Genistein and Selected Phytoestrogen-Containing Extracts Differently Modulate Antioxidant Properties and Cell Differentiation: an in Vitro Study in NIH-3T3, HaCaT and MCF-7 Cells.

During the last decades, plant extracts containing phytoestrogens have increasingly been used as an alternative to oestradiol hormone replacement therapy. The aim of the present study was to compare the effects of genistein with those of different phytoestrogen-containing plant extracts (from red clover flowers and soybeans) on the proliferation and differentiation of NIH-3T3, HaCaT and MCF-7 cells. Our results showed poor correlations between direct anti/pro-oxidant effects and cytotoxicity of the tested samples. In contrast, genistein showed a direct correlation between significant pro-oxidative effects at cytotoxic concentrations and almost no pro-oxidative effects at non-cytotoxic concentrations. Moreover, the tested red clover extract and genistein induced keratin-8 (luminal and prognostic marker in breast cancer) expression only in MCF-7 cells, but this effect was not seen following treatment with the soybean extract. From this point of view, the effect of consumption of phytoestrogens in oestrogen-positive breast cancer remains to be elucidated. In conclusion, our study demonstrates that various phytoestrogen- containing plant extracts and genistein are able to specifically modulate antioxidant properties and differentiation of studied cells.

[Constituents of Lilium candidum L. and their antioxidative activity]. / Obsahové látky Lilium candidum L. a ich antioxidacná aktivita.

The paper deals with the separation and identification of a flavonoid glycoside from the petals of Lilium candidum L. and the antioxidative properties of the ethanolic extract of the flowers and selected compounds isolated from this species. The isolated flavonoid glycoside was identified as isorhamnetin-3-O-rutinoside by acid hydrolysis, TLC comparison with authentic samples, and UV and mass spectra. Isorhamnetin rutinoside was isolated from Lilium candidum L. for the first time. The antioxidative activity of the ethanolic extract of the flowers and some isolated compounds were determined using DPPH assays.

[Ouabain: from an arrow poison to a new steroidal hormone]. / Ouabaín--od sípového jedu k novému steroidnému hormónu.

For more than 200 years, cardiotonic glycosides have been used for the treatment of congestive heart failure. Ouabain is a well-known arrow poison obtained from different Acokanthera or Strophanthus species. Much information has now accumulated that this plant toxin and its congeners are mammalian steroid hormones involved in the pathophysiology of cardiovascular diseases. There is an interesting fact that 50% patients with essential hypertension have elevated levels of endogenous ouabain. A better knowledge of the interactions of these compounds with the hormones of salt and water metabolism might help to improve the diagnosis and therapy of hypertension.

Potential carcinogenic and inhibitory activity of compounds isolated from Lilium candidum L.

This paper deals with determination of potential carcinogenic and inhibitory activity of 10 compounds isolated from the ethanolic extract of Lilium candidum L. perfonned by DC polarography. The series of investigated compounds consisted of two spirostanol saponins, two pyroline derivatives, jatropham and its glucoside, flavonol kaempferol, 2-fenylethyl-alpha-L-arabinopyranosyl-(1-->6)-beta-D-glucopyranoside, 2-phenylethylpalmitate and methylsuccinic acid. Carcinogenic, resp. mutagenic activity data for these compounds, with the exception of kaempferol, are not available in scientific literature. All tested compounds were reduced in N,N-dimethylformamide in one or two well defined steps. They also fonned a reversible complex with alpha-lipoic acid, a compound serving as an indicator of carcinogenic activity. The marginal diffuse current values of these complexes served as a criterion of a very low potential carcinogenicity. The inhibitory activity of isolates were studied in the presence of 12-O-tetradecanoylphorbol-13-acetate, a specific tumor promoter for an epidermal carcinogenesis, and in the presence of a polyaromatic substance 7,12-dimetylbenz(a)anthracene known as a carcinogen. The spirostanol saponins possessed the highest inhibitory activity (> 70%) and jatropham (66%). The inhibitory activity of jatropham glucoside was significantly lower (49%). Practically zero inhibitory activity was measured for the 2-phenylethylpalmitate and methylsuccinic acid.

TLC and HPLC characteristics of desacetylmatricarin, leucodin, achillin and their 8alpha-angeloxy-derivatives.

Five guaianolides, including two pairs of isomers, from a Hungarian taxon of the Achillea millefolium group were characterized analytically. Different chromatographic systems on TLC and HPLC were developed for the analysis of these compounds. TLC of leucodin, 8alpha-angeloxy-leucodin, achillin, 8alpha-angeloxy-achillin and desacetylmatricarin was performed on silica gel using dichloromethaneacetone and cyclohexane-ethylacetate mixtures as mobile phases. HPLC on stationary phases as LiChrospher RP2, LiChrospher RP8, LiChrospher RP18e, Hypersil BDS C18 and Aquasil C18 required isocratic and gradient systems with different methanol-water mixtures as mobile phases. The presented RF values and retention times allow the identification of the respective 2-oxo-guaianolides which are marker substances for certain non-proazulene containing species. Their TLC and HPLC fingerprints are compared to those of proazulene containing species and are relevant for quality control.

[Determination of selected secondary metabolites and extracts of Philadelphus coronarius L]. / Stanovenie vybraných sekundárnych metabolitov a extraktívnych látok vo Philadelphus coronarius L.

The paper deals with the determination of selected secondary metabolites in the branches and leaves of Philadelphus coronarius L. Colorimetric methods were employed to determine the content of flavonoids (0.63% and 0.01%, respectively) and phenolic substances (3.25% and 0.70%, respectively) in leaves or branches. Also the content of extractive substances was determined in ethanolic, aqueous-ethanolic (85% and 60% ethanol), and aqueous extracts. This yielded 9.56%, 22.46%, 24.93%, and 33.15% of extractive substances in leaves and 2.20%, 6.04%, 8.84%, and 10.39% of extractive substances in branches.

[Anti-yeast activity of ethanol extracts of Lilium candidum L]. / Protikvasinková aktivita etanolových extraktov Lilium candidum L.

The paper deals with anti-yeast activity of ethanolic extracts from the flowers and bulbs of Lilium candidum L., Liliaceae, as well as some compounds isolated from these extracts. Several different methods were used for the determination of anti-yeast activity: Lowry method of protein determination, dilution and cultivation method. The extract from the bulbs was shown to be more active than the extract from the flowers, while isolated compounds were inactive against the tested yeasts.

[Anticomplement activity of saponins from Cynara cardunculus L]. / Antikomplementová aktivita saponínov z Cynara cardunculus L.

Triterpene saponins of ursane and oleane type isolated from the involucral bracts of Cynara cardunculus L. (Asteraceae) were investigated in in vitro assays for their activity on the complement system. The anticomplementary activity of bidesmosidic saponins on the classical pathway activation of the complement was higher than the activity of monodesmosidic saponins, but esterification of the carboxylic group of glucuronic acid in the sugar residue resulted in a significant decrease in anticomplementary activity.

[Constituents of the leaves of Ligustrum delavayanum Hariot]. / Obsahové látky listov Ligustrum delavayanum Hariot.

From the chloroform part of Ligustrum delavayanum Hariot, leaves five constituents were isolated: tyrosol, esculetine, kempferol-7-O-rhamnoside, quercetine-3-O-glucoside, and quercetine. Further six constituents (probably malvidine, two secoiridoid glycosides, caffeic acid monosaccharide ester, quercetine monoglycoside (no hyperoside), and quercetine diglycoside (no rutinoside)) were isolated from the methanolic part of above mentioned drug.

[Sterols in Lilium candidum L]. / Steroly v Lilium candidum L.

From the butanolic extract of petals of Lilium candidum L., beta-sitosterol and beta-sitosterol glucoside were isolated. The isolated compounds were identified by spectroscopic means and by comparison with literature data; beta-sitosterol glucoside was isolated in genus Lilium L. for the first time.

[Apigenin-7-methylglucuronide from Cynara cardunculus L]. / Apigenín-7-metylglukuronid z Cynara cardunculus L.

From an ethanolic extract of the flower buds of Cynara cardunculus L. (Asteraceae), apigenin-7-methylglucuronide and chlorogenic acid were isolated. The isolated compounds were identified by spectroscopic means, by comparison with authentic samples and literature data.

[Monodesmosidic saponins in Cynara cardunculus L]. / Monodezmozidové saponíny z Cynara cardunculus L.

From an ethanolic extract of the flower buds of Cynara cardunculus L. (Asteraceae), two monodesmosidic saponins, cynarasaponin B and a new cynarasaponin K, were isolated. The isolated compounds were identified by spectroscopic means and by comparison with standards and the literature data.

[Nonpolar components of the leaves of Philadelphus coronarius L]. / Nepolárne obsahové látky listov Philadelphus coronarius L.

The paper deals with the isolation of constituents from light petrol and chloroform extracts of the leaves of Philadelphus coronarius L., Saxifragaceae. From the light petrol extract uvaol and 3 beta,28-dihydroxyoleanane-11(12),13(18)-diene were isolated, while coumarins (umbelliferone, scopolin), stigmasteryl-3 beta-D-glucoside and the alcane type carbohydrate-C29H60 were isolated from the chloroform extract. The isolated compounds were identified by spectroscopic means and by comparison with standards and the literature data.

Growth inhibitory effect of ethyl acetate-soluble fraction of Cynara cardunculus L. in leukemia cells involves cell cycle arrest, cytochrome c release and activation of caspases.

An extract of artichoke Cynara cardunculus L. (CCE) has been shown to exhibit antioxidant and antigenotoxic properties. In this study, the ability of CCE to inhibit the growth of L1210 and HL-60 leukemia cells was studied. Treatment of leukemia cells with a variety of concentrations of CCE (500-2500 microg/microL) for 24 h resulted in dose-dependent inhibition of leukemia cell growth. The cell growth inhibition was accompanied by G(0)/G(1) cell cycle arrest and by a loss of cells in S phase. Futhermore, apoptosis detected as a sub-G(0) cell population and apoptotic DNA fragmentation was observed. More detailed analyses of apoptosis induced by CCE in HL-60 cells revealed that apoptosis progressed through the caspase-9/-3 pathway, as release of cytochrome c, caspase-9/-3 activations and specific proteolytic cleavage of poly(ADP-ribose) polymerase. Taken together, the results suggest that CCE exerts an antiproliferative activity on leukemia cells and induces apoptosis of these cells through a mitochondrial/caspase dependent pathway.

Antigenotoxic effect of extract from Cynara cardunculus L.

The extract of artichoke Cynara cardunculus L. (CCE) was investigated for its potential antigenotoxic and antioxidant effects using four experimental model systems. In the Saccharomyces cerevisiae mutagenicity/antimutagenicity assay, CCE significantly reduced the frequency of 4-nitroquinoline-N-oxide-induced revertants at the ilv1 locus and mitotic gene convertants at the trp5 locus in the diploid Saccharomyces cerevisiae tester strain D7. In the simultaneous toxicity and clastogenicity/anticlastogenicity assay, it exerted an anticlastogenic effect against N-nitroso-N'-methylurea-induced clastogenicity in the plant species Vicia sativa L. On the contrary, despite CCE not being mutagenic itself, in the preincubation Ames assay with metabolic activation, it significantly increased the mutagenic effect of 2-aminofluorene in the bacterial strain Salmonella typhimurium TA98. In the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, CCE exhibited considerable antioxidant activity. The SC50 value representing 0.0054% CCE corresponds to an antioxidant activity of 216.8 microm ascorbic acid which was used as a reference compound. Although the mechanism of CCE action still remains to be elucidated, different possible mechanisms are probably involved in the CCE antigenotoxic effects. It could be concluded that CCE is of particular interest as a suitable candidate for an effective chemopreventive agent.

Triterpenoid cynarasaponins from Cynara cardunculus L. reduce chemically induced mutagenesis in vitro.

Saponins, steroid or triterpene glycosides, are known to have a broad spectrum of biological and pharmacological activities. Three different triterpenoid saponins, marked here as 1s, 2s and 3s, from involucral bracts of Cynara cardunculus L. were isolated and their antimutagenic effect was assessed. Using spectrophotometric method it was shown that all three substances, 1s, 2s and 3s, possess very good absorptive capability. The antimutagenic effect of these substances was estimated against acridine orange (AO)- and ofloxacin-induced damage of chloroplast DNA in Euglena gracilis assay. These cynarasaponins were experimentally confirmed to exhibit different, statistically significant activity in reducing damage of chloroplast DNA of the flagellate E. gracilis induced by AO and ofloxacin (p(t) < 0.05-0.01). Our findings suggest that the antimutagenic effect of 1s, 2s and 3s against AO-induced chloroplast DNA impairment could be a result of their absorptive capacity. As far as ofloxacin is concerned, a possible mechanism of the reduction of the chloroplast DNA lesion was not elucidated so far. To our knowledge, these data demonstrate for the first time the antimutagnic activity of saponins isolated from involucral bracts of C. cardunculus exerted through different mode of action.