Studies On Pharmacodynamics Of R.glycyrrhizae And Flavonoids From Gancaofuzi Decoction

Traditional Chinese Formula Gancaofuzi decoction is composed by Radix Aconiti Praeparata, R.glycyrrhizae, Cinnamomum cassia Presl and A. macrocephalae Rhizoma. In clinic, it is mainly used for treating rheumatic and rheumatoid arthritis. By the study on the animal model in vivo and macrophage in vitro, we observed the anti-inflammatory and analgesic activities of R.glycyrrhizae and flavonoids in the Gancaofuzi decoction and their immunomodulatory effects. At the same time, we observed the effects of the different drug pairs to R.glycyrrhizae and the different active ingredients to flavonoids on inflammatory, analgesic and immunomodulatory effects in the Gancaofuzi decoction. The project used UPLC/MS technique to analyze endogenous metabolite changes in adjuvant-induced arthritis rats between Gancaofuzi decoction and flavonoids in Gancaofuzi decoction. Our study aimed at the effective substances basis of the Gancaofuzi decoction. The study includes the following four aspects:1. Study on effects of R.glycyrrhizae and the coupled drug dispensing on pharmacodynamics of R.glycyrrhizaeFirstly, inflammatory and analgesic effects of Gancaofuzi decoction were better than single herb R.glycyrrhizae and its drugs pairs, indicating the all crude drugs in Gancaofuzi decoction together play a role in anti-inflammatory and analgesic effects, in line with traditional Chinese medicine to increase application compatibility pharmacodynamics theory. Through the summary of anti-inflammatory, analgesic and immune cells results of single herb R.glycyrrhizae, we found that R.glycyrrhizae played a primary role as an anti-inflammatory effect in Gancaofuzi decoction. When the single herb R.glycyrrhizaewas compatible with other herbs in Gancaofuzi decoction, the results showed all administration groups had a good inhibition both the secretion of TNF-α in immune cell and the serum levels of TNF-α in inflammatory model. While the single herb R.glycyrrhizawas mixed with Radix Aconiti Praeparata, the administration group exhibited to significantly inhibit the secretion of interleukin and enhance its analgesic effect. In terms of anti-inflammatory, the single herb R.glycyrrhiza mixed with Cinnamomum cassia Presl and A. macrocephalae Rhizoma showed a good inhibition of the acute and chronic inflammation.2. Study on effects of flavonoids and the compatibility of effective components on pharmacodynamics of flavonoids from Gancaofuzi decoction.Sencondly, the Gancaofuzi decoction and flavonoids had a good anti-inflammatory effect. When the flavonoids were mixed with alkaloids in Gancaofuzi decoction, it could be a certain extent to increase the application of flavonoids’ anti-inflammatory effects, but no significant difference. On the contrary, the compatibility of saponins and flavonoids or compatibility of the three kinds of constituents together in Gancaofuzi decoction reduced efficacy. The compatibility of flavonoids and saponins or compatibility of the three kinds of constituents together had a significant analgesic effect. Due to the great anti-inflammatory effects of flavonoids, flavonoids also had significant analgesic effectsin the pain caused by inflammation in the actual experiment.3. Metabolomic applied to study the adjuvant arthritis treated mechanisms of Gancaofuzi decoction and flavonoids.Thirdly, a metabonomics method based on the UPLC-Q-TOF-MS was used to analyze themetabolic changes in adjuvant-induced arthritis rats’ urine and find out the biomarkers. We identified 20 potential biomarkers with adjuvant arthritis-related including creatinine, uric acid, citric acid, pantothenic acid, xanthurenic acid, and taurine, etc. The changes of 13 endogenous metabolites were considered to be in response to therapeutic effects of Gancaofuzi decoction and flavonoids including homocysteine, taurine, p-cresol, kynurenicacid, xanthurenic acid, pyroglutamate, pantethine, purines, suberic acid, sebacic acid, phenethylamine, hippuric acid and indole. From the above identified biomarkers, we known Gancaofuzi decoction and flavonoids affected the metabolic pathways include: phenylalanine metabolism, tryptophan metabolism, citrate cycle, and pantothenate and Co A biosynthesis and taurine and hypotaurine metabolism. Fluctuations in the relative content of these metabolites showed Gancaofuzi decoction and flavonoids could mainly adjust the energy metabolism, amino acid metabolism and intestinal flora metabolism of rats with adjuvant arthritis.Compared with the flavonoids, Gancaofuzi decoction affected endogenous metabolites with adjuvant arthritis-related in a wider range, presumably because other active ingredients had a certain influence in relevant metabolic pathways of rats with adjuvant arthritis.4. Study on effects of the compatibility of effective components on metabolomics of flavonoids from Gancaofuzi decoctionFinally, we studied the effects of the treatment of adjuvant arthritis on metabolomics after flavonoids mixed with saponins and alkaloids. The results showed that the compatibility of three components together had a better regulation on the majority of biomarkers than flavonoids alone, which prominently improved the ability to regulate purine metabolism and taurine metabolism. When flavonoids were mixed with alkaloids, the regulations of most biomarkers were better than flavonoids alone. Three metabolites of tryptophan metabolic pathway were shown a great regulation, but the regulations of taurine and purine metabolism were weaker than the compatibility of three components together. When flavonoids were mixed with saponins, the regulations of most biomarkers were inferior to flavonoids alone, but which in tryptophan metabolism also showed a good regulation.Inconclusion, the pharmacodynamic and metabolomics studies defined the effective substance basisof Gancaofuzi decoction to treat the rheumatism arthralgia, which provided new ideas, new methods to study on pharmacodynamic material basis of Chinese medicine prescriptions.