Study On The Rapeutic Effect And Mechanism Of Huoxue Capsule On Qi Deficiency And Blood Stasis Rats

Chinese Herbal Compound Formula is the primary means of chinese clinical medicine treatment of disease, its efficacy and safety have already been proved by more than two thousand years of clinical experience. Metabolomics, which could combine so many Chinese herbal medicinal ingredients with complexity of the body, is a major tool for the effect and mechanism research of Chinese herb. This paper took Huoxue Capsules, which derived from Xuefuzhuyu decoction, as the object of study, and the qi-deficiency and blood-stasis model was made, and then the endogenous substances of plasma, urine and rat brain tissue under the pathophysiological and Huoxue capsules intervention were investigated by mean of HPLC-FLD method and HPLC-MS metabolomics, thus illustrating the effect and mechanisms of Huoxue capsules on qi-deficiency and blood-stasis. The main contents of this paper are listed as follows:1. Taking the content of amino acids in plasma as the index, the effect of Huoxue capsules on qi-deficiency and blood-stasis model rats was observed in this part of research. Through precolumn derivatization method with high performance liquid chromatography method established, amino acids in plasma of rats from the control group, model group and Huoxue capsules administrated group were determined. The results showed that:in comparison with normal group,serine and phenylalanine significantly increased, as well as aspartic acid, glutamic acid, glycine, tyrosine, tryptophan and isoleucine were significantly lower in model group (P<0.01);however,the five kinds of amino acids significantly increased,and serine and phenylalanine significantly decreased in high-dose group compared with model group(P<0.05);meanwhile,there was significant increase in aspartic acid, glutamic acid and decrease in serine, phenylalanine for middle-dose group compared to model group(P<0.01); and serine had dropped dramatically in low dose group compared to model group (P<0.01). We can infer that the amino acids changed have associated with qi-deficiency and blood-stasis syndrome from results above. Meanwhile, changes of amino acid content aforementioned could be rectified by Huoxue capsules, revealing that influence on certain metabolic pathways of amino acids may be a therapeutic approach of Huoxue capsule for qi-deficiency and blood-stasis syndrome,which is consistent with the literature report.2. Taking the inhibitory and excitatory amino acids in plasma as the index, the effect of Huoxue Capsules on qi-deficiency and blood-stasis model rats was observed. Through precolumn derivatization method with high performance liquid chromatography method established, amino acids in brain tissue of rats from the control group, model group and Huoxue capsules administrated group were determined.The results proved that:in comparison with normal group,aspartic acid, and glutamic acid significantly increased (P<0.05), as well as γ-aminobutyric acid、alanine and glycine were significantly decreased in model group (P<0.01); in comparing with the model group, there was significant decrease in aspartic acid and glutamic acid and increase in the rest of the three kinds of amino acid for high-dose group (P<0.05); there was significant decrease in γ-aminobutyric acid and increase in aspartic in middle-dose group and low-dose group in comparison with model group (P<0.01). We can infer that Huoxue Capsules could probably make the content of amino acids in brain of model rats return to the mormal lever.3. Based on LC-MS metabolomics, comparing the control group, model group and Huoxue capsules group differences in rat urine substance, looking qi-deficiency and blood-stasis syndrome associated with pathological markers and metabolic pathways involved in Huoxue capsules. The results showed that:comparing with the normal group, the content of4-hydroxyphenyl pyruvic acid, aspartic acid, histidine4-hydroxy-L-glutamic acid, uridine,3-methoxy adrenaline were changed remarkably in urine of model rats, which suggested qi-deficiency and blood-stasis syndrome was associated with amino acid metabolism pathways, purine, pyrimidine metabolic pathways, and these substances above can be deemed as the key metabolites of qi-deficiency and blood stasis syndrome. Furthermore homocysteine, ketoglutarate, galactose acid, uridine were significantly different between the model group and Huoxue capsules administrated group, which revealed that Huoxue capsules could affect metabolic pathways of methionine, histidine acid metabolic pathways, carbohydrate metabolism and pyrimidine metabolism pathways aiming to improve blood stasis symptoms.