RESUMO
Objective:To explore active components and mechanism of Dipsaci Radix in treating rheumatoid arthritis （RA） based on ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry （UPLC-QTOF-MS/MS） and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP）. Method:UPLC-QTOF-MS/MS with electrospray ionization （ESI） was used to qualitatively analyze the chemical components in methanol extract of Dipsaci Radix under positive and negative ion scanning modes. The mobile phase consisted of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-10 min， 0.2%-20%B； 10-20 min， 20%-40%B； 20-25 min， 40%-50%B； 25-33 min， 50%-98%B； 33-35 min， 98%-0.2%B）， and the scanning range was <italic>m</italic>/<italic>z</italic> 50-2 000. Based on TCMIP， candidate target groups of Dipsaci Radix， RA and syndrome with Yin deficiency of liver and kidney were obtained， and correlation analysis on "disease-syndrome-prescription" was used to enrich the main active components and key targets. Cytoscape 3.8.0 and STRING 11.0 database were used to construct protein-protein interaction （PPI） network diagram. Metascape platform was used to analysis gene ontology biological progress and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathways. Result:A total of 81 ingredients were identified by UPLC-QTOF-MS/MS. Based on TCMIP， 283 candidate targets corresponding to 81 ingredients， 7 RA related targets and 215 genes corresponding to syndrome with Yin deficiency of liver and kidney were collected. With further correlation analysis on "disease-syndrome-prescription"， 17 key active ingredients were predicted， mainly including saponins and fatty acids of Dipsaci Radix. It mainly involved 7 hub targets， namely tumor necrosis factor （TNF）， nuclear factor-<italic>κ</italic>B subunit 1 （NF-<italic>κ</italic>B<sub>1</sub>）， hepatocyte nuclear factor 4 alpha （HNF4A）， nuclear receptor subfamily 3 group C member 1 （NR3C1）， peroxisome proliferator activated receptor gamma （PPARG）， nuclear receptor subfamily 1 group H member 4 （NR1H4） and nuclear receptor coactivator 1 （NCOA1）. All of them were related to inflammation， and two of them were related to bile acid pathway. The 7 hub targets and 7 pathways played an important role in RA were screen out， including 4 bile acid related pathways and 3 inflammatory related pathways. Conclusion:UPLC-QTOF-MS/MS combined with TCMIP preliminarily elucidates the regulatory effect of multi-components in Dipsaci Radix on several pathways related to the inflammatory response and bile acid synthesis and metabolism， which lays a foundation for further study on the mechanism of Dipsaci Radix against RA.
RESUMO
Objective: To establish a quick method of ultra-performance liquid chromatography-quadrupole time-of-fight mass spectrometry (UPLC-Triple-TOF/MS) for the analysis of components of crude and sweated Dipsaci Radix. Methods: The separation was performed on the chromatographic column of Agilent Eclipse XDB-C18 (250 mm × 4.6 mm, 5.0 μm), and the mobile phase was 0.1% formic acid solution-methanol, with a gradient elution at a flow rate of 0.8 mL/min, the detection wavelength was 215 nm, the column temperature was 25 ℃. UPLC-Triple-TOF 5600+ time of flight liquid and mass spectrometer was used for mass spectrometry. Electrospray ion source negative ion mode was adopted, and the scanning range was m/z 100-1 500. The components of crude and sweated Dipsaci Radix were quickly identified according to the information obtained by high-resolution mass spectrometry combined with secondary mass spectrometry. Results: Fifty-two common components were identified or tentatively characterized based on the retention time and MS spectra. They were triterpenoid saponins, iridoids, phenolic acids etc. The crack rules of primary components were also analyzed. And comparing the components of crude and sweated Dipsaci Radix, it showed that the content of 20 components such as loganin acid, chlorogenic acid, loganin, isochlorogenic acid A, and asperosaponin VI was decreased after sweating, and caffeic acid, isochlorogenic acid, isochlorogenic acid C, and triplostoside A was increased. Conclusion: The types of components of crude and sweated Dipsaci Radix are identical, but there are differences in the content of the components. The content of the components of crude are higher than the sweated Dipsaci Radix. UPLC-Triple-TOF-MS technology was used to analyze the influence of “sweating” on the chemical composition of the Dipsaci Radix, so as to provide a theoretical basis for the study of the chemical constituents of sweated Dipsaci Radix and further research on the origin processing of Dipsaci Radix.
RESUMO
Through the records of herbal and market investigations,the samples collected from different market and origin were analyzed,and the appearance character indexes were determined in order to revise the commodity specification and grade standard of Dipsaci Radix combined with production practice. This is also to analyze association of the appearance with quality different of intrinsic components. The investigation results indicated the root's long and thick was better,and atrovirens of fracture surface,there are most of the market is dominated by the ungraded goods. Through principal component analysis,variance analysis and cluster analysis,combining with the feasibility of actual operation and herbal record,the length,middle diameter and phloem color were filtrated and the specification was divided into 2 ranks: the selection and ungraded goods,and the grade was divided into 2 ranks for the selection goods: big,small selected goods. Moreover,there were no significant correlation between the appearance and the intrinsic quality index of Dipsaci Radix,the content of extractum were significantly positive correlation with the moisture,but had extremely negative correlation with the total ash. Multiple comparisons indicated that the content of the moisture,extractum and asperosaponin VI of the selected goods were higher than the ungraded goods,while the total ash content were lower,and they hasn't significant difference. The commodity specification and grade standard of Dipsaci Radix as a basis provide commodity specification and grade standard of communities and standardizing market trade order.
Assuntos
Análise por Conglomerados , Dipsacaceae , Química , Medicamentos de Ervas Chinesas , Padrões de Referência , Raízes de Plantas , Química , Plantas Medicinais , Química , Análise de Componente PrincipalRESUMO
Objective: To screen the extraction process of Wuzi Yanzong plus and minus Recipe (WYR) by pharmacodynamics and long-term toxicity test, and optimize it by orthogonal experiment to determine the best extraction process of WYR. Methods Long-term toxicity experiments were carried out on different processed WYR samples. The penile erection latency and sex hormone levels in rats with kidney-yang deficiency were used as pharmacodynamic indicators. The results of long-term toxicity and efficacy experiments were combined to screen out the optimal process plan. Using the content of ingredients and the rate of ointment as indicators, the orthogonal test was used to screen the optimal level of different extraction processes and verified. Results Long-term toxicity test results showed that all mice survived healthily and no obvious toxicity was observed. The results of pharmacodynamic experiments showed that the extracts of Morindae Officinalis Radix, Eucommiae Cortex, Taxilli Herba, Dipsaci Radix, and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle were refluxed with 70% ethanol, and the remaining medicinal materials were extracted with 70% ethanol as the solvent to obtain the best extraction method of WYR. The suitable extraction process was as follows: Morindae Officinalis Radix, Eucommiae Cortex, Taxilli Herba, Dipsaci Radix, and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle were extracted three times with 10 times 70% ethanol for 2.0 h each time; The remaining medicinal materials were percolated with 10 times 70% ethanol at 1.5 mL/min. The medicinal herbs were firstly soaked for 16 h before percolation. Conclusion WYR can significantly improve the penile erection latency and sex hormone levels in rats with kidney yang deficiency. The optimal process conditions are reasonable and feasible, which provides a basis for the follow-up development of WYR.
RESUMO
Objective To explore the possible mechanism of Dipsaci Radix in the treatment of osteoporosis based on systems pharmacology method. Methods The drug components of Dipsaci Radix were obtained from TCMSP database to screen active ingredients and predict target protein. The target protein of osteoporosis was searched in the TTD and CTD database in order to build Drug-Target protein-Disease interaction Network. Then, the analysis on the role of core target protein pathways were performed by DAVID tool. The protein interaction network of primary signal pathways was built in String database. Results Seven active ingredients of Dipsaci Radix and their 63 target proteins were obtained, 118 targets of osteoporosis were selected, and 118 cross-acting proteins between Dipsaci Radix and osteoporosis were obtained. Then 24 signaling pathways related to cell proliferation and differentiation and immune function were enriched, wherein PI3K-AKT signaling pathway contained the largest number of related genes. String database analysis showed that AKT1, MAPK1, and MAPK3 protein appeared frequently in signal pathways. Conclusion Dipsaci Radix may affect the PI3K-AKT signaling pathway mainly through AKT1, MAPK1, and MAPK3 for the treatment of osteoporosis. This study provides a new idea for the further mechanism study on the treatment of osteoporosis of Dipsaci Radix.
RESUMO
Objective To investigate the effect of decoction and drug serum of sweated and crude Dipsaci Radix on the proliferation of human osteoblast-like cells (MG-63) and osteoblasts. Methods MG-63 cells and osteoblasts were co-cultured with decoction and rat serum containing Dipsaci Radix before and after “sweating”. The cell proliferation was detected by MTT method and the ALP activity of osteoblasts was detected by nitrophenyl phosphate method. Results Both decoction and drug-containing serum can significantly promote the proliferation of MG-63 cells and osteoblasts (P < 0.01), and significantly increase the ALP activity of osteoblasts (P < 0.01). Moreover, sweated group were generally better than or non-inferior to crude group at the same dose concentration of two administration methods. Conclusion Combined with component studies, it can be inferred that the changes in composition before and after "sweating" affect its role in promoting cell proliferation and differentiation, with view to laying the foundation for further research.
RESUMO
Objective: To analyze the composition and the content of volatile components in Dipsaci Radix and its wine broiled products. Methods: Headspace solid phase microextraction (HS-SPME) combined with gas chromatography-mass spectrometry ( GC-MS) was used to analyze the volatile components in Dipsaci Radix and its wine broiled products. The area normalization method was used to calculate the relative content. Results: Totally 31 ingredients were identified, which mainly included phenols, olefins, alcohols and aldehydes. Totally 25 peaks were detected out from the volatile components in Dipsaci Radix, and among them, 22 components were identified, which accounted for 94. 74% of the total volatile components. The content of 5-allylguaiacol was the highest, which accounted for 28. 37% of the total volatile components. Totally 23 peaks were detected out from the volatile components in wine-pro-cessed Dipsaci Radix, and among them, 17 ingredients were identified, which accounted for 85. 92% of the total volatile components. The content of nonanal was the highest, which accounted for 9. 66% of the total volatile components. Conclusion: The composition and the content of volatile components in Dipsaci Radix and its wine broiled products are different. The experiment provides scientific reference for the further research and development of the volatile components in Dipsaci Radix and its wine broiled products.
RESUMO
Objective To establish and compare HPLC fingerprint chromatograms of Dipsaci Radix decoction pieces, aqueous decoction and formula granules.Methods The HPLC analysis was carried out in Wondasil C18 column (4.6 mm × 250 mm, 5 μm) with a mobile phase of acetonitrile-0.1% phosphoric acid by gradient elution. The flow rate was 1.0 mL/min; the detection wavelength was set at 212 nm; the column temperature was kept at 30℃. Results The fingerprint chromatograms from 12 batches of Dipsaci Radix decoction pieces, aqueous decoction and formula granules were established respectively. 14 common peaks in the fingerprint chromatogram in the formula granules could be tracked in the aqueous decoction, and 13 common peaks in the fingerprint chromatogram could be tracked in the decoction pieces. 2 chemical compounds were identified, such as asperosaponinⅥ and chlorogenic acid.ConclusionThe method of HPLC fingerprint chromatograms is stable and with good repeatability. Dipsaci Radix decoction pieces, aqueous decoction and formula granules are basically the same chemical composition.
RESUMO
Objective: To establish a method for the determination of loganic acid, chiratin, and loganin in Dipsaci Radix and improve the quality evaluation by analyzing Dipsaci Radix from different habitats. Methods: The analysis was performed on Venusil MP C18 column (250 mm×4.6 mm, 5 μm) with mobile phase consisting of 0.1% H3PO4-acetonitrile for gradient elution: 0-10 min, 8%-9% B; 10-30 min, 9%-11% B; 30-35 min, 11%-100% B. The flow rate was 1 mL/min and the column temperature was 25℃. Results: The results showed that loganic acid, chiratin, and loganin were well separated with the good linearity in 18.4-368.2, 2.02-40.4, and 17.5-349.6 μg/mL, respectively. The average recoveries of the three iridoid glycosides were 99.34%, 99.19%, and 101.61%. Conclusion: Loganic acid, chiratin, and loganin are the main iridoid glycosides in Dipsaci Radix. The method can easily be applied to the content determination of loganic acid, chiratin, and loganin in Dipsaci Radix quickly. The content ranges of loganic acid, chiratin, and loganin are 20.4-186.8, 26.4-177.7, and 1.9-13.2 mg/g, respectively, and the sum of the three iridoid glycosides shows no significant differences in various Dipsaci Radix from different habitats, and proposes to evaluate the quality of Dipsaci Radix.
RESUMO
Objective: A high performance liquid chromatography-electrospray tandem mass spectrometry (HPLC-ESI-MS) method was developed for the component analysis of the crude and sweated Dipsaci Radix. Methods: Analyses were carried out on an Agilent Eclipse XDB-C18 (250 mm × 4.6 mm, 5.0 μm) column by gradient elution using (A) 0.1% aqueous formic acid and (B) acetonitrile. The column was maintained at 30℃, and the flow rate was 0.8 mL/min. Chemical characteristics of the crude and sweated Dipsaci Radix were analyzed by MS techniques with an ESI source, the quasi molecular ions of compounds in both negative and positive modes were observed for molecule mass information as [M-H]-, [M + HCOO]-, [M + H]+, and [M + Na]+. Results: An HPLC-ESI-MS spectrum of the crude and sweated Dipsaci Radix was established with 42 peaks respectively, and the potential structures of 31 characteristic components were identified by study on the MS of compounds and comparing with reference data and some of standards. After being sweated, the contents of some components in Dipsaci Radix decreased, such as loganic acid, 4-O-caffeoylquinic acid, chlorogenic acid, caffeic acid, loganin, isochlorogenic acid B, dipsanoside A, and dipsanoside B, but some ones increased, such as cantleyoside, dipsacoside X, dipsacoside VI, dipsacoside VII, dipsacoside B, dipsacoside XIII, and dipsacus saponin A, The amount of other ingredients are almost the same. Conclusion: The HPLC- ESI-MS spectrum could comprehensively control the quality of the crude and sweated Dipsaci Radix.
RESUMO
This study was aimed to optimize extraction technology of Asperosaponin VI from Dipsaci Radix by re-sponse surface methodology (RSM). A single-factor test and Box-Behnken design (BBD) were employed to optimize three extraction variables, including extraction time (A), ethanol concentration (B), and the ratio of ethanol to materi-al (C). The transfer rate of Asperosaponin VI was used as the evaluation index. The results showed that the optimized conditions were A of 33.13 min, B of 51.58 %, C of 23.39 mL·g-1. Under these conditions, the transfer rate was 88.39 ± 0.212% (n = 3), which was well matched with the predicted transfer rate. It was concluded that RSM can be used for extraction optimization of ultrasonic extraction process for Asperosaponin VI. This method had certain prac-tical values.
RESUMO
To optimize the extraction technology of the total saponins in Dipsaci Radix, the extraction efficiency of total saponins was investigated with respecting to three variables including time, ethanol concentration and liquid-to-solid ratio. On the basis of a series of one-factor-at-a-time experiments, a polynomial regression model equation was fitted by the combined use of central composite experimental design and regression analysis. By analyzing the regression model using response surface analysis, the optimum extraction conditions of total saponins from Dipsaci Radix were identified as follows: 7 times of 55% alcohol, refluxing 3 times, 2.8 hours every time, and the extraction efficiency of total saponins was up to 0.201 1 g·g-1 and the asperosaponin Ⅵ was up to 0.015 4 g·g-1 under the optimized conditions. Confirmatory experiment indicated the good prediction ability of the established model and provided the basis for further development and utilization of Dipsaci Radix.