Jolanta Nazaruk

Polyphenolic compounds and in vitro antimicrobial and antioxidant activity of aqueous extracts from leaves of some Cirsium species

Crude aqueous extracts from leaves of Cirsium arvense, C. oleraceum, C. palustre, C. rivulare and C. vulgare were investigated. The content of tannins in mentioned sources, determined by the weight method with hide powder, varied between 1 and 7.63%. Total phenolic content, analysed by using Folin–Ciocalteaus method, ranged between 54 and 96 mg g−1, was expressed as milligrams of gallic acid per gram of dry extract. Phenolic acids were identified by HPLC method. Antimicrobial activity of those extracts was examined. Cirsium palustre extract was the most active against investigated microorganisms. It was observed that the content of small-molecular phenolic compounds had greater influence on the activity of extracts than tannins. The total antioxidant activity indicated by radical cation 2,2′-azini-di-(3-ethylbenzthiazoline sulphonate) ABTS•+, expressed as total antioxidant status (TAS) ranged from 2.31 to 2.78 mM L−1.

The influence of selected flavonoids from the leaves of Cirsium palustre (L.) Scop. on collagen expression in human skin fibroblasts.

Ten flavonoids belonging to the subclasses of flavones, flavanones and aurones were isolated from methanolic extract of Cirsium palustre leaves after multistep chromatographic separation. Their structures were elucidated with spectroscopic methods. All compounds, except for luteolin 7‐O‐glucoside, were isolated for the first time. Four compounds—eriodictyol 7‐O‐glucoside (6), 6‐hydroxyluteolin 7‐O‐glucoside (11), scutellarein 7‐O‐glucoside (12) and pedalitin (14)—were tested for their effect on collagen expression in normal human dermal fibroblasts. Among them, compound 11 at 40 μM and compound 14, at all concentrations used (1, 20, 40 μM), significantly enhanced the level of total collagen secreted into the medium. Furthermore, compound 11 significantly stimulated type I collagen expression, whereas compound 14 activated type I and III collagen expression at the mRNA level, depending on concentration. MMP‐2 activity was inhibited by all study compounds, with the greatest effect recorded with compound 14 at 20 μM. The lack of effect on collagen content in the medium of compound 6‐ and compound 12‐treated cells, besides an increase in COL1A1 and COL1A2 expression, might be caused by diminished expression of HSP47 gene, resulting in decreased procollagen secretion. Future study of compounds 11 and 14 for their potential therapeutic use in conditions connected with collagen biosynthesis deficiency is required. Copyright

Determination of the total polyphenolic content in Cirsium palustre (L.) leaves extracts with manganese(IV) chemiluminescence detection.

It was found that weak chemiluminescence of manganese(IV)-hexametaphosphate-formaldehyde system was greatly enhanced by plant polyphenolic compounds. Based on this finding, a new flow injection chemiluminescence method (FI-CL) was developed for the determination of the total content of polyphenols in plant extracts. The calibration graph obtained for standard solutions of 6-hydroxyluteolin 7-O-glucoside (6OHLG) was linear in the range 0.001-0.8 μg mL(-1). The method was simple, rapid (203 samples h(-1)) and sensitive with a detection limit of 0.25 ng mL(-1). The FI-CL method was successfully applied to the determination of the total polyphenols content (as 6OHLG equivalents) in methanolic, ethyl acetate and aqueous extracts from leaves of Cirsium palustre (L.). Two types of solvent extraction methods (reflux and ultrasound assisted extraction) were used and compared in terms of extraction efficiency. A positive, significant linear correlation between the results obtained by FI-CL method and spectrophotometric methods was observed.

Chemical composition of the essential oils from the roots of Erigeron acris L. and Erigeron annuus (L.) Pers.

The chemical compositions of essential oils from the roots of Erigeron acris and Erigeron annuus were studied. The essential oils were obtained by hydrodistillation in 1.0% and 0.05% yield, respectively, and analyzed by GC, GC-MS. Fifty four and forty seven constituents were identified. Predominant constituents of both oils were poly-acetylene esters: (Z,Z)-matricaria ester (49.4% and 45.9%, respectively) and (Z)-lachnophyllum ester (37.2% and 27.5%, respectively), that were accompanied by their stereoisomers as well as appropriate lactones. Polyacetylenic compounds amounted to 92.1% of E. acris oil and 85.8% of E. annuus oil. Both oils contained the same monoterpene hydrocarbons, amounting to 4.2% and 5.8%, respectively, and traces of almost the same monoterpene oxygenated compounds. The dominant sesquiterpenes in E. acris were elemenes and tricyclic sesquiterpene hydrocarbons, while in E. annuus β-sesquiphellandrene and β-bisabolene dominated. After flash chromatography of essential oil from E. acris, fractions contained acetylene esters and acetylene lactones were obtained. The configuration about double bonds for these compounds has been elucidated on the basis of 1H- and 13C-NMR analysis.

Phytochemical profile and therapeutic potential of Viscum album L.

Viscum album L., the European mistletoe, is a common species from the Viscaceae family. This evergreen hemiparasitic shrub grows on various trees and contains diverse, biologically active substances. Its chemical composition may vary depending on the time of harvest, species of the host tree and the manufacturing process. Among well-described and most active phytochemicals identified in V. album are lectins and viscotoxins, which play substantial role in cancer treatment because of their apoptotic and cytotoxic effects. Another group of compounds found in mistletoe are phenolic acids, phenylpropanoids and flavonoids with antioxidant and anti-inflammatory activities, which decrease blood pressure. Other mistletoe components include, among others, triterpenes with cytotoxic and apoptotic properties, and phytosterols, oligo- and polysaccharides. Extracts from the plant, especially aqueous, are applied in traditional and official medicine, among others in treating hypertension or arthritis. Potentially, it can also be used as a hepatoprotective or a sedative drug.

Determination of polyphenolic compounds in Cirsium palustre (L.) extracts by high performance liquid chromatography with chemiluminescence detection

The first method for the simultaneous determination of polyphenolic antioxidants in extracts from leaves of Cirsium palustre based on high performance liquid chromatography combined with flow injection chemiluminescence detection (HPLC-FI-CL) has been developed. The extracts were prepared by using methanol as extraction medium and two types of extraction methods (reflux and ultrasound assisted extraction). The post-column CL determination of polyphenols was based on their enhancing effect on the chemiluminescence intensity generated in manganese(IV)-hexametaphosphate-formaldehyde system in a phosphoric acid medium. Main antioxidants determined in C. palustre leaves were eriodictyol-7-O-glucoside, luteolin-7-O-glucoside and 6-hydroxyluteolin-7-O-glucoside belonging to flavonoids, and chlorogenic acid belonging to phenolic acids. Chromatographic separation was carried out on a C18 column with gradient elution by using a mobile phase containing 0.25% (v/v) phosphoric acid in water (solvent A) and 100% methanol (solvent B). Under the optimized conditions of chromatographic separation and CL detection the validation of the method was performed. The calibration curves showed good linearity in the concentration range from 0.5 to 40 µg mL(-1). The HPLC-FI-CL method was successfully applied to the determination of four polyphenolic compounds in methanolic extracts from leaves of C. palustre. The accuracy of the developed method was confirmed by the comparison of the results with those obtained by an HPLC-PDA method. The relative error of determination does not exceed 6.1%. However, the HPLC-FI-CL method is characterized by 40-65 times higher sensitivity compared to the HPLC-PDA method.

Determination of the flavonoids/antioxidant levels in Cirsium oleraceum and Cirsium rivulare extracts with cerium(IV)–rhodamine 6G chemiluminescence detection

The determination of the sum of flavonoid compounds in extracts from inflorescences (expressed as mgL(-1) of apigenin) and leaves (expressed as mgL(-1) of linarin) of Cirsium oleraceum and Cirsium rivulare species by flow injection system with chemiluminescence detection (FI-CL) has been carried out. The method is based on the strong enhancement by polyphenols occurring in both plants of the CL signal generated by the reaction of cerium(IV) with rhodamine 6G in a sulfuric acid medium. Under the optimized conditions, the linear working ranges of 0.1-10 and 2.5-50μmolL(-1) were obtained for apigenin and linarin, respectively. The developed method is simple, sensitive with the detection limits of 38nmolL(-1) (apigenin) and 840nmolL(-1) (linarin) and offers high sample throughput (up to 300 samples per hour). The relative standard deviation was 0.62% and 3.75% for 10 measurements of 5μmolL(-1) apigenin and linarin, respectively. The proposed method has been successfully applied to determine the flavonoids/antioxidant levels in aqueous and methanolic extracts from inflorescences and leaves of C. oleraceum and C. rivulare. A possible mechanism of the enhancement of cerium(IV)-rhodamine 6G CL system by polyphenols was briefly discussed. For comparative studies, the antioxidant activity of C. oleraceum and C. rivulare extracts was also evaluated by spectrophotometric 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging method.

Enhancement of antibacterial effects of extracts from Cirsium species using sodium picolinate and estimation of their toxicity

In this study, antimicrobial properties and toxicity of extracts from Cirsium spp.: Cirsium arvense, C. oleraceum, C. palustre, C. rivulare and C. vulgare in combination with sodium picolinate (PS) or sodium benzoate (BS), were investigated. Three micro-organisms were used: Staphylococcus aureus, Bacillus subtilis and Pseudomonas aeruginosa. Minimum inhibitory concentration (MIC) of extracts was found at 1.56–50.0 mg mL−1. Unlike the case of BS, adding PS to extracts from flowers of C. palustre and C. arvense enhanced their antimicrobial effect on S. aureus (MIC from 6.25–12.5 mg mL−1 to 1.25–5.0 mg mL−1). An MTT test was used to study toxicity effects. The extracts from C. palustre or C. arvense mixed with PS had a concentration-dependent, slightly cytotoxic or stimulating effect on the viability of normal human skin fibroblasts. The total phenolic content (TPC) of samples varied from 44 to 178 mg gallic acid equivalent per 1 g of extract. The highest TPC was observed in C. palustre (l) and C. oleraceum (f). Our results did not show any correlation between antimicrobial activities and TPC. Cirsium palustre (f) and C. arvense (f) extracts were analysed by gas chromatography/mass spectrometry (GC/MS). About 30 compounds were found to be present in extracts from two Cirsium species in amounts of not less than 0.2% of TIC.

In vitro Antiproliferative and Antifungal Activity of Essential Oils from Erigeron acris L. and Erigeron annuus (L.) Pers.

Antiproliferative and antifungal activities of essential oils from Erigeron acris root and herb and from Erigeron annuus herb were investigated. The cell viability assay was performed in cultured fi broblasts, cancer cell lines (MCF-7 and MDA-MBA-231), and endometrial adenocarcinoma (Ishikawa) cells as well as colon adenocarcinoma (DLD-1) cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The essential oil from E. acris root showed the highest antiproliferative activity in the MCF-7 cell line with an IC50 value of 14.5 μg/mL. No effect of the essential oil on normal cells at that concentration was found. Antifungal activity against various strains of five Candida species, i.e. C. albicans, C. glabrata, C. tropicalis, C. krusei, and C. parapsilosis, was tested by the microdilution method. It was found that all examined oils can be useful as antifungal agents against the abovementioned species, but the essential oil of E. acris herb was the most active. Their minimum inhibitory concentrations (MIC) ranged from 30 to 0.4 μL/mL. The data presented suggest that essential oils from E. acris and E. annuus possess antifungal activity against Candida spp. and antiproliferative activity against breast cancer MCF-7 cells

Scutellarin-dependent inhibition of collagen biosynthesis in cultured fibroblasts.

The effects of the flavonoid compound scutellarin (SCUT) on collagen biosynthesis, prolidase activity, expression of β1 integrin, insulin-like growth factor-I (IGF-I) receptor and the transcription factor NF-κB were evaluated in human dermal fibroblasts. Confluent fibroblasts were treated with micromolar concentrations (10–30 µM) of SCUT for 24 h. It was found that a SCUT-dependent decrease in collagen biosynthesis was accompanied by an increase in prolidase activity. Since the IGF-I receptor (IGF-IR) is the most potent regulator of both collagen biosynthesis and prolidase activity, and prolidase is regulated by β1 integrin signalling, the effect of SCUT on IGF-IR and β1 integrin receptor expressions were evaluated. It was found that the exposure of the cells to SCUT contributed to an increase in IGF-IR and β1 integrin receptor expressions. This was accompanied by an increase in expression of NF-κB, the known inhibitor of collagen gene expression. These data suggest that the SCUT-dependent decrease of collagen biosynthesis in cultured human skin fibroblasts results from activation of NF-κB, which is responsible for the down-regulation of collagen gene expression.