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Immunologic enhancement of compound Chinese herbal medicinal ingredients and their efficacy comparison with compound Chinese herbal medicines.

Two compound Chinese herbal medicinal ingredients (cCIs) were prepared respectively with epimedium polysaccharide (EPS) plus propolis flavone (PF) and astragalus polysaccharide (APS) plus ginsenoside (GS). Also, two compound Chinese herbal medicines (cCMs) with the same ingredient content as corresponding cCIs were made with the extracts of epimedium plus propolis and astragalus plus ginseng. In rabbit immune trial, two cCIs, physiological saline in the control, were respectively injected to the rabbits vaccinated with inactivated rabbit hemorrhagic disease vaccine. On Days 3, 7, 10, 14, 21, 28, 35 and 42 after vaccination, the dynamic changes of serum antibody titers were determined by hemagglutination inhibition (HI) test. In chicken immune trial, all of cCIs and cCMs were mixed respectively with inactivated Newcastle disease vaccine virus to vaccinate chickens, taking oil-adjuvant and non-adjuvant vaccine as controls. On Days 7, 14, 21, 28, 35 and 42 after vaccination, the dynamic changes of peripheral lymphocyte proliferation and serum antibody titers were tested respectively by MTT method and HI test method. The results showed that both cCIs could significantly raise antibody titer in rabbits, which the effect of compound Chinese herbal medicinal ingredients 1 (cCIs 1) was better than that of compound Chinese herbal medicinal ingredients 2 (cCIs 2). All of cCIs and cCMs could markedly promote lymphocyte proliferation and enhance antibody titer in chickens, which was similar to oil adjuvant, the immunologic enhancement of cCIs were slightly superior to that of the cCMs.

Adjuvants, Immunologic↗

Anti-thrombic activity of Korean herbal medicine, Dae-Jo-Whan and its herbs.

The anti-thrombic properties of the Korean herbal medicine, Dae-Jo-Hwan (DJW), which is consisted of 11 herbs (indicated as concentrations) of Rehmanniae radix 24%, Hominis placenta 5%, Testudinis carapax 9%, Eucommiae cortex 9%, Asparagi radix 9%, Phellodendri cortex 9%, Achyranthis radix 7%, Liriopis tuber 7%, Angelicae sinensis radix 7%, Ginseng radix 6%, and Schizandrae fructus 3%, were investigated. The extracts of DJW and its 11 herbs, except G. radix, A. sinensis radix and S. Fructus, inhibited the endotoxin-induced hepatic venous thrombosis in high cholesterol diet-treated rats. Also the extract inhibited the endotoxin-induced decrease in blood platelets and fibrinogen, and endotoxin-induced increase in fibrin degradation products (FDP) on disseminated intravascular coagulation in normal rats. In in vitro experiments, the extract was shown to have inhibitory effect on collagen- and ADP-induced blood platelet aggregation, on thrombin-induced conversion of fibrinogen to fibrin and on the activity of plasminogen or plasmin. In conclusion, the protection of extracts of Korean herbs on the ischemic infarction induced artificially might be related to their inhibitory effects on DIC, platelet coagulation and thrombic action.

Adenosine Diphosphate↗

Ginsenosides may reverse the dexamethasone-induced down-regulation of glucocorticoid receptor.

The effects of glucocorticoid (GC) hormones are mediated via an intracellular receptor, the glucocorticoid receptor (GR). It has been established that glucocorticoid down-regulate GR. Ginsenosides (GSS) from extract of Panax ginseng have demonstrated glucocorticoid-like activities in homeostasis and regulation of immunity, etc. We hypothesize that ginsenosides might mediate some of their actions by binding to the GR. The present study is aimed to determine whether GSS can act like a GC analog in the activation of glucocorticoid response element-luciferase activity in HL7702 cells. We found that GSS alone had no effect on the expression of reporter gene, but it enhanced dexamethasone (Dex)-induced transcription of reporter gene. To further explore the effects of GSS, we examined the influence of GSS on the gene and protein expression as well as hormone binding activity of GR by semi-quantitative RT-PCR, Western blot, and radioligand-binding assay, respectively. GSS partially reversed the Dex-induced decrease in GR expression and hormone binding activity with an optimal dose of 25 microg/ml, implicating a positive regulatory effect of GSS on GR expression and binding activity. Therefore, our result suggests that GSS may reverse partially the dexamethasone-induced down-regulation of glucocorticoid receptor.

Blotting, Western↗

Phylogeny of Panax using chloroplast trnC-trnD intergenic region and the utility of trnC-trnD in interspecific studies of plants.

Sequences of the chloroplast trnC-trnD region and the internal transcribed spacer (ITS) regions of nuclear ribosomal DNA were obtained for all species of Panax L. (the ginseng plant genus, Araliaceae) to reconstruct phylogenetic relationships. The trnC-trnD phylogeny is congruent with the ITS phylogeny for the diploid taxa of Panax. This study is the first use of the trnC-trnD sequence data for phylogenetic analysis at the interspecific level. We evaluated this DNA region for its phylogenetic utility at the lower taxonomic level for flowering plants. The trnC-trnD region includes the trnC-petN intergenic spacer, the petN gene, the petN-psbM intergenic spacer, the psbM gene, and the psbM-trnD intergenic spacer. The petN and psbM genes are small, 90 and 104-114 bp across angiosperms, respectively, and have conserved sequences. We have designed universal amplification and sequencing primers within these two genes. Using these primers, we have successfully amplified the entire trnC-trnD region for a diversity of flowering plant groups, including Aralia L. (Araliaceae), Calycanthus L. (Calycanthaceae), Corylus L. (Betulaceae), Hamamelis L. (Hamamelidaceae), Hydrocotyle L. (Apiaceae), Illigera Blume (Hernandiaceae), Nelumbo Adans. (Nelumbonaceae), Nolana L. ex L.f. (Solanaceae), Prunus L. (Rosaceae), and Staphylea L. (Staphyleaceae). In Panax, the trnC-trnD region provides a similar number of informative phylogenetic characters as the ITS regions and a slightly higher number of informative characters than the chloroplast ndhF gene. We thus demonstrate the utility of the trnC-trnD region for lower-level phylogenetic studies in flowering plants.

Chloroplasts↗

Nonvitamin, nonmineral dietary supplements: issues and findings from NHANES III.

The Commission on Dietary Supplement Labels encourages nutrition professionals to become knowledgeable about all dietary supplements. The Dietary Supplement Health and Education Act of 1995 (DSHEA) expanded the definition of dietary supplements beyond essential nutrients while distinguishing them from drugs or food additives. In order to give practical advice to consumers and policymakers, dietetics professionals need to understand the implications resulting from this less-restrictive regulatory environment for supplements. Dietetics professionals must also become familiar with claims made by manufacturers, retailers, and others regarding popular nonvitamin, nonmineral (NVNM) supplements, as well as usage prevalence and trends. However, NVNM supplements currently are classified inconsistently, and information on the prevalence of use is limited. Sales data suggest that total intake is increasing, and garlic and ginseng are consistently among the most popular supplements. Reported use of NVNM supplements in the third National Health and Nutrition Examination Survey was highest for garlic and lecithin. The data suggest associations of NVNM supplement use with age and more healthful lifestyles; however, there is also a reported link with higher alcohol consumption and obesity. Associations with education, income, region, and urbanization are not evident from the sales data. Standardized survey procedures regarding question phraseology, referent time period, and supplement categorization--along with use of representative samples--will improve our ability to assess supplement use, prevalence, and trends.

Adolescent↗

Specific interaction between tetrandrine and Quillaja saponins in promoting permeabilization of plasma membrane in human leukemic HL-60 cells.

Spontaneous Ni2+ entry (leak), measured as fluorescence quench in fura-2-loaded HL-60 cells at the excitation wavelength of 360 nm, was strongly inhibited by tetrandrine (TET, 100 microM), a Ca2+ antagonist of Chinese herbal origin. Exposure of the cells for 5 min to saponins from Quillaja saponaria (QS, 30 microg/ml), surfactants well known to permeabilize the plasma membrane by complexing with cholesterol, promoted Ni2+ entry without causing fura-2 leak-out. Unexpectedly, TET caused an immediate (within 2.5 min) augmentation of QS-promoted Ni2+ entry; and a 5-min treatment with both TET and QS resulted not only in an enhanced Ni2+ entry, but also a fura-2 leak-out. Ginseng saponins (100 microg/ml) alone or together with TET did not cause such a permeabilization. Permeabilization induced by 1-3 microM digitonin, another cholesterol-complexing glycoside, could not be enhanced by TET. TET did not affect permeabilization induced by Triton X-100 (0.01%), a detergent which non-specifically disrupts the hydrophobic interaction at the plasma membrane. TET also did not enhance Ni2+ entry triggered by ionomycin (0.35 microM) or SK&F 96365 (20 microM). Further, it did not augment Ni2+ entry when the plasma membrane fluidity was modulated by changes of temperature (27-47 degrees C) or treatment with 5% ethanol. This QS-promoted Ni2+ entry could not be amplified by other lipophilic Ca2+ antagonists, such as diltiazem (100 microM) and verapamil (100 microM). The results hence indicate that TET enhanced Ni2+ entry (or permeabilization) elicited by QS treatment, but not other perturbations of the plasma membrane. We suggest that pore formation at the plasma membrane, a consequence of QS-cholesterol interaction, can be specifically enhanced by TET. Also, a comparative study of the effects of TET and its very close analogues, hernandezine and berbamine, reveals that the methoxyl group at the R2 position of TET appears to be crucial in enhancing QS-promoted Ni2+ entry.

Alkaloids↗

Cdk2 activity is associated with depolarization of mitochondrial membrane potential during apoptosis.

In this study we show that panaxadiol, a ginseng saponin with a dammarane skeleton, induces apoptotic cell death by depolarization of mitochondrial membrane potential in human hepatoma SK-HEP-1 cells. Sequential activation of caspases-9, -3, and -7, but not of caspase-8, occurs after mitochondrial membrane depolarization and cytochrome c release from the mitochondria of panaxadiol-treated cells. Moreover, Cdk2 kinase activity, but not Cdc2 kinase activity, is markedly upregulated in the early stages of apoptosis. Olomoucine or roscovitine, specific Cdks inhibitors, effectively prevent mitochondrial membrane depolarization as well as apoptotic cell death in panaxadiol-treated cells. Thus, panaxadiol-treatment induces cell death-dependent activation of Cdk2 kinase activity, which is functionally associated with depolarization of mitochondrial membrane potential and subsequent cytochrome c release.

Antineoplastic Agents↗

Reversal of P-glycoprotein-mediated multidrug resistance by ginsenoside Rg(3).

Multidrug resistance has been a major problem in cancer chemotherapy. In this study, in vitro and in vivo modulations of MDR by ginsenoside Rg(3), a red ginseng saponin, were investigated. In flow cytometric analysis using rhodamine 123 as an artificial substrate, Rg(3) promoted accumulation of rhodamine 123 in drug-resistant KBV20C cells in a dose-dependent manner, but it had no effect on parental KB cells. Additionally Rg(3) inhibited [3H]vinblastine efflux and reversed MDR to doxorubicin, COL, VCR, and VP-16 in KBV20C cells. Reverse transcriptase-polymerase chain reaction and immuno-blot analysis after exposure of KBV20C cells to Rg(3) showed that inhibition of drug efflux by Rg(3) was due to neither repression of MDR1 gene expression nor Pgp level. Photo-affinity labeling study with [3H]azidopine, however, revealed that Rg(3) competed with [3H]azidopine for binding to the Pgp demonstrating that Rg(3) competed with anticancer drug for binding to Pgp thereby blocking drug efflux. Furthermore, Rg(3) increased life span in mice implanted with DOX-resistant murine leukemia P388 cells in vivo and inhibited body weight increase significantly.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Effects of ginsenoside on G protein-coupled inwardly rectifying K+ channel activity expressed in Xenopus oocytes.

Recently, we provided evidence that ginsenoside, the active component of Panax ginseng, uses the pertussis toxin-insensitive Galpha(q/11)-phospholipase C-beta3 signal transduction pathway to increase Ca(2+)-activated Cl(-) currents in the Xenopus oocyte. Other investigators have shown that stimulation of receptors linked to the Galpha(q)-phospholipase C pathway inhibits the activity of G protein-coupled inwardly rectifying K(+) (GIRK) channels. In the present study, we sought to determine whether ginsenoside influenced the activity of GIRK 1 and GIRK 4 (GIRK 1/4) channels expressed in the Xenopus oocyte, and if so, the underlying signal transduction mechanism. In oocytes injected with GIRK 1/4 channel cRNA, bath-applied ginsenoside inhibited the high K(+) solution-elicited GIRK current (EC(50): 4.9+/-4.3 microg/ml). Pretreatment of the oocyte with pertussis toxin reduced the high K(+) solution-elicited GIRK current by 49%, but it did not alter the inhibitory effect of ginsenoside on the GIRK current. Prior intraoocyte injection of cRNA(s) coding Galpha(q), Galpha(11) or Galpha(q)/Galpha(11), but not Galpha(i2) or Galpha(oA), attenuated the inhibitory ginsenoside effect. Injection of cRNAs coding Gbeta(1)gamma(2) also attenuated the ginsenoside effect. Preincubation of GIRK channel-expressing oocytes with phospholipase C inhibitor, [1-[6-((17b-3-Methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl]-1H-pyrrole-2,5-dione] (U73122), or protein kinase C inhibitor, staurosporine or chelerythrine, blocked the inhibitory ginsenoside effect on the GIRK current. Intraoocyte injection of bis (o-aminophenoxy)ethane-N,N,N',N'-tetracetic acid (BAPTA), a free Ca(2+) chelator, had no significant effect on the action of ginsenoside. Taken together, these results suggest that ginsenoside inhibits the activity of the GIRK 1/4 channel expressed in the Xenopus oocyte through a pertussis toxin-insensitive and Galpha(q/11)-, phospholipase C- and protein kinase C-mediated signal transduction pathway.

Animals↗

Ginsenoside Rg1 attenuates dopamine-induced apoptosis in PC12 cells by suppressing oxidative stress.

In Parkinson's disease, neuroprotective therapy to rescue dopamine neurons has been proposed. Ginsenoside Rg1, one of the biologically active ingredients of ginseng, may be a candidate neuroprotective drug. In the present study, the mechanism underlying the neuroprotection provided by ginsenosde Rg1 was studied against apoptosis induced by exogenous dopamine in PC12 cells. Pretreatment with ginsenoside Rg1 markedly reduced the generation of dopamine-induced reactive oxygen species and the release of mitochondrial cytochrome c into the cytosol, and subsequently inhibited the activation of caspase-3. In addition, Rg1 pretreatment also reduced inducible nitric oxide (NO) synthase protein level and NO production. These results suggested that ginsenoside Rg1 may attenuate dopamine-induced apoptotic cell death through suppression of intracellular oxidative stress, and that it may rescue or protect dopamine neurons in Parkinson's disease.

Animals↗

Ginsenoside Rg3 mediates endothelium-dependent relaxation in response to ginsenosides in rat aorta: role of K+ channels.

The aim of the present study was to characterize the endothelium-dependent relaxation elicited by ginsenosides, a mixture of saponin extracted from Panax ginseng, in isolated rat aorta. Relaxations elicited by ginsenosides were mimicked by ginsenoside Rg1 and ginsenoside Rg1, two major ginsenosides of the protopanaxatriol group. Ginsenoside Rg3 was about 100-fold more potent than ginsenoside Rg1. The endothelium-dependent relaxation in response to ginsenoside Rg3 was associated with the formation of cycle GMP. These effects were abolished by N(G)-nitro-L-arginine and methylene blue. Relaxations in response to ginsenoside Rg3 were unaffected by atropine, diphenhydramine, [D-Pro2, D-Trp7,9]substance P, propranolol, nifedipine, verapamil and glibenclamide but were markedly reduced by tetraethylammonium. Tetraethylammonium modestly reduced the relaxation induced by sodium nitroprusside. These findings indicate that ginsenoside Rg3 is a major mediator of the endothelium-dependent nitric oxide-mediated relaxation in response to ginsenosides in isolated rat aorta, possibly via activation of tetraethylammonium-sensitive K+ channels.

Adrenergic beta-Antagonists↗

Use of alternative and complementary medicine in menopause.

OBJECTIVES: To review the clinical evidence available for the treatment of menopausal symptoms with alternative and complementary medicine. METHODS: The MEDLINE, PREMEDLINE and COCHRANE electronic databases for the years 1980-2002 were searched for articles concerning soy products, black cohosh, dong quai, acupuncture, ginseng and evening primrose oil. Studies pertaining to menopausal vasomotor symptoms, lipid profiles and bone mineral densities of postmenopausal women were included. The data from clinical trials were reviewed. RESULTS: Soy isoflavones slightly decrease total cholesterol and LDL levels. The clinical significance of this small change is yet to be determined. The synthetic isoflavone derivative ipriflavone increases bone mineral density in healthy peri- and postmenopausal women with moderate bone mineral densities. Although earlier reports have claimed that soy is beneficial for the improvement of vasomotor symptoms, recent data do not support this claim. There are insufficient data on the other alternative therapies for treating menopausal symptoms at this time. CONCLUSION: Alternative and complementary medicine may play a role in the management of menopause, however, well-designed large studies are still needed.

Angelica sinensis↗

Micelle-mediated extraction and preconcentration of ginsenosides from Chinese herbal medicine.

The feasibility of employing micelle-mediated extraction as an alternative and effective method for the solubilization, purification and/or preconcentration of active ingredients from herbal products is demonstrated for the first time using the root of American ginseng as a model. When compared to methanol and water, an aqueous surfactant solution containing 10% Triton X-100 yielded faster kinetics and higher recovery for the extraction of various ginsenosides. An experimental design approach (uniform design) was demonstrated as a novel and useful method for the optimization of experimental factors involved in the micelle-mediated extraction process. For the preconcentration of ginsenosides prior to chromatographic determination, a salting-out agent (sodium sulfate) was employed to make the efficient cloud point extraction of both hydrophobic and hydrophilic ginsenosides into the surfactant-rich phase possible, as well as to increase the preconcentration factor by reducing the volume of the surfactant-rich phase.

Chromatography, High Pressure Liquid↗

Simultaneous determination of ginsenosides and saikosaponins by high-performance liquid chromatography.

Octadecyl porous glass was used as the packing for reversed-phase high-performance liquid chromatography. A mixture of ginsenosides and saikosaponins (saponins of ginseng and bupleurum root, respectively) were analysed with detection at 203 nm. A well resolved chromatogram of ginsenoside Rb1, Rc, Rb2 and Rd and saikosaponin a, b2 and c was obtained with acetonitrile-water (25.5:74.5) as the mobile phase. The whole separation was achieved in 20 min with a flow-rate of 1.5 ml/min. Calibration graphs for ginsenoside Rb1, Rc, Rb2 and Rd and saikosaponin a and c were linear up to 5 micrograms. Rapid and accurate simultaneous determinations of the saponins are possible by the described method.

Chromatography, High Pressure Liquid↗

High-performance liquid chromatography of glycosides on a new type of hydroxyapatite column.

High-performance liquid chromatography on a column of newly developed hard spherical hydroxyapatite was applied to the separation of a variety of glycosides of tri- and diterpenes including Ginseng saponins and Stevia sweet glycosides, affording satisfactory results by using a simple solvent system, aqueous acetonitrile. This normal phase chromatography is superior to chromatography on a silica gel column for the separation of water-soluble glycosides.

Chromatography, High Pressure Liquid↗

Effects of Sheng-Mai injection on the PRPP synthetase activity in BFU-es and CFU-es from bone marrows of mice with benzene-induced aplastic anemia.

160 Kunming mice were divided at random into 3 groups. Group 1: normal control (40 mice). Group 2: aplastic anemia (AA) control (60 mice); benzene inhalation was carried out for 2.5 months and sterilized normal saline was injected i.p. for another 6 weeks. Group 3: treated AA (60 mice); benzene was administered by inhalation in a similar manner, Sheng-Mai Injection (SMI) was administered i.p. for 6 weeks after the AA models were established. SMI is a famous Chinese traditional prescription of Panax ginseng C.A. Meyer (0.1 g/ml), Ophiopogon japonicus (Thunb.) Ker-Gawl (0.312 g/ml) and Fructus Schisandrae (0.158 g/ml). Activities of phosphoribosylpyrophosphate (PRPP) synthetase in BFU-Es and CFU-Es were estimated by ion pair reversed phase HPLC (IPrHPLC). Accompanying the sharp drop in counts of erythroid progenitor cells, the PRPP synthetase activity in CFU-Es of AA mice was reduced significantly (P<0.01), whereas there were no remarkable changes of this enzyme activity in their BFU-Es compared with the control group. Both the counts of erythroid progenitor cells and PRPP synthetase activity in CFU-Es returned nearly to normal levels following treatment with SMI of mice in Group 3 (P<0.01). Our results suggest that the attenuation of PRPP synthetase activity in peripheral erythrocytes of AA patients may originate from the weakening of activity of this enzyme in CFU-Es from their bone marrow. The impairment of PRPP formation would explain ATP depletion and disorders of energy metabolism in AA erythrocytes. SMI can distinctly increase the reduced quantity of erythroid progenitor cells and promote rapid restoration of PRPP synthetase activity in CFU-Es of AA mice.

Administration, Inhalation↗

Ginsenoside Rf potentiates U-50,488H-induced analgesia and inhibits tolerance to its analgesia in mice.

In the present study, the effect of ginsenoside Rf (Rf), a trace component of Panax ginseng on U-50,488H (U50), a selective kappa opioid-induced analgesia and its tolerance to analgesia was studied using the mice tail-flick test. In addition, the possible mechanism by which Rf may affect U50-induced analgesia was investigated. Intraperitoneal administration of U50 (40 mg/kg) produced analgesia. Rf (10(-14)-10(-10) mg/kg) on co treatment dose-dependently potentiated the U50 (40 mg/kg)-induced analgesia. Rf (10(-12)-10(-2) mg/ml) did not alter the binding of [3H] naloxone, a opioid ligand and [3H]PN200-110, a dihydropyridine ligand to mice whole brain membrane. Twice daily administration of U50 (40 mg/kg) for six days induced tolerance to its analgesia. Chronic treatment (day 4-day 6) of Rf (10(-14)-10(-10) mg/kg) to U50-tolerant mice, dose-dependently inhibited the tolerance. The inhibition of tolerance to U50-induced analgesia by Rf was not altered by flumazenil (0.1 mg/kg), a benzodiazepine receptor antagonist and picrotoxin (1 mg/kg), a GABA(A)-gated chloride channel blocker on chronic treatment. In conclusion, these findings for the first time demonstrated that ginsenoside Rf potentiates U50-induced analgesia, inhibits tolerance to its analgesia, and suggests that Rf affects U50-induced analgesia via non-opioid, non-dihydropyridine-sensitive Ca(+2) and non-benzodiazepine-GABA(A)ergic mechanisms in mice.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗

Induction of nitric oxide synthase expression by Withania somnifera in macrophages.

Withania somnifera (ashwagandha, Indian ginseng) is an immunostimulant herbal medicine used to improve overall health and prevent diseases, particularly in the elderly. However, the mechanisms underlying its immunostimulant effect is poorly understood. To elucidate the mechanism of Withania somnifera, we investigated the effect of a methanolic extract from the root of Withania somnifera (WS) on nitric oxide (NO) production in J774 macrophages. We found that WS (1-256 microg/ml) produced a significant and concentration-dependent increase in NO production, an effect which was abolished by N(G)nitro-L-arginine methyl ester (L-NAME, 3-300 microM), a non-selective inhibitor of NO synthase (NOS), dexamethasone (10 microM), an inhibitor of protein synthesis and N(alpha-p)-tosyl-L-lysine chloromethyl ketone (TLCK, 0.01-10 microM), an inhibitor of nuclear factor-kappaB (NF-kappaB) activation. Dexamethasone did not have any effect on NO production once NOS had been induced (i.e. 12 h after WS). Moreover, western blot analysis showed that WS increased, in a concentration-dependent fashion, inducible NOS protein expression. These results demonstrate that WS may induce the synthesis of inducible NOS expression likely by acting at transcriptional level. The increased NO production by macrophages could account, at least in part, for the immunostimulant properties of Withania somnifera.

Animals↗