基于高通量检测技术的腮肿病未熟热证、成熟热证期标志物与机制研究

The febrile disease theory of Mongolian Medicine is a branch of knowledge focused on studying infectious and immunologic diseases. The theory of Mongolian Medicine regarding fever mechanism, syndrome conversion and diagnosis is original. Mongolian Medicine dissects the process of fever into several syndromes. The conversion of each syndrome is regular and manipulable. According to the febrile disease theory of Mongolian Medicine, clinical doctors can accurately determine each syndrome and the conversion of syndromes, which information can be used for accurate diagnosis and treatment of the disease. It has been proved that the febrile disease theory of Mongolian Medicine fits in with clinical needs. It is reliable and manipulable in clinical practice. However, studies on this theory are rare and the scientific basis of this theory is unclear, which prevent the application of this theory. The current research focuses on two syndromes of mumps, immature heat syndrome and mature heat syndrome. We will collect serum samples from patients with immature heat syndrome of mumps and mature heat syndrome of mumps. And then, we will use advanced mass spectrometer to compare serum proteomes of these two syndromes to identify differentially expressed proteins. Bioinformatic tools will be used for the selection of possible biomarkers for diagnosis and differentiation of these two syndromes. The possible biomarkers will also be confirmed by using antibody microarrays in more patient samples. We expect to study the conversion mechanism of these two syndromes in proteomic scope. Our study may provide molecular basis for the application of the febrile disease theory of Mongolian Medicine.

蒙医温病学是专门研究感染免疫性疾病的学科，该理论在发热机理、变证规律、诊疗等方面具有独到的论述。它把发热过程分为若干个证期，每个证期的变化都具有规律性和可控性，医生能够准确判定即时的证和变证去向进行精确诊疗。实践证明，该理论切合临床，操作性强，诊疗效果可靠，但缺乏现代科学研究，其技术内涵尚未阐明，从而阻碍其推广和发挥应有的作用。本研究拟选择蒙医腮肿病（腮腺炎）未熟热证期、成熟热证期病人为研究对象，采用高通量质谱技术，检测未熟热、成熟热二证期血浆差异表达蛋白，结合生物医学信息和网络技术进一步筛选和验证与变证的相关性和变证过程中的作用、意义，寻找二证期诊断指标和特异性标志物。这些标志物也将用芯片在更多的样本中进行验证。我们预期从蛋白质组学水平探讨和阐述二证期变证机制，并为临床辩证诊疗提供实验室依据。

收集符合条件的流感、麻疹、腮肿病共22例，健康人10例。采集每例病人蒙医未熟热、成熟热证期静脉血，采用高通量质谱检测技术检测三组病人未熟热、成熟热证期血浆蛋白质组，共检测到血浆低丰度蛋白质或多肽1499个，其中二证期差异表达共1323个，差异表达1.3倍以上共1077个，成熟热和未熟热与正常组相比变化趋势一致的共164个（其中上调95个，下调69个），其中三种疾病在二证期共同上/下调1.3倍以上共8个（上调6个，下调2个）。可见，不同证期显现差异表达率88.3%，差异表达1.3倍以上71.8%，说明蒙医证期转变过程中蛋白质的变化谱之广泛；成熟热和未熟热与正常组相比变化趋势一致的占10.9%，说明差异表达的规律性较强，提示不同病症可以经过相同证期；其中二证期共同差异表达1.3倍以上共8个，其上、下调比例为6:2。该8种蛋白具有很好的证期标志物意义，可进一步验证。未熟热证期是感染后发生病理变化的初始阶段，是所有热症均可经过的病理过程，是诊疗关键期。蒙医未熟热、成熟热理论技术疗效确切、操作简便。推广该理论技术将对感染性疾病防治带来广泛的应用前景，尤其对医疗条件较差的偏远地区的传染病防治工作提供合适的方案，研究阐明这一临床理论体系的科学内涵具有重要意义。