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Antioxidant and immuno-enhancing effects of Echinacea purpurea.

We studied the protective effects of Echinacea purpurea against radiation by evaluating changes in the peripheral blood cell count and peripheral blood antioxidant activity. E. purpurea administration had a suppressive effect on radiation-induced leukopenia, especially on lymphocytes and monocytes, and resulted in a faster recovery of blood cell counts. Mouse peripheral blood antioxidant activity was increased by E. purpurea, and a relationship between the suppressive effect on radiation-induced leukopenia and the antioxidant effect was suggested. Furthermore, we reviewed the evidence of augmentation of found in this study humoral immunity. The effects of immune activation by E. purpurea were investigated by measuring total immunoglobulin (IgG, IgM). The radioprotective effects of immune activation by E. purpurea were investigated by measuring T lymphocyte subsets in the peripheral blood of mice following whole-body irradiation. E. purpurea activates macrophages to stimulate IFN-gamma production in association with the secondary activation of T lymphocytes, resulting in a decrease in IgG and IgM production. Cytokines released from macrophages in mouse peripheral blood after E. purpurea administration activated helper T cells to proliferate. In addition, it is reported that activated macrophages in association with the secondary T lymphocyte activation increases IFN-gamma production and stimulates proliferation of cytotoxic T cells and suppressor T cells. We think that CD 4 and CD 8 subsets were more immunologically enhanced by E. purpurea than helper T cells and suppressor T cell these results reflect activation. In addition, we think that these results reflect cell-mediated immune responses.

Adjuvants, Immunologic↗

Does the extract of the plant Echinacea purpurea influence the clinical course of recurrent genital herpes?

An increasing proportion of the population perceive complementary medicine as a safer alternative for non-life threatening conditions such as genital herpes. The extract of the plant Echinacea purpurea (Echinaforce) has been shown to have immunomodulating properties and has been advocated in the lay press for the treatment of genital herpes. This study, a single centre, prospective, double blind, placebo-controlled cross-over trial set out to assess whether an extract of the plant and root of E. purpurea can prevent or decrease the frequency and severity of genital herpes recurrences. These were assessed using a detailed history and clinical review of symptoms. Visual analogue scales were used for documentation and haematological and immunological parameters were measured. Over a one-year period, 50 patients took part in the study receiving 6 months' placebo and 6 months' Echinaforce each. No statistically significant benefit could be detected in this study comparing placebo versus Echinaforce in the treatment of frequently recurrent genital herpes.

Adult↗

Isolation, culture, and plant regeneration from Echinacea purpurea protoplasts.

A plant regeneration system from the isolated protoplasts of Echinacea purpurea L. using an alginate solid/liquid culture is described in the chapter. Viable protoplasts were isolated rom 100 mg of young leaves of 4-wk-old seedlings in an isolation mixture containing 1.0% cellulase Onozuka R-10, 0.5% pectinase, and 0.3 mol/L mannitol. After isolation and purification, the mesophyll protoplasts were embedded into 0.6% Na-alginate at the density 1 x 10(-5) mL and cultured in modified Murashige and Skoog (MS) culture medium supplemented with 0.3 mol/L sucrose, 2.5 micromol/L benzylaminopurine (BA), and 5.0 micromol/L 2,4-dichlorophenoxyacetic acid (2,4-D). The visible colonies were present after 4 wk of culture. The protoplast-derived clones were transferred onto gellan gum-solidified basal medium supplemented with 1.0 micromol/L BA and 2.0 micromol/L indole-3-butyric acid (IBA) and formed compact and green calli. Shoot development was achieved by subculturing the calli onto the same basal medium supplemented with 5.0 micromol/L BA and 2.0 micromol/L IBA. Further subculture onto basal medium resulted in the regeneration of complete plantlets.

Cell Culture Techniques↗

Effects of oral administration of Echinacea purpurea (American herb) on incidence of spontaneous leukemia caused by recombinant leukemia viruses in AKR/J mice.

Four-week-old female AKR/J mice were given oral doses of powdered leaves from Echinacea purpurea three times weekly for 8 weeks (7.5 mg/mouse/week): controls received phosphate-buffered saline. Mean survival age of experimental AKR/J mice treated with the E. purpurea preparation was significantly prolonged and enlargement of thymic lymphoma in experimental mice was significantly suppressed compared with controls. In normal 3-week-old female AKR/J mice, mortality from thymic lymphoma was delayed markedly after injection into the thymus of cell-free extract of thymus from the experimental 28-week-old female AKR/J mice that received the oral E. purpurea preparation was injected directly into the thymus. Proliferation of endogenous recombinant murine leukemia viruses (MuLV) in the thymus was markedly inhibited after the first oral administration of the E. purpurea preparation as compared with untreated controls (final age, 28 weeks). Production of endogenous interferon (IFN)-gamma in AKR/J mice was also effectively augmented by the oral treatment with the E. purpurea preparation, however, the production of other cytokines such as tumor necrosis factor (TNF)-alpha and interleukin (IL)-12 was minimal. These results suggest that this suppressive effects on spontaneously occurring leukemia caused by endogenous recombinant MuLV in female AKR/J mice may depend on enhancement of nonspecific immune or cellular immune systems (or of both) by the E. purpurea preparation.

Animals↗

Bioavailability and pharmacokinetics of Echinacea purpurea preparations and their interaction with the immune system.

Echinacea is a widely used herbal remedy for the prevention and treatment of the common cold. Recently, many new insights concerning the molecular mode of action of the main lipophilic constituents, the alkamides, have renewed interest in this plant. In order to compare the bioavailability of alkamides from liquid and tablet preparations of E. purpurea (Echinaforce) in humans and to study the effects on ex vivo stimulated blood cells, a randomized, single-dose, crossover study with 10 (8 test, 2 placebo) volunteers has been performed. They received either 4 ml of the standardized E. purpurea (Echinaforce) tincture or 12 E. purpurea (Echinaforce) tablets or placebo. Both doses contained the same amount (0.07 mg) of the major alkamides, dodeca-2E,4E,8Z, 10E/Z-tetraenoic acid isobutylamides. Liquid chromatography electrospray ionization ion-trap mass spectrometry was used to determine the content of alkamides in serum. It was found that the arithmetic mean C(max) of dodeca-2E,4E, 8Z,10E/Z-tetraenoic acid isobutylamides absorbed after oral application of the Echinaforce tincture appeared after 30 min (0.40 ng/ml serum). In comparison, the t(max) of tablets was 45 min with a C(max) of 0.12 ng/ml. An ex vivo stimulation of blood by LPS was carried out to measure the influence of E. purpurea on the innate and adaptive immune system. Both E. purpurea preparations led to the same effects on the immune system according to the concentration of pro-inflammatory cytokines TNF-alpha and IL-8. 23 hours after oral application a significant down-regulation of TNF-alpha and IL-8 in LPS pre-stimulated whole blood was found. However, no significant changes in the concentration of IL-6 were observed. Although a quarter of the dodeca-2E,4E,8Z, 10E/Z-tetraenoic acid isobutylamides was absorbed from the tablets, the study shows that the formulations trigger the same effects on the measured immune parameters.

Adult↗

Deleterious effects of Echinacea purpurea and melatonin on myeloid cells in mouse spleen and bone marrow.

The neurohormone, melatonin, a product of the pineal gland, is a potent immune cell stimulant. Phytochemicals contained in root extracts of the plant species Echinacea purpurea are also potent as immune cell stimulants. Both agents are potent stimulants of T, B, and/or natural killer cells, but little is known of their effect on other hemopoietic cells, specifically granular leukocytes, also participants in a wide variety of disease defense processes. Given their current popularity and availability for amelioration of a) jet lag and sleep disorders (melatonin) and b) virus-mediated respiratory infections (E. purpurea), we investigated the effects of these agents on granular leukocytes and their precursors, myeloid cells. Mice received these agents daily for 7 or 14 days via the diet, thus mimicking human administration, after which spleens and bone marrow were removed and assessed for mature, differentiated granulocytes and their myeloid progenitors. The influence of these agents was directly related to the stage of cell maturity. Administration of both agents together resulted in significantly elevated levels of myeloid progenitor cells in both bone marrow and spleen and significantly reduced levels of mature, functional granulocyte progeny in both organs, suggesting a) increased precursor proliferation, b) antiapoptosis among the progenitors, and/or c) inhibition of precursor maturation-the latter readily explaining the paucity of mature granulocyte progeny. In conclusion, individual administration of either the herbal derivative and melatonin was either without effect (E. purpurea) or even advantageous (melatonin) to cells of this lineage, but when administered together, these agents significantly perturbed myelopoiesis.

Animals↗

Immunological activity of larch arabinogalactan and Echinacea: a preliminary, randomized, double-blind, placebo-controlled trial.

UNLABELLED: The immunomodulating effects of two Echinacea species, E. purpurea and E. angustifolia and larch arabinogalactan extracted from Larix occidentalis were examined in a randomized, double-blind, placebo-controlled, prospective four-week clinical trial at a naturopathic medical school research center. SUBJECTS/MATERIALS: Forty-eight healthy female volunteers (22-51 y) were randomly assigned to one of six groups: standardized extract of E. purpurea (EP); ultra-refined E. purpurea/E. angustifolia (urEPA); E. purpurea/E. angustifolia (EPA); E. purpurea/E. angustifolia plus larch arabinogalactan (EPALA); larch arabinogalactan (LA); or placebo. METHODS: Immunological tests with enumerative measurements, stool cultures for Lactobacillus acidophilus and yeast, and health-related quality of life (HRQoL) using the Medical Outcomes Study derived SF-36 self-administered questionnaire were assessed at baseline and at four weeks. RESULTS: Complement properdin increased by 21 percent in the EPA group (p<0.05) and by 18 percent in the EPALA group (p<0.05), compared to the placebo group (p>0.05). SF-36 showed improvements in overall physical health, vitality, and emotional health in the same two groups (EPA and EPALA). DISCUSSION: Volunteers in the EPA and EPALA groups had increased production of complement properdin after four weeks of intervention. The increased complement properdin may be an indication of one aspect of immune system stimulation in patients treated with either E. purpurea/E. angustifolia or E. purpurea/E. angustifolia plus larch arabinogalactan.

Adult↗

[Studies on water-soluble constituents of Echinacea prupurea].

AIM: To study the water-soluble constituents of Echinacea purpurea. METHODS: The compounds were isolated by chromatography method and their structures were identified on the basis of spectral analyses. RESULTS: Five compounds were identified as 2-(4-hydroxylphenyl)-ethyl-O-alpha-L-rhamnopy-ranoasyl-(1-->6)-beta-D- glucopyranoside, named echipuroside A 1); (6S, 9R)-6-hydroxy-3-one-alpha-inonol-9-O-beta-D-glucopyranoside 2), ampelopsisionoside 3); phenylmethyl-6-O-beta-D-xylopyranosyl(1-->6)-beta-D-glucopyranoside 4) and phyenylmethyl-beta-D-glucopyranoside 5). CONCLUSION: Compound 1 is a new compound, the others were isolated from this plant for the first time.

Cyclohexanones↗

Determination of phenolic compounds in dietary supplements and tea blends containing Echinacea by liquid chromatography with coulometric electrochemical detection.

Analytical methodologies with ultrasonic extraction and liquid chromatography (LC) were developed for the determination of phenolic compounds in dietary supplements containing Echinacea. The phenolic compounds determined by these methods included caftaric acid, chlorogenic acid, cynarin, echinacoside, and cichoric acid. Samples from tablets, capsules, and bags of tea blends were extracted by sonication for < or = 30 min with methanol-water (60 + 40). The extracts were centrifuged and filtered, and the filtrates were diluted and analyzed by LC using a reversed-phase column and coulometric electrochemical (EC) detection. The mobile phase was acetonitrile-ammonium formate buffer, pH 3.5 (15.3 + 84.7) containing tetrabutyl ammonium hydrogen sulfate as an ion-pairing reagent. Extraction conditions (e.g., composition of the extraction solvent and sonication time) were optimized for different types of samples. Intra- and interday analytical variations were determined, and intraday analyses were performed by 2 independent analysts using 2 different LC systems. Results were generally comparable. The LC method with EC detection showed better sensitivity and selectivity when compared with LC with ultraviolet detection, although results were similar for the 2 methods for major compounds, i.e., caftaric acid, echinacoside, and cichoric acid. The identities of these major compounds found in samples were confirmed by LC/electrospray ionization mass spectrometry.

Caffeic Acids↗

[Lipid compounds from Echinacea purpurea].

OBJECTIVE: To study the lipid constituents from Echinacea purpurea. METHOD: The compounds were isolated by chromatography method and the structures were identified on the basis of spectral analyses. RESULT: Five compounds were isolated and identified as, 1 beta, 6 alpha-dihydroxy-4(14)-eudesmene(1), (2E, 4E, 8Z, 10E)-N-isobutyl-2,4,8,10-dodecatetraenamide(2), (2E, 4E, 8Z, 10Z)-N-isobutyl-2,4,8,10-dodecatetraenamide(3), cerotic acid(4), hyxacosyl alcohol(5). CONCLUSION: Compounds 1,4 and 5 were obtained from the plant for the first time.

Echinacea↗

[Determination of cichoric acid in Echinacea purpuea].

OBJECTIVE: To analyze the natural change rule of active components of E. purpuea by measuring content of cichoric acid. METHOD: Reverse HPLC method was used. RESULT: The maximum cichoric acid content of the roots occured in seedling age of May, and that of the flowers occured in blooming stage of mid July, but cichoric acid in stems was generally low anyway. The maximum content of cichoric acid in the plant above ground occured in the blooming stage of mid July. CONCLUSION: The measuring method of content of cichoric acid is successful and reliable. The optimum stage of harvest in Echinacea purpuea should be guided by natural change rule of cichoric acid content.

Caffeic Acids↗

[Compensatory reactions of immune system and action of Purple Coneflower (Echinacea purpurea (L.) Moench) preparations].

UNLABELLED: The objective of the study was to investigate the mechanisms of action of Purple Coneflower (Echinacea purpurea (L.) Moench) preparations on the immune system and to define which part of the plant is the most effective as an immunostimulator. MATERIAL AND METHODS: The extracts from overground parts and roots of Purple Coneflower were injected into rabbits at the dose of 1 ml/kg. The stimulation of T lymphocytes in vitro was tested by incubation of rabbit blood with the different concentrations of the Purple Coneflower preparation (10, 50, 100 microg/ml) and assessed by the method of spontaneous rosettes. The number of lymphocytes, which bind to heterogenic erythrocytes, was counted. In order to determine the local effect, 30 ml of Purple Coneflower preparation (95 mg of dry material/1 ml) was used for the rinsing of mouth cavity in humans. The reaction of phagocytosis was assessed by using latex particles and calculating phagocytosis activity and phagocytosis index. RESULTS: Purple Coneflower extracts from roots were more effective phytoimmunostimulators than those from overground parts. They significantly increased in vivo the number of leucocytes and lymphocytes, especially T lymphocytes, in peripheral blood of rabbits as compared with the control group (p<0.001). The stimulation of T lymphocytes by Purple Coneflower preparation in vitro was the most pronounced at the concentration of 50 microg/ml (20.8+/-1.01% of spontaneous rosettes, p<0.001 with control). An increase in concentration till 100 microg/ml was followed by non-specific inhibition (9.16+/-1.6% of spontaneous rosettes, p>0.05 with control). Purple Coneflower preparations activated phagocytosis of neutrophils in vitro and in vivo. After rinsing mouth cavity with 30 ml of Purple Coneflower root preparation for 15 minutes, phagocytosis activity was significantly increased till 36% (p<0.05 as compared with the control (27%), where physiological solution was used for the rinsing). This local application of Purple Coneflower root preparation has significantly increased phagocytosis index up to 1.03 (p<0.02 as compared with the control index (0.65) after rinsing of the mouth cavity with physiological solution). CONCLUSIONS: Purple Coneflower preparations from roots activate the cellular immunity and stimulate phagocytosis of neutrophils in vitro, in vivo and after rinsing of mouth cavity.

Adjuvants, Immunologic↗

[Composite tincture from the tops of the purple coneflower (Echinacea purpurea) and the fruits of the dog rose (Rosa canina): technology of production and evaluation of quality].

This paper aimed to elaborate the technology of the production of a composite tincture with an immunostimulatory and adaptogenic effect, to select suitable methods of quality evaluation, and perform stability tests. The tops of the purple coneflower (Echinaceae herba) and fruits of the dog rose (Cynosbati fructus) in a ratio of 1:1 were employed to prepare the tincture. The tincture itself was prepared in a ratio of 1:5. The experiment revealed that ethanol 50% was the most suitable extracting agent. The optimal conditions for the obtaining of a quality tincture are formed when the drug is disintegrated into the required degree by grinding, percolation is used for extraction, and the rate of flowing of the extract from the percolator is 0.2 ml/min/100g of the drug. The determination of the dry residue, relative density, ethanol concentration and content of cichoric acid (spectrophotometrically), ascorbic acid (oxidimetrically), and tannins (manganometrically) served to evaluate the quality of prepared extracts. Stability tests (temperature 25+/-2 degrees C, relative humidity 60+/-5%, period of 2 years) have shown that the prepared tincture maintains its quality parameters for the given period.

Chemistry, Pharmaceutical↗

Application of purified polysaccharides from cell cultures of the plant Echinacea purpurea to mice mediates protection against systemic infections with Listeria monocytogenes and Candida albicans.

Purified polysaccharides from cell cultures of the plant Echinacea purpurea were investigated for their ability to enhance phagocytes' activities regarding nonspecific immunity in vitro and in vivo. Macrophages (M phi) from different organ origin could be activated to produce IL-1, TNF alpha and IL-6, to produce elevated amounts of reactive oxygen intermediates and to inhibit growth of Candida albicans in vitro. Furthermore, in vivo the substances could induce increased proliferation of phagocytes in spleen and bone marrow and migration of granulocytes to the peripheral blood. These effects indeed resulted in excellent protection of mice against the consequences of lethal infections with one predominantly M phi dependent and one predominantly granulocyte dependent pathogen, Listeria monocytogenes and C. albicans, respectively. Specific immune responses to sheep red blood cells (antibody production) and to listeria (DTH) were not affected by the polysaccharides. The possibility of clinical use is discussed.

Animals↗

Application of purified polysaccharides from cell cultures of the plant Echinacea purpurea to test subjects mediates activation of the phagocyte system.

Polysaccharides purified from large-scale cell cultures of the plant Echinacea purpurea were tested for their ability to activate human phagocytes in vitro and in vivo. These substances enhanced the spontaneous motility of PMN under soft agar and increased the ability of these cells to kill staphylococci. Monocytes were activated to secrete TNF-alpha, IL-6 and IL-1 whereas class II expression was unaffected. Intravenous application of the polysaccharides to test subjects immediately induced a fall in the number of PMN in the peripheral blood, indicating activation of adherence to endothelial cells. This fall was followed by a leukocytosis due to an increase in the number of PMN and a lesser increase of monocytes. The appearance of stab cells and some juvenile forms and even myelocytes indicated the migration of cells from the bone marrow into the peripheral blood. The acute phase C-reactive protein (CRP) was induced, probably due to activation of monocytes and macrophages to produce IL-6. In addition a moderate acceleration of the erythrocyte sedimentation rate was observed. Altogether, as in mice, the polysaccharides could induce acute phase reactions and activation of phagocytes in humans. The possibility of clinical use is discussed.

Adjuvants, Immunologic↗

Polysaccharides isolated from plant cell cultures of Echinacea purpurea enhance the resistance of immunosuppressed mice against systemic infections with Candida albicans and Listeria monocytogenes.

Polysaccharides (EP) isolated from large scale plant cell cultures of Echinacea purpurea, have been shown to activate human and murine phagocytes. In this study we investigated the influence of EP on the nonspecific immunity in immunodeficient mice. EP was effective in activating peritoneal macrophages isolated from animals after administration of cyclophosphamide (CP) or cyclosporin A (CsA). EP-treated macrophages exhibited increased production of tumor necrosis factor-alpha (TNF) and enhanced cytotoxicity against tumor target WEHI 164 as well as against the intracellular parasite Leishmania enrietti. After a CP-mediated reduction of leukocytes in the peripheral blood, the polysaccharides induced an earlier influx of neutrophil granulocytes as compared to PBS-treated controls. EP treatment of mice, immunosuppressed with CP or CsA, restored their resistance against lethal infections with the predominantly macrophage-dependent pathogen Listeria monocytogenes and predominantly granulocyte-dependent Candida albicans. Further, the effects of EP in allogeneic bone marrow chimeric mice are discussed. These findings may have therapeutical implications in prophylactic treatment of opportunistic infections.

Animals↗

Anti-inflammatory activity of Echinacea angustifolia fractions separated on the basis of molecular weight.

Five fractions of an aqueous extract obtained from the roots of Echinacea angustifolia were separated on the basis of molecular weight. The topical anti-inflammatory activity of the fractions has been evaluated in mice using the Croton oil ear test. The fraction with a molecular weight between 30,000 and 100,000 was the most active in inhibiting the oedema; it also reduced the infiltration of inflammatory cells. The activity of this fraction was comparable with that of a raw polysaccharidic extract obtained from E. angustifolia roots by differential solubility. The high-molecular weight polysaccharides are therefore proposed as the anti-inflammatory principles of the plant.

Animals↗

Gas chromatographic method for determination of uracil herbicides in roots of Echinacea angustifolia Moench (Asteraceae).

A GC/NPD method and a rapid screening TLC method were developed for the simultaneous determination of uracil herbicide residues (bromacil, lenacil, terbacil) in the roots of Echinacea angustifolia Moench (Asteraceae). The uracil herbicide residues were extracted into acetone. After evaporation of acetone from the acetone-water extract the residue was dissolved in water-methanol (5:1 v/v). Cyclohexane was used for removal of the non-polar co-extractives in the sample matrix. After separation of the cyclohexane phase the uracil herbicide residues were extracted into chloroform. This extract was purified on a Florisil column, and residues were eluted with dichloromethane-acetone (9:1, v/v). The cleaned up extract was analysed by the GC/NPD method on a capillary column DB-1 using atrazine as internal standard. A minimum recovery of 70% was attained for contamination levels of 0.02-0.40 mg kg(-1).

Asteraceae↗