Assessment of Photo-Induced Cytotoxic Activity of Cachrys sicula and Cachrys libanotis Enriched-Coumarin Extracts against Human Melanoma Cells.

Photochemotherapy is one of the most interesting current therapeutic approaches for the treatment of melanoma. Different classes of naturally occurring phytochemicals demonstrated interesting photoactive properties. The aim of this study was to evaluate the photocytotoxic potential of two Cachrys species from Southern Italy: C. sicula and C. libanotis (Apiaceae). The enriched-coumarin extracts were obtained from aerial parts through both traditional maceration and pressurized cyclic solid-liquid (PCSL) extraction using Naviglio extractor®. Qualitative and quantitative analyses of furanocoumarins were performed with GC-MS. The photocytotoxic effects were verified on C32 melanoma cells irradiated at a dose of 1.08 J/cm2. The apoptotic responses were also assessed. Moreover, phenolic content and the in vitro antioxidant potential were estimated. Xanthotoxin, bergapten, and isopimpinellin were identified. All the samples induced concentration-dependent photocytotoxic effects (IC50 ranging from 3.16 to 18.18 µg/mL). The C. libanotis sample obtained with Naviglio extractor® was the most effective one (IC50 = 3.16 ± 0.21 µg/mL), followed by C. sicula sample obtained with the same technique (IC50 = 8.83 ± 0.20 µg/mL). Both Cachrys samples obtained through PCSL induced up-regulation of apoptotic signals such as BAX (Bcl2-associated X protein) and PARP (poly ADP-ribose polymerase) cleavage. Moreover, these samples proved to be more photoactive, giving a greater upregulation of p21 protein in the presence of UVA radiation. Obtained results suggest that investigated species could be promising candidates for further investigations aimed to find new potential drugs for the photochemotherapy of skin cancer.

Fatty acids, coumarins and polyphenolic compounds of Ficus carica L. cv. Dottato: variation of bioactive compounds and biological activity of aerial parts.

Leaves, bark and woody part of Ficus carica L. cultivar Dottato collected in different months were examined to assess their chemical composition, antioxidant activity and phototoxicity on C32 human melanoma cells after UVA irradiation. The phytochemical investigation revealed different composition in the coumarin, fatty acid, polyphenol and flavonoid content. The second harvest of leaves and the first harvest of the bark possessed the highest antiradical activity with IC50 values of 64.00 ± 0.59 and 67.00 ± 1.09 µg/mL, respectively. Harvest III of leaves showed the best inhibition of lipid peroxidation (IC50 = 1.48 ± 0.04 µg/mL). Leaf samples of F. carica showed also the best antiproliferative activity in comparison with bark and woody part of F. carica.

Synthesis of a new bis(indolyl)methane that inhibits growth and induces apoptosis in human prostate cancer cells.

The synthesis and the antiproliferative activity against the human breast MCF-7, SkBr3 and the prostate LNCaP cancer cell lines of a series of bis(indolyl)methane derivatives are reported. The synthesis of new compounds was first accomplished by the reaction of different indoles with trimethoxyacetophenone in the presence of catalytic amounts of hydrochloric acid. A second procedure involving the use of oxalic acid dihydrate [(CO2H)2·2H2O] and N-cetyl-N,N,N-trimethylammonium bromide in water was carried out and led to better yields. Compound 5b significantly reduced LNCaP prostate cancer cell viability in a dose-dependent manner, with an IC50 of 0.64 ± 0.09 µM. To determine whether the growth inhibition was associated with the induction of apoptosis, treated cells were stained using DAPI. LNCaP cells treated with 1 µM of 5b showed the morphological changes characteristic of apoptosis after 24 h of incubation.

Fig latex (Ficus carica L. cultivar Dottato) in combination with UV irradiation decreases the viability of A375 melanoma cells in vitro.

Melanoma and nonmelanoma skin cancers are among the most prevalent cancers in the human population. In the present work latex of Ficus carica cultivar Dottato from Italy collected from fruits and leaves was examined to assess its free radical-scavenging activity with 1,1-diphenyl-2 picrylhydrazyl (DPPH) and its phototoxicity on A375 human melanoma cells. The latex obtained from the fruits of Ficus carica cv. Dottato showed the best antiradical activity with an IC50 value of 0.05 mg/ml while the latex obtained from the leaves showed the best antiproliferative activity with an IC50 value of 1.5 µg/ml on the human tumor cell line A375 (melanoma) after irradiation at a specific UVA dose (1.08 J/cm2). Control experiments with UVA light or drugs alone were carried out without significant cytotoxic effects. Polyphenolic content of the samples was also evaluated. This is the first study comparing F. carica latex of leaves and fruits. Plant derived natural products have long been and will continue to be an important source for anticancer drug development.

Changes in the phenolic and lipophilic composition, in the enzyme inhibition and antiproliferative activity of Ficus carica L. cultivar Dottato fruits during maturation.

Fruits of Ficus carica cultivar Dottato from Italy were examined to assess how the stage of ripeness influences their chemical composition, antioxidant activity, pancreatic lipase inhibition and antiproliferative properties on C32 melanoma cells. Fruits of the first harvest (June) showed a major content in furanocoumarins and pyranocoumarins whereas the fruits collected in September showed the highest polyphenolic content (11.9 mg/g of dried material). The total 70% ethanol extracts were portioned between methanol/water and n-hexane, dichloromethane and ethyl acetate, successively. Coumarins and fatty acid esters were the most abundant components of the n-hexane fractions. The dichloromethane fractions showed as major components 2 furanocoumarins (rutarenin and pimpinellin). The total extracts of F. carica cv. Dottato exhibited a significant dose-dependent antiradical and inhibition of lipid peroxidation activity, particularly fruits of the first harvest (June) that showed the highest activity with IC50 of 1.64 mg/mL and 0.004 mg/mL, respectively. Among single fractions, the ethyl acetate fraction from the second harvest (July) showed the highest antiradical activity with an IC50 value of 0.05 mg/mL while the dichloromethane fraction showed the best inhibition of lipid peroxidation with an IC50 value of 0.02 mg/mL. Dichloromethane fractions showed the highest photodynamic cytotoxicity with an IC50<5 µg/mL.

Chemical composition and protective effect of oregano (Origanum heracleoticum L.) ethanolic extract on oxidative damage and on inhibition of NO in LPS-stimulated RAW 264.7 macrophages.

The present study shows the chemical profile and the in vitro properties (antioxidant and inhibition of nitric oxide production) of the Origanum heracleoticum L. (Lamiaceae). The ethanolic extract of the aerial parts is characterized by terpenes and fatty acids. The extract, with high total phenol and flavonoid content, showed a significant radical-scavenging activity (IC(50) value of 12.8 µg/mL) using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test and an interesting antioxidant activity with the ß-carotene bleaching test (IC(50) values of 12.9 and 14.1 µg/mL at 30 and 60 min of incubation, respectively). The test for the inhibition of NO production, performed using the murine monocytic macrophage RAW 264.7 cell line, showed that the extract had significant activity with an IC(50) value of 108.5 µg/mL. The cytotoxic effect of O. heracleoticum extract in presence of lipopolysaccharide (LPS) (1 µg/mL) was evaluated but found to be negligible.

Phytochemical profile, antioxidant, anti-inflammatory and hypoglycemic potential of hydroalcoholic extracts from Citrus medica L. cv Diamante flowers, leaves and fruits at two maturity stages.

Since the past decade consumption of certain foods has been reported to have a positive effect on health. The object of the study was to determine for the first time the chemical composition and the antioxidant, anti-inflammatory and hypoglycemic potential of Citrus medica L. cv Diamante flowers, leaves and fruits (endocarp and mesocarp) at two maturity stages. Flowers and leaves were characterized by the highest total phenols and flavonoids content. A declining trend was observed during maturity of fruits for both phenols and flavonoids. The antioxidant activity evaluated by the ß-carotene bleaching test showed a strong activity for flowers and endocarp of mature fruits with IC50 values of 2.8 µg/mL and 3.5 µg/mL, respectively, after 30 min of incubation. Interestingly, the mature fruits endocarp (IC50 value of 426.0 µg/mL) could inhibit α-amylase with an IC50 value 2-fold higher than immature fruits. None of the tested extracts affected the proliferation of human skin fibroblasts 142BR. The obtained results suggest a potential use of C. medica L. cv Diamante as new valuable Citrus species with functional properties for food or nutraceutical product on the basis of high content of phytochemicals.

Chemical composition and bioactivity of Citrus medica L. cv. Diamante essential oil obtained by hydrodistillation, cold-pressing and supercritical carbon dioxide extraction.

The chemical composition of the essential oil of Citrus medica L. cv. Diamante peel obtained by hydrodistillation, cold-pressing and supercritical carbon dioxide extraction techniques was determined by GC/MS analysis. Forty-six components were fully characterised. Limonene and γ-terpinene were the major components of the oils obtained by hydrodistillation (HD) and cold-pressing (CP), while citropten was the major constituent in the oil obtained by supercritical carbon dioxide extraction (SFE). Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activities were evaluated. The essential oil obtained by hydrodistillation exerted the highest inhibitory activity against BChE (IC50 value of 154.6 µg mL⁻¹) and AChE (IC50 value of 171.3 µg mL⁻¹. Interestingly, the oil obtained by cold-pressing exhibited a selective inhibitory activity against AChE. The essential oils have also been evaluated for the inhibition of NO production in LPS induced RAW 264.7 macrophages. The oil obtained by hydrodistillation exerted a significant inhibition of NO production with an IC50 value of 17 µg mL⁻¹ (IC50 of positive control 53 µg mL⁻¹).

Protective effect of Hypericum calabricum Sprengel on oxidative damage and its inhibition of nitric oxide in lipopolysaccharide-stimulated RAW 264.7 macrophages.

The present study shows for the first time the phenolic composition and the in vitro properties (antioxidant and inhibition of nitric oxide production) of Hypericum calabricum Sprengel collected in Italy. The content of hypericins (hypericin and pseudohypericin), hyperforin, flavonoids (rutin, hyperoside, isoquercetrin, quercitrin, quercetin and biapigenin) and chlorogenic acid of H. calabricum, have been determined. The ethyl acetate fraction from the aerial parts of H. calabricum exhibited activity against the radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) with IC50 value of 1.6 jig/ml. The test for inhibition of nitric oxide (NO) production was performed using the murine monocytic macrophage cell line RAW 264.7. The ethyl acetate fraction had significant activity with an IC50 value of 102 jig/ml and this might indicate that it would have an anti-inflammatory effect in vivo.

Protective effect of Hypericum calabricum Sprengel on oxidative damage and its inhibition of nitric oxide in lipopolysaccharide-stimulated RAW 264.7 macrophages

The present study shows for the first time the phenolic composition and the in vitro properties (antioxidant and inhibition of nitric oxide production) of Hypericum calabricum Sprengel collected in Italy. The content of hypericins (hypericin and pseudohypericin), hyperforin, flavonoids (rutin, hyperoside, isoquercetrin, quercitrin, quercetin and biapigenin) and chlorogenic acid of H. calabricum, have been determined. The ethyl acetate fraction from the aerial parts of H. calabricum exhibited activity against the radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) with IC50 value of 1.6 jig/ml. The test for inhibition of nitric oxide (NO) production was performed using the murine monocytic macrophage cell line RAW 264.7. The ethyl acetate fraction had significant activity with an IC50 value of 102 jig/ml and this might indicate that it would have an anti-inflammatory effect in vivo.

Quantitative determination of Amaryllidaceae alkaloids from Galanthus reginae-olgae subsp. vernalis and in vitro activities relevant for neurodegenerative diseases.

In the present work the qualitative and quantitative analysis of Amaryllidaceae-type alkaloids in the aerial parts and bulbs of Galanthus reginae-olgae Orph. subsp. vernalis Kamari is presented for the first time using GC-MS analysis. The alkaloids galanthamine, lycorine, and tazettine were identified in both extracts while crinine and neronine were found only in the bulbs. The yield of alkaloid fraction from bulbs (36.8%) is very high compared to the yield from aerial parts (9.34%). Lycorine was the major component in both fractions. The antioxidant potential was determined by three complementary methods. The preparations to reduce the stable free radical DPPH to the yellow-colored 1,1-diphenyl-2-picrylhydrazyl with IC(50) values of 39 and 29 mug/mL for MeOH extracts from aerial parts and bulbs, respectively. The higher activity was given by EtOAc fraction of aerial parts with IC(50) of 10 mug/mL. This activity is probably due to the presence in EtOAc fraction of polar compounds such as polyphenols. The fraction exhibited a significant antioxidant capacity also in the beta-carotene-linoleic acid test system. A higher level of antioxidant activity was observed for EtOAc fraction from bulbs with IC(50) of 10 mug/mL after 30 min and 9 mug/mL after 60 min of incubation. In contrast, the fraction from bulbs performed poorly in the lipid peroxidation liposomes assay. Significant activity was obtained for dichloromethane fraction from aerial parts (IC(50) of 74 mug/mL). The major abundance of alkaloid in dichloromethane fraction may be responsible of the bulbs anti-cholinesterase highest activity (38.5%) at 0.5 mg/mL.

Protective effect of Pimpinella anisoides ethanolic extract and its constituents on oxidative damage and its inhibition of nitric oxide in lipopolysaccharide-stimulated RAW 264.7 macrophages.

The present study shows for the first time the chemical profile and the in vitro properties (antioxidant and inhibition of nitric oxide [NO] production) of the aromatic plant Pimpinella anisoides V Brig. (Family Apiaceae). The ethanolic extract of the fruits is characterized by 23 major components. Fourteen monoterpenes, two sesquiterpenes, one fatty acid, five methyl esters and one aldehyde were identified. Among them the most abundant components were the monoterpenes trans-anethole (54.5%), limonene (13.5%), and sabinene (4.4%). The extract showed significant antioxidant activity (50% inhibitory concentration [IC(50)], 3.02 mg/mL) using the 2,2-diphenyl-1-picrylhydrazyl test. The test for inhibition of NO production was performed using the murine monocytic macrophage cell line RAW 264.7. The ethanolic extract had significant activity with an IC(50) value of 72.7 microg/mL, and this might indicate that it would have an anti-inflammatory effect in vivo. Among the pure compounds that most effectively inhibited lipopolysaccharide-induced NO production were the most abundant constituents, trans-anethole and limonene, with IC(50) values of 102.7 microg/mL and 70.1 microg/mL, respectively. The cytotoxic effect of P. anisoides extract and pure compounds in the presence of lipopolysaccharide (1 microg/mL) was evaluated but found to be negligible.

Synthesis, inhibition of NO production and antiproliferative activities of some indole derivatives.

The synthesis and the biological evaluation of pyrano[3,2-e]indoles and their reaction intermediates are described. The compounds prepared were evaluated for their inhibition of NO production, antioxidant activity and also for their ability to inhibit in vitro the growth of four human tumor cell lines: large lung carcinoma (COR-L23), alveolar basal epithelial carcinoma (A549), amelanotic melanoma (C32) and melanoma (A375). The two reaction intermediates, 5a and 5b, showed the highest inhibition of NO production in murine monocytic macrophage (IC(50) = 1.1 microM and IC(50) = 2.3 microM respectively). Compound 5a was the most active against melanotic melanoma (IC(50) = 11.8 microM) while the other compounds exhibited weak cytotoxicity with IC(50) values >50 microM on all cell lines.

In vitro cytotoxic activity of Salsola oppositifolia Desf. (Amaranthaceae) in a panel of tumour cell lines.

The aim of the present study was to evaluate for the first time the in vitro cytotoxic activity of fractions and isolated flavonols from Salsola oppositifolia Desf. (Amaranthaceae). The n-hexane fraction demonstrated an effective cytotoxic activity on the large lung carcinoma and amelanotic melanoma cell lines with IC50 values of 19.1 microg/ml and 24.4 microg/ml, respectively. Also the dichloromethane fraction exhibited cytotoxic activity against COR-L23 (IC50 30.4 microg/ml) and C32 (IC50 33.2 microg/ml) cells, while the EtOAc fraction demonstrated a selective cytotoxic activity against MCF-7 cells (IC50 67.9 microg/ml). The major active constituents of this fraction were isorhamnetin-3-O-glucoside (1) and isorhamnetin-3-O-rutinoside (2), which showed an interesting activity against the cell line MCF-7 with IC50 values of 18.2 and 25.2 microg/ml, respectively. Compound 2 exhibited a strong activity against the hormone-dependent prostate carcinoma LNCaP cell line with an IC50 of 20.5 microg/ml. Constituents of S. oppositifolia were identified by GC-MS and NMR analyses.

Biological and pharmacological activities of iridoids: recent developments.

Iridoids represent a large group of cyclopenta[c]pyran monoterpenoids that occur wide-spread in nature, mainly in dicotyledonous plant families like Apocynaceae, Scrophulariaceae, Diervillaceae, Lamiaceae, Loganiaceae and Rubiaceae. Recently, more extensive studies revealed that iridoids exhibit a wide range of bioactivity, such as neuroprotective, antinflammatory and immunomodulator, hepatoprotective and cardioprotective effects. Anticancer, antioxidant, antimicrobic, hypoglycaemic, hypolipidemic, choleretic, antispasmodic and purgative properties were also reported. The aim of the present review is to discuss the recent developments on biological and pharmacological activities of iridoids, supporting the new therapeutic possibilities for the use of these compounds.

In vivo anti-inflammatory and in vitro antioxidant activities of Mediterranean dietary plants.

Five hydroalcoholic extracts of edible plants from Calabria region (Italy) used in local traditional medicine for the treatment of inflammatory diseases were evaluated for their in vivo topical anti-inflammatory activity (inhibition of croton oil-induced ear oedema in mice) and in vitro antioxidant and antiradical properties (inhibition of linoleic acid oxidation and bovine brain liposomes peroxidation, DPPH radical scavenging). All the extracts showed an anti-inflammatory effect: 300 microg/cm(2) provoked oedema reductions ranging from 21 to 27%. All the extracts exerted also radical scavenging and/or antioxidant properties, the most active plant being Mentha aquatica L. (Lamiaceae) which contained the highest amount of phenolics (337 mg/g) and of flavonoids (15.75 mg/g). Moreover, the content and the composition of sterols were assessed by GC-MS in the examined plants Borago officinalis L. (Boraginaceae) contained the highest number of sterols.

Biological properties of different extracts of two Senecio species.

This study reports on the antioxidant, anti-diabetic and cytotoxic properties of different extracts of two Senecio species, Senecio vulgaris and Senecio inaequidens. Leaves of the latter are used as food in specific populations of Southern of Africa. Both investigated species of Senecio were found to possess different biological activities. S. inaequidens extracts possess a greater antioxidant activity in comparison with S. vulgaris. Highly significant and dose-dependent performance was obtained with the methanolic and dichloromethane extracts of S. vulgaris and the n-hexane extract of S. inaequidens regarding cytotoxic activity on cancer cell lines, while none of the extracts demonstrated activity on normal cell line MRC-5. Both Senecio species also show appreciable alpha-amylase inhibition, particularly the methanolic and dichloromethane extracts, and this may confer anti-diabetic properties on the extract.

Potential antitumor agents: flavones and their derivatives from Linaria reflexa Desf.

The antiproliferative activity of several flavonoids isolated from Linaria reflexa Desf. (Scrophulariaceae) was evaluated in vitro by the SRB assay against the large cell lung carcinoma cell line COR-L23, hepatocellular carcinoma cell line HepG-2, renal adenocarcinoma cell line ACHN, amelanotic melanoma cell line C32, colorectal adenocarcinoma cell line Caco-2, and normal human fetal lung MRC5. Chemical modifications, that is acetylation, hydrolysis of rutinose unit, and hydrolysis of the terminal rhamnose unit, were performed on pectolinarin. Pectolinarin exhibited strong cytotoxic activity on COR-L23, Caco-2, and C32 cell lines with an IC50 of 5.03, 6.18, and 7.17 microM. Similar activities were recorded for the three natural monoacetyl pectolinarin derivatives linariin, isolinariin A, and isolinariin B. In contrast, peracetylpectolinarin displayed only marginal activity.

Antimicrobial activity and cytotoxicity of Cirsium tenoreanum.

The methanol and ethyl acetate extracts of the aerial parts of Cirsium tenoreanum, have been evaluated for their phytochemical constituents and antibacterial and cytotoxic properties. Three flavonoids, apigenin, quercetin-3-O-galactoside and kaempferol-3-O-ramnoside, were isolated. The ethyl acetate extract showed a good antiproliferative activity.