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

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

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

Hairy garlic (Allium subhirsutum) from Sicily (Italy): LC-DAD-MSn analysis of secondary metabolites and in vitro biological properties

Allium subhirsutum, known as hairy garlic, is a bulbous plant widespread in the Mediterranean area and locally used as a food and spice. In the present study, the chemical profile of the ethanolic extracts from bulbs (BE) and aerial parts (APE) were analyzed by HPLC-ESI-MSn, and antioxidant properties were evaluated by DPPH, ABTS and TEAC assays. The traditional use in the diet, and the well documented biological activity of Allium species suggest a potential as a new nutraceutical. For this reason, the potential usefulness of this food can be considered in the treatment and prevention of degenerative Alzheimer disease. For this reason, acetylcholinesterase inhibitory property was investigated. Furthermore, due to the observed presence of sulfur-containing and phenolic constituents, the cytotoxicity on tumor cells line was investigated. Results revealed significant AChE inhibitory activity for BE and APE. Both extracts exhibited also moderate antioxidant properties in the in vitro assays. Finally, limited cytotoxic activity was observed towards Human colon carcinoma and adenocarcinoma cell line, with differences between the individual parts tested. HPLC-ESI-MSn analysis showed that hairy garlic is a good source of sulphur compounds, flavonoids and phenylpropanoids derivatives, thus being a valid alternative to the common garlic (A. sativum). This work opens new opportunities for the application of A. subhirsutum as a health-promoting food

Antimicrobial, antioxidant, anti-inflammatory activities and phytoconstituents of extracts from the roots of Dissotis thollonii Cogn. (Melastomataceae)

Abstract Background Dissotis thollonii Cogn. belonging to the Malastomataceae family is used in the West Region of Cameroon for the treatment of inflammation, kidney diseases, pregnancy control and sinusitis. Despite the traditional use of this plant, no scientific report or information was found in the literature regarding neither its biological activity nor its chemical constituents. Aim of the study The present work was designed to determine the antimicrobial, antioxidant and anti-inflammatory activities of different extracts of the roots of D. thollonii Cogn. as well as the isolation and identification of the chemical constituents of this plant. Materials and methods The tests for antimicrobial, antioxidant and anti-inflammatory activities were performed over the MeOH, EtOAc, n-BuOH and aqueous extracts. Compounds were isolated from EtOAc and n-BuOH extracts of the roots of D. thollonii Cogn. through column chromatography and their structures were determined by means of NMR and MS analysis, and published data. Results According to the antimicrobial and antioxidant assays, the EtOAc and n-BuOH extracts were submitted to further separation and purification. This led to the isolation of twelve compounds identified as 3,3′-di- O -methylellagic acid 4′- O-β -D-xylopyranoside 1 , 3- O -methylellagic acid 4′- O-β -D-arabinopyranoside 2 , casuarinin 3 , betulinic acid 4 , β -sitosterol-3- O -D-glucopyranosyl-6′-mirystate 5 , cellobiosylsterol 6 , β -sitosterol 7 , β -sitosterol-3- O-β -D-glucopyranoside 8, arjunolic acid 9 , 3,3′-di- O -methylellagic acid 10 , ellagic acid 11 , and 3,3′-di- O -methylellagic acid 4′- O - β -D-glucopyranoside 12 . The EtOAc extract was the only antimicrobial active sample [diameter of the zone of inhibition (DZI) of 10 mm against Staphyloccocus aureus ] among all the tested extracts. The analysis of fractions of this extract revealed the presence of bioactive compounds with a described antimicrobial activity such as β -sitosterol, β -sitosterol-3- O-β -D-glucopyranoside and arjunolic acid. By using Trolox as the standard drug, all extracts showed antioxidant activity against DPPH in the following order of scavenging ability: Trolox > nBuOH > EtOAc > MeOH > WE (water extract). The ABTS •+ scavenging ability was similar to that found for the DPPH assay, being Trolox > n-BuOH > MeOH > EtOAc > WE. Along with the DPPH and ABTS assays, the FRAP assay showed the scale n-BuOH > MeOH > WE > EtOAc. The phytochemical study of the EtOAc and n-BuOH extracts revealed the presence of important known antioxidant compounds such as ellagic acid derivatives, arjunolic acid, betulinic acid and β -sitosterol. The anti-inflammatory properties of D. thollonii extracts were investigated using RAW 264.7 murine macrophage cells. The MeOH extract reduced the stimulated NO production in a concentration-dependent manner. 86% reduction was observed at the highest tested concentration of 100 μg/ml (IC 50 = 5.9 μg/ml). The n-BuOH extract showed higher dose dependent reduction of NO formation (IC 50 = 6.5 μg/ml) than the EtOAc extract (IC 50 = 18.1 μg/ml), whereas the water extract had no significant influence on the NO production. All the extracts did not have any influence on the macrophage viability. The phytochemical investigation of the EtOAc and n-BuOH extracts revealed that the main compounds identified do have potent anti-inflammatory properties. Conclusion The biological and phytochemical characterization of the root extracts of D. thollonii validates the use of this plant for the treatment of inflammation and sinusitis, thus providing evidence that this plant extracts, as well as some of the isolated compounds, might be potential sources of antioxidant and anti-inflammatory drugs

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)

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

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