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Objective: A network model of ulcerative colitis-associated colon cancer (UCRCC) and NF-κB signaling pathway was established, in order to predict the key inflammatory targets of UCRCC and identify the effect of GQD and composition of index components on these targets. Methods: HPLC method was used, the mobile phase was acetonitrile-0.2% phosphoric acid aqueous solution, and the gradient elution was carried out. The 10 indicators were determined. The literature of databases such as Pubmed and ScienceDirect were searched, the terms of the upstream and downstream proteins of UCRCC disease and NF-κB pathway were examined. The Cytoscape software was used to predict the key inflammatory targets of UCRCC. The UCRCC model was established by AOM/DSS method, and the effect of GQD and composition of index components on key inflammatory targets were identified by immunohistochemistry and ELISA. Results: The contents of puerarin, daidzin, glycyrrhizin, jatrorrhizine, baicalin, palmatine, berberine, baicalein, glycyrrhizic acid and wogonin in GQD were 1.677, 0.154, 0.159, 0.045, 0.448, 0.035, 0.095, 0.013, 0.111 and 0.006 μg/g, respectively. The top eight inflammatory factors of the key protein in the interaction network were NF-κBp65, iNOS, COX-2, Bcl-2, TNF-α, IL-1β, ICAM-1 and VCAM-1. Compared with the model group, the high-, medium- and low-dose groups of GQD, the composition of index components group and the 5-ASA group were able to down-regulate the relative expression levels of NF-κBp65, iNOS, Bcl-2, COX-2 in colonic tissues of UCRCC mice (P < 0.01), and down-regulate the expression levels of TNF-α, IL-1β, ICAM-1, VCAM-1 in serum of UCRCC mice (P < 0.05, 0.01). Conclusion: GQD and composition of index components can improve the pathological condition of colon tissue in UCRCC mice, down-regulate the expression of inflammatory factors in colon tissue of UCRCC mice, and have a certain intervention effect on UCRCC, which laid the foundation for determining the quality marker (Q-marker) of GQD.
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A sensitive and reliable liquid chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI-MS/ MS) method was established to simultaneously quantitate four categories of compounds (isoflavonoids, flavonoids, alkaloids and saponins) in Gegen-Qinlian decoction (GQD). These compounds were separated by a Shiseido CAPCELL PAK C18 column with a linear gradient consisting of 0.1% (v/v) formic acid in water (A) and 0.1% (v/v) formic acid in acetonitrile (B), and delivered at a flow rate of 0.3 mL/min. All the analytes were determined by electrospray positive ionization tandem mass spectrometry in a multiple reaction monitoring (MRM) mode. Linearity, accuracy, precision, recovery and stability of the method were evaluated with the validation over the range of 4.0-5385 ng/mL. The proposed method was applied to the analysis of a Chinese herbal preparation CQD successfully.
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A sensitive and reliable liquid chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI-MS/MS) method was established to simultaneously quantitate four categories of compounds (isoflavonoids, flavonoids, alkaloids and saponins) in Gegen-Qinlian decoction (GQD). These compounds were separated by a Shiseido CAPCELL PAK C18 column with a linear gradient consisting of 0.1% (v/v) formic acid in water (A) and 0.1% (v/v) formic acid in acetonitrile (B), and delivered at a flow rate of 0.3 mL/min. All the analytes were determined by electrospray positive ionization tandem mass spectrometry in a multiple reaction monitoring (MRM) mode. Linearity, accuracy, precision, recovery and stability of the method were evaluated with the validation over the range of 4.0-538 5 ng/mL. The proposed method was applied to the analysis of a Chinese herbal preparation GQD successfully.
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Object To study the HPCE fingerprinting of Gegen Qinlian Decoction (GQD). Methods A buffer was composed of 30 mmol/L sodium phosphate and 40 mmol/L borate solution. Capillary electrophoresis was performed using a 65 cm (43 cm to detector) ?50 ?m fused-silica capillary tube. Separation voltage was 22 kV, sampling time was 1 s, detected wavelength was 254 nm, and the temperature was maintained at 30 ℃. Results The 27 components in GQD were successfully separated. The observation of methodology was in keeping with quantitatine determination and qualitative study. Conclusion This method can be used for the quality control of the preparation of GQD with good precision.
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Object To set up the quantitative method for determining the puerarin in Gegen Qinlian Decoction (GQD) by RP-HPLC and to study the change of puerarin contents in various combinations of GQD. Methods L 8(2 7) orthogonal design and statistic analysis (SPSS 10.0) were used, the puerarin contents in the samples were determined by HPLC. Results Influence of Radix Scutellaria, Rhizoma Coptidis and Radix Glycyrrhizae Preparata on the puerarin content in GQD was insignificant. And interactions between two of three were insignificant too. In this experiment, puperarin in the sedimentation was found in the compatible decoction of Rhizoma Coptidis and Radix Puerariae. Conclusion The puerarin content is not markedly influenced by Radix Scutellaria, Rhizoma Coptidis and Radix Glycyrrhizae Preparata. Maybe, the change of pH is relevant to the sedimentation in the compatible decoction contained Rhizoma Coptidis. and Radix Puerariae
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Object To explore the regularity of recipe composition by observing different bacteriostasis among compositions of Gegen Qinlian Decoction (GQD) against Streptococcus pneumoniae. MethodsTo evaluate the minimal inhibitory concentration (MIC) of 15 decoctions through double fold dilution method. Results Not all of the decoctions are of the same MIC. Conclusion Not all of the 15 decoctions have the same bacteriostasis against S. pneumoniae. Among the four herbs of GQD Coptis chinensis Franch. has the strongest bacteriostasis and the next Scutellaria baicalensis Georgi. While Glycyrrhina wralensis Fisch and Pueraria lobata (Willd.) Ohwi have no bacteriostasis, but they can influence the bacteriostasis of C. chinensis and S. baicalensis when combinating with them.
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Object To set up the quantitative method of baicalin in Gegen Qinlian Decoction (GQD) by RP HPLC and to determine the change of baicalin contents in decoctions prepared from various combinations of GQD. Methods By L 8(2 7) orthogonal design and statistics analysis (SPSS10 0), eight decoctions with Radix Scutellariae either alone or incombination with one or more of the other three components were prepared or analyzed. Contents of baicalin in them were determined by RP HPLC. Results It was significant (P0 05) and interactions between two of three were insignificant either. In this experiment, the sedimentation was found in the decoctions contained Radix Scutellariae and Rhizoma Coptidis. Conclusion The content of baicalin is reduced by Radix Puerariae and Rhizoma Coptidis, The resolution of baicalin is not increased by Radix Glycyrrhizae Preparata. Maybe the change of pH leads to the sedimentation in decoctions contained Radix Scutellariae and Rhizoma Coptidis.