ABSTRACT
This study was designed to explore the antipyretic mechanism of Pueraria radix. The method of network pharmacology was used to determine the known ingredients corresponding to Pueraria radix, predict the drug-related gene /protein targets, and analyze the interplay between key ingredients and targets. Biological Information Annotation Databases (DAVID) was used to enrich the biological processes and pathways. The result of network analysis was validated by molecular docking. It was found that 49 active ingredients of Pueraria radix not only regulate 21 targets (e.g. PTGS2, EGFR), but also affect 11 biological processes (e.g. oxidation-reduction process, prostaglandin synthesis, positive regulation of fever generation and inflammatory response) and 7 metabolic pathways (arachidonic acid metabolism, serotonergic synapse and HIF-1, et al). Molecular docking results showed that more than 65% of the active ingredients could be well docked with key targets, and the relevant literatures indicated that the active components could inhibit the expression of PTGS2, which means the result has a high reliability. These results indicated that Pueraria radix may carry its pyretic action via a "multi-ingredients-multi-targets-multi-pathways" mode, which provides a scientific basis for further research and drug development.
ABSTRACT
OBJECTIVE:To establish the method for simultaneous determination of 14 kinds of isoflavones in Pueraria radix. METHODS:LC-MS/MS was adopted to detect 14 kinds of isaflavones in 14 batches of P. radix(P. radix:PL-1 to PL-7,P. thomsonii:PT-1 to PT-7). The determination was performed on Extend C18 with mobile phase consisted of methanol-water(gradient elution)at the flow rate of 0.8 mL/min. The column temperature was set at 22℃. The sample size was 5μL. Ion source was ESI source. The detection mode was negative ion detection. Scanning mode was MRM with jet voltage of -4500 V;ion source temperature was set at 600 ℃, and atomization gas was 60 psi. The auxiliary gas was 60 psi,collision gas was 7 psi,air curtain gas was 30 psi. SIMCA 13.0 software wasused for cluster analysis of above batches. RESULTS:The linear range of 14 kinds of isoflavones ranged 10-1000 ng/mL(puerarin of 10-5000 ng/mL,r>0.9900). RSDs of precision,stability and reproducibility tests were all lower than 5％. The recoveries were 95％-105％(RSD were 0.8％-4.5％,n=6). The total content of isoflavones were different significantly between P. radix and P. thomsonii. The contents of isoflavones in P. radix from different origins were different significantly. Among 14 kinds of isoflavones, the content of puerarin was the highest. Results of cluster analysis showed that 14 batches of P. radix could be clustered into 4 categories,including PL-2 as Ⅰ category,PL-3 and PL-4 as Ⅱ category,PL-5,PL-6 and PL-7 as Ⅲ category,other as Ⅳcategory. CONCLUSIONS:The method is simple and reproducible. It can be used for content determination of 14 kinds of isoflavones in P. radix and quality control.
ABSTRACT
Objective: To establish a method for simultaneously determination of five isoflavones (daidzein, daidzin, puerarin, mirificin, and 6″-O-xylosidepuerarin) in Pueraria Radix by single marker (QAMS), which is feasible and accurate. Methods: Puerarin was taken as internal standard substance to establish the relative correction factor (RCF) for quantitative analysis of multi-components with QAMS. Thus, the contents of mirificin, 6″-O-xylosidepuerarin, daidzin, and daidzein were calculated. The contents in 10 batches of samples were determined by external standard method and QAMS. The scientificalness and feasibility of the methods were evaluated by comparison of the quantitative results between external standard method and QAMS. Results: The reproducibility of RCF was perfect. The results calculated with QAMS were consistent with the results by the external standard method. Conclusion: QAMS is accurate and feasible to evaluate the quality of Pueraria Radix.
ABSTRACT
This study was designed to investigate the synergetic hepatoprotective effects from a mixture of Korean Red Ginseng and Pueraria Radix on carbon tetrachloride (CCl₄)-induced hepatotoxicity in mice. Liver toxicity was induced by intraperitoneal administration of CCl₄ (0.6 mg/kg) in 12 groups of ICR mice. The negative control group was given CCl₄ without test samples and the normal group was given no treatment. Among treatment groups, the RGAP treatment (Korean Red ginseng acetic acid extract : Pueraria Radix water extract, w/w, 38.4:57.6) decreased CCl₄-elevated ALT (101.60 IU/L), AST (833.89 IU/L), and LDH (365.02 IU/L) levels in the serum, and increased the SOD (11.03 unit/mg protein) and CAT (0.37 unit/mg protein) levels and the LPO levels (59.09 µM/g tissue) more than that in the mice group with CCl₄-induced control group hepatotoxicity. These results suggest that administration of a mixture of Korean Red ginseng and Pueraria Radix decreases CCl₄-induced liver damage and enhances antioxidant activity in mice and imply that administration of the mixture in a certain ratio is more effective than single administration of either Korean Red ginseng or Pueraria Radix alone.