Dose-dependent in vivo effect of Rhodiola and Echinacea on the mitogen-induced lymphocyte proliferation in mice.

Echinacea purpurea (EP) and Echinacea angustifolia (EA) are ones of the most important world's herbs with immunotropic activity. They were traditional medicinal plants used by North American Indians for the treatment of various illnesses. Now they are cultivated in many countries and are used mainly to treat respiratory tract infections. Rhodiola rosea (RR) and Rhodiola quadrifida (RQ) are medicinal plants originated from Asia and used traditionally as adaptogens, antidepressants, and anti-inflammatory remedies. We previously reported, that extracts of underground parts of RR and RQ exhibited immunotropic activity. We have demonstrated in pigs that in vitro RR or RQ supplementation of blood lymphocyte cultures stimulated T cell proliferative response to Con A in lower, and inhibited it in higher Rhodiola extract concentrations. The aim of this work was to evaluate the in vivo effect of these herbal remedies on the in vitro proliferative response of mouse splenic lymphocytes to another T-cell mitogen- Phaseolus vulgaris haemagglutinin (PHA). We have found significant stimulation of proliferative response, in comparison to the controls, in mice fed lower doses of tested remedies, and inhibition, no effect or lower stimulation, in mice fed higher doses of these drugs.

The effect of Chinese medicinal herb Rhodiola kirilowii extracts on cellular immunity in mice and rats.

UNLABELLED: Rhodiola kirilowii (RK) roots and rhizomes are traditionally used in China as a tonic, adaptogen, antimicrobial and anti-inflammatory drug. The aim of this work was to study the in vivo and in vitro effects of aqueous and 50% hydro-alcoholic extracts of RK rhizomes on some parameters of cellular immunity in H-2d mice and rats. We show for the first time that in vitro both extracts stimulated granulocyte activity and increased lymphocyte response to mitogens, and in vivo they enhanced the ability of lymphocytes derived from parental strain mice fed R. kirilowii aqueous and hydro-alcoholic extracts, to induce local cutaneous graft-versus-host reaction (GVH) in F1 hybrids. CONCLUSION: Rhodiola kirilowii extracts are cellular immunity enhancers.

The influence of Rhodiola quadrifida 50% hydro-alcoholic extract and salidroside on tumor-induced angiogenesis in mice.

UNLABELLED: The genus Rhodiola (Crassulaceae) consists of more than 100 species. They grow mainly in Tibet, China and Mongolia and are traditionally used as tonic, adaptogen, antidepressant and anti-inflammatory drugs. The best known is Rhodiola rosea (R. rosea) now cultivated also in Europe and North America, and present on the market as dietary supplement. Some authors reported anti-tumor activity of R. rosea extracts. Recently, we have published some data on immunomodulatory and antiangiogenic properties of R. rosea. Rhodiola quadrifida (R. quadrifida) belongs to the same family, but is almost not known in Europe, and there is no information about its possible anti-tumor as well as immunotropic and angiotropic activity. The aim of this study was to determine the influence of 50% hydro-alcoholic extract from rhizomes of R. quadrifida (Mongolian origin) and its main biologically active compound salidroside on tumor-induced angiogenesis. Angiogenesis was induced in the skin of Balb/c mice by grafting of syngeneic L-1 sarcoma cells. Mice were fed R. quadrifida extract or salidroside in daily doses 40, 200 and 400 microg, or 0.5, 1, 2, and 4 microg, respectively. After 72 hours, mice were sacrificed with lethal dose of Morbital. All newly formed blood vessels were identified and counted in dissection microscope. RESULTS: It was found that R. quadrifida extract and salidroside highly significantly decreased neovascular reaction in all doses applied.

The effect of Rhodiola quadrifida extracts on cellular immunity in mice and rats.

Rhodiola quadrifida (Rq) roots and rhizomes are traditionally used in Asia as a tonic, adaptogen, antidepressant and anti-inflammatory drug. The aim of this work was to study the in vivo effect of aqueous and 50% hydro-alcoholic extracts of Rq rhizomes on some parameters of cellular immunity in mice and rats. The metabolic activity of blood phagocyting cells was determined based on the measurement of intracellular respiratory burst after stimulation by PMA in RBA test. Potential bactericidal activity of phagocyting cells was determined in isolated blood leukocytes stimulated with killed microorganisms, according to the PKA test. Proliferative response of lymphocytes stimulated by mitogen concanavaline A (ConA) or lipopolysaccharide (LPS) were determined by MTT assay. Both extracts stimulated granulocytes activity in vitro and increased lymphocyte response to mitogens. The ability of parental strain mice lymphocytes to induce local cutaneous graft-versus-host reaction (GVH) in F1 hybrids was stimulated by 50% hydro-alcoholic extract only.

Phytochemical and analytical studies of extracts from Rhodiola rosea and Rhodiola quadrifida.

Column chromatography of hydrophilic extracts from Rhodiola rosea and Rodiola quadrifida led to the isolation of cinnamic alcohol, chlorogenic acid, rhodiooctanoside, rosiridin, rosavin and the phenolic compounds salidroside, rhodiolin and a novel compound consisting of viridoside with an attached arabinose unit (mongrhoside). HPLC analysis of plant material from different sources and from different collection periods showed a great variability in the composition and in the amount of pharmacologically active compounds contained.

The shikonin derivatives and pyrrolizidine alkaloids in hairy root cultures of Lithospermum canescens (Michx.) Lehm.

Hairy root cultures of Lithospermum canescens were established using three strains of Agrobacterium rhizogenes: ATCC 15834, LBA 9402 and NCIB 8196. Eight lines resulting from infection with A. rhizogenes ATCC 15834 demonstrated sufficient biomass increase and were submitted to further investigations. The contents of acetylshikonin (ACS) and isobutyrylshikonin (IBS) in transformed hairy roots made up ca. 10% of those observed in natural roots of L. canescens (24.35 and 14.48 mg g(-1) DW, respectively). One line, Lc1-D, produced the largest amounts of ACS (2.72 mg g(-1) DW) and IBS (0.307 mg g(-1) DW). Traces of pyrrolizidine alkaloids (PA), canescine and canescenine, were found in all lines of transformed hairy roots.

Immunomodulatory effect of shikonin derivatives isolated from Lithospermum canescens on cellular and humoral immunity in Balb/c mice.

The immunomodulatory activity of acetylshikonin (ACS) and isobutyrylshikonin (IBS) was studied in female and male inbred Balb/c mice, and in F1 hybrids (Balb/c x C3H). ACS and IBS were isolated from Lithospermum canescens Lehm. (Boraginaceae) roots. Splenocytes from mice fed 40 microg of ACS had higher proliferative potential in cultures with PHA than corresponding controls and also higher migratory in vitro activity than splenocytes obtained from control animals. ACS at a 40 microg daily dose stimulated G-v-H reaction but inhibited it at a 200 microg dose. IBS at a 40 microg dose significantly increased humoral response.

Detrimental effects of water extracts from surface and interior of Taxus baccata leaves on the two-spotted spider mite (Tetranychus urticae Koch).

Surface deposits on Taxus baccata needles removed by dipping in water of 96, 60 or 40 degrees C for 5 s caused changes in life history components of mites. Paclitaxel was among other peaks present in the removed fractions in concentrations between 0.017 and 0.170 microg/g of fresh weight (f.w.) increasing with temperature. Long extraction for 60 min at only 40 degrees C did not increase removable paclitaxel, but at 60 degrees C extraction rate was the highest (1.326 microg/g) suggesting that leakage from an interior of needles occurred. Mortality, developmental time, total fecundity, oviposition period and life history parameters of Tetranychus urticae Koch. were detrimentally affected.

Antimicrobial activity of Polyscias filicifolia cell biomass extracts.

Antibacterial activity of extracts of Polyscias filicifolia biomass from bioreactor and callus was determined using the agar disc-diffusion method. The microorganisms Staphylococcus aureus (three strains) showed the highest sensitivity to extracts of P. filicifolia biomass from a bioreactor. The values were comparable with nitrofurantoine used as a standard. Micrococcus flavus, Sreptococcus pyogenes and S. agalatiae were less sensitive. The effect of P. filicifolia callus extract on the above bacteria was less pronounced than that of extracts of biomass from a bioreactor.

In vitro propagation of Withania somnifera and isolation of withanolides with immunosuppressive activity.

Withania somnifera plantlets were produced in vitro from the shoot-tip of aseptically germinated seedlings. Culture conditions were optimized using different plant growth regulators which gave rise to 120 shoots from a single bud. The plantlets were then transferred to pots and maintained in greenhouse for 4 months. 90% of these in vitro propagated plantlets survived and showed normal growth. Leaves from these plants were used for isolation of the withanolides. Methanolic extract of leaves from plantlets growing in tissue culture and those transferred to the greenhouse were evaluated for immunomodulatory activity. While the extract from greenhouse samples showed potent immunosuppressive activity, those from tissue cultures samples did not show any activity. Fractionation and characterization of withanolides, using HPLC, NMR, MS methods revealed the presence of withaferin A in the greenhouse samples. Our results indicate that Withania species may require longer time and better differentiation and also natural environment for the production of withaferin A.

Polyprenols in hairy roots of Coluria geoides.

Long-chain polyisoprenoid alcohols built from several up to more than 100 isoprenoid units are common constituents of all living organisms. They were found mostly in plants, bacteria, yeasts and mammalian cells. In vitro hairy root culture of Coluria geoides was obtained from plants transformed with Agrobacterium rhizogenes. Growth was optimal at 0.75% (w/v) glucose and at 22 degrees C. Dry samples of roots were extracted and lipid content was analysed by HPLC. According to our estimation, polyprenols are accumulated in roots of C. geoides cultivated in vitro as a mixture of several prenologues with the dominating prenol composed of 16 isoprenoid units. The content of polyprenols in tissue was approx. 300 microg/g of dry weight.

Sample preparation for taxol and cephalomannine determination in various organs of Taxus sp.

Solid-phase extraction and preparative thin-layer chromatography were applied as sample preparation techniques for the purification of crude extracts from twigs and needles of various Taxus species as well as for the isolation of taxol and cephalomannine for further reversed phase high performance liquid chromatography analysis. Significant differences in the contents of taxanes examined were found. The preparative chromatographic methods used were compared and evaluated as routine and reproducible procedures for the rapid isolation and determination of taxol and cephalomannine in plant extracts.

Taxol on the surface of leaves and inside the needles of three species and two varieties of Taxus.

Leaves of three outside-growing Taxus species (Taxus baccata L.; T. baccava var. elegantissima; T cuspidata Sieb, et Zucc; T. media var. Hatfieldii; T media var. Hickii) and two varieties contained 26-890 µg/g of taxol (paclitaxel®), thus are substantial renewable resources for obtaining this drug. Taxol® was deposited on the surface of these needles in 0.1-1.29 µg/g which was removed with brief dipping in hot water amounting up to 0.58% of total concentration. Two other taxanes were identified (7-epi-10-deacetyltaxol; cephalomannine) but still many await identification. Thus, renewable resources of taxol were found both within and on the leaf surface of different Taxus species. The highest taxol concentrations were found to be in the T. media hybrids, higher than in the case of both original species.

[Searching for cytostatic substances in plant tissue of Maytenus molina by in vitro culture. III. Release of substances from active biological fractions from the callus extract of Maytenus wallichiana R. and B]. / Poszukiwanie substancji cytostatycznych w tkankach roslin z rodzaju Maytenus molina w kulturze in vitro. III. Wydzielenie substancji z aktywnych biologicznie frakcji ekstraktu kalusa Maytenus wallichiana R.et B.

Chromatographic studies controlled by biological evaluation (Tetrahymena test) showed some colour substances among them tingenone and beta-sitosterol. In order to identify the substances extraction and biologically controlled chromatography was carried out. Two substances were isolated, their properties corresponding with tingenone and beta-sitosterol.

[Investigating cytostatic substances in tissue of plants Maytenus Molina in in-vitro cultures. II. chromatographic test of extracts from callus of Maytenus wallichiana R. et B]. / Poszukiwanie substancji cytostatycznych w tkankach roslin rodzaju Maytenus Molina w kulturze in vitro. II. Badania chromatograficzne wyciagów z kalusa Maytenus wallichiana R. et B.

Maytenus wallichiana callus tissue was extracted. The extract was fractionated to achieve maytansine containing residue. In none of the fractions maytansine corresponding substance was found. However evaluation by Tetrahymena test indicated strong cytotoxicity (ID50 1.8 micrograms/cm3). Chromatography of biologically active fractions demonstrated the presence of colour chinone methide triterpenes (tingenone).

[The search for cytostatic substances in the tissues of plants of the genus Maytenus molina in in vitro culture. I. Callas culture and biological studies of its extracts]. / Poszukiwanie substancji cytostatycznych w tkankach roslin rodzaju Maytenus molina w kulturze in vitro. I. Kultura kalusa i badania biologiczne jego wyciagów.

Plant tissue cultures of Maytenus wallichiana Raju et Babu and Maytenus emarginata Ding Hou were initiated. Growth conditions of the callus and the optimum medium composition have been established. Increments of callus wet mass and dynamics of callus growth were determined. Morphological and microscopic observations were also performed. The most efficient growth of the callus, resulting in increments of its wet mass up to 6460%, was obtained on the modified Murashige and Skoog medium. Extracts of the callus were found to be inactive against microorganisms, but proved cytotoxic for lymphocytic leukaemia L 5178Y (ED50 18-48 micrograms/cm3) and the protozoon T. pyriformis (ID50 43-53 micrograms/cm3).

Regeneration of Plants by Embryogenesis with Callus Cultures of Carum carvi L.

This report describes the regeneration of plants from callus cultures of caraway, Carum carvi L. Callus of hypocotyl origin was maintained on medium containing Murashige and Skoog salts with Nitsch and Nitsch vitamins in the presence of 0.3 mg · l(-1) 2,4-D. Formation of somatic embryos was induced with suspension cultures once 2,4-D had been removed. Embryos developed into plantlets when subcultured on solid medium supplemented with 0.5 mg · l(-1) IBA and 10.0 mg · l(-1) adenine sulphate. Regenerated plantlets were transferred to soil.

Mutagenic effects of aqueous extracts of Symphytum officinale L. and of its alkaloidal fractions.

Symphytum officinale L. (Boraginaceae) is a medicinal plant widely used in therapy. It roots, described in the Polish Pharmacopoeia as Radix symphyti, are recommended as expectorants, especially for children. Aqueous solutions of three alkaloid fractions obtained from infusions of Symphytum officinale L. root were tested for their antimitotic and mutagenic activity in meristematic cells of the lateral roots of Vicia faba L., var minor. Lasiocarpine, a proven carcinogen, served as a positive control. Mutagenic effects were induced by lasiocarpine, by the alkaloidal fraction I and by diluted infusions from Radix symphyti. Fraction III had only antimitotic effect. The biological activity of the tested solutions is discussed in relation to the relevant literature.