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[Effect of Astragalus polysaccharides in promoting neutrophil-vascular endothelial cell adhesion and expression of related adhesive molecules].

OBJECTIVE: To explore the detoxication and tissue generation effect of Astragalus (As) in wound healing and its relation with inflammatory reaction, through observing the effect of Astragalus polysaccharides (AP) on neutrophil-endothelial cell adhesion and expression of related adhesive molecules. METHODS: Human polymorphonuclear leucocyte (PMN) or human umbilical vein endothelial cell (HUVEC) was treated separately with AP, AP plus interleukin 1 (IL-1) and tumor necrosis factor (TNF) to study the effect of AP on PMN adhesion with HUVEC by rose bengal staining, and that on expression of superficial adhesive factor by means of Cell-ELISA and APAAP method. RESULTS: When AP acted on HUVEC, it could significantly promote the adhesion of HUVEC with PMN, while when AP acted on PMN, the adhesion would not increase. When HUVEC was treated by AP plus IL-1, the IL-1 induced PMN adhesion with HUVEC could be strengthened, and the expression of HUVEC superficial adhesive factor ICAM-1 induced by IL-1 and TNF was strengthened also, but when PMN treated with AP, it showed no effect on the expression of adhesive factor CD18. CONCLUSION: AP promotes the adhesion between neutrophil and endothelial cell by way of promoting the expression of superficial I-CAM-1 on surface of endothelial cells, so as to improve the inflammatory reaction in the wound healing course, it possibly is one of the biological bases of the detoxication and tissue generation effects of AP.

Astragalus propinquus↗

[The effect of Astragalus polysaccharide on endotoxin-induced toxicity in mice].

Astragalus Polysaccharide (APS), an active component, was isolated from the radix of Astragalus membranaceus Bge var. mongholicus (Bge) Hsiao, and the effects of APS on E. coli endotoxin-induced liver damage were investigated in mice. The results showed that when mice were pretreated with APS (30, 60, 100 mg.kg-1.d-1 x 7d, ip), the survival rate of endotoxin intoxicated mice was increased the lowering of ATP levels and adenylate energy charge in mouse liver could be prevented. APS 100 mg.kg-1 could protect the mice from death on endotoxin (25 mg.kg-1) intoxication; and the level of ATP, the value of adenylate energy charge in the protected mouse liver were almost recovered to normal range. The effects of APS were shown to be dose-dependent. Concomitantly, the increase of MDA and decrease of GSH in mouse liver could be corrected by APS pretreatment. The results revealed that APS has an antioxidative action. An ultramicroscopic examination showed that the injury of the bio-membrane and the crest of mitochondria were ameliorated by APS pretreatment. These findings suggest that the protective effects of APS on E. coli endotoxin intoxicated mice may be due to its antioxidative action to protect the mitochondria bio-membrane, therefore, the adenylate metabolism is improved in mouse liver.

Adenosine Triphosphate↗

[Effects of Astragalus membranaceus injection on nitric oxide and endothelin concentration of intestinal mucosa after hemorrhage shock-reperfusion in rats].

OBJECTIVE: To observe the effects of Stragalus membranaceus injection on nitric oxide and endothelin levels of intestinal mucosa in reperfusion injury after hemorrhage shock. METHOD: 32 SD rats were randomly divided into four groups: normal group, model group, low dosage group, (treated with Astragalus membranaceus 10 g x kg(-1)); high dosage group (treated with Astragalus membranaceus 20 g x kg(-1)). Models of hemorrhagic shock for 60 minutes and reperfusion for 90 minutes were created. The animals were administrated 3 mL therapeutic solution before reperfusion. At the end of study, intestinal pathology was observed, and the concentration of lactic acid (LD), nitric oxide (NO), endothelin (ET) of intestinal mucosa were detected. RESULT: The intestinal pathology showed that intestinal mucosa epithelial cells damage in model group was severe, in low dosage group was medium, in high dosage group was slight, and no obvious damage was found in normal group. The concentration of LD and NO of small intestine mucous membrane in model group and low dosage group were significantly higher than those in high dosage group and normal group (P < 0.05), but there were no significant differences between high dosage group and normal group (P > 0.05). The concentration of ET of small intestine mucous membrane in model group was the highest of the four groups (P < 0.05). The concentration of ET in low dosage group was significantly higher than that in high dosage group and normal group (P < 0.05), but there were no significant differences between high dosage group and normal group (P > 0.05). CONCLUSION: Stragalus membranaceus injection can reduce small intestine mucous damage by protecting endothelium function in injury after hemorrhage shock-reperfusion.

Animals↗

[Standard operating procedure for Astragalus membranaceus].

In order to ensure the superior quality and safety of the raw materials of the Chinese herbal medicine, a standard operating procedure for astragalus was established on the base of the GAP of the Chinese herbal medicine as well as practice investigation and experiments. This standard operating procedure provides the technical requirements for astragalus's growing, field management, controlling of the diseases and pests, harvesting, processing, packing, storing, transporting and quality monitoring.

Astragalus propinquus↗

[Protoplast culture and plant regeneration of the methionine resistant variant of Astragalus cicer L].

A protoplast-to-plant system for the methionine resistant variant of Astragalus cicer L. has been developed. The friable calli induced from stem segments of variant plants were used as materials for protoplast isolation through enzyme digestion. The effects of different media and plating densities on protoplast divisions and plant regeneration were studied. Sustained cell divisions and colony formation from the protoplasts of the methionine resistant cell line of Astragalus cicer L. were obtained by a DPD medium containing 2.0 mg/L 2,4- dichlorophenoxyacetic acid (2,4-D), 0.2 mg/L 6 -benzylaminopurine(6-BA), 0.3 mol/L mannitol, 200 mg/L casein hydrolysate and 2% (W/V) sucrose at a plating density of 2x10(5) /ml. The division frequency was 38.3%. At the same time, different dividing types of protoplasts were found. Organogenesis and shoot formation from the protoplast-derived calli were induced on MS medium supplemented with 0.5 mg/L NAA, 10 mg/L KT and 2% (W/V) sucrose. The protoplast-derived calli still expressed resistance to methionine. The protoplast to plant regeneration protocol developed in this study might provide the foundation for the resistant cell line as a parent for somatic hybridization.

Astragalus Plant↗

[Effect of 5-fluorouracil in combination with Astragalus membranaceus on amino acid metabolism in mice model of gastric carcinoma].

OBJECTIVE: To study the effect of 5-fluorouracil-FU in combination with astragalus membranaceus(AM) on amino acid metabolism in mice model of gastric carcinoma induced by 3-methylcholanthrene(MC). METHODS: Mice gastric carcinoma models were established by 3-methylcholanthrene induction and randomly divided into different groups, and received 5-FU treatment (group A) 5-FU plus AM (group B), 5-FU plus a high dose of AM(group C), no treatment (group D). Normal mice were used as control (group N). Free amino acid in the tumor specimens were examined. RESULTS: The levels of free Valine, Methionine, Leucine, Arginine and cystine in the tumor specimens in group D were significantly higher than that in group N(P< 0.05). The levels of free serine in group A, B, C, D were significantly higher than that in group N. The levels of free glutamic acid in group A, B were significantly higher than that in group N(P< 0.05). The levels of free proline in group C, D were significantly higher than that in group P, N(P< 0.05). CONCLUSIONS: The increasing levels of free serine and proline in tumor specimens in gastric cancer mice model reveals metabolic disturbance of amino acid. 5-FU plus astragalus membranaceus can decrease the level of free glutamic acid in the mice models, and inhibit tumor growth.

Amino Acids↗

[Primary research on the clinical significance of ventricular late potentials (VLPs), and the impact of mexiletine, lidocaine and Astragalus membranaceus on VLPs].

The body surface signal-averaged electrocardiogram (SAECG) was recorded on 316 cardiac patients, 84% (266 patients) of whom were also taken 24-hour Holter ECG. VLPs were detected positively in 6.1%, 25%, 25.7%, 14.3% and 5.5% of patients with angina pectoris, myocardial infarction, myocarditis, cardiomyopathy and arrhythmia of unknown origin respectively, and the patients with complex ventricular arrhythmia had more VLPs positive determination definitely (P less than 0.05). The specificity of VLPs in the prediction of complex ventricular arrhythmia was greater than 80%, and the positive predictive accuracy was 70-80%. 5, 5 and 10 patients with VLPs were treated with Mexiletini Hydrochloridum, Lidocaini Hydrochloridum and Injection of Astragalus membranaceus respectively. As a result of treatment, the transfer of VLPs positive to negative was unsuccessful, but only the intervention of Astragalus membranaceus Injection made the duration of VLPs shortened significantly (39.8 +/- 3.3 ms versus 44.5 +/- 5.9 ms, P less than 0.01).

Adult↗

[Isolation and identification of chemical constituents of Astragalus root].

Ten constituents have been isolated from the alcoholic extract of Astragalus membranaceus var. monghlicum root. All of them were identified. Among them astraisoflavanin (3S-(-)-mucronulatol-7-O-D-glucopyranoside) is a new compound. Dimethyl 4,4'-dimethoxy-5,6,5',6'-dimethylenedioxybiphyenyl 1-2, 2'-dicarboxylate is a known synthetic compound, but it was first isolated from natural resource Astragalus root and identified by the authors.

Astragalus propinquus↗

[F3, a fractionated extract of Astragalus membranaceus, potentiates lymphokine-activated killer cell cytotoxicity generated by low-dose recombinant interleukin-2].

Success with rIL-2 immunotherapy of human cancer appears to depend on the administration of high doses which are frequently associated with excessive toxicity. Future use of rIL-2 will require certain modifications based on the use of lower doses of rIL-2 without significant loss of antitumor efficacy. The authors tested in vitro the possibility of potentiating the activity of rIL-2 in terms of LAK cell generation. The authors hypothesized that co-incubation of LAK cell precursors with a Chinese herbal extract (F3) of Astragalus membranaceus (an immune modulator currently under study in the authors' laboratory), along with a low concentration of rIL-2 would generate levels of LAK cell activity equivalent to those generated by high concentrations of rIL-2 alone. The authors found: (1) a 10-fold potentiation of rIL-2 activity manifested by tumor cell killing activity of 80% resulting from LAK cell generation with F3 plus 100 u/ml of rIL-2 versus 76% generated by 1000 u/ml of rIL-2 alone; (2) a significant reduction in the number of effector LAK cells required for equicytotoxic reaction following LAK cell generation with F3 plus rIL-2 compared to rIL-2 alone. The authors conclude that potentiation of antitumor activity mediated by rIL-2 in low concentrations is possible by the concomitant use of another immune modulator such as Astragalus membranaceus.

Astragalus propinquus↗

Immunotherapy with Chinese medicinal herbs. II. Reversal of cyclophosphamide-induced immune suppression by administration of fractionated Astragalus membranaceus in vivo.

A partially purified fraction (F3) with an estimated molecular weight of 20,000 to 25,000 derived from the traditional Chinese medicinal herb Astragalus membranaceus, was found to possess a potent immunorestorative activity in vitro. Its capacity to aborogate the local xenogeneic graft versus host reaction (XGVHR) following injection in vivo was further studied in a newly developed animal model designed for preclinical evaluation of various biological response modifiers. F3 was injected intravenously into cyclophosphamide-primed rats at varied concentrations and schedules prior to grafting of mononuclear cells from healthy normal donors. Maximal abrogation of the local XGVHR mounted by the mononuclear cells, was observed following injection of 5.55 mg of F3 daily for eight days. This abrogation of XGVHR indicates a reversal of the immunosuppressive effect of cyclophosphamide as manifested by a significant decline in the local XGVHR volume from 99.42 +/- 9.2 mm3 (positive control) to 39.78 +/- 8.3 mm3 (p less than 0.001). This reversal of cyclophosphamide-induced immunosuppression by the administration of F3 was complete, since the volume of the abrogated local XGVHR (39.78 +/- 8.3 mm3) was comparable to 34.79 +/- 5.69 mm3 (p greater than 0.1) in the negative control group (no cyclophosphamide-priming; saline injection only). These data indicate that F3 administration markedly enhances the rats' ability to reject the xenogeneic graft and therefore possesses a strong immune potentiating activity in vivo. These preclinical data also provide the rational basis for the use of extracts of Astragalus membranaceus in phase I clinical trials among patients suffering from iatrogenic or inherent immune deficiency states.

Adjuvants, Immunologic↗

Fractionated extract of Astragalus membranaceus, a Chinese medicinal herb, potentiates LAK cell cytotoxicity generated by a low dose of recombinant interleukin-2.

Success with rIL-2 immunotherapy of human cancer appears to depend on the administration of high doses which are frequently associated with excessive toxicity. Future use of rIL-2 will require certain modifications based on the use of lower doses of rIL-2 without significant loss of antitumor efficacy. We tested in vitro the possibility of potentiating the activity of rIL-2 in terms of LAK cell generation. We hypothesized that co-incubation of LAK cell precursors with a Chinese herbal extract (F3) of Astragalus membranaceus, (an immune modulator currently under study in our laboratory), along with a low concentration of rIL-2, would generate levels of LAK cell activity equivalent to those generated by high concentrations of rIL-2 alone. We found (1) a 10-fold potentiation of rIL-2 activity manifested by tumor cell-killing activity of 80% resulting from LAK cell generation with F3 plus 100 u/ml of rIL-2 versus 76% generated by 1,000 u/ml of rIL-2 alone; (2) a significant reduction in the number of effector LAK cells required for equicytotoxic reaction following LAK cell generation with F3 plus rIL-2 compared to rIL-2 alone. We conclude that potentiation of antitumor activity mediated by rIL-2 in low concentrations is possible by the concomitant use of another immune modulator such as Astragalus membranaceus.

Adjuvants, Immunologic↗

[Intervention of lidocaine and Astragalus membranaceus on ventricular late potentials].

16 and 22 patients with positive ventricular late potentials (LP) recorded on signal-averaged electrocardiograms (SAECG) were treated with lidocaine 100 mg iv. or Astragalus membranaceus 24 g iv. drip for 2 weeks respectively. As a result, the SAECGs of 2 (12.5%) and 3 (13.6%) of them normalized respectively. Compared with baseline, there were no significant changes in average HFQRSD, LAS and RMS 40 after treatment of lidocaine. HFQRSD and LAS were shortened significantly 115.9 +/- 29.9 vs 125.1 +/- 29.4 ms (P < 0.001); and 44.8 +/- 15.4 vs 52.8 +/- 15.4 ms (P < 0.001), and RMS 40 was enlarged 20.0 +/- 18.6 vs 12.8 +/- 19.0 microV (P < 0.05) only after treatment of Astragalus membranaceus. It is suggested that the duration of LP was shortened.

Adult↗

Liquid chromatography-electrospray ionization mass spectrometry study of the flavonoids of the roots of Astragalus mongholicus and A. membranaceus.

High-performance liquid chromatography-electrospray ionization mass spectrometry has been applied to analyze the flavonoids of Huangqi, the roots of Astragalus mongholicus and A. membranaceus. Eight flavonoids were identified as calycosin-7-O-beta-D-glucoside, calycosin-7-O-beta-D-glucoside-6"-O-malonate (2), ononin, (6aR,11aR)-3-hydroxy-9,10-dimethoxypterocarpan-3-O-bet a-D-glucoside, calycosin, (3R)-7,2'-dihydroxy-3',4'-dimethoxyisoflavan-7-O-beta-D-glucoside, formononetin-7-O-beta-D-glucoside-6"-O-malonate and formononetin by direct comparison with the isolated standards from Huangqi. The existence of (6aR,11aR)-3-hydroxy-9,10-dimethoxypterocarpan, (3R)-7,2'-dihydroxy-3',4'-dimethoxyisoflavan, astrapterocarpanglucoside-6'-O-malonate and astraisoflavanglucoside-6'-O-malonate was detected. This is the first report of flavonoid glycoside malonates in these two Astragalus species, and malonate 2 is a structurally completely identified new compound.

Chromatography, Liquid↗

Phylogenetic position of Mesorhizobium huakuii subsp. rengei, a symbiont of Astragalus sinicus cv. Japan.

The phylogenetic position of Rhizobium huakuii bv. renge, a symbiont of Astragalus sinicus cv. Japan (renge-sou), was studied. The following phylogenetic approaches were used: restriction fragment length polymorphism (RFLP)-polymerase chain reaction (PCR) analysis of a full-length 16S rDNA fragment, 16S rDNA analysis of the first 300-bp sequence, bacteriophage typing, and amplification of the genomic region by random primer. All the data suggest that strains of R. huakuii bv. renge should be classified into subspecies of the new genus Mesorhizobium (Jarvis et al., Inter. J. System. Bacteriol., 47, 895-898, 1997) and renamed M. huakuii subsp. rengei. All the strains fell into a tight cluster which included M. loti and M. huakuii. The strains isolated from root nodules on A. sinicus were divided into three groups as follows: group I, M. huakuii subsp. rengei B3, M. huakuii subsp. rengei My6, M. huakuii subsp. rengei My7, M. huakuii subsp. rengei My3, and M. huakuii subsp. rengei OUT30020; group II, M. huakuii subsp. huakuii CCBAU103(T), M. huakuii subsp. huakuii ACCC13005, M. huakuii subsp. huakuii 7653R, and Mesorhizobium sp. N-1; group III, Mesorhizobium sp. OUT30019. All the strains isolated in Japan except strains N-1 and OUT30019 were classified into group I. Strains in group I were sensitive to bacteriophage H1 which was isolated from rice-paddy soil in Japan. Strains in groups II and III except for M. huakuii subsp. huakuii 7653R were resistant to phage H1. Rhizobium sp. ACMP18, a native symbiont of Astragalus cicer that forms nodules on A. sinicus, showed close similarity to M. huakuii subsp. huakuii CCBAU103(T), and should thus be classified as a Mesorhizobium sp. Taken together, the results of the analyses indicate that M. huakuii subsp. rengei forms a subgroup which is distinct from M. huakuii subsp. huakuii strains isolated in China and that strain B3 is the type strain.

Journal Article↗

Selenium absorption by excised Astragalus roots.

Absorption of selenate and selenite by excised roots of Astragalus Crotalariae, a selenium accumulator, and of A. lentiginosus, a non-accumulator, was favored by CaCl(2) and a pH of 4.0. The uptake of selenate and possibly selenite, is metabolically linked. Roots of a number of Astragalus species were examined, and in all cases selenate entered the roots much faster than selenite. In these short-term experiments there was no relation between uptake of the 2 ions and classification of a species as selenium-accumulator or non-accumulator.

Absorption↗

Exclusion of selenium from proteins of selenium-tolerant astragalus species.

Protein fractions from three selenium-tolerant and three selenium-sensitive Astragalus species, grown in the presence of [(75)Se]selenate, were analyzed for their selenium content. Though tolerant species are known to accumulate considerably more selenium than do sensitive plants, protein fractions from the three selenium accumulators were found to contain significantly less selenium (0.46 to 0.57 picomoles selenium per milligram protein) than did protein fractions from the three nonaccumulators (4.17 to 5.02 picomoles selenium per milligram protein). Under similar conditions, seedlings of Vigna radiata (L.) Wilczek had taken up selenium (6.31 picomoles selenium per milligram protein) at levels comparable to those observed in the proteins of the nonaccumulator Astragali. These results establish that the ability to tolerate and to circumvent the toxic effects of selenium, characteristic of the accumulator species of Astragalus, is associated with a reduced incorporation of this element into protein.

Journal Article↗

An analysis of genetic variation in Astragalus cremnophylax var. cremnophylax, a critically endangered plant, using AFLP markers.

A recently developed molecular technique (amplified fragment length polymorphisms, AFLP) was used for characterizing genetic heterogeneity within and among populations of a critically endangered species of plant, Astragalus cremnophylax var. cremnophylax. Using AFLP, up to 50 polymorphic genetic markers per AFLP-PCR amplification were generated, and a total of 220 variable markers overall. This information was used first to assess genetic diversity within each of the three known populations of Astragalus cremnophylax var. cremnophylax from Grand Canyon National Park in Arizona, USA: North Rim (NR; n = 970), South Rim Site 1 (SR1; n = 500), and South Rim Site 2 (SR2; n = 2). Diversity in the form of average heterozygosity [symbol: see text] H [symbol: see text] and the proportion of polymorphic genes [symbol: see text] P [symbol: see text] was greatest in the NR population ([symbol: see text] H [symbol: see text] = 0.13 and [symbol: see text] P [symbol: see text] = 0.38) and least in the SR2 population ([symbol: see text] H [symbol: see text] = 0.02 and [symbol: see text] P [symbol: see text] = 0.04). Diversity was also quite low for the SR1 population ([symbol: see text] H [symbol: see text] = 0.04 and [symbol: see text] P [symbol: see text] = 0.10). In addition, substantial genetic differentiation among populations was indicated by both phenetic (AMOVA) and genetic analyses (overall corrected FST = 0.41). This finding was corroborated by the results of several multivariate analyses which utilized the genetic data, including a UPGMA cluster analysis and a principal coordinate analysis which revealed the existence of discrete groups corresponding to the populations. Population structure was further revealed within the NR population which was known to consist of four spatially separated groups of plants. Several recommendations for the future management of the species are discussed.

Genetic Markers↗

Genetic diversity of rhizobial symbionts isolated from legume species within the genera Astragalus, Oxytropis, and Onobrychis.

The genetic diversity of 44 rhizobial isolates from Astragalus, Oxytropis, and Onobrychis spp. originating from different geographic locations was evaluated by mapped restriction site polymorphism (MRSP) analysis of 16S rRNA genes and by PCR DNA fingerprinting with repetitive sequences (REP-PCR). A comparison of tree topologies of reference strains constructed with data obtained by MRSP and by 16S rRNA gene sequence analyses showed that the topologies were in good agreement, indicating that the MSRP approach results in reasonable estimates of rhizobial phylogeny. The isolates were distributed into 14 distinct 16S rRNA gene types clustering into three major groups which corresponded with three of the genera within the legume symbionts. Most of the isolates were within the genus Mesorhizobium. Five were identified with different genomic species nodulating Lotus spp. and Cicer arietinum. Three Astragalus isolates were classified as Bradyrhizobium, one being similar to Bradyrhizobium elkanii and another being similar to Bradyrhizobium japonicum. Six of the isolates were related to species within the genus Rhizobium. Two were similar to Rhizobium leguminosarum, and the remainder were identified as Rhizobium gallicum. DNA fingerprinting by REP-PCR revealed a high level of diversity within single 16S ribosomal DNA types. The 44 isolates were distributed into 34 REP groups. Rhizobial classification at the genus and probably also the species levels was independent of geographic origin and host plant affinity.

DNA Fingerprinting↗