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Enhancement of the immune response in mice by Astragalus membranaceus extracts.

Astragalus membranaceus extracts injected into normal mice or mice immunodepressed by cyclophosphamide or radiation treatment or by aging are able to enhance the antibody response to a T-dependent antigen. Optimal conditions for maximum effect have been established in normal C3H/HeJ and (C57B1/10 x DBA/2)F1 mice. Enhancement of the antibody response was associated with increase of Th cell activity in normal and immunodepressed mice. The biological activity of endotoxin-free extracts is dependent on the carbohydrate content.

Adjuvants, Immunologic↗
PubMed2005

A novel homodimeric lectin from Astragalus mongholicus with antifungal activity.

A novel lectin (AMML) was isolated from a Chinese herb, i.e., the roots of Astragalus mongholicus, using a combination of ammonium sulfate fraction and ion exchange chromatographies. The molecular mass of intact AMML was determined to be 66,396 Da by MALDI-TOF mass spectrometry and 61.8 kDa by gel filtration, respectively. AMML was a dimeric protein composed of two identical subunits each with a molecular mass of 29.6 kDa. The lectin was a glycoprotein with a neutral carbohydrate content of 19.6%. The purified lectin hemagglutinated both rabbit and human erythrocytes, and showed preference for blood types O (native) and AB (trypsin-treated). Among various carbohydrates tested, the lectin was best inhibited by D-galactose and its derivatives with pronounced preference for lactose (3.13 mM). N-terminal amino acid sequence of AMML was determined as ESGINLQGDATLANN. The optimal pH range for lectin activity was between pH 4.5 and 7.5, and the lectin was active up to 65 degrees C. It also exerted antifungal activity against Botrytis cincerea, Fusarium oxysporum, Colletorichum sp., and Drechslera turcia but not against Rhizoctonia solani and Mycosphaerella arachidicola.

Amino Acid Sequence↗
PubMed2007

In vitro and in vivo anti-tumor effects of Astragalus membranaceus.

Astragalus membranaceus, a commonly used Chinese medicinal plant, has been shown to be capable of restoring the impaired T cell functions in cancer patients. In this study, the in vitro and in vivo anti-tumor effects of A. membranaceus were investigated. Five bioactive fractions were isolated from the root of A. membranaceus, the fraction designated as AI was found to be the most potent among the five fractions with respect to its mitogenicity on murine splenocytes. Besides investigating the cytostatic effect of AI, its activities on macrophage function, tumor necrosis factor production, induction of lymphokine-activated killer cell and tumor cell differentiation were also examined. The macrophage-like tumors and the myeloid tumors were found to be more sensitive to the cytostatic activity of AI, whereas the fibroblast-like tumors and the mouse Ehrlich ascites tumor appeared to be relatively resistant. Moreover, AI could effectively suppress the in vivo growth of syngeneic tumor in mice. Results showed that murine macrophage pretreated with AI had increased in vitro and in vivo cytostatic activities towards MBL-2 tumor. AI could also act as a priming agent for tumor necrosis factor production in tumor-bearing mice. Preincubation of mouse splenocytes with AI could induce in vitro lymphokine-activated killer-like activity towards WEHI-164 cell. Furthermore, AI was able to induce monocytic differentiation of both human and murine cells in vitro. AI administered in vivo could even partially restore the depressed mitogenic response in tumor-bearing mice. Collectively, the results showed that A. membranaceus could exhibit both in vitro and in vivo anti-tumor effects, which might be achieved through activating the anti-tumor immune mechanism of the host.

Animals↗
PubMed2004

Preparative isolation and purification of isoflavan and pterocarpan glycosides from Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao by high-speed counter-current chromatography.

(3R)-(-)-7,2'-Dihydroxy-3',4'-dimethyl isoflavan-7-O-beta-D-glucopyranoside and (6aR, 11aR) 9,10-di-methoxypterocarpan-3-O-beta-D-glucopyranoside were separated from the ethyl acetate extract of the root of Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao by high-speed counter-current chromatography (HSCCC). A two-phase system composed of ethyl acetate-ethanol-acetic acid-water (4:1:0.25:5, v/v) was selected by analytical HSCCC. Preparative HSCCC yielded, from 100 mg of the partially purified extract, 50 mg of isoflavan glycoside and 10 mg of pterocarpan glycoside each at over 95% purity by high-performance liquid chromatography (HPLC) analysis. Their structures were identified by MS, 1H NMR and 13C NMR.

Astragalus Plant↗
PubMed2005

Astrapterocarpan isolated from Astragalus membranaceus inhibits proliferation of vascular smooth muscle cells.

The inhibitory effects of astrapterocarpan, formononetin, and calycosin isolated from Astragalus membraneceus on platelet-derived growth factor (PDGF)-BB-induced proliferative response in rat vascular smooth muscle cells (A10 cells) were investigated. Astrapterocarpan significantly inhibited PDGF-BB-induced cell proliferation and DNA synthesis in a concentration-dependent manner. This inhibition was not attributed to toxicity. In contrast, formononetin and calycosin had no effect. We next examined the effect of astrapterocarpan on PDGF-BB signal transduction. Astrapterocarpan inhibited PDGF-BB-induced phosphorylation of extracellular signal-regulated kinase 1/2 (ERIC1/2) mitogen-activated protein (MAP) kinase. However, this compound had no effect on phosphorylation of PDGF-beta-receptor, Akt kinase and p38 MAP kinase. These results indicated that astrapterocarpan inhibits PDGF-BB-induced vascular smooth muscle cell proliferation and that this effect may be mediated, at least in part, by inhibition of the ERK1/2 MAP kinase cascade.

Animals↗
PubMed2005

Selenite precipitation by a rhizospheric strain of Stenotrophomonas sp. isolated from the root system of Astragalus bisulcatus: a biotechnological perspective.

A bacterial strain (SeITE02), related to the species Stenotrophomonas maltophilia and resistant to selenite (SeIV) up to 50 mM in the growth medium, was isolated from rhizospheric soil of a selenium hyperaccumulator plant, the legume Astragalus bisulcatus. The influence of SeIV on the active growth of this Se-tolerant bacterial strain has been investigated in oxic conditions, along with the isolate's ability to reduce selenite to elemental selenium (Se(0)). Interestingly, concentrations of 0.5 mM SeIV were wholly reduced by strain SeITE02 in liquid culture within 52 h. Moreover, 87% of SeIV added to the growth medium at the initial concentration of 2.0 mM underwent again reduction in 120 h. Actually, a selenite-mediated induction of a sort of adaptive response to detrimental SeIV effects magnified the efficiency of SeITE02 in reducing this toxic oxyanion. Furthermore, the SeIV influence on cell morphology of strain SeITE02 was evidenced by phase-contrast and electron microscopy analyses. In particular, transmission electron microscopy (TEM)-energy-dispersive X-ray (EDX) analysis of S. maltophilia strain SeITE02, grown in presence of SeIV, showed electron-dense Se(0) granules either in the cell cytoplasm or in the extracellular space. Therefore, the capability of strain SeITE02 to quickly reduce soluble and harmful SeIV to insoluble and unavailable Se(0) may be looked at as a promising exploitable option for the setup of low-cost biological treatments tailored to manage contamination in selenium-laden effluents.

Astragalus Plant↗
PubMed2005

Macrophage activation by polysaccharide isolated from Astragalus membranaceus.

We show that APS, a polysaccharide isolated from the roots of Astragalus membranaceus, significantly induces nitric oxide (NO) production and inducible NO synthase (iNOS) transcription through the activation of nuclear factor-kappaB/Rel (NF-kappaB/Rel). In vivo administration of APS induced NO production by peritoneal macrophages of B6C3F1 mice. APS also dose-dependently induced the production of NO in isolated mouse peritoneal macrophages and RAW 264.7, a mouse macrophage-like cell line. Moreover, iNOS protein and mRNA transcription were strongly induced by APS in RAW 264.7 cells. To further investigate the mechanism responsible for the induction of iNOS gene expression, we investigated the effect of APS on the activation of transcription factors including NF-kappaB/Rel and Oct, whose binding sites were located in the promoter of iNOS gene. Treatment of RAW 264.7 cells with APS produced strong induction of NF-kappaB/Rel-dependent reporter gene expression, whereas Oct-dependent gene expression was not affected by APS. Nuclear translocation and DNA binding activity of NF-kappaB/Rel was significantly induced by APS. The treatment with NF-kappaB SN50, an inhibitor of NF-kappaB/Rel nuclear translocation, effectively inhibited the activation of NF-kappaB/Rel binding complexes and NO production. In conclusion, we demonstrate that APS stimulates macrophages to express iNOS gene through the activation of NF-kappaB/Rel.

Active Transport, Cell Nucleus↗
PubMed2006

Regulation on maturation and function of dendritic cells by Astragalus mongholicus polysaccharides.

Astragalus mongholicus polysaccharides(ASP) isolated from one of the Chinese herbs-A. mongholicus which are known to have a variety of immunomodulatory activity. However, little is known about their immunomodulatory effects on murine bone marrow (BM)-derived dendritic cells (DC). DC are professional antigen presenting cells, which are pivotal for initiation of primary immune response. In this study, the regulatory effects of ASP on maturation and function of cultured murine BM-derived DC were investigated in vitro. ASP (10, 50, 100, 250 microg/ml) could increase the co-expression of CD-11c and MHC class II molecules on DC surface, and the 100 microg/ml is the optimal dose. The ability of unstimulated DC to uptake FITC-dextran was higher than that of ASP- or LPS-treated DC. We analyzed the concentration of IL-12 secreted by DC using ELISA. ASP-treated DC secreted a higher level of IL-12 than untreated DC. And ASP- or LPS-treated DC displayed a more mature morphology, with long protrusions, while untreated-DC displayed shorter protrusions than stimulated DC.

Animals↗
PubMed2005

Isoflavonoids from Astragalus mongholicus protect PC12 cells from toxicity induced by L-glutamate.

lsoflavonoids, formononetin, 9,10-dimethoxypterocarpan 3-O-beta-D-glucoside, ononin, calycosin 7-O-glc and calycosin, were isolated from the roots of Astragalus mongholicus Bunge (Leguminosae). The neuroprotective roles and direct antioxidant effects of these isoflavonoids were investigated by using PC12 cell model and DPPH (1,1-diphenyl-2-picrylhydrazyl) assay. Formononetin, ononin and calycosin were found inhibiting glutamate-induced cell injury, with an estimated 50% effective concentration (EC50) of 0.027 microg/ml, 0.047 microg/ml and 0.031 microg/ml, respectively. Pretreatment with them increased the activities of antioxidant enzymes, including superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and prevented the release of lactate dehydrogenase (LDH) in glutamate-injured PC12 cells. On the other hand, calycosin 7-O-glc and calycosin showed more scavenging activity to DPPH radicals than formononetin in the cell-free system. The inconsistency between the neuroprotective capabilities of isoflavonoids and their directly scavenging activity to DPPH radicals indicated that formononetin, ononin and calycosin probably depended on increasing endogenous antioxidant and stabilizing the cells' membrane structures to inhibit the cell damage induced by glutamate.

Animals↗
PubMed2006

A novel nodule-enhanced gene encoding a putative universal stress protein from Astragalus sinicus.

A nodule-enhanced gene, AsD243, was identified from infected roots of Astragalus sinicus using suppressive subtractive hybridization (SSH). It encodes a 20-kD protein related to the bacterial universal stress protein family (Usp). Sequence analysis showed that AsD243 is highly similar to the bacterial MJ0577-type of ATP-binding Usp proteins, which have been proposed to function as a molecular switch. Expression analyses revealed that AsD243 was transcribed in all plant organs, and progressively during all stages of nodulation. Its transcripts increased significantly at 7 days after inoculation, which is 2 days later than the onset of leghemoglobin expression in A. sinicus nodules. AsD243 was expressed more strongly in mature roots than in young roots regardless of inoculation status. We suggest that the AsD243 may have other functions in plant processes besides nodulation.

Amino Acid Sequence↗
PubMed2026

Astragalus membranaceus therapy in patients with spinocerebellar ataxia type 3.

BACKGROUND AND AIM: Spinocerebellar ataxia type 3 (SCA3) is the most common form of hereditary cerebellar ataxia. The insulin/insulin-like growth factor 1 (IGF1) system (IIS) has been proposed as a potential target for disease-modifying therapy. Astragalus membranaceus (AM) may modulate the IIS pathway and reduce neurodegeneration, as indicated by plasma neurofilament light chain (NfL), a biomarker of disease progression. EXPERIMENTAL PROCEDURE: A randomized, triple-blind, placebo-controlled crossover trial was conducted in 32 patients with SCA3. Participants received AM or placebo for three months, followed by a one-month washout, then crossed over to the alternate treatment. Plasma NfL and IIS-related markers were measured. Twenty-three participants completed the trial. RESULTS AND CONCLUSION: Intention-to-treat analysis revealed a modest within-treatment reduction in NfL levels (27.6 ± 10.7 vs. 25.6 ± 9.8 pg/mL, P = 0.040) and increased insulin (12.5 ± 15.2 vs. 21.1 ± 15.5 μIU/mL, P = 0.004) after AM. However, the adjusted regression model showed no significant between-treatment difference in NfL (P = 0.127), and changes in NfL were not correlated with insulin. No significant changes were observed in IGF1 or IGF-binding proteins. Exploratory subgroup analyses suggested larger biomarker changes in later-stage patients. No carryover effects or demographic differences were noted. Overall, AM administration was associated with modest within-treatment biomarker changes in NfL and insulin, although the adjusted between-treatment difference in NfL was not statistically significant. These findings should be interpreted cautiously as preliminary and hypothesis-generating observations rather than confirmatory evidence of therapeutic efficacy.

Astragalus membranaceus↗
PubMed2004

Kahiricosides II-V, cycloartane glycosides from an Egyptian collection of Astragalus kahiricus.

Four cycloartane-type saponins, kahiricosides II-V (1-4), were isolated from the aerial parts of Astragalus kahiricus of Egyptian origin. Their structures were established as 9beta,19-cyclolanost-24E-ene-3beta,6alpha,16beta,27-tetraol-3-O-beta-D-glucopyranoside, 9beta,19-cyclolanost-24E-ene-3beta,6alpha,16beta,27-tetraol-3-O-(2'-O-acetyl)-beta-D-glucopyranoside, 9beta,19-cyclolanost-24E-ene-3beta,6alpha,16beta,27-tetraol-3-O-(6'-O-acetyl)-beta-D-glucopyranoside, and 9beta,19-cyclolanost-24E-ene-3beta,6alpha,16beta,27-tetraol-3-O-beta-D-glucopyranosyl-27-O-beta-D-glucopyranoside based on chemical and spectral evidences. All compounds exhibited very weak cytotoxicity against the A2780 ovarian cancer cell line.

Antineoplastic Agents, Phytogenic↗
PubMed2005

Leishmanicidal cycloartane-type triterpene glycosides from Astragalus oleifolius.

Two new cycloartane-type glycosides oleifoliosides A (1) and B (2) were isolated from the lower stem parts of Astragalus oleifolius. Their structures were identified as 3-O-[beta-xylopyranosyl-(1 --> 2)-alpha-arabinopyranosyl]-6-O-beta-xylopyranosyl-3beta,6alpha,16beta,24(S),25-pentahydroxycycloartane and 3-O-[beta-xylopyranosyl-(1 --> 2)-alpha-arabinopyranosyl]-6-O-beta-glucopyranosyl-3beta,6alpha,16beta,24(S),25-pentahydroxycycloartane, respectively, by means of spectroscopic methods (IR, 1D and 2D NMR, ESI-MS). Three known cycloartane glycosides cyclocanthoside E (3), astragaloside II (4) and astragaloside IV (5) were also isolated and characterized. All five compounds were evaluated for in vitro trypanocidal, leishmanicidal and antiplasmodial activities as well as their cytotoxic potential on primary mammalian (L6) cells. Except for the compound 5, all compounds showed notable growth inhibitory activity against Leishmania donovani with IC50 values ranging from 13.2 to 21.3 microg/ml. Only weak activity against Trypanosoma brucei rhodesiense was observed with the known compounds astragaloside II (4, IC50 66.6 microg/ml) and cyclocanthoside E (3, IC50 85.2 microg/ml), while all compounds were inactive against Trypanosoma cruzi and Plasmodium falciparum. None of the compounds were toxic to mammalian cells (IC50's > 90 microg/ml). This is the first report of leishmanicidal and trypanocidal activity of cycloartane-type triterpene glycosides.

Animals↗
PubMed2005

Haemolytic activities and adjuvant effect of Astragalus membranaceus saponins (AMS) on the immune responses to ovalbumin in mice.

In this study, the haemolytic activities of Astragalus membranaceus saponins (AMS) and its adjuvant potentials on the cellular and humoral immune responses of ICR mice against OVA were evaluated. We determined the haemolytic activity of AMS using 0.5% rabbit red blood cell. AMS showed a slight haemolytic effect, with its haemolytic percent being 0.66% at the concentration of 500 microg/ml. Furthermore, the adjuvant potentials of AMS at three dose levels on the cellular and humoral immune responses of ICR mice against ovalbumin (OVA) were investigated. ICR mice were immunized subcutaneously with OVA 100 microg alone or with OVA 100 microg dissolved in saline containing Alum (200 microg), QuilA (10 and 20 microg) or AMS (50, 100 or 200 microg) on Day 1 and 15. Two weeks later (Day 28), concanavalin A (Con A)-, lipopolysaccharide (LPS)- and OVA-stimulated splenocyte proliferation and OVA-specific antibodies in serum were measured. AMS significantly enhanced the Con A-, LPS-, and OVA-induced splenocyte proliferation in the OVA-immunized mice especially at a dose of 100 microg (P<0.05 or P<0.001). OVA-specific IgG, IgG1 and IgG2b antibody titers in serum were also significantly enhanced by AMS compared with OVA control group (P<0.01 or P<0.001). Moreover, no significant differences (P>0.05) were observed between enhancing effect of AMS and QuilA on the OVA-specific IgG, IgG1 and IgG2b antibody responses to OVA in mice. In conclusion, the results suggest that AMS could be safely used as adjuvant with low or non-haemolytic effect.

Adjuvants, Immunologic↗
PubMed2003

Preparative isolation and purification of two isoflavones from Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao by high-speed counter-current chromatography.

Two isoflavones, calycosin-7-O-beta-D-glycoside and formononetin-7-O-beta-D-glycoside, were separated from n-butanol extract of the root of Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao by high-speed counter-current chromatography in two steps using two different solvent systems composed of ethyl acetate-ethanol-n-butanol-water (30:10:6:50, v/v) and ethyl acetate-ethanol-water (5:1:5, v/v). From 200 mg of crude extract, calycosin-7-O-beta-D-glycoside (12 mg) and formononetin-7-O-beta-D-glycoside (10 mg) were isolated at over 95% purity by HPLC analyses, and their structures were identified by MS, 1H NMR and 13C NMR.

Astragalus propinquus↗
PubMed1994

Cycloartane triterpene glycosides from the hairy root cultures of Astragalus membranaceus.

Agroastragaloside II, a new astragaloside was isolated from the hairy root culture of Astragalus membranaceus. Its structure was established as 3-O-beta-(2'-O-acetyl)-D-xylopyranosyl-6-O-beta-D-glucopyranosyl-(24S)- 3 beta,6 alpha,16 beta,24,25-pentahydroxy-9,19-cyclolanostane on the basis of spectroscopic data. Three known astragalosides, astragaloside II, isoastragaloside I and 3-O-beta-D-xylopyranosyl-cycloastragenol were also isolated.

Astragalus propinquus↗
PubMed2001

Genetic diversity of rhizobia isolated from Astragalus adsurgens growing in different geographical regions of China.

The genetic diversity among 95 isolates from Astragalus adsurgens was investigated using molecular biological methods. All of the isolates and 24 reference strains could be differentiated by AFLP, REP-, ERIC- and BOX-PCR fingerprinting analysis. By cluster analysis of the data, 31 AFLP and 38 Rep-PCR genomic groups were delineated, indicating considerable genetic diversity among the isolates. Fifty-four representative strains were further analyzed by RFLP of PCR-amplified 16S and 23S rDNA, revealing 26 rDNA genotypes among the isolates. The phylogenetic relationship of the isolates was determined by partial sequencing of 16S rRNA genes of 16 strains. The results suggest that the A. adsurgens rhizobia belong to the genera Agrobacterium, Mesorhizobium, Rhizobium and Sinorhizobium.

Astragalus Plant↗
PubMed2001

Secondary metabolites from the roots of Astragalus zahlbruckneri.

Four new phenolic glycosides, beta-apiofuranosyl-(1-->2)-beta-glucopyranosides (1-4), along with the cycloartane triterpenes 20(R),25-epoxy-3beta,6alpha,16beta,24alpha-tetrahydroxycycloartane (5) and 20(R),24(S)-epoxy-3beta,6alpha,25-trihydroxycycloartan-16-one (6) were isolated from roots of Astragalus zahlbruckneri. The structure elucidation of all compounds was based on their (1)H and (13)C NMR spectral data including 1D-TOCSY, DQF-COSY, HSQC, and HMBC experiments.

Astragalus Plant↗