Phytochemical Analysis, Antioxidant, Antimicrobial, and Cytotoxic Activity of Different Extracts of Xanthoparmelia stenophylla Lichen from Stara Planina, Serbia.

The aim of this study was to identify some of the secondary metabolites present in acetonic, methanolic, and hexanic extracts of lichen Xanthoparmelia stenophylla and to examine their antioxidant, antimicrobial, and cytotoxic activity. Compounds of the depsid structure of lecanoric acid, obtusic acid, and atranorin as well as usnic acid with a dibenzofuran structure were identified in the extracts by HPLC. The acetone extract was shown to have the highest total phenolic (167.03 ± 1.12 mg GAE/g) and total flavonoid content (178.84 ± 0.93 mg QE/g) as well as the best antioxidant activity (DPPH IC50 = 81.22 ± 0.54). However, the antimicrobial and antibiofilm tests showed the best activity of hexanic extract, especially against strains of B. cereus, B. subtilis, and S. aureus (MIC < 0.08, and 0.3125 mg/mL, respectively). Additionally, by using the MTT method, the acetonic extract was reported to exhibit a strong cytotoxic effect on the HeLa and HCT-116 cell lines, especially after 72 h (IC50 = 21.17 ± 1.85 and IC50 = 21.48 ± 3.55, respectively). The promising antioxidant, antimicrobial, and cytotoxic effects of Xanthoparmelia stenophylla extracts shown in the current study should be further investigated in vivo and under clinical conditions.

Chelidonium majus crude extract induces activation of peripheral blood mononuclear cells and enhances their cytotoxic effect toward HeLa cells.

The aim of the study was to examine the immunomodulatory effect of crude Chelidonium majus L ethanolic extract on ex vivo harvested peripheral blood mononuclear cells (PBMNCs). PBMNCs were isolated by density gradient centrifugation. The PBMNC cytotoxicity assay was performed with HeLa tumor cells as target cells. MTT assay was used to estimate the proliferation effect of extract and cytotoxic efficiency of treated PBMNCs. Flow cytometric analysis was used for immunophenotyping. Treatment induced moderate proliferative response, perturbation in PBMNC ratios, and the emergence of some unconventional subpopulations. The percentage ratio of double positive CD4+ and CD8+ T lymphocytes and monocytes, ratio of T and B lymphocytes expressing CD14, and percentage of NK cells expressing CD57 increased after treatment, indicating activation of PBMNC subpopulations. Cytotoxic activity against HeLa cells was enhanced. Activation of PBMNCs and enhancement of their cytotoxic effect toward HeLa cells indicate the immunostimulatory effect of Ch. majus ethanolic extract.

Antioxidant, Antigenotoxic and Cytotoxic Activity of Essential Oils and Methanol Extracts of Hyssopus officinalis L. Subsp. aristatus (Godr.) Nyman (Lamiaceae).

Hyssopus officinalis L. is a well-known aromatic plant used in traditional medicine and the food and cosmetics industry. The aim of this study is to assess the antioxidant, genotoxic, antigenotoxic and cytotoxic properties of characterized hyssop essential oils and methanol extracts. Chemical composition was analyzed by gas chromatography - mass spectrometry (GC-MS) and liquid chromatography with diode array detection and mass spectrometry (LC-DAD-MS), respectively. Antioxidant activity was examined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing/antioxidant power (FRAP) tests; genotoxic and antigenotoxic activity were examined by the comet assay, while cytotoxicity was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide dye (MTT) test against tumor cell lines (SW480, MDA-MB 231, HeLa) and non-transformed human lung fibroblast cell lines (MRC-5). The essential oils were rich in monoterpene hydrocarbons (e.g., limonene; 7.99-23.81%), oxygenated monoterpenes (1,8-cineole; 38.19-67.1%) and phenylpropanoids (methyl eugenol; 0.00-28.33%). In methanol extracts, the most abundant phenolics were chlorogenic and rosmarinic acid (23.35-33.46 and 3.53-17.98 mg/g, respectively). Methanol extracts expressed moderate to weak antioxidant activity (DPPH IC50 = 56.04-199.89 µg/mL, FRAP = 0.667-0.959 mmol Fe2+/g). Hyssop preparations significantly reduced DNA damage in human whole blood cells, induced by pretreatment with hydrogen peroxide. Methanol extracts exhibited selective and potent dose- and time-dependent activity against the HeLa cell line. Results of the current study demonstrated notable H. officinalis medicinal potential, which calls for further investigation.

Naphthoquinone rich Onosma visianii Clem (Boraginaceae) root extracts induce apoptosis and cell cycle arrest in HCT-116 and MDA-MB-231 cancer cell lines.

In the present study, five root extracts of Onosma visianii Clem were investigated for their in vitro cytotoxic activity. On the basis of HPLC-PDA analysis, these extracts have proved to be a rich source of naphthoquinones as natural colourants for food and cosmetic industry. All investigated root extracts contain acetylshikonin, isobutyrylshikonin and α-methylbutyrylshikonin as major compounds. As the most abundant source of active compounds for antitumour therapy, acetone, chloroform and ethyl acetate extracts showed strong cytotoxic activity towards HCT-116 and MDA-MB-231 cancer cell lines. Also, these extracts induced apoptosis and cell cycle arrest in HCT-116 and MDA-MB-231 cancer cell lines.

Chelidonium majus crude extract inhibits migration and induces cell cycle arrest and apoptosis in tumor cell lines.

ETHNOPHARMACOLOGICAL RELEVANCE: Chelidonium majus L (Papaveraceae) is widely used in alternative medicine for treatment of various disorders. Antitumor activities of alkaloids isolated from this plant have been reviewed, while there are only a few studies that examine properties of the whole extract. AIM OF THE STUDY: The aim of the present study was to investigate direct cytotoxic effects, as well as indirect antitumor effects of Chelidonium majus ethanolic extract against different tumor cell lines,. MATERIALS AND METHODS: MTT and SRB assays were performed to estimate cytotoxic effects of Chelidonium majus extract against human tumor cell lines A549, H460, HCT 116, SW480, MDA-MB 231 and MCF-7 and peripheral blood mononuclear cells from healthy individuals. Cell cycle analysis was performed by flow cytometry. Type of cell death induced by extract was determined by flow cytometry and cell morphology assessment. Inhibitory effect on migration of cancer cells was assessed by wound healing assay. RESULTS: Chelidonium majus extract showed selective time- and dose-dependent increase of cytotoxicity in all six cell lines, with individual cell line sensitivities. Extract promoted cell cycle arrest and induced apoptosis. Cotreatment with doxorubicin enhanced cytotoxicity of the drug. Also, inhibitory effect on migration was shown with non-toxic extract concentration. CONCLUSIONS: These results indicate possible usefulness of Chelidonium majus crude extract in antitumor therapy, whether through its direct cytotoxic effect, by prevention of metastasis, or as adjuvant therapy.

Cytotoxic effect of Potentilla reptans L. rhizome and aerial part extracts.

Potentilla reptans L. belongs to the least studied of the plants from Rosaceae family, Potentilla genus. There are no data on cytotoxicity of P. reptans extracts, though traditionaly it was used as antiinflammatory and antiinfective. The aim of these studies was to investigate potential antitumor activity of aqueous extracts (rhizome and aerial parts) of P. reptans on 4T1 mouse breast cancer cell line. _Aqueous extracts of rhizome and aerial parts of P. reptans were tested for cytotoxicity by the MTT colorimetric assay on 4T1 cancer cell line in concentration range 100-800 microg/mL. Aqueous extracts of P. reptans rhizome and aerial parts show concentration dependent cytotoxic effect in the range of tested concentrations. ICE50 value of P. reptans rhizome extract was 280.51 +/- 1.16 microg/mL. IC50 value of P. reptans aerial parts extract was 310.79 +/- 1.22 microg/mL. The significant difference in cytotoxicity among tested concentrations was observed. Aqueous extracts of P. reptans rhizome and aerial parts demonstrated weak cytotoxic activity on 4T1 mouse breast cancer cell line, which is in correlation with current cytotoxicity data for aqueous herbal extracts. Rhizome extract of P. reptans has slightly higher antitumor activity than aerial parts extract. The results represent the first report on cytotoxicity for this plant and further research on human cell lines is indicated.

Peptides with antimicrobial and anti-inflammatory activities that have therapeutic potential for treatment of acne vulgaris.

The pathogenesis of acne vulgaris is multifactorial involving infection of the pilosebaceous unit with Propionibacterium acnes and a cytokine-mediated inflammatory response. Five frog skin-derived antimicrobial peptides ([D4k]ascaphin-8, [G4K]XT-7, [T5k]temporin-DRa, brevinin-2GU, and B2RP-ERa), chosen for their low hemolytic activity against human erythrocytes, were assessed for their effects on the growth of clinical isolates of P. acnes and on the release of pro-inflammatory and anti-inflammatory cytokines from peripheral blood mononuclear (PBM) cells. All peptides inhibited the growth of P. acnes with the highest potency exhibited by [D4k]ascaphin-8 (minimum inhibitory concentration, MIC=3-12.5 µM). Release of TNF-α from concanavalin A (ConA)-stimulated PBM cells was significantly reduced by [D4k]ascaphin-8, [G4K]XT-7, brevinin-2GU, and B2RP-ERa (1 and 20 µg/ml) and by [T5k]temporin-DRa (20 µg/ml). Release of IFN-γ from unstimulated PBM cells was significantly reduced by [D4k]ascaphin-8 and brevinin-2GU (1 and 20 µg/ml). No peptide showed significant effects on Il-17 release. Release of the anti-inflammatory cytokines TGF-ß, IL-4, and IL-10 from both unstimulated and ConA-treated PBM cells was significantly increased by [T5k]temporin-DRa and B2RP-ERa (1 and 20µg/ml). The potent activities of [D4k]ascaphin-8 and [T5k]temporin-DRa in inhibiting the growth of P. acnes and the release of pro-inflammatory cytokines, and in stimulating the release of anti-inflammatory cytokines suggest a possible therapeutic role in the treatment of acne vulgaris.