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[Determination of six metal elements in plant Bupleurum scorzoneri folium Willd. by flame atomic absorption spectrophotometry (FAAS)].

The objective in this paper was to determine the amount of metal elements in the Plant Bupleurum scorzoneri folium Willd. by flame atomic absorption spectrophotometry (FAAS). FAAS method was established for the determination of Ca, Mg, Fe, Cu, Zn and Pb in the Plant Bupleurum scorzoneri folium Willd. after digestion with HNO3-HClO4 (V : V= 4 : 1) at 90-95 degrees C and at normal pressure. In the meantime, the optimum parameters of FAAS and the effects of solution medium on the results were studied. The analytical results of Ca, Mg, Fe, Cu and Zn were 5,588.9, 1,790.5, 869.3, 78.4, 44.3 microg x g(-1) respectively, and Pb was not checked out in the samples. The results showed that the recovery of standard addition was 99.57%-102.10%, the relative standard deviation (n= 9) was 0.18%-2.26%. The method is quick, simple and convenient, and the results are satisfactory.

Bupleurum↗

Structure and actions of saikosaponins isolated from Bupleurum falcatum L. I. Anti-inflammatory action of saikosaponins.

Anti-inflammatory action of saikosaponins isolated from the root of Bupleurum falcatum L were examined using female albino rats. The anti-exudative action by granuloma pouch method and the antigranulomatous action by cotton pellet method were demonstrated with i.m. and oral administrations of saikosaponins. The oral administration of saikosaponins in 10 times the dosage of i.m. injection showed almost the same effect. Among saikosaponins isolated from Bupleurum falcatum, saikosaponins a and d, not c, were demonstrated to have anti-inflammatory action. The relationship between structure and action of saikosaponins was discussed. No changes in body weight, adrenal weight, plasma-11-OH-corticosteroid level and hematocrit value were observed.

11-Hydroxycorticosteroids↗

[Chemical constituents of the aerial parts of six species of Bupleurum genus medicinally used in south-west region of China].

The chemical constituents of the aerial parts of six species of Bupleurum genus (B. kunmingense, B. polyclonum, B. wenchuanense, B. longicaule var. franchetii, B. rockii and B. chaishoui) medicinally used in south-west region of China have been investigated with the results of obtaining eight flavonoid compounds and six triterpenoid saponins. The aglycones of these eight flavonoid compounds are quercetin and isorhamnetin, which are different from the aglycones of the flavonoid compounds isolated from the aerial parts of B. chinense, the official species in Chinese Pharmacopoeia. In the latter case, the aglycone is kaempferol. This difference may be of chemotaxonomic value in Bupleurum genus.

Antiviral Agents↗

[Chemical constituents of the roots of Bupleurum longicaule Wall. ex DC. var. franchetii de Boiss and B. chaishoui Shan et Sheh].

From the roots of Bupleurum longicaule var. franchetii, B. chaishoui and the official species B. chinense, ten saikosaponins have been isolated and identified as saikosaponin a, saikosaponin d, saikosaponin c, saikosaponin e, saikosaponin b2, 2"-O-acetylsaikosaponin a, 3"-O-acetylsaikosaponin a, 3"-O-acetylsaikosaponin d, 6"-O-acetylsaikosaponin a and 6"-O-acetylsaikosaponin d respectively. It has been shown that the chemical constituents of these three Bupleurum plants are very similar, therefore, the medicinal value of B. longicaule var. franchetii and B. chaishoui is further verified.

Apiaceae↗

Effect of saikosaponin-A, a triterpenoid glycoside, isolated from Bupleurum falcatum on experimental allergic asthma.

The separation and isolation of an extract of Bupleurum falcatum were performed based on antiallergic activities in preliminary studies with higher plants. The final active compound was identified as saikosaponin-A (SSA), a triterpenoid glycoside. SSA at more than 1 mg/kg (i.v.) significantly inhibited the passive cutaneous anaphylaxis reaction in rats in a dose-dependent manner, attaining a maximum inhibition of approximately 60% with 10 mg/kg. SSA at 3 and 10 mg/kg also suppressed asthmatic bronchoconstriction in sensitized guinea-pigs. SSA possesses a weak inhibitory activity on histamine-induced tracheal contraction in guinea-pigs and on histamine release induced by A-23187 in rat mast cells. These results indicate that SSA has an inhibitory activity against allergic asthma. This activity seems to derive from both antagonism of the histamine action and inhibition of allergic mediators. Additional mechanisms may also be involved.

Animals↗

Effect of the antiulcer polysaccharide fraction from Bupleurum falcatum L. on the healing of gastric ulcer induced by acetic acid in rats.

An antiulcer polysaccharide fraction (BR-2) from Bupleurum falcatum L. was examined for its effect on the healing of chronic ulcers induced by acetic acid in rats. When BR-2 was administered orally to the rats, it was shown to be effective in the healing of acetic acid-induced chronic ulcer. This result suggests that the use of herbal prescriptions containing B. falcatum L. may prove useful for the treatment of peptic ulcers.

Acetic Acid↗

Antiangiogenic activity of Bupleurum longiradiatum on human umbilical venous endothelial cells.

The ethyl acetate fraction of Bupleurum longiradiatum was found to have an inhibitory effect on the tube-like formation of human umbilical venous endothelial (HUVE) cells. The active compounds, isolated from the fraction, were identified as acetylbupleurotoxin (P1) and bupleurotoxin (P2). The compounds P1 and P2 completely inhibited the tube-like formation of HUVE cells at 30 microg/ml, below the cytotoxic concentration. But, they did not exhibit antitumor activity on BDF1 mice bearing Lewis lung carcinoma cells despite their antiangiogenic activity.

Angiogenesis Inhibitors↗

Determination of saikosaponins a, c, and d in Bupleurum Chinese DC from different areas by capillary zone electrophoresis.

A fast capillary zone electrophoresis (CZE) method was developed for the determination and separation of saikosaponins a, c, and d in Chinese herbal extracts of Bupleurum Chinese DC from different areas. Detection at 214 nm with a system containing sodium borate buffer and mono-3-phenylcarbamoyl-beta-CD was found to be the most suitable approach for this analysis. Saikosaponins a, c, and d could be easily determined within 8 min. The effect of the concentration of mono-3-phenylcarbamoyl-beta-CD, the concentration of the running buffer and buffer pH value on the migration behavior of the saikosaponins is discussed.

Borates↗

Genetic characterization of asymmetric somatic hybrids between Bupleurum scorzonerifolium Willd and Triticum aestivum L.: potential application to the study of the wheat genome.

In this paper, we describe how Bupleurum scorzonerifolium/Triticum aestivum asymmetric somatic hybrids can be exploited to study the wheat genome. Protoplasts of B. scorzonerifolium Willd were irradiated with ultraviolet light (UV) and fused with protoplasts of common wheat (T. aestivum L.). All cell clones were similar in appearance to those of B. scorzonerifolium, while the regenerated plantlets were either intermediate or B. scorzonerifolium-like. Genotypic screening using isozymes showed that 39.3% of cell clones formed were hybrid. Some of the hybrid cell clones grew vigorously, and differentiated green leaves, shoots or plantlets. DNA marker analysis of the hybrids demonstrated that wheat DNA was integrated into the nuclear genomes of B. scorzonerifolium and in situ karyotyping cells revealed that a few wheat chromosome fragments had been introgressed into B. scorzonerifolium. The average wheat SSR retention frequency of the RH panel was 20.50%, but was only 6.67% in fusions with a non-irradiated donor. B. scorzonerifolium chromosomes and wheat SSR fragments in most asymmetric hybrid cell lines remained stable over a period of 2.5-3.5 years. We suggest the UV-induced asymmetric somatic hybrids between B. scorzonerifolium Willd and T. aestivum L. have the potential for use in the construction of an RH map of the wheat genome.

Bupleurum↗

Partial structure of an anti-ulcer pectic polysaccharide from the roots of Bupleurum falcatum L.

Methylation analysis of a pectic polysaccharide (Bupleuran 2IIc) with anti-ulcer activity, isolated from the roots of Bupleurum falcatum L., revealed (1----4)-linked alpha-GalA together with small proportions of 2,4- and 3,4-linked GalA, and variously linked neutral sugars. Digestion of Bupleuran 2IIc with endo-alpha-(1----4)-polygalacturonase gave mainly galacturono-oligosaccharides (PG-4) and small proportions of enzyme-resistant regions (PG-1-3). PG-1 contained the sequence----4)GalA- (1----2)-Rha-(1----4)-GalA-(1----, and partial acid hydrolysis gave GalA-(1----4)-Rha, GlcA-(1----4)-Rha, and several di- and oligosaccharides consisting variously of Xyl, Glc, Gal, and Man. PG-2 and PG-3 each contained Rha, Fuc, Ara, Xyl, Man, Gal, Glc, GalA, GlcA, 2-Me-Fuc, 2-Me-Xyl, apiose (Api), aceric acid (AceA), and 3-deoxy-D-manno-2-octulosonic acid (Kdo). PG-4 contained (1----4)-linked alpha-galacturono-di-to-penta- saccharides and GalA. The galacturono-tetra- and -penta-saccharides had one and three methyl-esterified GalA units, respectively, and some of the galacturono-oligosaccharides contained 2,4- or 2,3-linked GalA.

Anti-Ulcer Agents↗

Structural studies of endopolygalacturonase-resistant fragments of an antiulcer pectin from the roots of Bupleurum falcatum L.

The endo-alpha-(1-->4)-polygalacturonase-resistant fractions (PG-1, PG-2, and PG-3) from an antiulcer pectin (Bupleuran 2IIc), isolated from the roots of Bupleurum falcatum L., were further analysed by lithium degradation. The results indicated that PG-1 contained a small proportion of long, branched arabinosyl chains and a large proportion of short, neutral oligosaccharide chains. GLC-MS analysis showed that, after methylation the short, neutral oligosaccharide fraction consisted of at least 22 kinds of di- to tetra-saccharide alditols, such as Gal-(1-->4)-Rha-ol (a major component), Ara-(1-->4)-Rha-ol, Glc-(1-->4)-Rha-ol, Ara-->Ara-->Ara-ol, and Ara-->Ara-->Ara-->Ara-ol (minor components) in addition to heteroglycosyl alditols. After deesterification, PG-2 and PG-3 were digested with endo-alpha-(1-->4)-polygalacturonase again, and the enzyme-resistant intermediate size fraction (PG-2') was purified. Component sugar analysis indicated that PG-2' contained 2-Me-Fuc, 2-Me-Xyl, apiose (Api), aceric acid (AceA), 3-deoxy-D-lyxo-heptulosaric acid (Dha), and 3-deoxy-D-manno-2-octulosonic acid (Kdo) in addition to Rha, Fuc, Ara, Xyl, Man, Gal, Glc, GalA, and GlcA. Lithium degradation of PG-2' gave mainly a pentosyl-->6-deoxyhexosyl-->6-deoxyhexosyl-->pentitol fragment, with some neutral di- and tri-saccharide alditols, including a pentosyl-->deoxyhexitol. Methylation analysis of these degradation products indicated that they contained terminal Rha, Araf, Fuc, Xyl, and Gal, 4-linked Rha, 3-linked Fuc, 3-linked Ara, and 3'-linked Api. Bupleuran 2IIc was eluted as essentially a single peak on gel filtration on Sepharose CL-6B. The neutral sugar content of the successive fractions increased with increasing molecular weight, but each fraction also contained, in addition to Rha, Ara, and Gal, 2-Me-Fuc, 2-Me-Xyl, and Api.

Animals↗

Anti-proliferative activity of Bupleurum scrozonerifolium in A549 human lung cancer cells in vitro and in vivo.

Nan-Chai-Hu, the root of Bupleurum scorzonerifolium, is a traditional Chinese herb used in treatment of liver diseases such as hepatitis and cirrhosis. We recently reported that the acetone extract of B. scorzonerifolium (BS-AE) could inhibit proliferation and induce apoptosis in A549 human lung cancer cells. We further examined its anti-proliferative mechanisms and in vivo anticancer effect. Our results showed that BS-AE had the ability to cause cell cycle arrest in G2/M phase, inducing tubulin polymerization, and activating caspase-3 and -9 in A549 cells. BS-AE-induced apoptosis could be blocked by the broad caspase inhibitor z-VAD-fmk in majority. The result of in vivo study showed that BS-AE could suppress growth in A549 subcutaneous xenograft tumors. These results indicate that BS-AE exerts antiproliferative effects on A549 cells in vitro and in vivo, and prompted us to further evaluate and elucidate the chemical composition profile of BS-AE.

Animals↗

Bupleurum kaoi inhibits Coxsackie B virus type 1 infection of CCFS-1 cells by induction of type I interferons expression.

Coxsackie B virus type 1 (CVB1) infection is known to cause high morbidity and mortality in children, however, there is no effective drug for treating this disease. The present study aimed to examine the antiviral activity of Bupleurum kaoi (BK), a popular herbal drug for treating viral and bacterial infections, against CVB1 infection and its mechanisms of action. Our data showed that BK neutralized the CVB1-induced cytopathic effect in human neonatal foreskin fibroblast cell line (CCFS-1/KMC), with IC50 and EC50 values around 12.38 microg/ml and 50.93 microg/ml, respectively. Its CC50 and SI values were 883.56 microg/ml and 17.34, respectively. These results suggest that BK possessed anti-CVB1 activity, and showed no effect on CCFS-1 cell viability and growth at concentration 250 microg/ml. The time-of-addition studies showed that BK (50, 100 and 200 microg/ml) added at various time of preinfection (-1 to -3 h), coinfection (0 h) and postinfection (1-3 h) could inhibit CVB1 infection. Interestingly, BK also showed an inhibition on viral replication through the induction of IFN-alpha/beta expression. In conclusion, BK possessed antiviral activity against CVB1 infection. It interfered the early stage of viral replication and viral replication after infection through the induction of type I interferon expression.

Antiviral Agents↗

Dual inhibition of cyclooxygenase-1 and 5-lipoxygenase by aerial part of Bupleurum fruticescens methanol extract.

The effect of the methanol extract from aerial parts of Bupleurum fruticescens on the release of eicosanoids and hydrolytic enzymes was determined on in vitro cell systems. The extract had a significant effect on 5-lipoxygenase (5-LOX) activity, inhibiting both LTB4 and 5(S)-HETE production with IC50 values of 112 microg/ml and 95 microg/ml, respectively. At concentrations of 200 microg/ml, the extract also inhibited cyclooxygenase-1 (90%) and elastase activities (54%). The 12-LOX activity in intact platelets was not affected; a fact, which suggests that phospholipase A2 (PLA2) activity, is not modified by the extract.

Animals↗

Isolation and characterization of an anti-complementary polysaccharide D3-S1 from the roots of Bupleurum smithii.

The preliminary data from hemolytic assays indicated that the hot-water extract of the roots of Bupleurum smithii had anti-complementary activity. Further bioactivity-guided fractionation led to the isolation of D3-S1, a homogeneous form of acidic polysaccharide. D3-S1 was a branched polysaccharide with average molecular weight about 2,000,000 Da, composed of Ara, Gal and GalA in the ratio of 2.6:1.0:1.2, along with trace of Rha, Glc, Xyl and Man. Methylation analysis and NMR identified the linkages of the residues of D3-S1. Functional analysis showed that D3-S1 inhibited complement activation on both the classic and alternative pathways with CH(50) value of 0.34+/-0.02 mg/ml and AP(50) value of 0.081+/-0.003 mg/ml, respectively. Preliminary mechanism studies by using complement component depleted-sera indicated that D3-S1 selectively interacts with C1s, C3 and C4, but not C1q, C1r, C2, C5 and C9. The results suggested that D3-S1 could be of potential benefits in treatment of the complement-associated diseases.

Animals↗

Requirement for ERK activation in acetone extract identified from Bupleurum scorzonerifolium induced A549 tumor cell apoptosis and keratin 8 phosphorylation.

We previously demonstrated that the crude acetone extract of Bupleurum scorzonerifolium (AE-BS) 60 microg/ml has anti-proliferation activity and apoptosis effects to A549 human lung cancer cells. They can also cause tumor cell arrest in G2/M phase. To better understand its target protein in A549 cell, two-dimensional electrophoresis and liquid chromatography-tandem mass spectrometry were applied. The modification of keratin 8 was identified. By immunoblot, the expression of phosphorylated keratin 8 at Ser-73 was increased from 2.0 to 3.0-fold after AE-BS treatment 24 to 48 hr respectively as compared with untreated A549 control cells. Furthermore, the A549 cells were pretreated with 50 microM PD98059, a specific inhibitor of the upstream regulator of ERK1/2, or with the p38 kinase inhibitor 20 microM SB203580 or JNK inhibitor 20 microM SP600125 for 30 min, followed by 24 h of incubation with AE-BS, PD98059 can inhibit K8-Ser-73 hyperphosphorylation and prevented cell apoptosis which was induced by AE-BS significantly. By immunoblot, AE-BS also can induce ERK 1/2 phosphorylation. In conclusion, our data indicate that the AE-BS induced tumor apoptosis in A549 cells was related to ERK 1/2 activation. The molecular mechanism of hyperphosphorylation of K8 on Ser-73 was associated with ERK 1/2 activation rather than JNK and p38 kinase. The apoptosis induced by AE-BS may be related to K8 phosphorylation.

Anthracenes↗

Immunosuppressive flavones and lignans from Bupleurum scorzonerifolium.

Two lignans, isochaihulactone and chaihunaphthone, together with eleven known compounds were isolated from the root of Bupleurum scorzonerifolium. Their structures were established on the basis of spectral evidence. In biological testing, eugenin and saikochromone potently inhibited CD28-costimulated activation of human peripheral blood T cells.

Antineoplastic Agents, Phytogenic↗

Haemolytic activities and adjuvant effect of Bupleurum chinense saponins on the immune responses to ovalbumin in mice.

In this study, the haemolytic activities of Bupleurum chinense saponins (BCS) and its adjuvant potentials on the immune responses of ICR mice against ovalbumin (OVA) were evaluated. BCS showed a slight haemolytic effect, with its haemolytic percents being 3.32% and 1.19% at the concentrations of 500 and 250 microg/ml, respectively. ICR mice were immunized subcutaneously with OVA 100 microg alone or with OVA 100 microg dissolved in saline containing aluminium hydroxide gel (Alum, 200 microg), QuilA (10 and 20 microg) or BCS (50, 100 or 200 microg) on Days 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. BCS 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 levels in serum were also significantly enhanced by BCS 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 BCS and QuilA on the OVA-specific IgG2b antibody responses to OVA in mice. In conclusion, the results suggest that BCS could be safely used as adjuvant with low or non-haemolytic effect.

Adjuvants, Immunologic↗