Studies On Substance Identification Strategies And Knowledge Correlation Network Of The Analogous Formulae From Zhang Zhongjing's Formulae

Formula is the common application form to treat diseases in clinical practice of traditional Chinese medicine.Efficacy of the formula reflects the moderating effects of certain formula on syndromes.What is the origin of the formula's efficacy?Is there a chemical substance group in the formula that is related to a certain efficacy?These are the most important scientific questions to reveal the scientific connotation of TCM formulaeTo clarifying the TCM formulae's efficacy substances group,we must first identify the chemical substances in the TCM formulae.Recently,liquid chromatography-mass spectrometry(LC-MS)has replaced traditional phytochemical isolation and analysis with its efficient separation capabilities and extensive structural information,becomes a mainstream technology in the field of TCM formulae substances identification.However,there are still several technical defects for the LC-MS research on the TCM formulae including the difficulty for LC-MS data processing and low efficiency of identification of those active ingredients,both of which are limited LC-MS application scope and identification efficiency.On the other hand,even if we achieved comprehensive substances identification of a certain formula,we still canno correlate the chemical subatance,activity and indication information together to clarify the relations between these knowledge units.If we want to investigate the TCM formulae's efficacy substances group,we must combine these information units together and employ the correlation analysis.Thus,how to improve the efficiency of substances identification of TCM formulae and achive integrated analysis of each knowledge units have become key technology bottlenecks during the study of TCM formulae's efficacy substances group.Zhang Zhongjing's formulae(Treatise on Cold Damage and Concise Essential of the Golden Cabinet)is the most famous and ancient TCM formulae group,which is also the source of various other formulae during the centuries long clinical application history of TCM formulae.The analogous formula is one of the most common phenomenon among Zhang Zhongjing's formulae,and becomes the breach to reveal the scientific connotation of Zhang Zhongjing's formulae.This thesis is using the famous analogous formulae from Zhang Zhongjing's formulae,Xiao-Qing-Long decoctions formulae group and Wu-Ling-San formulae group as research targets,and developed three new approaches for the substances identification of TCM formulae,and constructed three knowledge networks for the correlation research of multiple knowledge units.Based on the developed techniques,this thesis has revealed the Xiao-Qing-Long decoctions formulae's efficacy substances group(Xuanfei Pingchuan,Zhuyang Jiebiao).This thesis including two major parts,the main contents and academic contributions are summarized as follows:The first part is focused on the bottlenecks of substance identification of TCM formulae:1.Developed a pre-classification strategy for identification of compounds in TCM analogous formulae.By constructing three-dimensional ion maps,pre-classified peaks in analogous formulae by their chemical structure similarity is achieved.This study systematically identified 85 and 37 compounds of Xiao-Qing-Long decoctions and Wu-Ling-San analogous formulae,respectively.This approch reduced the difficulty of LC-MS data processing and improved the identification efficacy.2.Developed parallel chromatography coupled with on-line mass spectrometry and microplate based assays platform.Developed ultrafiltration combined with LC-MS.Both approaches are employed for active compounds screening from TCM formulae.These studies rapidly screened anti-oxidant,anti-tumor,anti-inflammatory and lipase inhibition active components from herbal or TCM formulae.The second part of this thesis is focused on the correlation analysis of knowledge units of TCM formulae and applied on the reaearch of TCM formulae's efficacy substances group study:3.Based on the developed substance identification technique,this study constructed three knowledge networks of Zhang Zhongjing's herbal formulae by integrating analysis of herbal,chemical substances,TCM symdrome,clinical efficacy.4.By correlation analysis of multi knowledge units of TCM analgous formulae,this study revealed the Xiao-Qing-Long decoctions formulae's efficacy substances group(Xuanfei Pingchuan,Zhuyang Jiebiao)and found 44 efficacy substances,and validated these results are validated by the analysis of the same efficacy TCM that are commonly used in clinical,and western blot experiment.