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Astragali Radix, a medicinal herb for invigorating Qi, has anti-aging, anti-tumor, immunoregulatory, blood sugar-and lipid-lowering, anti-fibrosis, anti-radiation and other pharmacological effects. This article reviewed the studies about the chemical components and pharmacological effects of Astragali Radix. According to the theory of quality markers(Q-markers) of Chinese medicinal materials, we predicted the Q-markers of Astragali Radix from traditional efficacy, chemical component validity, measurability, plant phylogeny, and pharmacokinetis. The results showed that total polysaccharides, flavonoids(e.g., calycosin-7-O-β-D-glucoside, formononetin, calycosin, quercetin, and ononin), and saponins(e.g., astragalosides Ⅱ, Ⅲ, and Ⅳ) can be taken as the main Q-markers. This review lays a foundation for regulating the quality research and standard establishment of Astragali Radix, and benefits the control and quality supervision of the production process of Astragali Radix and its related products.
Subject(s)
Astragalus Plant , Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/pharmacology , Flavonoids , Plant RootsABSTRACT
Astragali Radix is one of the most commonly used medicinal materials. In recent years, its cultivated varieties and a variety of adulterants have flooded the market, which makes its quality uneven, and the development of quality control methods has become a research hotspot. Therefore, figuring out the quality markers of Astragali Radix is of great significance for its comprehensive evaluation. In this study, the fingerprints of 15 batches of Astragali Radix were established by HPLC, and the main components causing intergroup differences were screened out by PLS-DA. On the basis of literature review and network pharmacology analysis, the targets and pathways of active ingredients were obtained from SwissTargetPrediction, PubChem Compound and other databases, and then the "component-target-pathway" network was constructed with Cytoscape 3.7.1 for the prediction of potential quality markers. Twenty-eight common peaks were identified in the established fingerprint, and three differential components were selected as potential quality markers for Astragali Radix, which were astragaloside Ⅳ, calycosin-7-O-β-D-glucoside and ononin. The proposed method based on HPLC fingerprint of Astragali Radix is convenient and feasible, facilitating the improvement in its quality control.
Subject(s)
Astragalus Plant , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Plant Roots , Quality ControlABSTRACT
OBJECTIVE: To investigate the anti-breast cancer effect of ononin and its mechanism in vitro. METHODS: After treating with different concentration of ononin, the MCF-7 cells viability was determined by MTS, EdU and Hoechst were used to measure the proliferative ability and apoptotic rate of MCF-7, respectively. The invasion and migration of MCF-7 were determined by Transwell assays. The protein expression including PCNA, apoptosis-related proteins (Bax and Bcl-2) and MMP-9 were evaluated by Western-blotting. RESULTS: Ononin inhibited the cells viability of MCF-7 in a dose-and time-dependent manner. The EdU positive cells were decreased, Hoechst positive cells were increased, invasive and migration cells were decreased after treating with ononin in MCF-7. Moreover, ononin up-regulated Bax expression and down-regulated Bcl-2, PCNA as well as MMP-9 expression. CONCLUSION: Ononin could inhibit the MCF-7 cells proliferation, promote cells apoptosis, decrease cells invasion and migration, which may involve the regulative effect on PCNA, Bax /Bcl-2 and MMP-9 expression.
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Objective To investigate the effects of calycosin, formononetin, calycosin-7-glucoside and ononin on PC 12 cells differentiation. Methods PC 12 cells were cultured and treated with different concentrations of nerve growth factor (NGF), calycosin, formononetin, calycosin-7-glucoside and ononin for 5 days, once a day, 3 times in a row. The neurite outgrowth of PC 12 cells was observed and the expression of β III-tubulin were measured by immunofluorescence. Results Compared with the vehicle group, neurite outgrowth and the expression of β III tubulin in PC 12 cells had not promoted by calycosin, formononetin, calycosin-7-glucoside and ononin (0.01-10.00 μmol/L). Conclusion PC 12 cells differentiation could not be induced by calycosin, formononetin, calycosin-7-glucoside and ononin.
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Objective: To investigate the effects of five kinds of flavonoids (calycosin, formononetin, ononin, isoliquiritigenin, and medicarpin) from Hedysari Radix on promoting osteogenic differentiation of rat bone marrow stromal cells (rBMSCs) and rat calvarial osteoblasts (ROBs). Methods: rBMSCs were isolated according to plastic adherence. ROBs were isolated by enzyme digestion method. The proliferation of rBMSCs and ROBs were detected by MTT assay. ALP activity and calcium content of rBMSCs and ROBs cells were detected by alkaline phosphatase kit and calcium kit. Mineralized nodule formation was detected by alizarin red staining. Results: The five components could promote proliferation, increase ALP activity, increase calcium content, and increase the area and number of calcified nodules of rBMSCs and ROBs (P < 0.05). Among them, calycosin had the best effect on promoting the osteogenic differentiation of rBMSCs, and medicarpin promoted the osteogenic differentiation of ROBs with the best effect, followed by calycosin. Conclusion: Five flavonoids promoted the improvement of osteogenic function, while calycosin has better osteogenic activity on rBMSCs and ROBs and can be used as an excellent osteoinductive factor.
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Objective: To study the chemical constituents of flavonoids from Glycyrrhizae Radix et Rhizoma. Methods: The compounds were isolated and purified by column chromatography over HP-20 macroporous resin, silica gel, Sephadex LH-20, and preparative RP-HPLC. Their structures were elucidated by physicochemical properties and spectral analyses. Results: Ten flavonoids were isolated and identified as 4’,6,7-trihydroxy-2’-methoxyl-chalcone (1), 3’,4’,5,7-tetrahydroxy-8-(3-hydroxy-3- methylbutyl)-isoflavone (2), isoliquiritigenin (3), isoliquiritin (4), echinatin (5), orobol (6), ononin (7), 2(S)-3’,5’,7-trihydroxy- flavanone (8), 2(S)-naringenin-4’-O-β-D-glucopyranoside (9), and 4’,7-dihydroxyflavone (10). Conclusion: Compounds 1 and 2 are new compounds named isolicochalcone B and licoisoflavone G, while compound 9 is isolated from the genus for the first time.
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Objective To study the correlation of anti-osteoporosis effect of Hedysarum polybotrys and HPLC fingerprint. Methods Osteoporosis model was established through ovary resection of female rats to select the best part after performing ig administration with different extracts of H. polybotrys. The fingerprints of ethanol extract of ten batches of H. polybotrys from different habitats were established by HPLC. The partial least squares regression (PLSR) analysis was used to explore the correlation between peak areas of HPLC fingerprints and efficacy. Moreover, the effects of active ingredient of H. polybotrys on the function of osteoblasts were observed. Results The ethanol extract of H. polybotrys was the most effective part. Nine common peaks of the active fraction were selected. Adenosine, calycosin and ononin in ethanol extract of H. polybotrys have been identified separately, among which adenosine and calycosin were positively correlated with the pharmacodynamic data, and ononin was negatively correlated with the effect of drug. Additionally, calycosin can increase osteoblast ALP activity. Conclusion The results indicate that anti-osteoporosis effect of H. polybotrys is related to various components, among which adenosine and calycosin have a certain contribution to this effect, and calycosin can also stimulate the differentiation of osteoblasts in vitro.
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Objective To investigate the chemical constituents from Disporum cantoniense. Methods Compounds were isolated and purified by macroporus resin, ods, Sephadex LH-20, MCI-gel CHP20 resin column chromatography and preparative HPLC. The structures were identified by spectral analysis. Results Twenty compounds were isolated and identified as 2-hydroxybenzyl alcohol (1), p-hydroxybenzaldehyde (2), 4-hydroxyacetophenone (3), 3,5-dimethoxy-4-hydroxy-benzaldehyde (4), 4-(4-hydroxyphenyl)-2- butanone (5), neoliquiritin (6), 2,3,5,4’-tetrahydroxy stilbene-2-Ο-β-D-glucoside (7), (E)-1-(4’-hydroxyphenyl)-but-1-en-3-one (8), isoquercitrin (9), 3-hydroxy-1-(4-hydroxy-3,5-dimethoxy-phenyl)-1-propanone (10), icariol A2 (11), ecdysterone (12), glansreginic acid (13), hesperidin (14), ononin (15), quercetin (16), isorhamnetin-3-O-β-D-glucoside (17), (E)-4-(4-hydroxy-3-methoxyphenyl) but-3-en-2-one (18), (-)-secoisolariciresinol (19), and luteolin (20). Conclusion Compounds 1, 3-8, 10-15, and 17-19 are isolated from the genus of Disporum for the first time.
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Objective To establish a method for simultaneous determination for the contents of four flavones (ononin, calycosin, genistein and formononetin) of Hedysari Radix in Gansu Province with quantitative analysis of multi-components by single-marker (QAMS); To prove its feasibility and accuracy. Methods Calycosin was taken as internal standard substance. Relative correction factors (RCF) of ononin, genistein and formononetin to calycosin were established. The contents of ononin, calycosin, genistein and formononetin were determined to realize QAMS. Results RCF was with good repeatability. The results of QAMS were consistent with the results of the external standard method. Conclusion The method that determines the contents of ononin, genistein and formononetin with calycosin as internal standard substance, can be used for quantitative analysis of Hedysari Radix.
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Objective: To study the correlation of HPLC fingerprint of Hedysari Radix and efficacy of improving immunity of normal mice, and provide the basis for finding bioactive components and selecting the index composition of quality control. Methods: The mice delayed type hypersensitivity (DTH) model induced by dinitoflruorobenzene (DNFB) was used to study the effect and then to perform ig administration with different extracts of Hedysari Radix for selecting the best part. To study the correlation between fingerprint and efficacy and find out the effective substances foundation of improving immunity, the peak area of each common peak from HPLC fingerprint was associated with the date of improving immunity in the method of partial least square regression (PLS). Results: In ear swelling experiment, water extract of Hedysari Radix was the most effective part which could improve the immunity function. PLS was used to evaluate the correlation of effect of improving immunity and components of the water extract including adenosine, ononin, genistein, formononeti, and medicarpin, the index of compounds to characterize the quality of Hedysari Radix, showed the negative correlation with the effectiveness. Conclusion: A new method is established to evaluate the activity of enhancing immunity of Hedysari Radix. It is of practical significance as an effective approach for controlling quality and exploring the material basis for efficacy of traditional Chinese medicine compounds.
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OBJECTIVE:To establish a method for the simultaneous determination of flavonoids components in Astragali Ra-dix,and to explore the relationship among flavonoids components,varieties,origins and planting patterns. METHODS:HPLC was performed on the column of Venusil ASB with mobile phase of acetonitrile-0.3% formic acid (gradient elution) at a flow rate of 1.0 ml/min,detection wavelength was 260 nm,and column temperature was 25 ℃. Medicinal material quality of Astragalus mem-branaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao and A. membranaceus (Fisch.) Bge of wild and cultivated from different province was compared. RESULTS:The linear range of the mass concentration was 0.008 9-2.224 mg/ml for calycosin glucoside (r=0.999 5),0.005 2-1.3 mg/ml for ononin(r=0.999 6),0.002 8-0.697 6 mg/ml for calycosin(r=0.999 9)and 0.002-0.5 mg/ml for formononetin (r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 1%;recoveries were 99.52%-100.74%(RSD=0.41%,n=6)for calycosin glucoside,98.84%-100.60%(RSD=0.60%,n=6)for ononin ,98.47%-101.74%(RSD=1.08%,n=6)for calycosin,100.10%-101.59%(RSD=0.32%,n=6)for formononetin. In terms of varieties,the contents of calycosin glycosides,ononin and flavonoids in A. membranaceus(Fisch.)Bge. var. mongholicus(Bge.)Hsiao were higher than those of A. membranaceus (Fisch.)Bge,but the contents of calycosin and formononetin were less than those of A. membranaceus (Fisch.)Bge;in terms of origins,calycosin glycosides and flavonoids of Inner Mongolia and Shanxi held the highest contents,fol-lowed by those of Northeast China and Gansu,and lowest in Shandong,Anhui and Shaanxi;in terms of planting patterns,the con-tents of calycosin glycosides,ononin and flavonoids of wild Astragali Radix were higher than those of cultivated varieties,and the contents of calycosin and formononetin of cultivated varieties were higher than those of wild ones. CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the simultaneous determination of flavonoids components in Astragali Radix. The flavonoids components show great differences in Astragali Radix from different origins,and they are affected by varieties,ori-gins and planting patterns.
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Objective: To study the chemical constituents from the roots and rhizomes of Glycyrrhiza uralensis. Methods: The roots and rhizome of G. uralensis were extracted with 95% EtOH. Then the extract was partitioned with petroleum ether, ethyl acetate, and n-butanol successively. The n-butanol fraction was isolated by silica gel and ODS column chromatography and preparative HPLC. The structures of compounds were identified by spectroscopic methods (UV, MS, 1D-NMR, and 2D-NMR). Results: Fourteen compounds were obtained from the n-butanol fraction. They were macedonoside E (1), 22β-acetyl-uralsaponin C (2), glycyrrhizic acid (3), uralsaponin F (4), licorice-saponin G2 (5), 22β-acetoxyl-glycyrrhaldehyde (6), 3β-O-[β-D-(6-methyl)-glucuronopyranosyl-(1→2)-β-D-glucuronopyranosyl]-glycyrrhizic acid (7), liquiritigenin (8), naringenin (9), isoliquiritigenin (10), ononin (11), liquiritin (12), isoviolanthin (13), and liquiritin apioside (14). Conclusion: Macedonoside E (1) and 22β-acetyl-uralsaponin C (2) are new compounds.
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OBJECTIVE: To establish an UPLC method for simultaneous determination of six flavonoid active components (campanulin, ononin, quercetin, calycosin, kaempferol, and formononetin) of the roots of Astragalus membranaceus (Fisch.) Bge.var.mongholicus (Bge.) Hsiao collected from the main producing areas (Gansu Province, Shaanxi Province, and Inner Mongolia). METHODS: Separation was performed at 30°C on an ACQUITY UPLC BEH C18 column (2.1 mm ×100 mm, 1.7 μm) with a gradient elution system of water containing 0.1% formic acid (mobile phase A) and acetonitrile - isopropanol (7:3) containing 0.1% formic acid (mobile phase B). The flow rate was 0.25 mL·min-1. The detection wavelength was 254 nm, and the sample injection volume was 2 μL. RESULTS: The six flavonoid active components showed good linearity in the selected concentration ranges (r≥0.999), with average recoveries of 99.60%-101.2% (n=6) and RSDs of 1.2%-2.0%. CONCLUSION: This study for the first time establishes an UPLC method for simultaneous determination of six flavonoid active components (campanulin, ononin, quercetin, calycosin, kaempferol, and formononetin) of the roots of Astragalus membranaceus (Fisch.) Bge.var.mongholicus (Bge.) Hsiao. This method is specific, reproducible, controllable, and can be used for the quality control of Radix Astragali.
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OBJECTIVE:To develop an HPLC method for simultaneous determination of calycosin-7-O-?-D-glycoside,ono-nin,calycosin and formononetin in Radix Astragali from eleven different habitats and to explore the internal factors of geoherbalism based on aspect of contents of active constituents. METHODS:The sample was separated on Zorbax Eclipse XDB-C18(250 mm?4.6 mm,5 ?m)column. The mobile phase consisted of acetonitrile-0.2% formic acid solution (gradient elution). The UV detection wavelength was set at 280 nm. RESULTS:The linear ranges of calycosin-7-O-?-D-glycoside,ononin,calycosin and formononetin were 0.005 1～0.510 mg?mL-1,0.005 0～0.300 mg?mL-1,0.004 9～0.294 mg?mL-1,0.004 6～0.276 mg?mL-1,respectively(r=0.999 9).Calycosin-7-O-?-D-glycoside and ononin took up a big proportion in Radix Astragali from Inner Mongolia and Shanxi; calycosin and formononetin took up a big proportion in Radix Astragali from Anguo,Chicheng of Hebei province and Dingxi of Gansu province. CONCLUSION:The method is simple,rapid and accurate. Results of study are in line with textual research on Radix Astragali.