Irma Podolak

The influence of LTS-4, a saponoside from Lysimachia thyrsiflora L., on human skin fibroblasts and human melanoma cells.

We investigated the effect of a triterpene saponoside from Lysimachia thyrsiflora L. upon the viability, proliferation, morphology and cell motility of human melanoma HTB-140 cells and human skin fibroblasts (HSFs). The compound, denoted LTS-4, decreased the viability and rate of cell growth of both cell types in a time-and dose-dependent manner, and proved cytotoxic against cancer cells at significantly lower concentrations than for fibroblasts. LTS-4 also affected the morphology of the examined cells, causing vacuolisation and actin cytoskeleton disintegration, and had an inhibitory effect on the tumour cell motility.

Triterpene saponosides from Lysimachia ciliata differentially attenuate invasive potential of prostate cancer cells.

Neither androgen ablation nor chemotherapeutic agents are effective in reducing the risk of prostate cancer progression. On the other hand, multifaceted effects of phytochemicals, such as triterpene saponins, on cancer cells have been suggested. A promising safety and tolerability profile indicate their possible application in the treatment of advanced prostate cancers. We analyzed the specificity, selectivity and versatility of desglucoanagalloside B effects on human prostate cancer cells derived from prostate cancer metastases to brain (DU-145 cells) and bone (PC-3 cells). Prominent growth arrest and apoptotic response of both cell types was observed in the presence of sub-micromolar desglucoanagalloside B concentrations. This was accompanied by cytochrome c release and caspase 3/7 activation. A relatively low cytostatic and pro-apoptotic response of cancer cells to a desglucoanagalloside B analog, anagallosaponin IV, illustrated the specificity of the effects of desglucoanagalloside B, whereas the low sensitivity of normal prostate PNT2 cells to desglucoanagalloside B showed the selectivity of its action. Inhibition of cancer cell motility was observed in the presence of both saponins, however only desglucoanagalloside B attenuated cancer cell invasive potential, predominantly through an effect on cell elastic properties. These data demonstrate the versatility of its effects on prostate cancer cells. In contrast to PNT2 cells, cancer cells tested in this study were relatively resistant to mitoxantrone. The multifaceted action of desglucoanagalloside B on basic cellular traits, crucial for prostate cancer progression, opens perspectives for elaboration of combined palliative therapies and new prostate cancer prophylaxis regimens.

Usnic acid and atranorin exert selective cytostatic and anti-invasive effects on human prostate and melanoma cancer cells

OBJECTIVES AND METHODS Lichens are an interesting source of potential anti-tumor compounds, among which usnic acid and atranorin seem to be the most promising, but their impact on invasive potential of tumor cells has not yet been comprehensively addressed. The aim of the study was focused on the impact of the two lichen metabolites, on the viability (by Trypan blue test and fluoresceine diacetate and ethidium bromide assay), proliferation (cell counting in a Bürkers chamber), apoptosis (flow cytometry analysis and Western blot) and motile activity (cell movement recording and image analysis) and actin cytoskeleton organization (immunofluorescent staining) of melanoma HTB-140, prostate cancers DU-145 and PC-3, normal human skin fibroblasts and prostate epithelial PNT2 cells, with special emphasis to their selectivity and versatility. RESULTS Both compounds exerted strong inhibitory effects on cancer cell proliferation, migration and actin cytoskeleton organization, while their effect on apoptosis process was less relevant. The impact of usnic acid on the examined cancer cells was found more efficient in comparison to atranorin. Also, selective effect of both agents on tumor cells was observed. SIGNIFICANCE The ability of usnic acid and atranorin to inhibit cancer cells motility may have future implications for development of new therapeutic strategies targeted at the interference with the metastatic cascade.

A new cytotoxic triterpene saponin from Lysimachia nummularia L.

A new glycosylated triterpene 1 (named nummularoside) was isolated from the underground parts of Lysimachia nummularia L. Its chemical structure was elucidated as 3-O-β-{{[β-D-xylopyranosyl-(1→2)]-[β-D-xylopyranosyl-(1→4)]-β-D-glucopyranosyl-(1→4)}-[β-D-glucopyranosyl-(1→2)-]-α-L-arabinopyranosyl]}, protoprimulagenin A on the basis of extensive NMR and MS spectral data. The saponin showed significant activity against prostate cancer cells DU145 and PC3 (EC50 1.2 and 7.4 μg/mL, respectively), while it did not affect normal cells (EC50 30 μg/mL), in contrast to the reference compound (mitoxanthrone, EC50 0.45 μg/mL). Glioblastoma cells were also significantly affected by the tested saponin (EC50 6.0 μg/mL), whereas the activity against melanoma cells was moderate (EC50 17.5-23.2 μg/mL).

Qualitative and quantitative analysis of diosmin in tablets by thin-layer chromatography with densitometric UV detection

Conditions have been established for determination of diosmin in pharmaceutical formulations, for example ‘Diosminin’, by TLC with densitometric detection. Chromatographic separation on silica gel was performed with chloroform—methanol—water, 23 + 12 + 2 (v/v), as mobile phase and the spots were analyzed by densitometry at λ = 344 nm. Under the experimental conditions established repeatable and accurate results were obtained. The method is characterized by high sensitivity — the limit of detection was 20 ng — and satisfactory recovery — from 99.8 to 103.0%. Response was linearly dependent on concentration in the range 5 to 50 µg mL−1.

Densitometric analysis of diosgenin in methanolic extracts of Allium ursinum collected at different times during plant development

Diosgenin in methanolic extracts of fresh leaves and bulbs of Allium ursinum collected at different times during plant development has been analyzed quantitatively by densitometric TLC. After acid hydrolysis of the extracts, TLC was performed on silica gel plates with n-hexane-acetone 4:1 (v/v) as mobile phase. Bands were visualized with 25% H2SO4 in methanol, and diosgenin was quantified densitometrically at 540 nm. Variation in diosgenin content with development time was noted.

Densitometric determination of genistin and daidzin in different cultivars of soy ( Glycine max )

Conditions have been established for identification and quantification of genistin and daidzin in extracts of different soy cultivars by densitometric thin-layer chromatography. Good resolution of the genistin and daidzin peaks was achieved by chromatographic separation on silica gel F254 HPTLC plates with chloroform—metha-nol—water, 23 + 8 + 1 (v/v), as mobile phase. Under these analytical conditions there was no interference from other extract components.

Densitometric determination of embelin in Lysimachia punctata

Athin layer chromatographic method with densitometric UV detection at λ = 285 nm has been developed for quantification of embelin in Lysimachia punctata. TLC analysis was performed on aluminumbacked silica gel 60 F254 plates with n-hexane—ethyl acetate, 7 + 3 (v/v), as mobile phase. Under these experimental conditions the method was highly sensitive (the limit of detection was 12.5 ng) and recovery was satisfactory (from 89.45% to 92.28%). Good results obtained during validation of the method were confirmed in quantification of embelin, because high precision and accuracy were achieved.

In vitro anti-denaturation and anti-hyaluronidase activities of extracts and galactolipids from leaves of Impatiens parviflora DC

The in vitro anti-denaturation and anti-hyaluronidase activities of Impatiens parviflora extracts and isolated galactolipids (MGDG-1, DGDG-1) were investigated. This is the first report on these compounds in I. parviflora. All extracts showed anti-hyaluronidase activity, but only methanolic extract from fresh leaves exhibited significant activity against heat-induced denaturation of BSA in a dose-dependent manner. At 500 μg/mL, the extract and the reference drug showed 79.05% and 99.81% inhibition of protein denaturation, respectively. These results indicate that fresh leaves of I. parviflora may be beneficial in inflammatory conditions, especially those associated with protein denaturation, such as rheumatoid arthritis. The study revealed that only MGDG-1 showed weak activity in anti-denaturation assay but both galactolipids were potent inhibitors of hyaluronidase. MGDG-1 completely inhibited the enzyme activity at the concentration of 127.9 μg/mL. These results indicate the potential of galactolipids in the treatment of diseases associated with the loss of hyaluronic acid.

Quantification of saponins in different plant parts of Lysimachia L. Species by validated HPTLC-densitometric method

Quantitative high-performance thin-layer chromatography (HPTLC) analysis of triterpene saponins in different parts of eight Lysimachia L. species is described. For separation, silica gel F254 plates were used with chloroform-methanol-water (8:7:1, v/v) as mobile phase 1 and with n-butanol-acetic acid-water (6:1:3, v/v) as mobile phase 2. The method was validated for selectivity, sensitivity, accuracy, precision, and indirect precision. Saponins were most abundant in the underground parts of all analyzed species, where their content ranged from 0.12% in Lysimachia clethroides to 1.18% in Lysimachia ephemerum. The saponin levels in aerial parts of the plant were generally lower, the only exception being L. ephemerum leaf (1.57%) and Lysimachia thyrsiflora stem (2.26%). The method is sensitive with good precision and accuracy and may be suitable for comparison of the differences in saponin content among samples from various related species and genera.