Investigation On The Material Basis Of Harmonious Effection Of Gancao In Herb-pair ’Honghua-Gancao’

The Traditional Chinese medicines have much unique advantages in treatment for cerebral-cardio vascular diseases. However, it’s rather difficult to study the compatibility of Chinese herb formulas, owning to its complex component and flexibility in clinical application. As the basic unit of Chinese herb compound formulas, herb-pair usually play a pivotal role in Chinese herb compound formulas, thus research on herb-pair has already to be the key point in the research of Chinese herb compound formulas, Therefore, under the guidance of the theories of Xiangshi’relationship,’Carthamus tinctorius-Glycyrrhiza radix’was selected as the object of study, energy metabolism was investigated by HPLC analytical method. Meanwhile, HPLC/MS metabonomics technology was employed in systematic studying the metabolic network of Carthamus tinctorius and’ Carthamus tinctorius-Glycyrrhiza radix’on blood stasis model rats. Therefore, the material basis of harmonious effection of Glycyrrhiza radix was revealed from the perspective of energy metabolism and metabolomics. This dissertation has three chapters, and the main contributions of the author are listed as follows:1. Under the guidance of the theories of’Qiqinghehe’in’Shen Nong Materia Medica’ we selected’Carthamus tinctorius-Glycyrrhiza radix’, which is common herb-pair treated for blood stasis syndrome, as the research object.2. Study on energy metabolism.1) A high performance liquid chromatographic method was established for simultaneously determining adenosine phosphate compositions (ATP, ADP and AMP) in rats’tissues (heart, liver, spleen, lung, kidney and cerebrum). The method showed a good linear relationship in the ranges of1.25-250.0μg/mL for ATP,2.0-400.0μg/mL for ADP,2.5-500.0μg/mL for AMP, and the limit of detection was0.028μg/mL,0.036(μg/mL and0.025μg/mL, respectively. The results for the assay of precision, stability and reproducibility conformed to the standard requirement of detecting biological samples. The result showed that this analytical method could apply to studying the quantity of three adenosine phosphate compositions in rats’tissues.2) Making hemorheological disorder of blood stasis model rat, optimized analytical method in this study was applied to simultaneously determining adenosine phosphate compositions between normal and model rats. The result showed that the contents of adenosine phosphate in model rats after administration of Carthamus tinctorius and’Carthamus tinctorius-Glycyrrhiza radix’were higher than those in model control group, which showed significant difference. Moreover, The result also manifest that Carthamus tinctorius could improve the energy metabolism in vivo, and herb-pair of Carthamus tinctorius-Glycyrrhiza radix’’was superior to improving the energy metabolism disorder with hemorheological disorder of blood stasis syndromes.3. Study on metabolomics. Urine of normal and model rats was investigated so as to acquire the flag of pathology based on HPLC-MS metabonomics. After long-term intragastric administration of Carthamus tinctorius, Carthamus tinctorius-Glycyrrhiza radix, Carthamus tinctorius-Glycyrrhetinic acid and Carthamus tinctorius-Prednisoloe, the similarities and differences in endogenous substances and their corresponding metabolic pathways were investigated based on the urinary metobonomics to explore the harmonic function of Glycyrrhiza radix. It was proved that the four drug-pairs aforementioned could improve the symptoms of the model rats. Meanwhile, comparing with the normal group, the histidine metabolic pathway, tyrosine metabolic pathway, tryptophan metabolic pathway, arginine and proline metabolic pathway, valine, leucine and isoleucine metabolic pathway, purine metabolic pathway, pyrimidine metabolic pathway, riboflavin metabolic pathway and citrate cycle were abnormal in model rats, and it can be inferred that the lever of norepinephrine and glucocorticoid were up-regulation so that the system activity was strengthened owning to the long-time cold stimulation. After fifteen days’respective treatment of Carthamus tinctorius, Carthamus tinctorius-Glycyrrhiza radix, Carthamus tinctorius-Glycyrrhetinic acid and Carthamus tinctorius-Prednisolone, it was found that symptoms of the model rats was relieved by same responsive substances and metabolic pathway which the four drug-pairs effected.