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PubMed2006

In vitro and in vivo studies of a novel potential anticancer agent of isochaihulactone on human lung cancer A549 cells.

We previously demonstrated that the crude acetone extract of Bupleurum scorzonerifolium (BS-AE) 60 microg/ml has anti-proliferation activity and apoptotic effects on A549 non-small cell lung cancer (NSCLC). A novel lignan, isochaihulactone (4-benzo[1,3]dioxol-5-ylmethyl-3(3,4,5-trimethoxyl-benzylidene)-dihydro-furan-2-one), was isolated from BS-AE and identified from spectral evidence ((1)H NMR, (13)C NMR, IR, and MS) and by comparison with authentic synthetic standards. Isochaihulactone was cytotoxic (IC(50)=10-50 microM) in a variety of human tumor cell lines. In in vitro and in vivo microtubule assembly assays, it inhibited tubulin polymerization in a concentration-dependent manner. As determined by flow cytometry, isochaihulactone caused G2/M phase arrest and apoptosis in a time- and concentration-dependent manner. G2/M arrest was correlated with increased p21/WAF1 levels, downregulation of the checkpoint proteins cyclin B1/cdc2 and mobility shift of cdc25C. Moreover, isochaihulactone (30 and 50 mg/kg) inhibited the growth of non-small cell lung carcinoma A549 xenograft in nude mice. These findings indicate isochaihulactone is a promising new antimitotic anticancer compound with potential for clinical application in the future.

4-Butyrolactone↗

PubMed2006

The presence of natural human antibodies reactive against pharmacologically active pectic polysaccharides from herbal medicines.

Direct ELISA was performed using normal human sera and human colostrum, to analyse the presence of antibodies which react with pharmacologically active pectic polysaccharides isolated from plants used in traditional Japanese herbal (Kampo) medicine. All sera and colostrum were shown to contain IgM, IgG, IgA and secretory IgA class antibodies which react with the active pectic polysaccharides to different degrees. The reacting IgG antibody in normal human serum recognized the ramified regions (rhamnogalacturonan core with carbohydrate side-chains) of the pharmacologically active pectic polysaccharides as the active sites for complement-activating activity. Correlation analysis indicated that a significant and positive correlation was observed between reactivity with the reacting antibody of IgG class and the degree of complement-activating activity of the active polysaccharides. The reacting IgG class antibody, which was purified from normal human serum by affinity chromatography on bupleuran 2IIc (a pharmacologically active pectic polysaccharide from the roots of Bupleurum falcatum)-immobilized Sepharose, showed cross-reactivity not only with some other pharmacologically active pectic polysaccharides from other medicinal herbs but also with autoantigens such as single-strand DNA, myosin and tublin from mammals.

Angelica↗

PubMed2004

Seasonal branch nutrient dynamics in two Mediterranean woody shrubs with contrasted phenology.

BACKGROUND AND AIMS: Mediterranean woody plants have a wide variety of phenological strategies. Some authors have classified the Mediterranean phanaerophytes into two broad phenological categories: phenophase-overlappers (that overlap resource-demanding activities in a short period of the year) and phenophase-sequencers (that protract resource-demanding activities throughout the year). In this work the impact of both phenological strategies on leaf nutrient accumulation and retranslocation dynamics at the level of leaves and branches was evaluated. Phenophase-overlappers were expected to accumulate nutrients in leaves throughout most of the year and withdraw them efficiently in a short period. Phenophase-sequencers were expected to withdraw nutrients progressively throughout the year, without long accumulation periods. METHODS: To test this hypothesis, variations in phenology and leaf NPK in the crown of a phenophase-overlapper Cistus laurifolius and a phenophase-sequencer Bupleurum fruticosum were monitored monthly during 2 years. KEY RESULTS: Changes in nutrient concentration at the leaf level were not clearly related with the different phenologies. Nitrogen and phosphorous resorption efficiencies were lower in the phenophase-overlapper, and accumulation-retranslocation seasonality was similar in both species. Changes in the branch nutrient pool agreed with the hypothesis that the phenophase-overlapper accumulated nutrients from summer until the bud burst of the following spring, recovering a large nutrient pool during massive leaf shedding. The phenophase-sequencer did not accumulate nutrients from autumn until early spring, achieving lower nutrient recovery during spring leaf shedding. CONCLUSIONS: It is concluded that phenological demands influence branch nutrient cycling. This effect is easier to detect by assessing changes in the branch nutrient pool rather than changes in the leaf nutrient concentration.

Bupleurum↗

PubMed2005

Gene expression profiling predicts liver responses to a herbal remedy after partial hepatectomy in mice.

The aim of this study was to demonstrate that regenerating liver responses to a herbal remedy could be presented by gene expression profiling. Compositions of the ingredients in the remedy containing Scutellaria baicalensis Georgi and Bupleurum scorzonerifolfium Wild (S/B remedy) were analyzed and quantified by high performance liquid chromatography. By using a 70% partial hepatectomy in BALB/c mice as an in vivo model, the effects of high dose (50 mg/kg) and low dose (1 mg/kg) S/B remedy were evaluated by cDNA microarray, followed by RT-PCR and real-time PCR confirmation. Factors affecting proliferative activities of mouse hepatocytes were measured by DNA flow cytometry, BrdU incorporation assay and serum interleukin-6 (IL-6) level. Based on global gene expression profiles, the results showed that the low dose S/B remedy down-regulated expression of immediate early genes and cell cycle-related genes, whereas the high dose had opposite effects. The gene expression was further verified by real-time RT-PCR. Proliferative activities, in terms of synthetic phase fractions and G2/M phase fractions, in vehicle, low dose, and high dose groups were 18.45+/-2.56%, 14.65+/-1.06%; 9.27+/-0.85%, 7.80+/-0.11%; and 18.90+/-2.17%, 22.95+/-0.25%, respectively. The serum IL-6 level was also dose-dependent in both low and high dose S/B remedy-treated mice. We conclude that in vivo gene expression profiling correlates with liver responses to a herbal remedy, which provides a new direction for pharmaceutical studies on human diseases.

Animals↗

PubMed2005

[Identification of the fumatory Radix Bupleuri with sulfur by static headspace GC-MS].

OBJECTIVE: To identify whether Radix Bupleuri (Bupleurum chinense) was fumed with sulfur. METHOD: A static headspace GC-MS method was used to detect sulfur in the fumatory Radix Bupleuri, the authentic samples free of sulfur was detected as reference. RESULT: Sulfur was detected in six samples from nine samples collected in different locations. CONCLUSION: The method can be used to detect sulfur rapidly in the fumatory Radix Bupleuri with sulfur.

Bupleurum↗

PubMed2006

[Effect of saikosaponins on glial fibrillary acidic protein expression in hippocampal astrocytes of pentetrazole-induced chronic kindling rats].

OBJECTIVE: To study the effect of saikosaponins, the active ingredients of Bupleurum chinense DC, on glial fibrillary acidic protein (GFAP) expression in hippocampal astrocytes of chronic kindling rats induced by pentetrazole (PTZ). METHODS: Forty-eight healthy Sprague-Dawley rats were randomized into 6 equal groups, namely the blank control group (Group A), normal saline group (Group B), sodium valproate group (Group C), and 3 saikosaponins groups of high, medium and small doses (Groups D, E, and F, respectively). The rats (except those in Group A) received intraperitoneal injection of PTZ to induce chronic kindling 1 h after the respective agents as indicated were administered intragastrically on a daily basis for 4 consecutive weeks. Upon completion of the treatment course, the rats were sacrificed and the brain tissues were sampled, sliced and stained for immunohistochemical examination. The results were analyzed to calculate the positive cell count, cross-sectional area of the cells and the gray scale. RESULTS: In group B, the positive cell population and cross-sectional area of the positive cells were the greatest among the groups (P<0.01), but the positive cell gray scale of the CA1 and CA2 regions and the dentate gyrus (DG) of the hippocampus was the lowest. The CA1 region of Group B was significantly different from that of groups A, C and D (P<0.01), and the CA2 region different from groups A, C, D and E (P<0.05), while the DG different from group F (P<0.05) and groups A, C, D and E (P<0.01). CONCLUSION: In chronic kindling rats induced by PTZ, GFAP overexpression can be inhibited by saikosaponins, which suppress the abnormal activation of hippocampal astrocyte of the kindling rats.

Animals↗

PubMed2006

An HPLC/MS method for identifying major constituents in the hypocholesterolemic extracts of Chinese medicine formula 'Xue-Fu-Zhu-Yu decoction'.

In this paper, HPLC-ESI-MS technique was used to analyze the hypocholesterolemic extracts of 'Xue-Fu-Zhu-Yu decoction', a traditional Chinese medicine consisting of six crude drugs (i.e. Paeonia lactiflora, Ligusticum chuanxiong, Citrus aurantium, Carthamus tinctorius, Prunus persica and Bupleurum falcatum). A total of 17 compounds were identified and their plant derivations were also distinguished. Nine compounds among them were unambiguously determined as oxypaeoniflorin, amygdalin, albiflorin, paeoniflorin, ferulic acid, naringin, hesperidin, senkyunolide I and neohesperidin by comparing the retention times (tR), UV spectra and m/z values with those of the reference compounds. The other eight compounds were tentatively identified as prunasin, 6-hydroxy-kaempferol 3,6-diglucoside, 6-hydroxykaempferol 3-rutinoside-6-glucoside, galloylpaeoniflorin, 6-hydroxy-kaempferol 3-glucoside, anhydrosafflor yellow B, narirutin and kaempferol 3-rutinoside by MS2 spectra and the comparison of their UV spectra and MS spectra with literature data.

Anticholesteremic Agents↗

PubMed1994

Characterization of the immunoregulatory action of saikosaponin-d.

The immunoregulatory action of saikosaponin-d (SSd), which was isolated from the root of Bupleurum falcatum L. and has a steroid-like structure, was examined on splenic T lymphocytes of C57BL/6 mice. SSd displayed a definite action in vitro to bidirectionally control the growth response of T lymphocytes stimulated by concanavalin A, anti-CD3 monoclonal antibody, and calcium ionophore A23187 plus phorbol 12-myristate 13-acetate. Low concentrations (1-3 micrograms/ml) of SSd upregulated the responses to suboptimum stimuli of agonists, particularly during the relatively late stage of the responses, whereas it downregulated the responses to supraoptimal stimuli. Under appropriate experimental conditions, SSd promoted interleukin-2 (IL-2) production and IL-2 receptor expression. It also accelerated c-fos gene transcription, but it did not modulate the level of tyrosine phosphorylation of cellular proteins. We concluded from these results that SSd uniquely modulates T lymphocyte function and that at least one target of the action of SSd is located at or before the step of c-fos gene transcription and after T-cell receptor/CD3-mediated protein tyrosine kinase activation.

Adjuvants, Immunologic↗

PubMed1986

Effects of saikosaponin-d on aminonucleoside nephrosis in rats.

The effects of saikosaponin-d extracted from the roots of Bupleurum falcatum L. on aminonucleoside nephrosis were studied in rats. Urine protein excretion in rats receiving aminonucleoside alone was significantly elevated on the 2nd day after the last injection of aminonucleoside and reached a peak on the 11th day. Urinary protein on the 11th day was reduced by 48 and 46%, respectively in animals treated with saikosaponin-d from the 2nd and 8th day after the last injection of aminonucleoside. Electron microscopically, the degree of abnormality, e.g. loss or fusion of foot processes, in the glomerular epitherial cells was significantly lower in the rats treated with saikosaponin-d after aminonucleoside injection than in the rats treated with aminonucleoside alone. It is concluded that saikosaponin-d prevents the development of proteinuria induced by aminonucleoside in the rat.

Animals↗

PubMed1993

Determination of acidic saponins in crude drugs by high-performance liquid chromatography on octadecylsilyl porous glass.

High-performance liquid chromatographic (HPLC) analysis on octadecylsilyl porous glass was investigated for acidic saponins in ginseng, bupleurum root and senega. The acidic saponins, malonyl-ginsenosides, malonyl-saikosaponins and senegins, as well as neutral saponins in the crude drugs were separated rapidly by HPLC on this column with aqueous acetonitrile containing KH2PO4 as the mobile phase at room temperature.

Carbohydrate Sequence↗

PubMed1995

Cell type-oriented differential modulatory actions of saikosaponin-d on growth responses and DNA fragmentation of lymphocytes triggered by receptor-mediated and receptor-bypassed pathways.

We examined the immunoregulatory action of saikosaponin-d (SSd), which was isolated from the root of Bupleurum talcatum L. and had a steroid-like structure, on murine thymocytes, and compared the action with that on spleen cells. Constitutive DNA synthesis or the growth response stimulated with anti-CD3mAb of thymocytes were down-regulated by 3 micrograms/ml SSd, whereas with spleen cells these were up-regulated by the same concentration of SSd. On the other hand, 3 micrograms/ml of SSd greatly up-regulated the growth response and interleukin 2 (IL-2)/interleukin 4 (IL-4) production induced through a receptor-bypassed pathway by calcium ionophore A23187 plus phorbol 12-myristate 13-acetate (PMA) in thymocytes, whereas it only slightly up-regulated them in spleen cells. Moreover, the same concentration of SSd inhibited DNA fragmentation in thymocytes induced by A23187 or PMA. These results suggest a unique cell type-dependent immuno-modulatory action of SSd.

Animals↗

PubMed1991

Effects of saikosaponin-d on the functions and morphology of macrophages.

The effects of saikosaponin-d, isolated from Bupleurum Radix, on phagocytosis and spreading of mouse peritoneal macrophages were investigated. Macrophages from saikosaponin-d-treated mice showed a significant increase in spreading activity followed by an increase in phagocytic activity. An intense distribution of microfilaments and microtubules was also observed in these macrophages by immunofluorescence microscopy.

Animals↗

PubMed1991

Effect of saikosaponin on the immune responses in mice.

The in vivo effects of saikosaponin, isolated from Bupleurum radix, on the immune responses are still poorly understood. We have already shown that saikosaponin-d increases phagocytic activities of murine peritoneal macrophages such as spreading activity, phagocytosis, lysosomal enzyme activity and intracellular killing activity of living yeast. This work extends these observations by showing that treatment with saikosaponin also increased the antibody response in plaque-forming cell numbers after in vivo immunization with sheep red blood cells (SRBC) and an augmentation of spleen cell proliferation responses to stimulation with T- or B-cell mitogens both before and after immunization. Furthermore, after SRBC immunization, the macrophages from mice treated with saikosaponin-d revealed significant increases in spreading activity and lysosomal enzyme activity. The chemiluminescences of the macrophages from mice treated with saikosaponin-d stimulated by opsonized zymosan and PMA were enhanced and interleukin-1 production by the cells was increased in a dose-dependent manner. These results demonstrate that saikosaponin-d may stimulate in vivo immunological lymphocyte functions, partly by activating some macrophage functions.

Animals↗

PubMed1991

Inhibition of platelet activation and endothelial cell injury by flavan-3-ol and saikosaponin compounds.

The effects of flavan-3-ol and saikosaponin compounds on platelet aggregation, platelet thromboxane biosynthesis and H2O2-induced endothelial cell injury were studied. Seven flavan-3-ol compounds isolated from Camellia sinensis L. var sinensis O. Kuntze (Theaceae) and three saikosaponin compounds isolated from Bupleurum falcatum L. (Umbelliferae) were used. Among the 10 compounds tested, only epigallocatechin and saikosaponin a significantly inhibited human platelet aggregation induced by ADP, and the potency of inhibition was comparable with aspirin. Both of epigallocatechin and saikosaponin a dose-dependently inhibited the platelet thromboxane formation from exogenous and endogenous arachidonic acid. In the prevention of H2O2-induced endothelial cell injury in culture, only gallocatechin-3-0-gallate and epicatechin-3-0-gallate were effective. The inhibitory effect of epigallocatechin and saikosaponin a on platelet activation and the cytoprotective effect of gallocatechin-3-0-gallate and epicatechin-3-0-gallate on H2O2-induced endothelial cell injury could give evidence of explaining the possible role of flavan-3-ol and saikosaponin compounds in maintaining vascular homeostasis.

Animals↗

PubMed2005

Saikosaponin-d inhibits T cell activation through the modulation of PKCtheta, JNK, and NF-kappaB transcription factor.

The effects of saikosaponin-d, a triterpene saponin derived from Bupleurum falcatum L. (Umbelliferae), on the signaling pathways of T cell activation were examined. The results showed that saikosaponin-d potently suppressed both early (CD69) and late (CD71) expressions of mouse T cells stimulated with Con A or PMA. It interfered with PKCtheta translocation from cytosol to membrane fraction and inhibited the phosphorylations of IkappaBalpha and JNK, but not ERK, in PMA-activated mouse T cells. Additionally, it inhibited PMA and ionomycin-stimulated IL-2 production in mouse T cells. In summary, these results indicate that the mechanism by which saikosaponin-d inhibits T cell activation would involve the suppression of CD69 and CD71 expressions and IL-2 production, and the modulation of PKC pathway through PKCtheta, JNK, and NF-kappaB transcription factor. This may herald a novel approach for further studies of saikosaponin-d as a candidate for use in the treatment of inflammatory and autoimmune diseases.

Animals↗

PubMed2005

Cholesteryl hemisuccinate as a membrane stabilizer in dipalmitoylphosphatidylcholine liposomes containing saikosaponin-d.

In the present study, cholesteryl hemisuccinate (CHEMS) was evaluated for use as a membrane stabilizer in dipalmitoylphosphatidylcholine (DPPC) liposomes. Differential scanning calorimetry (DSC) and a calcein release study showed that CHEMS was more effective than cholesterol (CHOL) in increasing DPPC membrane stability. The findings of Fourier transform infrared spectroscopy (FT-IR) also suggested that CHEMS interacts with DPPC via both hydrogen bonding and electrostatic interaction. More importantly, CHEMS did not interact with saikosaponin-d (SSD), a triterpene saponin from Bupleurum species, unlike CHOL. SSD-containing liposomes with DPPC, CHEMS and DSPE-PEG could greatly decrease the hemolytic activity of SSD. This study demonstrated that CHEMS has more stabilization ability than CHOL since CHEMS may exhibit both hydrogen bond interaction and electrostatic interaction with DPPC membrane while CHOL only has hydrogen bond interaction, resulting in stable and low-hemolytic SSD-liposomes.

1,2-Dipalmitoylphosphatidylcholine↗

PubMed2004

Preventive effect of traditional herbal formulae against experimental hypercholesterolemia in rats with special reference to blood lipoprotein cholesterol levels.

To investigate the effect of traditional herbal formulae (Zhong Yao) against experimental hypercholesterolemia, we assayed serum lipoprotein levels and hepatic function in rats administered herbal formulae under cholesterol rich diet. Two kinds of herbal formulae; prescription 1 (Bupleurum chinense and other 9 kinds of herbs) and 2 (Camellia sinensis and other 5 kinds of herbs) were prepared. All of 35 rats were separated into 4 groups, group P fed under cholesterol rich diet for 15 days and administered the prescriptions for 2 days, group Q administered the prescriptions for 17 days under the same diet, group R fed under the same diet but no prescriptions, and group S fed under regular diet. In rats administered prescription 1, serum low density lipoprotein cholesterol levels in group Q were preserved as well as standard group S. Such a preservation was not observed under administration of prescription 2. Regarding choleretic function, serum total bilic acid levels measured in group P were extremely increased when prescription 1 was administered for only 2 days after stopping the cholesterol rich diet. In study on telomerase activity of liver extracts to assess hepatocyte function, significantly higher level of activity was noted in group Q administered prescription 1. From these results, it was suggested that prescription 1 had much preventive effect against hypercholesterolemia in rats, probably due to its choleretic function.

Animals↗

PubMed2005

In vitro anti-Helicobacter pylori action of 30 Chinese herbal medicines used to treat ulcer diseases.

Infection by Helicobacter pylori has been ascertained to be an important etiologic impetus leading usually to chronic active gastritis and gastric ulcer with growing incidences worldwide. Utilizing as the test pathogen a standard and five clinic strains of Helicobacter pylori, the antibacterial action was assessed in vitro with ethanol extracts of 30 Chinese herbal medicines which have been frequently prescribed since ancient times for treating gastritis-like disorders. Among the 30 tested materials, the ethanol extracts of Abrus cantoniensis (Fabaceae), Saussurea lappa (Asteraceae) and Eugenia caryophyllata (Myrtaceae) were strongly inhibitory to all test strains (MICs: approximately 40 microg/ml), and Hippophae rhamnoides (Elaeagnaceae), Fritillaria thunbergii (Liliaceae), Magnolia officinalis and Schisandra chinensis (Magnoliaceae), Corydalis yanhusuo (Papaveraceae), Citrus reticulata (Rutaceae), Bupleurum chinense and Ligusticum chuanxiong (Apiaceae) substantially active with MICs close to 60.0 microg/ml. As to antibacterial actions of the aqueous extracts of the same drugs, those derived from Cassia obtusifolia (Fabaceae), Fritillaria thunbergii and Eugenia caryophyllata were remarkably inhibitory against all the six Helicobacter pylori strains (MICs: approximately 60 microg/ml). The work compared almost quantitatively the magnitude of the anti-Helicobacter pylori actions of the 30 most prescribed gastritis-treating Chinese herbal drugs, and located as well some source plants where potent anti-Helicobacter pylori phytochemicals could be characterized.

Anti-Bacterial Agents↗