Study Of The Essence Of The Spleen QI Deficiency Based On Metabolomics Of Urine

Objective: The study of spleen qi deficiency essence is an important element in syndrome essence. Traditional Chinese Medicine believes that spleen qi deficiency could lead to dysfunction of transportion, and stagnation of body fluid which can produce dampness, phlegm, drink as pathological products, thus triggering a variety of diseases. As the theory goes, " internal injury of spleen and stomach can cause a lot of diseases" "spleen is the source of sputum crudum ". In modern medicine, the spleen is a unit of multi-systems, multi-functional organs, which is based on the digestive system, including energy metabolism and water metabolism, and is closely connected with blood, endocrine, immune and nervous system. Metabolic syndrome(MS) is a disease which is seen as central task of central obesity, it involves a variety of metabolic abnormalities, including blood pressure, blood sugar, blood lipids, and it is associated with cardiovascular disease closely. The main etiology of MS is obesity. Phlegm-dampness is an important pathological factor in obesity, which is also the center of MS, so the phlegm-dampness is the central pathology factor in the main syndromes of MS.This shows that the spleen deficiency which causes the dysfunction of transportion is important in the pathogenesis of MS. Therefore the morbidity of MS is inseparable with the spleen. Metabolomics is an emerging science and is applied widely in the medicine domain. It measures small molecules of the metabolite of body fluid and tissue qualitatively or quantitatively by modern instrumental analysis methods such as nuclear magnetic resonance (NMR), chromato- graph(GC), mass spectrum(MS), reveals the physiological and pathological state in disease process systematically, however this systematic character- ristic observation is similar to holistic concept in the Traditional Chinese Medicine. At present, metabolomics has already been seen as a new technology to apply in diagnosis of clinical disease. We investigated the small molecular metabolite in urine of Metabolism syndrome patients as the foundation of spleen qi deficiency using method of disease and syndrome with combination and liquid chromatography-mass spectrometry technology. We expect to discover the change pattern or characteristic peak in metabolome through comprehensive and integrated analysis, attempting to express qualitatively or quantitatively in diagnosing spleen qi deficiency of Traditiongal Chinese medicine or evaluation index. Then further reveal metabolome characteristic in spleen qi deficiency and also bring to light the modern research target of spleen essence in Chinese medicine.Methods:The cases come from physical examination center of second hospital affiliated with Dalian Medical University.The age is from 40 to 65. Collection of physical examination personnel's early morning urine or second morning urine , then lay them aside in -20℃. Diagnosis standard for metabolism syndrome was promulgated by international diabetes alliance (IDF) on April, 2005.Traditional Chinese medicine New Medicine Clinical research Guiding principle" promulgated in 2002 is taken to diagnose spleen qi deficiency. The cases were divided into three groups:including cont group, metabolism syndrome group, and metabolism syndrome combined spleen qi deficiency group. The urine were storaged at -20℃for one week,and then were storaged at -80℃.The ultra performance liquid chromatogram-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF- MS)is utilized to analysis ingredient in urine, and to express the different metabolite qualitatively. Partial least squares-discrmininant analysis (PLS-DA) is used to process the data.Results:After analysis of metabolomics, from the PLS-DA schema chart we discovered that metabolism outline had the satisfactory discri- mination in every two groups of urine, and we obtained the corresponding latent designated object: 130.048m/z unknown compound and 310.198m/z meat alkali are latent tracer in urine of Metabolism syndrome group,the density in urine decreased compared to cont group; the density of 229.137 m/z leucylproline was increased, but the density of 265.115m/z pheny- lacetylglutamine was decreased in urine of metabolism syndrome and spleen- qi-deficiency compared to normol and metabolism syndrome.Conclusions:1. Urine metabolism profiling has the satisfactory discrimination between the spleen qi deficiency connect with metabolism syndrome and the normal group.2. We had discovered some latent tracers of spleen qi deficiency, such as leucylproline and phenylacetylglutamine, this is helpful for clinical doctor to practice the standar of western medicine's microscopic diagnosis of spleen qi deficiency, and it played a standard role in quantification of the Chinese medicine spleen qi deficiency card.3. The systematic characteristic of metabolomics observed is similar to holistic concept in traditional Chinese medicine., this can unifies the syn- drome essence of traditional Chinese medicine and the modern research better.