Hedysarum Polybotrys Research Articles

Immunomodulatory Effects of Hedysarum polybotrys Extract in Mice Macrophages, Splenocytes and Leucopenia

Astragali Radix (Huang-Qi) is a popular herbal medicine commonly used as a constituent in tonic herbal preparations. Hedysarum polybotrys Handel-Mazzetti is one species used of Astragali Radix. In this study, the immunomodulatory properties of H. polybotrys were explored by LPS-activated and SNP-treated RAW 264.7 cells and splenocytes and, daunoblastina-induced leucopenia BALB/c mice. Formononetin was used as the bioactive marker to monitor the quality of the H. polybotrys extracts. H. polybotrys was extracted with hot-water and methanol, and MeOH extract partitioned with H2O (M-H) and ethyl acetate (M-EA) to yield four different fractions. M-EA had the highest formononetin and total proanthocyanidin content and showed stronger inhibitory effects on the production and expression of NO, PGE2, iNOS and COX-2 in LPS-activated RAW 264.7 cells and splenocytes than the other fractions. In addition, M-EA significantly stimulated the proliferation of LPS-activated RAW 264.7 cells and splenocytes, enhanced NO radicals scavenging and attenuated NO-induced cytotoxicity. Furthermore, M-EA also significantly increased the rate of recovery of white blood cells level in daunoblastina-induced leucopenia mice. These evidences suggest that this traditional Qi-tonifying herb has potential effects in clinical conditions when immune-enhancing and anti-inflammatory effect is desired.

Characterization and Antitumor Activity of a Glucan Structure-Activity Relationship Analysis

A water-soluble polysaccharide named as HPS-1 was isolated from the roots of Hedysarum polybotrys Hand.-Mazz by hot water extraction, anion-exchange and gel-permeation chromatography and tested for its antitumor activity. Its structural characteristics were investigated by FTIR, HPLC, NMR spectroscopy, GLCMS, methylation analysis, Periodate oxidation and Smith degradation. Based on the data obtained, HPS-1 was found to be an α- (14)-D-glucan, with a single α-D-glucose at the C-6 position every nine residue, on average, along the main chain. The glucan has a weight-average molecular weight of about 9.4×104 Da. MTT assay revealed that HPS-1 significantly inhibited the proliferation of Human hepatocellular carcinoma HEP-G2 cells and human gastric cancer MGC-803 cells in vitro, indicating HPS-1 could have a possible cancer therapeutic potential.

Phytochemicals and biological studies of plants in genus Hedysarum.

In China, several species (Hedysarum polybotrys Hand.-Mazz., Hedysarum limprichtii Hlbr., Hedysarum vicioider Turcz. var. Taipeicum Hand.-Mazz. Liu, Hedysarum smithianum, et al.) of genus Hedysarum have a long history of use in traditional Chinese medicine (TCM). In TCM, these plants are used to increase the energy of the body. To date, 155 compounds, including flavonoids, triterpenes, coumarins, lignanoids, nitrogen compounds, sterols, carbohydrates, fatty compounds, and benzofuran, have been isolated from plants of the genus Hedysarum. Various chemical constituents contribute to the antioxidant, anti-tumor, anti-aging, anti-diabetic, and anti-hypertensive properties of these plants. Hedysarum species are used to treat infestation with gastrointestinal nematodes and may support the immune system and peripheral nervous system. In the present review, we summarize the research on the phytochemistry and pharmacology of Hedysarum species, which will be useful for better utilization of these important species in TCM.

Molecular identification of Astragali Radix and its adulterants by ITS sequences

To explore a new method for identification Astragali Radix from its adulterants by using ITS sequence. Thirteen samples of the different Astragali Radix materials and 6 samples of the adulterants of the roots of Hedysarum polybotrys, Medicago sativa and Althaea rosea were collected. ITS sequence was amplified by PCR and sequenced unidirectionally. The interspecific K-2-P distances of Astragali Radix and its adulterants were calculated, and NJ tree and UPGMA tree were constructed by MEGA 4. ITS sequences were obtained from 19 samples respectively, there were Astragali Radix 646-650 bp, H. polybotrys 664 bp, Medicago sativa 659 bp, Althaea rosea 728 bp, which were registered in the GenBank. Phylogeny trees reconstruction using NJ and UPGMA analysis based on ITS nucleotide sequences can effectively distinguish Astragali Radix from adulterants. ITS sequence can be used to identify Astragali Radix from its adulterants successfully and is an efficient molecular marker for authentication of Astragali Radix and its adulterants.

Effects of different salt-alkaline stress on seed germination and physiological characteristics of Hedysarum polybotrys

In order to get the method for improving the salt resistance of Hedysarum polybotrys seeds and seedlings under different salt-alkaline stress, the seed germination and physiological characteristics of H. polybotrys seedlings were studied. Several physiological indexes of H. polybotrys seeds under different salt-alkaline stress, such as the germination vigor, germination rate, relative germination rate, relative salt damage rate were measured. And others indexes of the seedlings like chlorophyll contents, soluble protein contents, the permeability of plasmalemma, the activities of POD and SOD were also measured. Different salt-alkaline stress decreased the germination rate, vigor of germinate, germination index, while relative salt damage rate increased. With the increased salt-alkaline concentration, the adverse effects became more obvious. The strength of the salts: Na2CO3 > Na2SO4 > NaCl. With the increase of the salt-alkaline concentration, the chlorophyll contents and the soluble protein contents decreased, but the permeability of plasmalemma increased. The change trend of SOD and POD activity was similar, it is increased firstly, and then decreased as the stress intensity extended, the most significant increase of Na2SO4 and Na2CO3 in the concentration of salt-alkaline was 25 mmol x L(-1), but NaCl was 50 mmol x L(-1). The seeds and seedlings inhibition of the salts was Na2CO3 > Na2SO4 > NaCl.

Investigation of Two Species of Huang-qi (Aatragalusmembranaceus and Hedysarum polybotrys) by HPLC, ITS,Microscopic Morphology and Antioxidant Activities

Hedysarum polybotrys is commonly substituted for Astragalus membranaceus in Taiwan for many years. This study described several different ways, including microscopic morphology, ITS (internal transcribed spacer) sequencing and HPLC fingerprint methods to identify these two species. Furthermore, the antioxidant activities of the two plants were evaluated and H. polybotrys had higher antioxidant activity than A. membranaceus. All the methods gave consistent conclusions that test samples KRA2, 3 and 8 belonged to A. membranaceus, while KRA1, 4, 5, 6 and 7 belonged to H. polybotrys. The results revealed that all the above methods may successfully discriminate between A. membranaceus and H. polybotrys.

Protection of ultra-filtration extract from Danggui Buxue Decoction (当归补血汤) on oxidative damage in cardiomyocytes of neonatal rats and its mechanism

To investigate whether the administration of the ultra-filtration extract from Danggui Buxue Decoction (EDBD) was able to protect cardiomyocytes from oxidative injury of rats induced by hydrogen peroxide (H(2)O(2)) and its potential mechanism. Myocardial cells from 1- to 2-day-old neonatal rats were cultured in Dulbecco's modified Eagle's medium low-glucose and Ham's F12 medium (1:1), and the cellular injury was induced by H(2)O(2). The ultra-filtration extract mixture from Angelica sinensis and Hedysarum polybotrys was given in three doses of 3.75, 7.5, and 15 mg/mL. Morphological changes of cardiomyocytes were observed by microscope. Survival rate of myocardial cells was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The cardiomyocyte damages were estimated by detecting lactate dehydrogenase (LDH) and creatine kinase (CK) releases in the medium, superoxide dismutase (SOD) activities, and intracellular malondialdehyde (MDA) and myeloperoxidase (MPO) contents. The levels of caspase-3 and heat shock protein 70 (hsp70) mRNA expression in cardiomyocytes were measured by reverse transcription polymerase chain reaction. The EDBD could protect the cardiomyocytes from H(2)O(2) injury in a dosedependent manner (3.75, 7.50, and 15.00 mg/mL). The EDBD could significantly decrease LDH and CK leakages and intracellular MDA and MPO contents, increase SOD activity, up-regulate hsp70 expression, and down-regulate caspase-3 expression. The EDBD has protection on cardiomyocytes injured by H(2)O(2) through improving cell antioxidant ability, up-regulating hsp70 expression, and inhibiting caspase-3 activity.

Chemical constituents from the roots of Hedysarum polybotrys

From the roots of Hedysarum polybotrys, ten compounds were isolated and identified as ferulic hexadecyl ester(1), ferulic(2), formononetin(3), formononetin-7-O-beta-D-glucoside(4), calycosine(5), calycosine-7-O-beta-D-glucoside(6), isoliquiritigenin(7), 7,4'-di-hydroxylflavone(8), liquiritigenin(9) and uracil(10). Compounds 2, 8, 9 and 10 were isolated from the genus Hedysarum for the first time, while compounds 6 and 7 were isolated from H. polybotrys for the first time.

Anti-tumor and immunoregulation activities of hedysari polysaccharide

Hedysari polysaccharide (HPS) is the main component of hedysarum polybotrys. HPS can inhibit tumor cell proliferation, block the cell cycle in the G2-M period, modulate apoptosis via the mitochon drial pathway, and improve the red blood cell innate immunity and T lymphocyte immune function. HPS can relieve low immunity caused by chemotherapeutics. HPS plays anti-tumor effects by these ways mentioned above. Key words: Neoplasms; Hedysari polysaccharide; Immunomodulatory

Structural characterization of a sulfated glucan isolated from the aqueous extract of Hedysarum polybotrys Hand.-Mazz.

A water-soluble sulfated glucan, SHG, was isolated from Hedysarum polybotrys Hand.-Mazz using anion-exchange and gel-permeation chromatography. Elemental analysis indicated that SHG was a sulfated polysaccharide containing small amount of sulfate groups (1.47mass%). The molecular weight was estimated to be 1.72×105Da by high-performance gel permeation chromatography (HPGPC) and size exclusion chromatography-multi angle laser light scattering (SEC-MALLS). SHG took random coil compact conformation in 0.1M NaNO3. Compositional analysis revealed that SHG was composed of glucose only. On the base of partial acid hydrolysis, methylation analysis, gas chromatography-mass spectrometry (GC-MS), Fourier transform infrared (FT-IR) spectra, nuclear magnetic resonance (NMR) spectroscopy of 1H, 13C, β-correlation spectroscopy (β-COSY), total correlation spectroscopy (TOCSY), heteronuclear single quantum coherence (HSQC), and heteronuclear multiple bond correlation (HMBC), the results showed that SHG was composed of α-d-(1→4)-linked glucopyranosyl residues, with branches at C-6 consisting of non-reducing terminal approximately every eight residues. Sulfate groups may attach to the backbone at O-6, occasionally occurring per thirty-eight anhydrous glucose units.

Antidiabetic properties of purified polysaccharide fromHedysarum polybotrys

Hedysarum polybotrys polysaccharide (HPS) is the principal active fraction responsible for the antidiabetic properties of this species. The aim of this study was to determine the antidiabetic properties of 4 purified fractions of different molecular weight range HPSs (HPS1, HPS2, HPS3, HPS4). HPS3 was selected for examination of its hypoglycemic mechanism because of its significant hypoglycemic effect in alloxan-induced diabetic mice. The changes in blood glucose levels and oral glucose tolerance tests (OGTT) showed that hypoglycemia was more pronounced in HPS3-treated groups than in the diabetes mellitus model (DM) control group. The interleukin-6, tumor necrosis factor-alpha, leptin, and free fatty acid levels were significantly lower in the HPS3-treated groups and HPS3 + metformin (HPS3+MET) group than in the DM control group, while plasma insulin, hepatic glycogen, superoxide dismutase, and nitric oxide synthetase activity were significantly higher. Treatment with HPS3 or HPS3+MET also significantly lowered malonaldehyde levels compared with the DM control group, while it elevated the nitric oxide and total antioxidant capacity. HPS3 altered the plasma lipid levels by lowering cholesterol and triglyceride concentrations, while elevating the plasma high-density lipoprotein cholesterol level. Therefore, these results suggest that HPS3 may partly ameliorate hyperglycemia and hyperlipidemia associated with type 2 diabetes through increased insulin secretion, inhibition of lipid peroxidation, promotion of sensitivity to insulin, suppression of gluconeogenesis and reduction in the biosynthesis fatty acid, cholesterol and cell cytokines related to insulin resistance, and it could be a useful adjunct therapy to a proven first-line therapy for type 2 diabetes using metformin.

A New Phenylpropionate Derivative from the Rhizomes of Hedysarum polybotrys

Aim To study the chemical constituents from the rhizome of Hedysarum polybotrys. Methods Chemical constituents were isolated by chromatographic methods and the structures were elucidated by NMR spectra and physicochemical properties. Results A new phenylpropionate derivative was isolated from the rhizome of Hedysarum polybotrys. Its structure was established as ( E)-3-(4-hydroxy-2-methoxyphenyl)-propenoic acid 4-hydroxy-3-methoxyphenyl ester. Conclusion The compound is a new phenylpropionate derivative.

Characterization and anti-tumor activity of a polysaccharide from Hedysarum polybotrys Hand.-Mazz

A water-soluble polysaccharide named as HPS-1 was isolated from the roots of Hedysarum polybotrys Hand.-Mazz by hot water extraction, anion-exchange and gel-permeation chromatography and tested for its anti-tumor activity. Its structural characteristics were investigated by FTIR, HPLC, NMR spectroscopy, GLC–MS, methylation analysis, periodate oxidation and Smith degradation. Based on the data obtained, HPS-1 was found to be an α-(1 → 4)- d-glucan, with a single α- d-glucose at the C-6 position every nine residue, on average, along the main chain. The glucan has a weight-average molecular weight of about 9.4 × 10 4 Da. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay revealed that HPS-1 significantly inhibited the proliferation of human hepatocellular carcinoma HEP-G2 cells and human gastric cancer MGC-803 cells in vitro, indicating HPS-1 could have a possible cancer therapeutic potential.

A novel compound from Hedysarum polybotrys

A polyhydroxyl constituent (1), named as polybotrin, along with two known compounds, were isolated from the roots of Hedysarum polybotrys. Their structures were identified based on chemical and spectroscopic evidence.

Saponins from the roots of Hedysarum polybotrys

Saponins from the roots of Hedysarum polybotrys

Structural determination of saponins from Hedysarum polybotrys

Four saponins (1-4) were isolated from the roots of Hedysarum polybotrys Hand.-Mazz. and obtained from Hedysarum (Tourn.) L for the first time. Their structures were identified on the basis of spectroscopic data. MS, 1D and 2D NMR including HMQC, HMBC, 1H-1H COSY and NOESY permitted correct assignments of all 1H and 13C signals of 1 and 2, and the 13C NMR data of 2 were reported for the first time.

The effect of medicinal plants used in Chinese folk medicine on RANTES secretion by virus-infected human epithelial cells

The accumulation of inflammatory cells in the infective sites has been reported to play a crucial role in the progression of chronic inflammation and multiple sclerosis after viral infection. In the present study, nine ethanol extracts of Forsythia suspensa Vahl. (Oleaceae), Lonicera japonica Thunb. (Caprifoliaceae), Isatis indigotica Fort. (Cruciferae), Strobilanthes cusia (Ness.) O. Kuntze (Acanthaceae), Astragalus membranaceus (Fisch.) Bge. (Leguminosae), Hedysarum polybotrys Hand.-Mazz. (Leguminosae), Andrographis paniculata (Burm. f.) Ness. (Acanthaceae), Glycyrrhiza uralensis Fischer. (Leguminosae) and Ligusticum wallichii Franch. (Umbelliferae), medicinal plants traditionally used in China for treating conditions likely to be associated with inflammation and viral infection, were screened for their effect on RANTES secretion by influenza A virus (H1N1)-infected human bronchial epithelial cells (A549). With exception of Lonicera japonica, Isatis indigotica, Astragalus membranaceus and Hedysarum polybotrys, all plants tested at concentration of 200 μg/ml possessed more than 50% suppressing effect on RANTES secretion by H1N1-infected A549 bronchial epithelial cells. Among the plants tested, Andrographis paniculata showed the most promising property to inhibit RANTES secretion with an IC 50 of 1.2 ± 0.4 μg/ml while the next two were Glycyrrhiza uralensis and Forsythia suspensa (IC 50 ranging from 35 to 48 μg/ml).

Species identification of Radix Astragali (Huangqi) by DNA sequence of its 5S-rRNA spacer domain

About 300 species and varieties of Astragalus are identified in China, making the identification of the origin of a particular Astragalus species on the consumer market difficult. A molecular genetic approach was developed to identify various species of Astragalus. Although the 5S-rRNA coding sequence is conserved in higher eukaryotes, the spacer domain of the 5S-rRNA gene has great diversity among different species. The 5S-rRNA spacer domain was amplified by polymerase chain reaction (PCR) from the isolated genomic DNA, and the PCR products (∼300 bp) covering the 5S-rRNA spacer domain were sequenced. The nucleotide sequences of Astragalus membranaceus, A. membranaceus var. mongholicus, A. lehmannianus, A. hoantchy, and of one closely related species Hedysarum polybotrys (Hongqi), were determined. Diversity in DNA sequence and restriction enzyme mapping among various species was found in their 5S-rRNA spacer domains. This is the first report on the detection of 5S-rRNA spacer region sequence of Astragalus, and the results could be used for genetic identification of Huangqi.

RAPD Analysis of Astragalus Medicines Marketed in Taiwan

The genetic variability of Astragalus medicine materials sold by twenty randomly selected stores in Taiwan was investigated using RAPD analysis in order to obtain available primers which could clearly differentiate among them. Total DNA isolated from the rhizomes of the samples were used as templates, and sixty 10 mer arbitrary primers were used in the analysis. The aim of the present study is to construct an identification model of molecular biotechniques applicable to Chinese herbal medicines in RAPD analysis. Three of the primers, OPT-03, OPT-13, and OPT-17, revealed polymorphic RAPD fingerprints among the samples of Astragalus membranaceus, and between Astragalus membranaceus and Hedysarum polybotrys samples. SSCP analysis was also conducted on PCR products from the ITS-1 region of ribosomal DNA in order to differentiate the two species.

Studies on the constituents of Hedysarum polybotrys Hand.-Mazz.

Thirteen phenolic compounds were isolated from Hedysarum polybotrys HAND.-MAZZ., and their structures were determined by spectroscopic studies. Among the thirteen, 5-hydroxy-2-(2-hydroxy-4-methoxyphenyl)-6-methoxybenzofuran (X) is a new compound, and 6-hydroxy-2-(2-hydroxy-4-methoxyphenyl)-benzofuran (VIII) was isolated from this plant for the first time.