Mariangela Marrelli

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.

Antiproliferative activity against human tumor cell lines and toxicity test on Mediterranean dietary plants.

Sixteen edible plants from Southern Italy were evaluated for their in vitro antiproliferative properties, using the sulforodamine B (SRB) assay, on four human cancer cell lines: breast cancer MCF-7, prostate cancer LNCaP, amelanotic melanoma C32 and renal adenocarcinoma ACHN. After 48 h of incubation the most antiproliferative plant extract was Cynara cardunculus ssp. cardunculus on C32 and ACHN cell lines with IC(50) of 21 and 18 microg/ml, respectively. Mentha aquatica showed a selective antiproliferative activity on breast cancer while significant activity was exerted by Cichorium intybus on melanoma. These species contained the highest amount of phenolics. The acute toxicity of the hydroalcohol extracts from all the plants were evaluated by using the Microtox acute toxicity test. This bacterial test measures the decrease in light emission from the marine luminescent Vibrio fischeri bacteria when exposed to organic extracts. This inhibition test was revealed to be highly sensitive, cost effective and easy to operate, requiring just 15 min to predict the sample toxicity. All the extracts analyzed resulted to give values very less than a limit of 20% value, demonstrating so an irrelevant toxicity for the human health. In contrast, Echium vulgare and Malva sylvestris showed bioluminescence inhibition values of 19.42% and 17.32%, respectively, just under the established limit.

Acetylcholinesterase and butyrylcholinesterase inhibition of ethanolic extract and monoterpenes from Pimpinella anisoides V Brig. (Apiaceae)

Ethanolic extract from the fruits of Pimpinella anisoides, an aromatic plant and a spice, exhibited activity against AChE and BChE, with IC(50) values of 227.5 and 362.1 microg/ml, respectively. The most abundant constituents of the extract were trans-anethole, (+)-limonene and (+)-sabinene. trans-Anethole exhibited the highest activity against AChE and BChE with IC(50) values of 134.7 and 209.6 microg/ml, respectively. The bicyclic monoterpene (+)-sabinene exhibited a promising activity against AChE (IC(50) of 176.5 microg/ml) and BChE (IC(50) of 218.6 microg/ml).

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.

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.

In vitro investigation of the potential health benefits of wild Mediterranean dietary plants as anti-obesity agents with α-amylase and pancreatic lipase inhibitory activities.

BACKGROUND Inhibition of digestive enzymes is one of the most widely studied mechanisms used to determine the potential efficacy of natural products as anti-obesity agents. In vitro studies reported here were performed to evaluate the inhibitory activity of formulations of edible plants from Italy on amylase and lipase by monitoring the hydrolysis of nitrophenyl caprilate and the hydrolysis of glycoside bonds in digestible carbohydrate foods. RESULTS The formulation obtained from Capparis sicula exhibited the strongest inhibitory effect on pancreatic lipase (IC50 = 0.53 mg mL(-1) ) while the Borago officinalis formulation exhibited the strongest inhibitory effect on α-amylase (IC50 = 31.61 µg mL(-1) ). In order to characterise the extracts, high-performance thin-layer chromatography analysis of the formulations was performed, revealing the predominance of (±)-catechin in Mentha aquatica formulation, rutin in C. sicula, and caffeic acid and chlorogenic acid in Echium vulgare. CONCLUSION The results obtained indicated that the extracts of C. sicula and B. officinalis could be good candidates for further studies to isolate pancreatic lipase and α-amylase inhibitors, respectively.

Wild Mediterranean Dietary Plants as Inhibitors of Pancreatic Lipase

Lipids are essential compounds for all living organisms. Agents that inhibit fat digestion are of theoretical benefit in the treatment of obesity. A total of 18 species (21 hydroalcoholic extracts) of edible plants from Calabria region (Italy) were evaluated for their in vitro pancreatic lipase inhibitory activity. Lipase activity was measured by monitoring the hydrolysis of p‐NPC, which releases the yellow chromogen, p‐nitrophenol. The aqueous ethanol extracts of Portulaca oleracea (leaves) and Silene vulgaris (leaves) exhibited the strongest inhibitory effect on lipase. The amounts of total phenolics, measured by Folin‐Ciocalteu method, varied widely in the different analysed extracts and ranged from 29 to 482 mg/g of extract. In this study, the findings do not show any relationship between lipase inhibitory activity and total phenolic content. Copyright

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 (IC50 = 1.1 μM and IC50 = 2.3 μM respectively). Compound 5a was the most active against melanotic melanoma (IC50 = 11.8 μM) while the other compounds exhibited weak cytotoxicity with IC50 values >50 μM on all cell lines.

Hypericum perforatum: Influences of the habitat on chemical composition, photo-induced cytotoxicity, and antiradical activity.

Abstract Context: Hypericin, isolated from Hypericum perforatum L. and about another 300 Hypericum species (Guttiferae), is one of the most powerful photosensitizers found in nature. Objective: The aim of this study was to assess the variability of chemical composition and biological activities of four H. perforatum samples, collected at different altitudes in the South Apennine of Italy. Materials and methods: MTT assay was used to evaluate the antiproliferative activity of different samples concentrations (0.6–100 µg/mL) after irradiation at 365 nm. The inhibition of nitric oxide production was evaluated after 24 h of incubation using the macrophage cell line RAW 264.7 and sample solutions ranging from 12.5 to 1000 µg/mL. Antioxidant activities were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and β-carotene bleaching test (ranges were 12.5–1000 and 1–400 µg/mL, respectively). Chemical composition was evaluated through HPTLC, and different contents of hypericin and rutin have been observed. Results: The most phototoxic sample was collected from Zumpano (no. 1 at 370 m), with IC50 values of 24.61 ± 0.02 μg/mL. Sample no. 1 showed also the best radical scavenging activity (IC50 value of 9.18 ± 0.03 μg/mL) and the best antioxidant activity (IC50 value of 10.04 ± 0.03 μg/mL after 30 min of incubation). Best activity of extract no. 1 was well in accordance with chemical data, including the phenolic total content and particular metabolome profile. Discussion and conclusion: This paper confirms the usefulness in maintaining the exploration of H. perforatum activities, in order to confirm its potentiality as a multipurpose plant.

Allelopathic potential of Artemisia arborescens: Isolation, identification and quantification of phytotoxic compounds through fractionation-guided bioassays

The aerial part of Artemisia arborescens L. (Asteraceae) was extracted with water and methanol, and both extracts were fractionated using n-hexane, chloroform, ethyl acetate and n-butanol. The potential phytotoxicity of both crude extracts and their fractions were assayed in vitro on seed germination and root growth of lettuce (Lactuca sativa L.), a sensitive species largely employed in the allelopathy studies. The inhibitory activities were analysed by dose–response curves and the ED 50 were estimated. Crude extracts strongly inhibited both germination and root growth processes. The fraction-bioassay indicated the following hierarchy of phytotoxicity for both physiological processes: ethyl acetate ≥ n-hexane > chloroform ≥ n-butanol. On the n-hexane fraction, GC–MS analyses were carried out to characterise and quantify some of the potential allelochemicals. Twenty-one compounds were identified and three of them, camphor, trans-caryophyllene and pulegone were quantified.