Total flavonoids of Astragalus membranaceus protect against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced neurotoxicity in mice by inhibiting ferroptosis through SLC7A11/GPX-4 signaling pathway

Parkinson’s disease(PD)is a common neurodegenerative disorder with no cure.Astragalus membranaceus is used in Chinese culture as a food supplement to boost immunity.The present study aimed to explore the neuroprotective effects of total flavonoids extracted from A.membranaceus(TFA)and their protective mechanisms.TFA offered neuroprotection against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)in the mouse model of Parkinsonism,by improving behavior performance in the gait analysis and pole test,and inhibiting the decline of tyrosine hydroxylase(TH)positive neurons and TH protein expression in substantia nigra of mice.TFA also prevented 1-methyl-4-phenylpyridinium(MPP+)induced neurotoxicity in SHSY5Y cells,by increasing GSH and GSH/GSSG ratio,and reducing reactive oxygen species.In addition,the neuroprotective effects of TFA were associated with its ability to restore MPTP/MPP+induced downregulation of SLC7A11 and glutathione peroxidase 4(GPX-4).In conclusion,we demonstrated that TFA exerted significant neuroprotection against MPTP/MPP+induced neurodegeneration by inhibiting ferroptosis through the regulation of SLC7A11/GPX-4 axis,suggesting the use of TFA as a possible food supplement in the prevention of PD.

Effects of Astragalus membranaceus on Energy Metabolism and Expression of CNTF Protein in Skeletal Muscle of Exercise-induced Fatigue Rats

[Objectives]This study was conducted to investigate the effects of Astragalus membranaceus in different groups on energy metabolism and CNTF protein expression in skeletal muscle of exercise-induced fatigue rats.[Methods]Thirty-five clean male SD rats were randomly divided into a normal group,and low-,meddle-and high-dose groups of A.membranaceus aqueous solution,with 7 rats in each group.The low-dose,medium-dose and high-dose groups were given by gavage at 0.65,1.3 and 2.6 g/kg,respectively,while the normal group and the model group were given normal food and water.The weight of rats was observed.The contents of serum urea,lactate,muscle glycogen,liver glycogen and CNTF expression were detected.[Results]After modeling,compared with the normal group,the serum lactate and urea contents of rats in the model group significantly increased(P<0.01),while the muscle glycogen content(P<0.01)and liver glycogen content(P<0.05)of the skeletal muscle significantly decreased.Compared with the model group,the low-,meddle-and high-dose groups of A.membranaceus significantly reduced the levels of lactate and urea in serum(P<0.01),while the levels of muscle glycogen and liver glycogen in the skeletal muscle significantly increased(P<0.01,P<0.05).[Conclusions]This study provides a good research foundation for the treatment of exercise-induced fatigue using traditional Chinese herb A.membranaceus in modern clinical practice.

ISOFLAVONES FROM ASTRAGALUS MEMBRANACEUS

Two new isoflavones (8, 3'-dihydroxy-7,4'-dimethoxyisoflavone, odoratin-7-0-[3-D-glu-copyranoside) and four known isoflavones (formononetin, 7,3'-dihydroxy-8,4'-dimethoxyisoflavone, calycosin, calycosin-7-0-(3-D-glucopyranoside) were isolated from the roots of Astragalus mem-branaceus (Fisch.) Bunge. Their structures were established by spectral analysis.

Study on Pollinating Insects of Astragalus membranaceus (Ficsh) Bunge

[Objective The aim was to study species and pollinating characters of Astragalus membranaceus(Ficsh)Bunge pollinating insects and lay a theory foundation for the breeding of Astragalus membranaceus(Ficsh)Bunge.[Method] With Astragalus membranaceus(Ficsh)Bunge as research object,the species of pollinating insect and pollination behavior were investigated.[Result] There were 16 pollinating insect species,among which,Bombus ignitus,Bombus lucoru,Apis sp.,Betasyrphus serarius(wiedemann)and Colias erate(Esper)we...

Comparative analysis of twenty-five compounds in different parts of Astragalus membranaceus var.mongholicus and Astragalus membranaceus by UPLC-MS/MS

As a traditional Chinese medicine,the root of Astragalus membranaceus var.mongholicus(AMM) or A.membranaceus(AM) has been widely used in China and other Asian countries for thousands of years.Till now,the flavonoids,phenolic acids and saponins are considered as the main active components contributing to their therapeutic effect in these plants.In order to clarify the distribution and contents of these compounds in different organs of these plants,a rapid and sensitive analytical method for simultaneous determination of 25 active compounds including seven types(i.e.dihydroflavones,isoflavane,isoflavones,flavones.pterocarpans,phenolic acid and saponins) within 10 min was established using ultra-pressure liquid chromatography coupled with tandem mass spectrometry(UPLC-MS/MS).Then,the established method was fully validated and successfully applied to the determination of the contents of these analytes in different parts(root,rhizome,stem,leaf and flower) of AMM and AM.The results indicated that the contents of the same type of compounds in two different species plants were significantly different.Moreover,the obvious differences were also found for the distribution and contents of different type of compounds in five organs of the same species.The present study could provide necessary information for the rational development and utilization of AMM and AM resource.

Study on Tissue Culture and Radiation Mutation of Astragalus membranaceus Bge. var. mongholicus(Bge.) Hsiao

[Objective] The research aimed to study the tissue culture technology and callus induction by radiation mutation of A. membranaceus Bge. [ Method ] With the different parts of Astragalus membranaceus Bge. var. mongholicus （ Bge. ） Hsiao aseptic seedling as explants （ leaves, cotyledons, hypocotyls） induced callus, and cotyledon and hypocotyls taken by the method of radiation mutation were studied. [ Result]The results showed that the three explants had relatively high callus induced rate in the medium which respectively made up of MS ＋6-BA 2.0 mg/L ＋ NAA2.0 mg/L, LS ＋6-BA2.0 mg/L ＋NAA0.1 mg/L, MS ＋ 6-BA2.0 rng/L ＋ NAA2.0 rag/L; the optimum mutation time of hypocotyls and cotyledons was 15 minutes; the growth of the callus induced from hypocotyls would be better as the mutation time increased, but when it reached a certain time the growth would be weaken, the induction rate also would be reduced. [ Conclusion] This study will provide the scientific reference in tissue culture and mutation breeding of A. membranaceus Bge.

Protective effect of Astragalus membranaceus on intestinal mucosa reperfusion injury after hemorrhagic shock in rats

AIM： To study the protective effect of Astragalus rnernbranaceus on intestinal mucosa reperfusion injury and its mechanism after hemorrhagic shock in rats. METHODS： A total of 32 SD rats were randomly divided into four groups （n = 8, each group）： normal group, model group, low dosage group （treated with 10 g/kg Astragalus membranaceus） and high dosage group （treated with 20 g/kg Astragalus membranaceus）. The model of hemorrhagic shock for 60 min and reperfusion for 90 min was established. Therapeutic solution （3 mL） was administrated before reperfusion. At the end of the study, the observed intestinal pathology was analyzed. The blood concentrations of lactic acid （LD）, nitric oxide （NO）, endothelin-1 （ET-1）, malondialdehyde （MDA） and the activity of superoxide dismutase （SOD）, glutathione peroxidase （GSH-PX） in intestinal mucosa were determined. RESULTS： The intestinal mucosa pathology showed severe damage in model group and low dosage group, slight damage in high dosage group and no obvious damage in normal group. The Chiu＇s score in low dose group and high dose group was significantly lower than that in model group. The content of MDA in model group was higher than that in low and high dose groups, while that in high dose group was almost the same as in normal group. The activity of SOD and GSH-PX was the lowest in model group and significantly higher in high dose group than in normal and low dose groups. The concentrations of LD and ET-1 in model group were the highest. The concentrations of NO in model group and low dose group were significantly lower than those in high dose group and normal group. CONCLUSION： High dose Astraga/us membranaeus has much better protective effect on hemorrhagic shockreperfusion injury of intestinal mucosa than low dose Astragalus membranaceus. The mechanism may be that Astragalus membranaceus can improve antioxidative effect and regulate NO/ET level during hemorrhagic reperfusion.

Amelioration of experimental colitis by Astragalus membranaceus through anti-oxidation and inhibition of adhesion molecule synthesis

AIM: To investigate the protective effects of Astragalus membranaceus(Am) against hapten-induced colitis in male Sprague-Dawley rats as well as its underlying mechanism.METHODS: Experimental colitis was induced in rats by enema administration of 2,4-dinitrobenzene sulfonic acid (DNBS). Rats were either pretreated with Am extract (2 or 4 g/kg, p.o. once daily) starting from 10 d before DNBS enema, or received Am post-treatment (2 or 4 g/kg, p.o.twice daily) on the three consecutive days following DNBS administration. Colonic lesion area and histological damage were determined, while the activities of myeloperoxidase (MPO) and xanthine oxidase, as well as reduced glutathione (GSH) content were measured in the excised colonic tissues. Besides, protein expression of inducible nitrite oxide synthase (iNOS), intercellular adhesion molecule-1 (ICAM-1) and P-selectin was also detected by Western blot analysis.RESULTS: Our findings had shown that both macroscopic lesion area and histological colonic damage induced by DNBS were significantly reduced by both Am pre- and post-treatments. These were accompanied by attenuation of the elevated colonic MPO activity and downregulation of the iNOS, P-selectin, and ICAM-1 protein expression.Besides, deprivation of colonic GSH level under colitis condition was also preserved.CONCLUSION: These results demonstrate that Am possesses both preventive and therapeutic potential in experimental colitis. The anti-inflammatory actions involve anti-oxidation along with inhibition of adhesion molecule synthesis in the colonic tissues.

Two new saponins from the aerial part of Astragalus membranaceus var. mongholicus

Two new saponins named mongholicoside A （1） and mongholicoside B （2） were isolated from the aerial part of Astragalus membranaceus var mongholicus. Their structures were determined by 1D and 2D NMR, ESI-MS techniques and chemical methods.

UGPase of Astragalus membranaceus : cDNA Cloning, Analyzing and Expressing in Escherichia coli

On the basis of sequences of UGPase from plants, a cDNA encoding the enzyme was isolated from the hairy root of Astragalus membranaceus (Fisch.) Bunge. The cDNA consisted of 1 831 bp and encoded a polypeptide of 471 amino acid residues with a calculated molecular weight of 51.5 kD and a deduced isoelectric point of 6.01. Then the open read frame of the cDNA was ligated into pET28(a) + vector and expressed in E. coli BL21. SDS_PAGE showed that the expressed protein was ca. 40% in the total bacterial protein. Enzyme activity assay demonstrated that the UGPase activity in the transformed bacteria was 0.50-3.27 times higher than that of the control. Northern blotting revealed that ugp was expressed in the leaf, stem, root and hairy root of A. membranaceus , with a higher level in root and hairy root.

Protective effects of combined Astragalus membranaceus and magnesium ions on global brain ischemia

The traditional Chinese herb Astragalus membranaceus is a well-known treatment for neurological diseases and is considered to exhibit anti-dementia properties.This study investigated the synergistic effects of magnesium ions and Astragalus membranaceus on global brain ischemia in rats.4＇-6-diamidino-2-phenylindole staining demonstrated that the number of living neurons was significantly greater in the rat hippocampus after administration of a combination of Astragalus membranaceus and magnesium,compared with a vehicle group,an Astragalus membranaceus alone group,and a magnesium alone group.Western blot assay revealed that cleaved Caspase-3 protein expression was significantly reduced in the rat hippocampus in the combined Astragalus membranaceus and magnesium group compared with the Astragalus membranaceus alone group and the magnesium alone group.The results suggested that the combination of Astragalus membranaceus and magnesium exhibits a stronger neuroprotective effect on global brain ischemia in rats compared with Astragalus membranaceus or magnesium alone.This effect was associated with decreased Caspase-3 expression.

Chemistry Behind the Immunomodulatory Activity of Astragalus membranaceus

Huang Qi(黄芪Astragalus membranaceus)is a well-known and widely used herb in traditional Chinese medicine(TCM)tonic preparations.It has been used for many ailments over the last 2000 years.Flavonoids,saponins,and polysaccharides have been shown to be the main compounds responsible for the biological and pharmacological activities,especially the immunomodulatory properties,of such tonic preparations.This review summarizes the published data on Astragalus extracts and fractions and the natural compounds responsible for the immunomodulatory activity with special reference to the modulation of nuclear factor-kappa B and related pathways(e.g.,Nrf2).In addition,this review highlights the importance of Astragalus membranaceus in TCM for treating patients with diseases related to immunocompromised conditions,such as cancer and diabetes.

A new isoflavane from processed Astragalus membranaceus

A new isoflavane named astraganoside, together with five known compounds had been isolated from the processed Astragralus membranaceus. The structure of the novel compound was elucidated as (3R, 4R)-3-(2-hydroxy-3,4-dimethoxyphenyl)chroman-4,7-diol-7-O-β-d-glucopyranoside (1) based on spectroscopic methods including UV, IR, ESI-MS, 1D NMR and 2D NMR techniques.

Isolation and Identification of Root Rot Fungus of Astragalus membranaceus

The perennial root of Astragalus membranaceus is used as a medicine, while root rot is a main factor causing reduction of quality and commodity value of A. membranaceus . The screening and research of the pathogenic species and their characteristics could provide theoretical and practical basis for the control of this disease. A pathogenic strain was isolated and purified from the root part of four-year-old A. membranaceus , and identified by morphological and molecular biological methods as Fusarium oxysporum . This study will provide a theoretical basis for the research of the biological characteristics and control of F. oxysporum .

THE IN VITRO POTENTIATION OF LAK CELL CYTOTOXICITY IN CANCER AND AIDS PATIENTS INDUCED BY F3 A FRACTIONATED EXTRACT OF ASTRAGALUS MEMBRANACEUS

The in vitro induction of LAK cell activity was studied in cancer and AIDS patients. F3, an immuno regulatory component of Astragalus membranaceus was shown capable of potentiating LAK cell activity induced by rIL-2. The LAK cells killing activity against Hs294T melanoma cell line induced by 50 U/ml rIL-2 in the Presence of F3 (55 μg/ml) reached 64%, which was comparablc to that (60%) induced by 500 U/ml of rIL-2alone. With F3 and rIL-2, the effcctor to target ratio could be reduced to one-half in order to obtain an equivalent level of cytotoxicity induced by rIL-2 alone.In some patients whose Peripheral blood Iymphocytes were relatively inert of rIL-2, F3 could make them responsive to rIL-2 induction. These results imply that F3 may be useful to potentiate LAk cell activity, reduce the dosage of rIL-2 and thus minimize the later's toxic side effects when used in vivo.

The Effects of Astragalus Membranaceus Extract on Sperm Quality in vitro

This study is to explore the effects on sperm quality and fertilizing ability by using astragalus membranaceus extract as additive among infertile males. Human sperms were cultured in the concentrated astragalus membranaceus extract for 1 h, then semen parameters were measured by the equipment of computer assisted semen analyzer (CASA). The results are as follows: compared with the original semen, the sperm motility and velocity, the percentage of grade A sperms and the swaying frequency of sperm heads were improved markedly in the semen treated with astragalus membranaceus extract (P<0.05). Meanwhile, the astragalus membranaceus aqueous extract was analyzed by atomic absorption spectrophotometer. It was found there exist several metal elements and a certain amount of trace elements in the extract (10 mg/ml). The concentration of K +, Ca 2+ , Zn 2+ were 25.1 μg/ml, 7.28 μg/ml, and 0.47 μg/ml respectively. The influence of this additive on artificial insemination was also discussed. We considered that the studied Chinese herbal medicine additive has good effects on sperm preparation in vitro, and this might be a potential method in assisted reproductive technology.

Amino Acid Analysis of Astragalus Membranaceus (Fisch.)Bge

Amino acid composition of Astragalus Membranaceus Bge(AMB)was analysed by automatic analyzer.The results showed that the amount of combined amino acids was 8.05%of total weight,the free amino acids was 0.38%and the whole amino acids was 8.43%.AMB contains all of the essential amino acids except Trp xysshhant.Besides the therapy effect,the AMB has significant nutrient effect.

Bioinformatics analysis of Astragalus membranaceus on the treatment of prostate cancer

Objective:Prostate cancer(PCa)is the second most common male malignancy tumor in the world,and the occurrence rate and the mortality of PCa keeps increasing nowadays,exploration of accurate biomarker and predict clinical outcome of PCa is of great significance.Traditional Chinese medicine Huangqi(Astragalus membranaceus,AM)has been widely used for PCa treatment in China,while the pharmacological mechanisms are still unclear.The current paper intend to perform a network pharmacology analysis to reveal the mechanism of the effects of AM in PCa.Methods:Traditional Chinese Medicine Systems Pharmacology Database(TCMSP)was used to obtain chemical constituents of AM.PCa-related target genes were collected via OMIM and GeneCards databases.PCa-AM common target protein interaction network was established by the STRING database.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were performed to further explore the PCa mechanism and therapeutic effect of AM.The network diagrams of the active component-action target and protein-protein interaction(PPI)networks were constructed using Cytoscape software.Results:A total of 20 active ingredients contained and 462 putative identified target genes were screened from AM,of which 46 overlapped with the targets of PCa and were considered potential therapeutic targets.The analysis of the network results showed that the AM activity components quercetin,kaempferol,isorhamnetin,3,9-di-O-methylnissolin and 7-O-methylisomucronulatol have a good binding activity with top ten screened targets,such as MYC,AKT1,CCND1,MAPK1,CASP3,EGFR,ESR1,VEGFA,ERBB2 and IL-6.GO and KEGG analyses revealed that these targets were associated with proteoglycans in cancer,virus infection,microRNAs in cancer,PI3K-Akt signaling pathway,and other biological process.Conclusion:This study demonstrated the multicomponent,multitarget,and multichannel characteristics of AM,which provided a novel approach for further research of the mechanism of AM in the treatment of PCa.

A New Isoflavone Glucoside from Astragalus membranaceus

A new isoflavone glucoside, 3'-methoxy-5'-hydroxy-isoflavone-7-O-beta-D-glucoside, was isolated from the roots of Astragalus membranaceus and its structure was elucidated by spectroscopic data.

The mechanism of Astragalus membranaceus in treating peritoneal fibrosis by intervening the key syndrome and pathology based on Q-marker theory

Objective:To evaluate the mechanism of Astragalus membranaceus(AM)by intervening peritoneal mesothelial cells(PMCs),epithelial-mesenchymal transition(EMT),and spleen deficiency syndrome(SDS)in peritoneal fibrosis(PF),we base on employing the strategy of Q-marker theory combination network pharmacology method.Methods:First,we obtained the Q-markers of AM by searching the relevant literature and its pharmacological information was collected based on SwissADME.The SwissTargetPrediction and pharmmaper were employed to predict its potential target.Secondly,GeneCards,DisGeNET,and OMIM databases were employed to search the related targets of EMT,SDS,and PF.VENNY2.1 tool was employed to obtain the intersection targets of AM and the three;then the“AM potential target-SDS-EMT-PF”Venn diagram was constructed.The common targets of AM,EMT and SDS were uploaded to the STRING database and obtained the PPI protein interaction network map.Cytoscape 3.7.2 was employed to evaluate the core target of PPI network.PATHER and Metascape databases were used to analyze protein type,GO biological process,and KEGG pathway.Finally,a network diagram of the“TCM-component-disease target-pathway”was drawn.Results:A total of 10 AM Q-makers were screened out,corresponding to 335 targets of AM,2,728 targets of SDS,373 of EMT,and 612 PF targets were found.Among them,there are 155 common AM targets related to SDS and EMT.Key targets such as ALB,AKT1,VEGFA,TNF,EGFR,CASP3,SRC,STAT3,HSP90AA1,and ESR1 were obtained.The core drug include quercetin,astragalosideIII,Calycosin-7-O-beta-D-glucoside,astragalosideIV,etc.The types of PPI proteins include protein modification enzymes,metabolite transferases,transmembrane signal receptors,etc.Biological processes include the regulation of kinase activity,the positive regulation of transferase activity,and the regulation of kinase activity.The key pathways may include PI3K-Akt signaling pathways,the non-smad pathway of the TGF-βsignaling pathway,and the AGE-RAGE signaling pathway.Conclusion:AM could prevent and treat PF through a multi-component multi-target-multi-path mechanism.Astragalus saponins may be the main component types of AM intervening EMT pathology by strengthening the spleen and nourishing Qi.AstragalosideIV and astragalosideIII may be the constituents that can invigorate the spleen and replenish Qi.The results of this study contribute to a more comprehensive understanding of the critical components and mechanisms of AM by intervening SDS and EMT in the treatment of PF.