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.

Network pharmacology and molecular dynamics study of the effect of the Astragalus-Coptis drug pair on diabetic kidney disease

BACKGROUND Diabetic kidney disease(DKD)is the primary cause of end-stage renal disease.The Astragalus-Coptis drug pair is frequently employed in the management of DKD.However,the precise molecular mechanism underlying its therapeutic effect remains elusive.AIM To investigate the synergistic effects of multiple active ingredients in the Astragalus-Coptis drug pair on DKD through multiple targets and pathways.METHODS The ingredients of the Astragalus-Coptis drug pair were collected and screened using the TCMSP database and the SwissADME platform.The targets were predicted using the SwissTargetPrediction database,while the DKD differential gene expression analysis was obtained from the Gene Expression Omnibus database.DKD targets were acquired from the GeneCards,Online Mendelian Inheritance in Man database,and DisGeNET databases,with common targets identified through the Venny platform.The protein-protein interaction network and the“disease-active ingredient-target”network of the common targets were constructed utilizing the STRING database and Cytoscape software,followed by the analysis of the interaction relationships and further screening of key targets and core active ingredients.Gene Ontology(GO)function and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichments were performed using the DAVID database.The tissue and organ distributions of key targets were evaluated.PyMOL and AutoDock software validate the molecular docking between the core ingredients and key targets.Finally,molecular dynamics(MD)simulations were conducted to simulate the optimal complex formed by interactions between core ingredients and key target proteins.RESULTS A total of 27 active ingredients and 512 potential targets of the Astragalus-Coptis drug pair were identified.There were 273 common targets between DKD and the Astragalus-Coptis drug pair.Through protein-protein interaction network topology analysis,we identified 9 core active ingredients and 10 key targets.GO and KEGG pathway enrichment analyses revealed that Astragalus-Coptis drug pair treatment for DKD involves various biological processes,including protein phosphorylation,negative regulation of apoptosis,inflammatory response,and endoplasmic reticulum unfolded protein response.These pathways are mainly associated with the advanced glycation end products(AGE)-receptor for AGE products signaling pathway in diabetic complications,as well as the Lipid and atherosclerosis.Molecular docking and MD simulations demonstrated high affinity and stability between the core active ingredients and key targets.Notably,the quercetin-AKT serine/threonine kinase 1(AKT1)and quercetin-tumor necrosis factor(TNF)protein complexes exhibited exceptional stability.CONCLUSION This study demonstrated that DKD treatment with the Astragalus-Coptis drug pair involves multiple ingredients,targets,and signaling pathways.We propose a novel approach for investigating the molecular mechanism underlying the therapeutic effects of the Astragalus-Coptis drug pair on DKD.Furthermore,we suggest that quercetin is the most potent active ingredient and specifically targets AKT1 and TNF,providing a theoretical foundation for further exploration of pharmacologically active ingredients and elucidating their molecular mechanisms in DKD treatment.

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...

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.

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.

膜荚黄芪(Astragalus membranaceus (Fisch) Bunge)组织培养再生体系的建立

本研究以膜荚黄芪的子叶、下胚轴、幼茎、幼叶为外植体,诱导愈伤组织及再生芽,建立膜荚黄芪组织培养再生体系。结果表明,子叶是最佳的外植体。在6-BA与2,4-D或NAA培养基组合中,以MS+6-BA 0.5mg/L+NAA 0.5mg/L为最佳的愈伤组织及芽诱导培养基。黄芪再生植株较难生根,在IAA、IBA和NAA激素培养基组合中,以1/2MS+IBA 2.0mg/L或1/2MS+IBA 1.0mg/L+NAA 1.0mg/L为最佳的试管苗生根培养基,生根率50%左右。

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.

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.

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.

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.

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.

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.

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 .

Predict the anti-cancer mechanism of astragalus membranaceus based on network pharmacology

Objective: To explore the anti-cancer mechanism of active ingredients of Astragalus membranaceus (AM) through network pharmacology. Methods: TCMSP, PubChem, STICTH and GeneCards databases were used to predict and screen the main active ingredients and anti-cancer targets of AM. Active ingredient-target-disease network was constructed by Cytoscape 3.7.0 software, and protein interaction network was constructed by STRING platform. KEGG signaling pathway and GO biological process of targets were analyzed by Bioconductor database. Results: Twenty-four active ingredients were screened from AM, which acted on 106 cancer targets such as PTGS, NCOA2, ADRB2, PRSS1, NOS2, NOS3, GABRA1. Through these targets, the anti-cancer effect of AM mainly acts on small cell lung cancer, colorectal cancer, thyroid cancer, breast cancer, non-small cell lung cancer, hepatocellular carcinoma, pancreatic cancer, gastric cancer, endometrial cancer, enriched in chemical carcinogenesis, Platinum drug resistance, Epstein-Barr virus infection, TNF signaling pathway, Toll-like receptor signaling pathway, p53 signaling pathway, VEGF signaling pathway, NF-kappa B signaling pathway, and PI3K - Akt signaling pathway. Conclusion: This study found that the main anti-cancer compounds of AM are kaempferol, quercetin, 7-O-methylisomucronulatol, formononetin, isorhamnetin, Calycosin, 3,9-di-O-methylnissolin. The main targets include PTGS, PTGS1, NCOA2, ADRB2, PRSS1, NOS2, NOS3, GABRA1, F2. The mechanisms involved in anticancer could be summarized as following: blocking the chemical carcinogenesis, reversing the platinum drug resistance, anti - Epstein - Barrvirus infection, and inhibiting cell proliferation related signaling pathways, such as TNF signaling pathway, Tolllike receptor signaling pathway, p53 signaling pathway, VEGF signaling pathway, NF-kappa B signaling pathway, PI3K - AKT signaling pathway.

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.