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ObjectiveTo establish a method based on specific polymerase chain reaction （PCR） that can accurately and rapidly identify Astragalus membranaceus var. mongholicus （AMM） seeds and A. membranaceus （AM） seeds. MethodThe Chloroplast Genome Information Resource （CGIR） and IdenDSS were used to obtain the characteristic DNA fragments of AMM and AM， and the specific single nucleotide polymorphism （SNP） sites of AMM and AM were screened out， on the basis of which the specific primers MG-F/MG-R of AMM and MJ-F/MJ-R of AM were designed. The specific PCR method for identifying AMM and AM was established and optimized， and the specificity and applicability of the method were investigated. ResultThe specific PCR conditions for the identification of AMM were primers MG-F/MG-R， annealing at 62 ℃， and 28 cycles. After PCR amplification and gel electrophoresis， the specific band appeared at about 220 bp， with no band for the seeds of AM or adulterants. The specific PCR conditions for identifying the AM were primers MJ-F/MJ-R， annealing at 58 ℃， and 28 cycles. After PCR amplification and gel electrophoresis， the band appeared at about 150 bp， with no band of AMM or adulterants. ConclusionThe specific PCR method established in this study can accurately and quickly identify the seeds of AMM and AM， which provides a basis for the classification and accurate identification of Astragalus seeds and adulterants.
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Astragalus, which was first documented in Shennong Bencao Jing, is the dried root of Astragalus membranaceus (Fisch.) Bge. or Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao. The active ingredients astragalus membranaceus saponins (AMS), astragalus polysaccharides (APS) and astragalus flavonoids (AFS) have pharmacological effects such as anti-tumor properties, lowering blood sugar, regulating lipid metabolism, cardiovascular protection, anti-oxidation, bone protection, anti-fibrosis, etc. Fibrosis affects almost all organs, particularly vital organs such as the lungs, liver, heart and kidneys. The primary pathological changes of fibrosis involve abnormal increase of myofibroblasts and excessive deposition of extracellular matrix (ECM) components, which lead to the formation of scar tissue, ultimately resulting in fibrosis and even functional loss or failure of organs, which seriously threatens human health and life. Recent, studies have shown that Astragalus membranaceus has a good therapetuic effect on organ fibrosis. This article reviews the current advances of Astragalus in the prevention and treatment of fibrosis of lungs, liver, heart, kidneys and other important organs.
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italic>Astragalus is a commonly used Chinese medicinal material in traditional Chinese medicine (TCM), and with the increase of planting area in recent years, the damage of Astragalus root rot has worsened year by year, which seriously affecting its quality and yield. Fusarium oxysporum is one of the main pathogens causing root rot in astragalus. In this study, UPLC-Q-TOF-MS based metabolomic approach combined with multivariate statistical analysis were used to analyze the metabolite changes of Astragalus in response to F. oxysporum infection. The results showed that 62 metabolites in the Astragalus had significant changes after inoculation of F. oxysporum. Polar metabolites included 40 flavonoids, 8 saponins, 2 nucleosides, 1 vitamin, 1 organic acid, 1 amino acid; while lipid metabolites included 3 fatty acids, 1 diradylglycerols, 2 lysophosphatidylcholine, 1 lysophosphatidylglycerol, 1 phosphatidylinositol, 1 sterol lipid. Among these differential metabolites, the relative content of flavonoids, vitamin B2, tryptophan and salicylic acid were increased, while the relative content of saponins were decreased. Correlation analysis showed that the flavonoids were positively correlated with each other, and positively correlated with most lipids, but negatively correlated with most saponins. In addition, studies have shown that F. oxysporum infection is not an influencing factor for the generation of malonyl substitution of flavonoid. This study elucidates the effect of F. oxysporum infection on Astragalus from the perspective of plant metabolism, which provides a basis for exploring the interaction mechanism between the Astragalus and F. oxysporum and further promoting molecular breeding.
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Root rot severely restricts the sustainable development of Astragalus membranaceus var. mongholicus (AMM) industry. Resistance breeding is an economical and environmentally safe way to manage the disease and its key lies in the obtaining of resistance indicators. This study aimed to quickly and accurately screen the resistance-related (RR) metabolites so as to provide reference for the screening of indicators of AMM breeding for resistance. LC-MS-based targeted metabolomics and real-time quantitative PCR technology were employed, in combination with multivariate statistical analysis, in analyzing the dynamic changes of phenylpropanoid metabolites in AMM in response to root rot pathogen Fusarium solani (FS) infection and identifying the differential metabolites. The LC-MS method established showed high sensitivity; each metabolite had a good linear relationship (R2 ≥ 0.968 9) in the corresponding linear range of the respective standard curve; the recoveries and the relative standard deviations (RSDs) (n = 6) ranged from 70% to 107% and from 1.2% to 9.9%, respectively. Obvious disturbances were observed in the changes of the targeted metabolites in AMM infected by FS. These metabolites, compared with the mock-inoculated (CK) group, showed different up or down regulation with time series. Calycosin-7-O-β-D-glucoside, ononin, calycosin and formononetin were identified as differential metabolites, and they all belong to flavonoids. The first three compounds were significantly negatively correlated (r ≤ -0.97, P < 0.05) with the content of FS in the root of AMM. As potential RR metabolites, they are helpful in obtaining promising resistance indicators for AMM against FS infection.
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Astragalosides are the main active constituents of traditional Chinese medicine Huang-Qi, of which cycloastragenol-type glycosides are the most typical and major bioactive compounds. This kind of compounds exhibit various biological functions including cardiovascular protective, neuroprotective, etc. Owing to the limitations of natural sources and the difficulties encountered in chemical synthesis, re-engineering of biosynthetic machinery will offer an alternative and promising approach to producing astragalosides. However, the biosynthetic pathway for astragalosides remains elusive due to their complex structures and numerous reaction types and steps. Herein, guided by transcriptome and phylogenetic analyses, a cycloartenol synthase and four glycosyltransferases catalyzing the committed steps in the biosynthesis of such bioactive astragalosides were functionally characterized from Astragalus membranaceus. AmCAS1, the first reported cycloartenol synthase from Astragalus genus, is capable of catalyzing the formation of cycloartenol; AmUGT15, AmUGT14, AmUGT13, and AmUGT7 are four glycosyltransferases biochemically characterized to catalyze 3-O-xylosylation, 3-O-glucosylation, 25-O-glucosylation/O-xylosylation and 2'-O-glucosylation of cycloastragenol glycosides, respectively. These findings not only clarified the crucial enzymes for the biosynthesis and the molecular basis for the structural diversity of astragalosides in Astragalus plants, also paved the way for further completely deciphering the biosynthetic pathway and constructing an artificial pathway for their efficient production.
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OBJECTIVE To optimize extraction technology of couplet medicinals of Astragalus membranaceus-Puerariae lobatae. METHODS With contents of puerarin，daidzin，calycosin-7-O-β-D-glucopyranoside，daidzein，calycosin and formononetin and the yield of dry extract as index，the analytic hierarchy method was used to determine the weight coefficient of each index and calculate the comprehensive score. The effects of solid-liquid ratio， extraction times and extraction time on the comprehensive score were investigated by single factor test. The level of each factor was determined. By multi-index comprehensive scoring method， using comprehensive scores of above 7 indexes as indexes，the extraction technology of couplet medicinals of A. membranaceus-P. lobata was optimized by orthogonal experiment，and the validation tests were conducted. RESULTS The weight coefficient for the contents of puerarin，daidzin，calycosin-7-O-β-D-glucopyranoside，daidzein，calycosin and formononetin and the yield of dry extract were respectively 0.304 7，0.065 2，0.185 8，0.185 8，0.107 8，0.107 8 and 0.042 7. The optimal extraction technology was determined as follows： solid-liquid of 1∶8（g/mL），extracting 3 times and for 1 h each time. RSD of each evaluation index in the validation test results was lower than 3.00% （n＝3）. CONCLUSIONS The optimized extraction technology for A. membranaceus-P. lobata is stable and feasible.
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@#Objective To investigate the effect of astragalus membranaceus（AM）injection on apoptosis and autophagy of human gastric epithelial cell line（GES⁃1）induced by enterovirus 71（EV71）. Methods GES⁃1 cells were cultured in vitro and divided into infected group（EV71 infected at a MOI of 3 and control group（no virus infected）. The morpho⁃logical changes of EV71 infected cells were observed by inverted microscope. The level of VP1 in GES⁃1 cells infected with EV71 was detected by Western blot；CCK⁃8 assay was used to detect the viability of GES⁃1 cells infected with EV71；Nuclear staining with DAPI was used to observe the morphological changes of nuclear apoptosis infected with EV71. GES⁃1 cells were divided into control group（without virus infection），infection group and AM intervention group with final concentration of 1，2. 5，5 and 10 μg／mL，respectively. Western blot was used to detect the effect of AM intervention on the expression of apoptosis⁃related proteins Caspase⁃3，PARP and autophagy⁃related proteins LC3 and P62 in GES⁃1 cells infected withEV71. CCK⁃8 method was used to detect the effect of AM intervention on the viability of GES⁃1 cells infected with EV71. Results GES⁃1 cells were round，shrunken with nuclear pyknosis and uneven size；VP1 level increased（t = 41. 56，P < 0. 01），cell viability decreased（t = 19. 07，P < 0. 01），Caspase⁃3 and PARP proteins were cut off（t = 35. 29 and 3. 648， P < 0. 01 and 0. 021 8，respectively），LC3Ⅱ／LC3Ⅰ ratio increased（t = 10. 16，P = 0. 000 5）and P62 protein was degraded（t = 68. 68，P < 0. 01）；AM inhibited the degradation of Caspase⁃3，PARP and P62 proteins induced by EV71 （t = 52. 66，59. 60 and 40. 22，respectively，each P < 0. 01）and increased the ratio of LC3Ⅱ／LC3Ⅰ（t = 5. 521，P = 0. 005 3），andreducedtheinhibitoryeffectofEV71ontheviabilityofGES⁃1cells（t =4. 420，P =0. 0115）. Conclusion EV71 infection induced apoptosis of GES⁃ 1 cells and AM intervention inhibited EV71 induced apoptosis by inhibiting EV71 induced autophagy.
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The primary chemical components of Astragalus membranaceus include polysaccharides, saponins, flavonoids, and amino acids. Recent studies have shown that Astragalus membranaceus has multiple functions, including improving immune function and exerting antioxidative, anti-radiation, anti-tumor, antibacterial, antiviral, and hormone-like effects. Astragalus membranaceus and its extracts are widely used in clinical practice because they have obvious therapeutic effects against various autoimmune diseases and relatively less adverse reaction. Multiple sclerosis (MS) is an autoimmune disease of central nervous system (CNS), which mainly caused by immune disorder that leads to inflammatory demyelination, inflammatory cell infiltration, and axonal degeneration in the CNS. In this review, the authors analyzed the clinical manifestations of MS and experimental autoimmune encephalomyelitis (EAE) and focused on the efficacy of Astragalus membranaceus and its chemical components in the treatment of MS/EAE.
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Animals , Humans , Astragalus propinquus/chemistry , Multiple Sclerosis/drug therapy , Encephalomyelitis, Autoimmune, Experimental/metabolism , Drugs, Chinese Herbal/chemistry , PolysaccharidesABSTRACT
OBJECTIVE To study the mitigation effect and its possible mechanism of Astragalus membranaceus polysaccharide on the pulmonary hypertension induced by monocrotaline in rats. METHODS One hundred SD male rats were randomly divided into normal control group ，monocrotaline group ，A. membranaceus polysaccharide low-dose and high-dose groups. In addition to the normal control group, rats in other groups were injected with monocrotaline by single intraperitoneal injection of 60 mg/kg. On days 2 to 28 after administration ，rats in the A. membranaceus polysaccharide low-dose and high-dose groups were intraperitoneally injected with A. membranaceus polysaccharide of 200 mg/kg and 400 mg/kg，respectively，once a day. There were 25 rats in each group，and 15 rats were taken for index detection. The mean pulmonary artery pressure （mPAP）and right heart hypertrophy index （RVHI）were detected ，and morphology changes of pulmonary artery and cardiomyocytes were monitored . mRNA and protein expression of IL- 17 in their lung tissues were detected. RESULTS Compared with normal control group ，mPAP and RVHI of monocrotaline group and A. membranaceus polysaccharide groups were increased significantly （P＜0.01）；mRNA and protein expression of IL- 17 in lung tissues were significantly increased （P＜0.01），and there were obvious pathological changes in pulmonary artery and cardiomyocytes. Compared with monocrotaline group ，mPAP and RVHI were significantly decreased in A. membranaceus polysaccharide groups （P＜0.01），while mRNA and protein expression of IL- 17 in lung tissue were decreased significantly （P＜0.01）；pathological changes in pulmonary artery and cardiomyocytes were improved. Compared with A. membranaceus polysaccharide low-dose group ，above indexes and pathological changes were improved significantly in high-dose group. CONCLUSIONS A. membranaceus polysaccharide can reduce monocrotaline-induced pulmonary hypertension ，improve pulmonary artery structure and myocardial remodeling in rats ， the mechanism of which is presumably related to the down-regulation of IL- 17 expression in lung tissue of rats.
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Objective: To investigate the effect and the mechanism of Astragalus membranaceus (Huangqi in Chinese, HQ) extract on the intestinal absorption of six alkaloids of Aconitum carmichaelii (Fuzi in Chinese, FZ) in rats with spleen deficiency and provide novel insights into the application of HQ on modulating intestinal barrier. Methods: Four-week-old male Sprague-Dawley rats were fed with Xiaochengqi Decoction to induce the spleen deficiency model for 40 d. Single-pass intestinal perfusion model were used to study the effects of HQ extract on the absorption of alkaloids. Protein expression and mRNA levels of MRP2 and BCRP and tight junction proteins (TJ, including Claudin-1, Occludin and ZO-1) were measured using Western blot and real-time PCR, respectively. The location and expression of TJ protein was also investigated by the immunofluorescence method. Results: Compared with the normal group, the protein expression of MRP2, BCRP and TJ proteins in the model group were significantly down-regulated. After oral administration of HQ, the alkaloid absorption in intestinal villi was inhibited, MRP2, BCRP and TJ proteins were up-regulated, the green fluorescence staining of Claudin-1, Occludin, and ZO-1 was enhanced, and a thick layer of mucus was deposited on the surface of the epithelium of the intestinal cavity. Conclusion: HQ as an intestinal barrier modulator improves the physiological changes of the intestinal environment of spleen deficiency to reduce the absorption of toxic components, leading to a decrease in the absorption of drug-like molecules.
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Seven compounds were isolated from Astragalus membranaceus of northern shaanxi by silica gel and Sephadex LH-20 column chromatographies. Their chemical structures were identified on the basis of their physical and chemical properties. These compounds were elucidated as astragaloside IV (1), formononetin (2), calycosin (3), 1-(4-hydroxyphenyl)-4-(2,4-hydroxyphenyl)-2-hydroxy-1,4-but anedione (4), (E)-4-methylcinnamic acid (5), quercetin (6), and uridine (7). Compound 4 is a new compound and compound 5 was isolated from the plants of Astragalus Linn. for the first time. The results of in vitro antitumor activity assay showed that compound 4 could inhibit the proliferation of A549 with IC50 values of 11.41 μmol·L-1.
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Objective:To study the effects of 24-epibrassinolide （EBR） at different concentrations on seed germination， seedling growth and key enzyme activities for flavonoid biosynthesis in <italic>Astragalus membranaceus </italic>var.<italic> mongholicus</italic> under polyethylene glycol （PEG）-induced drought stress， in order to provide theoretical reference for standardizing the drought-resistant cultivation techniques of <italic>A. membranaceus </italic>var.<italic> mongholicus</italic>. Method:The seeds of <italic>A. membranaceus </italic>var.<italic> mongholicus </italic>were soaked in EBR solution at different concentrations（0.001， 0.01， 0.1， 1， 10 μmol·L<sup>-1</sup>） followed by foliar spraying to explore their effects on seed germination， seedling growth， photosynthesis， chlorophyll and malondialdehyde （MDA） contents， and key enzyme activities for flavonoid biosynthesis under drought stress induced by 20% polyethylene glycol （PEG）-6000. Result:Compared with the control（CK）group， PEG-induced drought stress led to an obvious decrease in germination potential， germination rate， germination index， vigor index， relative germination rate， plant height， root length， aboveground and root dry weight， net photosynthetic rate （<italic>Pn</italic>）， stomata conductance （<italic>Gs</italic>）<italic>， </italic>transpiration rate （<italic>Tr</italic>）， chlorophyll contents and chlorophyll a/b ratio， while a significant increase in intercellular carbon dioxide concentration （<italic>Gi</italic>）， MDA， flavonoids contents， and key enzyme activities for flavonoid biosynthesis such as phenylalanine ammonia lyase （PAL）. The treatment with exogenous EBR solution at the suitable concentration significantly enhanced the adaptation of <italic>A. membranaceus </italic>var.<italic> mongholicus </italic>seeds and seedlings to PEG-induced drought stress， manifested as significantly elevated germination potential， germination rate， germination index， vigor index， relative germination rate， plant height， root length， aboveground and root dry weight， <italic>Pn</italic>， <italic>Gs， Tr</italic>， chlorophyll a/b ratio， chlorophyll and flavonoids contents and key enzyme activities for flavonoid biosynthesis like PAL while lowered <italic>Ci</italic> and MDA contents. The optimal concentration of EBR solution was 0.1 μmol·L<sup>-1</sup>. Conclusion:Exogenous EBR solution at the suitable concentration ameliorates the inhibitory effect of 20% PEG stress against seed germination and seedling growth of <italic>A. membranaceus </italic>var.<italic> mongholicus</italic>， reduces the oxidative damage in leaves， and improves the stress resistance to a certain extent by up-regulating the key enzyme activities and promoting flavonoid synthesis.
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A quantitative analytical method based on HPLC coupled with the charged aerosol detector (CAD) for quantitative analysis of multi-components with a single marker (QAMS) was established for simultaneous determinations of astragaloside Ⅰ, astragaloside Ⅱ, astragaloside Ⅳ, calycosin-7-O-β-D-glucoside, formononetin and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan in Astragalus membranaceus. The separation was performed on an Agilent SB-C18 (150 mm×4.6 mm, 3.5 μm), with gradient elution using the mobile phase consisting of 0.05% formic acid solution and 0.05% formic acid acetonitrile at the flow rate of 1.0 mL·min-1. The column temperature was 35 ℃, and the injection volume was 20 μL. For CAD, the drift tube temperature was at 50 ℃. The contents of six components in A. membranaceus were determined by both external standard method (ESM) and QAMS, and then were compared. The results showed that chromatographic peaks were separated well and the linear ranges of astragaloside Ⅰ, astragaloside Ⅱ, astragaloside Ⅳ, calycosin-7-glucoside, formononetin and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan were 0.113-2.250 mg·mL-1, 0.012-0.240 mg·mL-1, 0.004-0.080 mg·mL-1, 0.065-1.300 mg·mL-1, 0.005-0.100 mg·mL-1 and 0.007-0.150 mg·mL-1, respectively. The content ranges of astragaloside Ⅰ, astragaloside Ⅱ, astragaloside Ⅳ, calycosin-7-glucoside, formononetin and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan were 0.306-0.922 mg·g-1, 0.053-0.183 mg·g-1, 0.015-0.092 mg·g-1, 0.069-0.823 mg·g-1, 0-0.098 mg·g-1 and 0.020-0.107 mg·g-1 in 20 batches of A. membranaceus, respectively. Using astragaloside Ⅱ as an internal reference, the relative correlation factors of astragaloside Ⅰ, astragaloside Ⅳ, calycosin-7-O-β-D-glucoside, formononetin, and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan were calculated as 0.561, 0.835, 0.299, 0.796, and 0.799, respectively. The results were compared with those obtained by the external standard method to verify the feasibility, rationality and repeatability of QAMS method, and there was no significant difference in assay results between the two methods. In conclusion, the QAMS method is accurate and feasible, and could be used to determine the contents such as astragaloside Ⅰ, astragaloside Ⅱ, astragaloside Ⅳ, calycosin-7-glucoside, formononetin and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan, and it can be used for quality control of A. membranaceus.
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OBJECTIVE：To study metabonomics of secondary components of Astragalus membranaceus injection on leukopenia model mice. METHODS ：The mice were divided into normal group ，model group ，A. membranaceus injection group （0.004 mL/g），secondary components low-dose ，medium-dose and high-dose groups （0.004，0.008，0.016 mL/g），with 7 mice in each group. Except for normal group ，other groups were given cyclophosphamide （80 mg/kg） intraperitoneally to induce leukopenia model. After modeling ，administration groups were given relevant medicine intraperitoneally ，and normal group and model group were given constant volume of sterile water for injection intraperitoneally ，once a day ，for consecutive 7 days. During the experiment ，the changes of body weight of mice were measured every 2 days. After the last administration ，the blood routine indexes [white blood cell （WBC），neutrophil（NE），lymphocyte（LY）and monocyte （MO）counts] of mice were detected by animal blood analyzer. UPLC-Q Exactive Orbitrap-HRMS combined with multivariate statistical analysis were used to analyze the metabonomics of mice serum. RESULTS ：Compared with model group ，body weight of mice in the secondary component low-dose group increased significantly on the 4th and 8th day of administration （P＜0.05），and the counts of WBC ，NE，LY and MO in serum of mice in secondary component groups increased significantly （P＜0.05 or P＜0.01）. Metabonomic analysis showed that the secondary components of A. membranaceus injection could significantly regulate the contents of 8 endogenous metabolites in serum， including spermidine ， uric acid ， citric acid ， nicotinamide， eicosapentaenoic acid ， linoleic acid ， erucamide and sphingosine-1-phosphate. Their effects involved linoleic acid metabolism pathway ，nicotinic acid and nicotinamide metabolism pathway and tricarboxylic acid cycle pathway. CONCLUSIONS ：The secondary components of A. membranaceus injection possess the effect of increasing white blood cells in leukopenia model mice ，the mechanism of which may be related to intervention of linoleic acid metabolism pathway ， nicotinic acid and nicotinamide metabolism pathway ，and tricarboxylic acid cycle pathway.
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Astragalus membranaceus (A. membranaceus) is a widely used traditional herb in China and Korea. A. membranaceus polysaccharides (AMP), which make up a major part of the root extract, have been shown to modulate immune modulations, especially activation of bone marrow-derived dendritic cells (BMDCs) and T cells. However, the immune stimulatory effect of AMP in the mouse in vivo and human peripheral blood DCs (PBDCs) has not been well investigated. In this study, we found that intravenous (i.v.) injection of AMP in C57BL/6 mice induced remarkable elevations in co-stimulatory and MHC class I and II molecule levels in the splenic DCs and its subsets. The stimulatory effect of DCs by AMP was elevated 6 h after treatment, which rapidly decreased 18 h after injection. Furthermore, AMP promoted intracellular production of pro-inflammatory cytokines in spleen DC subsets, which contributed elevation of serum cytokine levels. Finally, the AMP promoted PBDC activation. Thus, these results demonstrate that AMP can be used as an immune stimulatory molecules in human and mouse.
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This work is to establish the fingerprint of Astragalus membranaceus var. mongholicus by HPLC-ELSD method, and to analyze the simulated wildness degree of A. membranaceus var. mongholicus in the genuine region of Inner Mongolia, Ningxia and Gansu. Compared with wild A. membranaceus var. mongholicus, the quality differences of A. membranaceus var. mongholicus in the genuine region were analyzed by identification of chromatographic peaks and similarity evaluation, cluster analysis(CA), principal components analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA). HPLC fingerprints of A. membranaceus var. mongholicus in different genuine regions are established. The qualitative analysis of mass spectrometry identified 18 components. The similarity evaluation shows that the similarity of 32 batches of A. membranaceus var. mongholicus samples was 0.688-0.993. Among them, the similarity of samples in Shanxi, Inner Mongolia, Ningxia is 0.688-0.993, 0.835-0.989, 0.934-0.988, respectively and the similarity of samples in Gansu is 0.729-0.876 except No. 25 sample. The results of CA show that the samples of A. membranaceus var. mongholicus can be grouped into four categories according to the production area except the No. 11 and No. 25 samples. The results of PCA indicate that 32 batches of A. membranaceus var. mongholicus samples can be clustered according to quality and origin, and the quality of A. membranaceus var. mongholicus in Inner Mongolia is the closest to the wild breed. The results of OPLS-DA indicate that there are six components that can distinguish the wild and domestic A. membranaceus var. mongholicus, which are malonylastragaloside Ⅰ, astragaloside Ⅰ, calycosin-7-O-β-D-glycoside-6″-O-malonate, calycosin-7-O-β-D-glycoside, formononetin-7-O-β-D-glycoside-6″-O-malonate, and astrapterocarpan-3-O-β-D-glycoside-6″-O-malonate. The established method can be used to analyze differences between A. membranaceus var. mongholicus origin and planting environment, and can provide references for the protection and replacement of wild A. membranaceus var. mongholicus resources, and the cultivation, processing and production of A. membranaceus var. mongholicus.
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Astragalus propinquus , ChinaABSTRACT
OBJECTIVE：To establish a method for simultaneous determination of calycosin glucoside ，ononin，calycosin， formononetin，astragaloside Ⅳ，isoastragaloside Ⅱ，cycloastragenol and isoastragaloside Ⅰ in Astragalus membranaceus before and after bidirectional solid fermentation with Cordyceps kyushuensis ，and to investigate the effects of fermentation on the contents of above 8 components in A. membranaceus . METHODS ：HPLC-DAD-ELSD was adopted. The determination was performed on Agilent 5 TC-C18 column with mobile phase consisted of 0.1％ formic acid aqueous solution-acetonitrile （gradient elution ）at the flow rate of 1 mL/min. The column temperature was set at 30 ℃. DAD detection wavelength was set at 260 nm，ELSD evaporation tube temperature was 100 ℃，atomizer temperature was 80 ℃，carrier gas flow rate was 1.6 L/min；injection volume was 15 μL. RESULTS：The eight components had a good linear relationship within their respective ranges of concentration （all R2＞0.999 0）； RSDs of precision ，stability and repeatability tests were all less than 3％（n＝3 or n＝6）；the recoveries was 97.88％-101.32％， and RSDs were 1.22％-2.39％（n＝6）. Setting the content of components in unfermented A. membranaceus as 100％，after bidirectional solid fermentation with C. kyushuensis ，the change rates of 8 components were －98.51％，－96.41％，－94.74％， －96.40％，289.20％，20.25％，－75.05％，562.46％，respectively. CONCLUSIONS ：After fermentation with C. kyushuensis ，the contents of active components as astragaloside Ⅳ，isoastragaloside Ⅰ and isoastragaloside Ⅱ can be increased significantly in A. membranaceus .
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OBJECTIVE：To provide referenc e for intuitively and c onveniently cognizing the research focus of Astragalus membranaceus in the treatment of diabetes. METHODS ：Using the method of bibliometrics ，the literatures about the treatment of diabetes by A. membranaceus were retrieved from CNKI full-text database during Jan. 1st，2000-Jun. 30th，2020. Bibliometric method was used to statistically analyze the number of articles ，authors and their research directions ，keywords，distribution of research institutions ，source journals ，frequency of citations ，fund funding ，etc. VOSviewer software was used for visual display. RESULTS：A total of 2 960 literatures were included ，and the number of literatures showed an increasing trend ；the author with the most literatures was Fan Ying from Liaoning University of TCM ，whose research direction was the effect of different effective parts of A. membranaceus on serum insulin and adiponectin in diabetic animal models ；in the keyword frequency analysis ，diabetic nephropathy（845 times），A. membranaceus （615 times），diabetes（587 times），A. membranaceus injection（350 times），diabetic peripheral neuropathy （303 times），qi tonifying drug （202 times）were hot topics. There were 10 institutions with more than 20 literatures，of which Nanjing University of TCM has published the most literatures （58 pieces）. Shaanxi Journal of Traditional Chinese Medicine ，Chinese Journal of Integrated Traditional and Western Nephropathy and Inner Mongolia Journal of Traditional Chinese Medicine were the journals with a large number of； literatures；Research Progress on Chemical Constituents and Pharmacological Effects of Astragalus membranaceus published from Chen guohui and others was cited most frequently，for 690 times；there are many projects funded by national scientific research funds （including National Natural Science Foundation of China and the National Key Basic m Research Special Foundation of China ）. CONCLUSIONS ：In the past 20 years， the level of researches about A. membranaceus in the treatment of diabetes have been steadilyimproved. However ，the distribution of literature is uneven and there are few high-quality core journals. Mainly ，there are relatively few clinical trials. It is suggested that researchers should explore the methods and ideas of A. membranaceus in the treatment of diabetes from the perspectives of macro clinical efficacy ，micro pharmacology ，A. membranaceus genetic resource bank and traditional Chinese medicine formulations.
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OBJECTIVE：To study “Qi-invigorating”effect and its possible mechanism of total saponins of Astragalus membranaceus on rats with Qi-deficiency ，and to provide reference for elucidating the material basis of “Qi-invigorating”effect of A. membranaceus . METHODS ：Forty male Wistar rats were randomly divided into normal group ，model group ，positive control group [Buzhong yiqi pills ，4.5 g/（kg·d）]，A. membranaceus total saponins high-dose and low-dose groups [ 252，28 g/（kg·d），by the amount of total saponins] according to body weight ，with 8 rats in each group. Except for normal group ，the model of Qi-deficiency was made in other groups by the method of “diet disorder+fatigue ”. At the same time ，administration groups were given relevant medicine intragastrically ，and normal group and model group were given constant volume of water，once a day ，for consecutive 21 days. After last administration ，the general situation of rats was observed ；the body weight ，spleen index and thymus index of rats were detected ；weight-bearing swimming time was recorded ；the levels of spleen T lymphocyte subsets CD 3 and CD 4，the levels of ATP and ADP in liver tissue ，serum levels of ALB ，RBC and HBG in blood as well as the serum levels of SOD，MDA，lactate，LDH，CK，IL-2，IL-12 and TNF-α were all detected. RESULTS：Compared with normal group ，body weight，thymus index ，spleen index ，weight-bearing swimming time ，the level of spleen T lymphocyte subsets CD 3，ATP，ADP， ALB，IL-2 and IL- 12 were decreased or shortened significantly in model group （P＜0.05 or P＜0.01）. The levels of MDA ， lactate，CK and TNF-α were increased significantly （P＜ 0.05）. Compared with model group ，body weight ，spleen index，weight-bearing swimming time ，the level of spleen T lymphocyte subsets CD 3 and the levels of ATP ，ADP，ALB， RBC and IL- 2 were increased significantly or prolonged（P＜0.05）；while the levels of MDA ，lactate，CK and TNF-α were decreased significantly in A. membranaceus total saponins high-dose group（P＜0.05 or P＜0.01）.Weight-bearing swimming time ，the levels of ATP ，ADP and IL- 2 in A. membranaceus total saponins low-dose group were increased significantly or prolonged （P＜0.05 or P＜0.01），while the levels of MDA ，lactate，CK and TNF-α were decreased significantly （P＜0.05 or P＜0.01）. Compared with positive control group ，spleen index ，spleen T lymphocyte subsets CD 3，weight-bearing swimming time and ATP level of A. membranaceus total saponins high-dose group were increased significantly or prolonged （P＜0.05 or P＜0.01），while MDA levels of A. membranaceus total saponins high-dose and low-dose groups were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：A. membranaceus total saponins can reduce the body ’s accumulation of blood lactic acid ，the activity of CK ，the level of lipid peroxide and regulate immunity to tonify Qi ，delay fatigue and improve exercise ability.
ABSTRACT
Objective::Astragalus membranaceus var. mongholicus and A. membranaceus are medicinal Astragalus, which are closely related and similar in composition, but with unclear medicinal value. Water-soluble protein profiles for A. membranaceus var. mongholicus and A. membranaceus were established to explore the differences between the two kinds of Astragalus Radix. Method::The water-soluble protein components were obtained through water ultrasonic extraction and acetone precipitation. After digested with trypsin, the obtained peptides were analyzed by nano ESI-LC-MS/MS method. Proteome Discovery 1.4 software was used to identify the proteins by comparing with the legume protein database, and the different expression water-soluble proteins were analyzed by the label-free quantitative software SIEVE. Finally, relevant information for common expression proteins, including classification, molecular function, involved biological process and signaling pathway, were analyzed by bioinformatics. Result::There were 920 and 717 specific proteins identified for A. membranaceus var. mongholicus and A. membranaceus, respectively. Totally 472 proteins were found to be co-expressed, in which 21 were differentially expressed, such as PR-10 protein, NDK-1 protein, glutelin A2, and phospholipase D. There were 14 highly expressed proteins in A. membranaceus var. mongholicus and 7 highly expressed proteins in A. membranaceus. Conclusion::There are significant differences in water-soluble protein profiles for two kinds of Astragalus Radix. Specific proteins, differentially expressed proteins and common expressed proteins can provide references for the identification of A. membranaceus var. mongholicus and A. membranaceus. It also can be used to define pharmacological mechanisms and search for drug action targets.