ABSTRACT
Objective: To investigate the active ingredients and molecular mechanism of Astragalus membranaceus var. mongholicus, Angelica sinensis, Angelicae dahurica, and Gleditsia sinensis in Tuoli Xiaodu Powder in promoting of diabetic wound healing. Methods: UPLC-Q-TOF/MS in positive and negative ion modes was applied to analyze the components in the ethanol extract from Tuoli Xiaodu Powder. Molecular docking technology was used to predict the targets proteins of these components. The function and pathway annotations of target proteins were performed through relevant databases such as Uniprot and KEGG. The drug components-target-function diagram was constructed using Cytoscape software. Results: Twenty-eight compounds containing flavonoids, saponins, coumarins, alkaloids, and triterpenoids were identified in positive and negative ion modes. Among these compounds, 17 compounds could interact with 17 target proteins, and there were 210 pairs of component-target relationships by analyzing the results of molecular docking. Among them, five targets were related to immune regulation, six targets were related to antibacterial and anti-inflammatory effects, six targets were related to cell differentiation, 10 targets were related to cell migration, six targets were related to angiogenesis, two targets were related to stimulation of epithelial growth factor, six targets were related to vasodilation, and two targets were related to estrogen. Conclusion: The flavonoids, saponins, coumarins, steroids, and triterpenoids contained in the simplified formula possess many biological effects such as antibacterial, anti-inflammatory, immune regulation, and angiogenesis. These functions may be related to its modulation of NF-κB, PI3K/Akt/eNOS, and MAPK pathway through regulating NF-κB, MAPK, PI3K, and ERK2 targets.
ABSTRACT
Objective To analyze and evaluate the chemical components from flowers of Astragalus membranaceus var. mongholicus, and to investigate the potential value of the medicinal plant resources. Methods UV-Vis spectrophotometry was used to determine the total contents of polysaccharides and water-soluble protein. HPLC-PDA/ELSD method was used to determine monosaccharides and oligosaccharides and GC-MS method was used to determine volatile components and the fatty acids in the flowers of A. membranaceus var. mongholicus. UPLC-TQ-MS method was used to analyze the nucleosides and amino acids. Results The flowers of A. membranaceus var. mongholicus contain abundant polysaccharides (47.02 mg/g), water-soluble protein (470.66 mg/g), fructose (45.46 mg/g), glucose (8.71 mg/g), and sucrose (1.05 mg/g). There were 32 kinds of volatile components detected in the flowers of A. membranaceus var. mongholicus, in which oxy-derivatives were the main components. In addition, six nucleosides and 15 amino acids were detected in the flowers of A. membranaceus var. mongholicus, and their total contents were 2.77 mg/g and 6.52 mg/g, respectively. Eight fatty acids in the flowers of A. membranaceus var. mongholicus were also detected, in which myristic acid, palmitic acid, and oleic acid were the main components. Conclusion This study investigated the composition and content of various nutritional components of the flower of A. membranaceus var. mongholicus, which provides a scientific basis for its utilization and development.
ABSTRACT
Objective To establish a stable and rapid separation and purification method for Astragalus membranaceus (Am) pathogenesis-related protein-10 (AmPR-10) using an automatic intelligent protein purification system AKTA Avant 25, and analyze its physiochemical and biological activity. Methods Am was extracted by Tris-HCl buffer. The crude extract was captured by anion exchange chromatography, and finely separated by hydrophobic chromatography and gel filtration chromatography. The relative molecular weight of AmPR-10 was measured by MALDI-TOF/TOF mass spectrometry, the protein identification was determined by mass spectrometry and MS/MS Ion Search, the glycoprotein identification was estimated by periodic acid-Schiff method, and the ribonuclease activity and effect factors were analyzed by agarose gel electrophoresis. Results The electrophoretically pure AmPR-10 was obtained by three-step purification of Q Sepharose Fast Flow, Butyl Sepharose High Performance and SuperdexTM 75 10/300 GL from the crude extraction. The relative molecular weight of AmPR-10 was 16 801. AmPR-10 was highly homologous to PR-10 and has no carbohydrate chains. Incubated at 56 ℃ for 30 min, AmPR-10 exhibited significant ribonuclease activity to total RNA of mammalian cells. The activity was insensitive to NaCl, pH value and mental ions, and weekly inhibited by 0.5 mol/L NaCl, pH 9.0, Mg2+ and Co2+. The activity was the same at EDTA as high as 20 mmol/L. Conclusion The three-step method of exchange chromatography-hydrophobic chromatography-gel filtration chromatography, a stable and rapid separation and purification method of AmPR-10, can be applied for other Chinese herbs. AmPR-10 might play an important role in resistance against virus.
ABSTRACT
Objective: To study the quality differences of medical material, raw decoction pieces, and processed products of Astragalus membranaceus var. mongholicus. Methods: The raw decoction pieces and processed products were obtained from genuine medicinal materials of A. membranaceus var. mongholicus. The reasons for the quality differences were analyzed by comparing the contents of astragaloside IV, calycosin-7-O-β-D-glucopyranoside, formononetin, total polysaccharides, saponins, and flavones. Results: With content analysis, the sequence was found as follows: astragaloside IV (medical material > raw decoction piece > honey-fried piece > alcohol-fried piece > salt-fried piece > fried piece); calycosin-7-O-β-D-glucopyranoside, formononetin, and total polysaccharides (medical material > raw decoction pieces > alcohol-fried piece > salt-fried piece > honey-fried piece > fried piece), total flavones (medical material > alcohol-fried piece > raw decoction pieces > salt-fried piece > honey-fried piece > fried piece), total saponins (medical material > honey-fried piece > raw decoction pieces > alcohol-fried piece > salt-fried piece > fried piece). Conclusion: The temperature and supplementary material may play the main roles for quality differences of A. membranaceus var. mongholicus.
ABSTRACT
Objective: To optimize the extraction technology of immune active glycoproteins AmPR10-16kD and HQGP-2 from Astragalus membranaceus var. mongholicus (AMM). Methods: The optimized extraction temperature conditions were investigated by circular dichroism of water-soluble protein involving in AmPR10-16kD and HQGP-2 with secondary structure from AMM. The optimized extraction technology was investigated using single factor test and orthogonal test with gray value of water-soluble protein AmPR10-16kD and HQGP-2 as the index which was determined by Image of gel graphical analysis software. In this study, the effects of temperature, solid-liquid ratio, time, solvent, granularity, and times on gray value were investigated, for which the inhibitory effect of water-soluble protein was determined as an evidence by CCK-8 method, and the content of water-soluble protein is determined as an evidence by BCA method. Results: The optimized extraction technique for proteins AmPR10-16kD and HQGP-2 in AMM was established, that was 5.0 g powder of AMM over the No.4 sieve, olvent Tris-HCl, solid-liquid ratio 1:16 and 60 min for extraction at the temperature of 40 ℃ and being mixed under 100 r/min. The water-soluble protein extract rate in the orthogonal test analysis was 65 mg/g, of which inhibitory effect was 90.90% at a concentration of 90 μg/mL. Conclusion: The optimal extraction conditions could accurately reflect the relative amounts of AmPR10-16kD and HQGP-2 maximum extraction rate, providing a stable, reasonable, and feasible extraction process for further study of the bioactive substance of AMM.
ABSTRACT
The HPLC-ELSD method was used in the content determination of astragaloside, astragalosideⅠ, astragalosideⅡand astragalosideⅢ in Mongolia Radix Astragali (Astragalus membranaceus(Fisch.) Bge. var. mongholicus(Bge.) Hsiao) among 16 batches from various habitats. The DIKMA Diamonsil C18 (150 mm× 4.6 mm, 5μm) was adopted with acetonitrile and water as the mobile phase at a gradient mode program. The flow rate was 1.0 mL·min-1. And the column temperature was 30℃. The ELSD detector parameters were the drift tube temperature at 90℃, and the air flow rate of 2.8 L·min-1. The SPSS 16.0 software was used in the cluster analysis of content determination. The results showed that when the injection volume was within the range of 0.093 2-1.02μg (r = 0.999 5), 0.789-8.78μg (r = 0.999 7), 0.506-3.13μg (r = 0.999 6), and 0.016 1-1.38μg (r = 0.999 2) for astragaloside, astragalosideⅠ, astragalosideⅡ and astragalosideⅢ, respectively, the average recoveries were 97.55%, 98.61%, 99.68%, 98.58%with RSD of 1.2%, 1.3%, 1.3%, 1.2%, respectively. The results of cluster analysis showed that the single using of astragaloside as index was unable to differentiate Mongolia Radix Astragali from various habitats. However, the simultaneous determination of 4 types of astragalosides as indexes can differentiate Mongolia Radix Astragali from various habitats. It was concluded that the method was simple, quick and accurate, which can directly reflect the quality status of Mongolia Radix Astragali from different origins. It also provided new ideas for the quality control of Mongolia Radix Astragali.
ABSTRACT
Objective: To determine the contents of astragaloside and flavonoids in four ecotypes of A. membranaceus, which are whip pole type, taproot type, binary type, and chicken feet type, and analyze the correlation between ecotypes and quality. Methods: The contents of astragaloside and flavonoids in A. membranaceus were determined by UPLC, and the measured data were analyzed by principal component analysis (PCA) and cluster analysis (CA). Results: The order of contents of astragaloside and flavonoids in A. membranaceus was whip pole type > taproot type > binary type > chicken feet type; PCA revealed that four ecotypes of ecotypes A. membranaceus could be separated significantly; CA showed that each ecotype of A. membranaceus could be clustered into one class perfectly. Conclusion: The quality of A. membranaceus is closely associated with ecotypes. The quality merits of the order of four ecotypes of A. membranaceus is whip pole type > taproot type > binary type > chicken feet type.
ABSTRACT
OBJECTIVE: To study the chemical constituents of roots of Astragalus membranaceus (Fisch) Bge. var. mongholicus (Bge) Hsiao. METHODS: The chemical constituents of Astragalus membranaceus (Fisch) Bge. var. mongholicus (Bge) Hsiao were isolated and purified by D101 column chromatography and preparation HPLC. Their structures were identified by physicochemical properties and spectral analysis methods. RESULTS: Seven compounds were obtained and identified as 4, 4′-dimethyl-6′-hydroxychalcone (1), 4-methoxy-4′, 6′-dihydroxychalcone(2), 7, 4′-dihydroxyflavonone(3), 4, 4′, 6′-trihydroxychalcone(4), 4′-hydroxyflavonone-7-O-β-D-glucoside(5), 2′-hydroxy-3′, 4′-dimethylisoflavan-7-O-β-Z)-glucoside(6), and 3, 2′-dihydroxy-3′, 4′-dimethylisoflavan-7-O-β-D-glucoside (7). CONCLUSION: All these compounds were isolated from this plant for the first time.
ABSTRACT
Objective: To compare the chemical constituents in traditional planted and two-year-old cultivated Astragali Radix, and to discuss the chemical differences. Methods: The HPLC-ELSD fingerprint of Astragali Radix was collected, and the contents of astragaloside, calycosin-7-O-β-D-glucoside, total polysccharide, and extract were also determined. The similarity analysis, metabolomic multivariate statistical analysis, and quality comparison were performed. Results: The contents of calycosin-7-O-β-D-glucoside and total polysaccharide in traditional planted Astragali Radix were obviously higher than those in two-year-old cultivated Astragali Radix. But the contents of astragaloside and extract showed no difference. The two kinds of Astragali Radix could be distinguished by HPLC-ELSD metabolomic analysis. The differential metabolites were calycosin-7-O-β-D-glucoside, ononin, peaks 4, 5, 13, astragalosides I and II. Conclusion: HPLC-ELSD metabolomic approach combined with the content determination of index components could be used for the quality evaluation of traditional planted and two-year-old cultivated Astragali Radix.
ABSTRACT
Objective: To establish the method of purifying astragaloside. Methods: Astrageloside was determined by HPLC fingerprinting to compare macroporous resin absorbing method with extraction refine by n butyl alcohol. Results: The HPLC fingerprints of each method were difference. Conclusion: AB 8 macroporous resin is better than the others for purifying astrageloside.