基于LC/MS、NMR分析方法的犊牛腹泻中兽医证候本质的代谢组学研究

Syndrome, which is an important concept in Traditional Chinese Veterinarian theory, is the starting point and cornerstone of diagnosis and treatment. But as a result of systematic, holistic and dynamic characteristics of syndrome, syndrome diagnostic criteria and the essence has always been a problem in application and development of Traditional Chinese Veterinarian Medicine(TCVM) .This project intends to explore a method for studying the syndrome essence of Traditional Chinese Veterinarian based on the Metabolomics theory and methods and systems biology methods. First refer to classic works and related literature, The diagnostic criteria of damp-heat type and cold type calf diarrhea will be established based on cluster analysis, factor analysis and other multivariate statistical methods. Then endogenous small molecule metabolites spectrum in serum and urine of calf with damp-heat type and cold type diarrhea will be established by the methods of LC-MS and NMR. The biological marker characterization associated with damp-heat type and cold type diarrhea syndrome will be determined by pattern recognition analysis ways of the Metabonomics, and the syndrome essential of damp-heat type and cold type diarrhea will be revealed. The results of this research will explore a method for building diagnostic criteria and revealing the essence of Chinese veterinarian syndrome.

证侯是中兽医学理论体系中的重要概念，是辨证论治的起点和基石。但由于证侯的系统性、整体性和动态性特征，证侯诊断标准的客观界定以及症候的实质研究，一直是困扰中兽医药研究应用与发展的难题。本项目拟利用代谢物组学的理论及方法和系统生物学观点，为中兽医证侯本质的认识进行方法探索。项目首先明确湿热型犊牛腹泻和虚寒型犊牛腹泻的诊断标准，以临床自然腹泻犊牛病例为受试对象，采集血液和尿液，借助高效液相色谱-质谱联用、核磁共振现代分析技术手段，建立湿热型腹泻、虚寒型腹泻犊牛血清、尿液中内源性小分子代谢物谱，通过代谢组学模式识别分析，确定出特异性地关联湿热型腹泻和虚寒型腹泻犊牛的生物标记物，阐明犊牛腹泻中兽医证侯的微观本质。本研究结果将为中兽医证侯的诊断客观化和中兽医证侯本质的认识进行方法探索。

证侯是中兽医学理论体系中的重要概念，是辨证论治的起点和基石。但由于证侯的系统性、整体性和动态性特征，证侯诊断标准的客观界定以及症候的实质研究，一直是困扰中兽医药研究应用与发展的难题。本项目利用代谢组学的理论及方法和系统生物学观点，为中兽医证侯本质的认识进行方法探索。项目组对犊牛腹泻感染的病原学进行了研究，结果西北奶业主产区内的犊牛腹泻病原主要是细菌性病原；而在东北和华北奶业主产区则分别是轮状病毒和寄生虫，其中寄生虫病原主要包括隐孢子球虫和肠兰伯氏鞭毛虫。建立了湿热型犊牛腹泻诊断标准和虚寒型犊牛腹泻诊断标准，并利用基于HILIC UHPLC-Q-TOF MS技术的非靶向代谢组学方法分别对湿热型犊牛腹泻和虚寒型犊牛腹泻样本进行了代谢轮廓变化分析。共检测到虚寒型犊牛腹泻和湿热型犊牛腹泻差异代谢物26个，共涉及16个代谢通路，尤其对Protein digestion and absorption，Central carbon metabolism in cancer，ABC transporters，Aminoacyl-tRNA biosynthesis和Mineral absorption代谢通路影响较大。发表论文4篇，获得兰州市科技进步一等奖1项，甘肃省农牧渔业丰收奖一等奖1项，培养研究生1名。