrome " and "different diseases with the same TCM syndrome "， makes a series of studies on the profiles of salivary proteome expression, validation of differentially expressed proteins， physical and chemical nature, structure and function of the newly discovered proteins in the patients with Spleen deficiency (Spleen- Qi Deficiency，spleen- yang deficiency) and kidney deficiency (kidney -qi deficiency, kidney-yin and kidney-yang)；and establishs novel noninvasive molecular diagnostic models combined with disease and TCM syndrome using bioinformatics methods. The study has important significance for revealing material basis of the new theory hypothesis of "saliva protein associated with the Spleen and Kidney of TCM", elucidating the modern life science connotation of the TCM theory of "saliva is the liquid metaplasiaed by the Spleen and Kidney", innovating and developing related TCM theory, and promoting the modernization of TCM diagnosis.

近年来，安全无创便捷的唾液诊断研究已引起人们极大的兴趣，唾液被认为是具有极大科研和临床潜力的生物液，而蛋白组学技术的崛起为其突破提供了强大武器。本项目基于"脾为涎,肾为唾"、唾液为脾肾化液的传统中医理论，结合现代相关研究成果，首次提出"唾液-脾肾蛋白质关联"新假说，并运用SELDI-TOF-MS 结合iTRAQ定量蛋白质组学新技术，选择慢性溃疡性结肠炎、慢性胃炎、胃癌、慢性肾炎、2型糖尿病等5种常见相关疾病，以同病异证、异病同证为纲分组，进行脾虚（脾气虚和脾阳虚）、肾虚（肾气虚、肾阴虚和肾阳虚）及病证结合唾液蛋白质组差异表达、差异蛋白验证及新发现蛋白的理化性质、结构和功能系列研究，并运用生物信息学方法建立脾肾虚及病证结合新型无创伤分子诊断模型。该项目对于揭示唾液为脾肾化液理论及"唾液-脾肾蛋白质关联"新假说的物质基础与科学内涵，创新发展相关中医学术理论，推进中医诊断现代化均具有重要意义。

唾液被认为是具有极大科研和临床潜力的生物液，而蛋白组学技术的崛起为其突破提供了强大武器。本项目基于“脾为涎,肾为唾”、“唾液为脾肾化液”的传统中医理论，结合现代相关研究成果，首次提出“唾液-脾肾蛋白质关联”新假说，研究采集慢性胃炎、慢性肾炎、2型糖尿病、胃癌、肠癌、肝癌、甲状腺癌以及乳腺癌441例以及正常对照者180例，根据西医病种和中医异病同证、同病异证进行分组，采用SELDI-TOF-MS技术进行蛋白指纹图谱检测，寻找疾病组与对照组明显差异的蛋白峰，运用生物信息学方法建立了多个病证结合的唾液蛋白无创性新型分子诊断模型；同时，运用iTRAQ标记定量蛋白组学先进技术对慢性胃炎、胃癌、慢性肾炎、2型糖尿病、肝癌以及肠癌进行病证结合的唾液蛋白质组学定量研究，找到了多个疾病和证候相关的特异表达蛋白，并进行基因本体研究、相关通路分析及蛋白质交互作用研究，进一步对差异蛋白进行临床验证，发现了多个有诊断价值的特异性唾液蛋白。本研究发表相关论文18篇，获得省部级成果奖2项，申请国家发明专利13项，揭示了“唾液为脾肾化液理论”及“唾液-脾肾蛋白质关联”新假说的物质基础与科学内涵，创新发展了相关中医学术理论，推动了中西医结合分子诊断学与微观辨证学的发展，对于中医诊断现代化具有积极现实意义。

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