Biological activity of ethanol extract from leaves of Rosmarinus eriocalyx

Aging or senescence is a complex and inevitable process, which is not only attributed to individual genetic variation but also to external factors such as environmental conditions, nutrition, alcohol, and diseases [1]. The most widely accepted theory, that have been proposed to explain aging, is the free radical theory [2]. Aging and related diseases result from accumulated oxidative damage to cell constituents and tissues caused by excessive exposure to free radicals. Antioxidants, which mediate the imbalance between intracellular antioxidant defenses and oxidative damage by reducing the reactive oxygen species (ROS) levels, are believe to be able to reduce stress-induced premature senescence or slow down replicative senescence [3]. Rosmarinus eriocalyx (Jord. & Fourr.) is an aromatic evergreen bush belonging to Lamiaceae family and endemic to Algeria, Morocco and Spain, where it is used as a condiment to flavor soup and meat and as a traditional remedy [4]. The plant volatile fraction is characterized by the monoterpene ketone camphor, whereas its ethanolic extracts are rich sources of phenolic acids and diterpenes such as rosmarinic acid, carnosic acid and carnosol that are the main responsible for the noteworthy antioxidant activity [5]. In this setting, we aimed to evaluate R. eriocalyx biological activity in order to propose the plant as an anti-aging agent. For this purpose, we determined the cytotoxic activity of polar extracts obtained from leaves, flowers, and stems of R. eriocalyx on human fibroblast and human tumor cell lines (A375, MDA-MB 231, and T98G) by MTT assay [6]. Results showed that the ethanolic extract of leaves resulted the most active against A375 human melanoma cell line (IC50 value of 17.8 µg/ml). The total phenolic content values reported for R. eriocalyx ethanolic and aqueous extracts showed slight differences and free radical scavenging activity was stronger for ethanolic extracts than aqueous ones. On this basis, we selected the R. eriocalyx ethanolic extract to determine the antioxidant activity on human fibroblast by measuring its ability to prevent oxidation in cells using a ROS fluorescent probe (DCFH-DA) [7]. Results showed a remarkable activity in preventing oxidation of cells induced by 2.2’-azobis -2-amidinopropane (ABAP). Afterwards, we tested the same extract on the H2O2-induced premature senescence in young fibroblast cells where -galactosidase (SA--gal) activity was used to measure cell senescence [8]. Preliminary data showed a reduction of H2O2 stress-induced premature senescence indicating the potential of R. eriocalyx leaf extract to be formulated as an anti-aging agent. References [1] N. Getoff. Anti-aging and aging factors in life. The role of free radicals. Radiat. Phys. Chem. 2007, 76,1577-1586. [2] D. Harman. Aging: a theory based on free radical and radiation chemistry. J. Gerontol. 1956, 11, 298-300. [3] D. Fusco, G. Colloca, M.R. Lo Monaco, M. Cesari. Effects of antioxidant supplementation on the aging process. Clin. Interv. Aging. 2007, 2, 377-387. [4] M.S. Bendeddouche, H. Benhassaini, Z. Hazem, A. Romane. Essential oil analysis and antibacterial activity of Rosmarinus tournefortii from Algeria. Nat. Prod. Commun. 2011, 6, 1511-1514. [5] H. Bendif, M. Boudjeniba, M. Djamel Miara, L. Biqiku, M. Bramucci, G. Caprioli, G. Lupidi, L. Quassinti, G. Sagratini, L.A. Vitali, S. Vittori, F. Maggi. Rosmarinus eriocalyx: An alternative to Rosmarinus officinalis as a source of antioxidant compounds. Food Chem. 2017, 218, 78-88. [6] L. Quassinti, G. Lupidi, F. Maggi, F. Papa, S. Vittori, A. Bianco, M. Bramucci . Antioxidant and antiproliferative activity of Hypericum hircinum L. subsp. majus (Aiton) N. Robson essential oil. Nat. Prod. Res. 2013, 27, 862-868. [7] K.L. Wolfe, R.H. Liu. Cellular antioxidant activity (CAA) assay for assessing antioxidants, foods, and dietary supplements. J. Agric. Food Chem. 2007, 55, 8896-8907. [8] D.J. Kurz, S. Decary, Y. Hong, J.D. Erusalimsky. Senescence-associated -galactosidase reflects an increase in lysosomal mass during replicative ageing of human endothelial cells. J. Cell Sci. 2000, 113, 3613–3622

Bradykinin is not involved in angiotensin converting enzyme modulation of ovarian steroidogenesis and prostaglandin production in frog Rana esculenta

Angiotensin converting enzyme (ACE) was demonstrated to modulate the production of 17beta-estradiol, progesterone and prostaglandin E2 (PGE2) in frog ovary of Rana esculenta. However, the activity was not mediated by angiotensin II (Ang II). In an attempt to identify the peptide involved in the pathway modulated by ACE, bradykinin, another physiological substrate of ACE, was chosen and incubated in the presence of the membrane suspension purified from the frog ovary homogenate. The hydrolytic products were analysed by reverse-phase high-pressure liquid chromatography (HPLC) analysis and the results showed that bradykinin was metabolized by membrane suspension. The presence of the protease inhibitors in the incubation mixture indicated ACE and neutral endopeptidase as being responsible for the bradykinin hydrolysis. Frog ovary was incubated in vitro in the presence of bradykinin (10 microM), bradykinin receptor antagonist NPC 567 (1 mg mL-1), bradykinin fragment (1-7) (10 microM), ACE (2.5 mU mL-1), captopril (0.1 mM) and lisinopril (0.1 mM). The results showed no modulating activity by bradykinin on ovarian 17beta-estradiol and PGE2 production, thus demonstrating that it was not involved in the ACE-modulated pathway

DNA binding and oxidative DNA damage induced by climacostol\u2013copper(II) complexes: Implications for anticancer properties

Climacostol is a natural toxin isolated from the freshwater ciliated protozoan Climacostomum virens and belongs to the group of resorcinolic lipids. Climacostol exerts a potent antimicrobial activity against a panel of bacterial and fungal pathogens. In addition it inhibits the growth of tumor cell lines in a dose-dependent manner by inducing programmed cell death via intrinsic pathway. In this work, we investigated the possibility that climacostol exerts a prooxidant effect, inducing plasmid DNA strand breakage and eukaryotic DNA damage in presence of Cu(II) ions. Inhibition of DNA breakage using SOD, catalase and neocuproine confirmed the involvement of reactive oxygen species and Cu(I) ions in the DNA damage. UV\u2013visible absorption changes and mass spectrometric analysis identified a product of reaction as a deprotonated form of climacostol. Study of the interaction with DNA, using fluorescence spectroscopic techniques, showed that climacostol binds with DNA. Given the structure\u2013activity relationship of this compound and the mechanism of its prooxidant effect, we propose that the Cu(II)-mediated oxidative DNA damage by climacostol could explain its antimicrobial and antiproliferative activity

Antioxidant activity and cytotoxicity on tumor cells of the essential oil from Cedronella canariensis var. canariensis (L.) Webb & Berthel. (Lamiaceae)

Cedronella canariensis is a lemon-scented species of the family Lamiaceae endemic to the Canary Islands where it is used in the traditional medicine to prepare infusions or inhalations for anti-catarrhal, tonic, diuretic, hypoglycaemiant, hypotensive, antiinflammatory and decongestant of the respiratory tract. In this work we investigated for the first time the antioxidant activity of the essential oil and its inhibitory effects on tumour cells (A375, MDA-MB-231, HCT 116) proliferation by DPPH, ABTS, FRAP and MTT assays, respectively. The oil, analysed by GC-ionisation flame detector and GC–MS, was characterised by pinocarvone (58.0%) and b-pinene (10.8%) as the major constituents, being typical of the chemotype ‘canariensis’. Noteworthy was the cytotoxic activity of the oil against the tumour cells examined, with IC50 values of 4.3, 7.3 and 11.4 µg/mL on A375, MDA-MB-231 and HCT 116 tumour cells, respectively, as well as the scavenging activity against the ABTS radical (IC50 of 10.5 µg/mL)

antiproliferative activities of artemisia herba alba ethanolic extract in human colon cancer cell line hct116

Artemisia herba-alba (AHE) is a plant commonly used in traditional medicine for the treatment of various ailments. Here, we investigated the antioxidant and antitumor activity of the aqueous and ethanol extracts of AHE in human colon cancer HCT116 cells. The antioxidant activity was measured by DCFH assay, while antitumor effects were assessed by cell viability assays, cell cycle progression by flow cytometry, and DNA fragmentation analysis in addition to investigating the expression of key cell cycle and apoptotic proteins. While the aqueous extract had no antineoplastic effects, the ethanol extract significantly decreased HCT116 viability (IC50 of 51mg/mL at 24 h) and inhibited the production of reactive oxygen species (ROS). Treatment of HCT116 cells with the ethanol extract also caused dramatic increase in the PreG1 population with concomitant decrease in cycling cells, provoked DNA fragmentation, significant increase in the expression levels of p53 and Bax proteins and activated pro-apoptotic caspase-3. The results obtained suggest that the ethanol extract of AHE could be used as an easily accessible source of natural antioxidants and as potential phytochemicals against colon cancer.</p

Blue honeysuckle fruit (Lonicera caerulea L.) from eastern Russia: phenolic composition, nutritional value and biological activities of its polar extracts

In the present work we conducted a comprehensive chemical analysis of blue honeysuckle (Lonicera caerulea) spontaneously growing in eastern Russia. HPLC-DAD-ESI/MS analysis showed cyanidin-3-glucoside as the major constituent among phenolics, while nutritional analysis revealed fibre, protein, calcium and magnesium as the most important macro- and micronutrients, respectively. Fatty acid composition was dominated by polyunsaturated fatty acids, linoleic acid being the most abundant. Furthermore, we evaluated several in vitro biological activities such as antioxidant, antimicrobial, antiproliferative, wound healing and immunomodulatory effects of blue honeysuckle aqueous and ethanolic extracts that are often incorporated in food and nutraceutical preparations. While the fruit extracts were revealed to be potent radical scavengers with significant inhibition of ABTS radical, thus confirming the literature data, their inhibitory effects against microbial pathogens and tumor cell lines were negligible. The fruit aqueous extract did not show toxicity to human fibroblasts, but 24 h treatment with 150–200 μg per mL of extract slightly enhanced the cell migration when tested by scratched wound assay. Worth mentioning was the inhibitory effect displayed by the blue honeysuckle fruit aqueous extract on human lymphocytes

Plasmodium transmission blocking activities of Vernonia amygdalina extracts and isolated compounds

BACKGROUND: Medicinal plants are a validated source for discovery of new leads and standardized herbal medicines. The aim of this study was to assess the activity of Vernonia amygdalina leaf extracts and isolated compounds against gametocytes and sporogonic stages of Plasmodium berghei and to validate the findings on field isolates of Plasmodium falciparum. METHODS: Aqueous (Ver-H2O) and ethanolic (Ver-EtOH) leaf extracts were tested in vivo for activity against sexual and asexual blood stage P. berghei parasites. In vivo transmission blocking effects of Ver-EtOH and Ver-H2O were estimated by assessing P. berghei oocyst prevalence and density in Anopheles stephensi mosquitoes. Activity targeting early sporogonic stages (ESS), namely gametes, zygotes and ookinetes was assessed in vitro using P. berghei CTRPp.GFP strain. Bioassay guided fractionation was performed to characterize V. amygdalina fractions and molecules for anti-ESS activity. Fractions active against ESS of the murine parasite were tested for ex vivo transmission blocking activity on P. falciparum field isolates. Cytotoxic effects of extracts and isolated compounds vernolide and vernodalol were evaluated on the human cell lines HCT116 and EA.hy926. RESULTS: Ver-H2O reduced the P. berghei macrogametocyte density in mice by about 50% and Ver-EtOH reduced P. berghei oocyst prevalence and density by 27 and 90%, respectively, in An. stephensi mosquitoes. Ver-EtOH inhibited almost completely (>90%) ESS development in vitro at 50 μg/mL. At this concentration, four fractions obtained from the ethylacetate phase of the methanol extract displayed inhibitory activity >90% against ESS. Three tested fractions were also found active against field isolates of the human parasite P. falciparum, reducing oocyst prevalence in Anopheles coluzzii mosquitoes to one-half and oocyst density to one-fourth of controls. The molecules and fractions displayed considerable cytotoxicity on the two tested cell-lines. CONCLUSIONS: Vernonia amygdalina leaves contain molecules affecting multiple stages of Plasmodium, evidencing its potential for drug discovery. Chemical modification of the identified hit molecules, in particular vernodalol, could generate a library of druggable sesquiterpene lactones. The development of a multistage phytomedicine designed as preventive treatment to complement existing malaria control tools appears a challenging but feasible goal

Antioxidant and antiproliferative activities of different varieties of cauliflower (Brassica oleracea var. botrytis) after cooking processes.

Cauliflowers are well-known for their health benefits and they are becoming increasingly popular as a fresh vegetable significant source of nutritional antioxidants, such as vitamins and carotenoids, or biologically active dietary components, such as the polyphenols and glucosinolates [1]. Cauliflowers are also a rich source of nutrients such as calcium, zeaxanthin and lutein which have a protective action in eye health and they can also help in the prevention of cancer through the flavonoids known as quercetin or phytonutrient as sulforaphane [2]. Cultivation of coloured cauliflowers (Brassica oleracea var. botrytis) is spreading in Italy and this is also a consequence of the significant genetic improvement on the white type “Tardivo di Fano”, the green types “Verde di Macerata” and the violet type “Violetto di Catania” [3]. Recently, in the framework of a genetic improvement programme, pure lines of “orange type” were obtained [4]. Cooking as a domestic processing method has a great impact on food nutrients. Most vegetables are mainly consumed after being cooked, and cooking considerably affects their health-promoting compounds (glucosinolates, phenolic compounds, phytochemicals, and vitamin C). The cooking process is more important also to determine the quality and recovery of biological active components that characterized the different variety of cauliflowers selected. The aim of this work was to study the influence of these conditions during boiling water and microwave cooking on some properties of coloured cauliflowers. Total polyphenols, antioxidant activity, and antiproliferative activity on human breast adenocarcinoma cell line MDA-MB 231 were evaluated after cooking processes. The results showed that the antioxidant activity, tested using ABTS assay, was reduced during the cooking in all cauliflower varieties. An increase of antioxidant activity in orange and violet varieties was observed when tested with DPPH and FRAP assays. Furthermore, the phenolic substances increase in methanol extracts after cooking processes, especially with microwave treatment. The antiproliferative activity on MDA-MB 231 tumour cell line shows an increase in orange and violet cauliflower aqueous extracts cooked with microwave. In conclusion, the change of properties results lower for orange and violet cauliflowers respect to white or green varieties after cooking processes. In some case the cooking processes increase the antioxidant and antiproliferative activity. The best cooking method that preserves the healthy properties of all varieties of cauliflower is the microwave treatment. [1] Raiola A, Errico A, Petruk G, Monti DM, Barone A, Rigano MM. Bioactive Compounds In Brassicaceae Vegetables With A Role In The Prevention Of Chronic Diseases. Molecules. 2017;23(1), pii: E15. [2] Rosa EAS, Heaney RK, Fenwick GR, Portas CAM. Glucosinolates in Crop Plants, in Horticultural Reviews, Volume 19 (ed J. Janick), 2010, John Wiley & Sons, Inc., Oxford, UK. [3] Acciarri N, Sabatini E, Rotino GL, Ciriaci T, Pulcini L, Della Campa M, Maestrelli A. Breeding In Progress In New Typologies Of Orange Cauliflowers (Brassica oleracea var. botrytis) Proceedings of 5th International Symposium on Brassicas and the 16th Crucifer genetic Workshop. Lillehammer, Norway. 8-12 September 2008. [4] Lu S,Van Eck J, Zhou X, Lopez AB, O’Halloran DM, Cosman KM, Conlin BJ, Paolillo DJ, Garvin DF, Vrebalov J, Kochian, LV, Kupper H, Earle ED, Cao J, Lia L. The Cauliflower Or Gene Encodes A Dnaj Cysteine-Rich Domain-Containing Protein That Mediates High Levels Of β-Carotene Accumulation. Plant Cell 2006;18:3594-3605