Cellular Stress and p53-Associated Apoptosis by Juniperus communis L. Berry Extract Treatment in the Human SH-SY5Y Neuroblastoma Cells.

Plant phenolics have shown to activate apoptotic cell death in different tumourigenic cell lines. In this study, we evaluated the effects of juniper berry extract (Juniperus communis L.) on p53 protein, gene expression and DNA fragmentation in human neuroblastoma SH-SY5Y cells. In addition, we analyzed the phenolic composition of the extract. We found that juniper berry extract activated cellular relocalization of p53 and DNA fragmentation-dependent cell death. Differentially expressed genes between treated and non-treated cells were evaluated with the cDNA-RDA (representational difference analysis) method at the early time point of apoptotic process when p53 started to be activated and no caspase activity was detected. Twenty one overexpressed genes related to cellular stress, protein synthesis, cell survival and death were detected. Interestingly, they included endoplasmic reticulum (ER) stress inducer and sensor HSPA5 and other ER stress-related genes CALM2 and YKT6 indicating that ER stress response was involved in juniper berry extract mediated cell death. In composition analysis, we identified and quantified low concentrations of fifteen phenolic compounds. The main groups of them were flavones, flavonols, phenolic acids, flavanol and biflavonoid including glycosides of quercetin, apigenin, isoscutellarein and hypolaetin. It is suggested that juniper berry extract induced the p53-associated apoptosis through the potentiation and synergism by several phenolic compounds.

Effect of Bergenia crassifolia L. extracts on weight gain and feeding behavior of rats with high-caloric diet-induced obesity.

The objective of this study was to evaluate the feeding behavior and weight gain in rats with high-calorie diet-induced obesity that are treated with Bergenia crassifolia black and fermented leaves extracts. The daily dietary intake of all treated animals was reduced to 40% compared with the control group on day 22 of the experiment. A significant improvement in glucose tolerance was noted after 7 days of treatment with the Bergenia extracts. In rats treated with an extract of black leaves for 7 days, a significant reduction in the serum triglyceride level, 45% (p<0.05), compared with the control group was observed. However, the treatment did not affect the cholesterol level. Our results provide evidence for the potential use of B. crassifolia as an appetite and energy intake suppressant.

Antioxidant, pro-oxidant and cytotoxic properties of parsley.

Parsley (Petroselinum crispum) leaves were macerated with a mixture of methanol: water: acetic acid to produce a crude extract which was then defatted with (40°-60°) petrol. Antioxidant activity of the extract was evaluated using a battery of in vitro assays, viz., iron(iii) reduction, iron(ii) chelation and free radical scavenging assays. Evaluation of the pro-oxidant activity of the extract was based upon its effects upon DNA fragmentation and protein carbonylation. Cytotoxicity and apoptotic effects of the extract were determined in non-cancerous CV1-P fibroblast and cancerous A375 melanoma cells using MTT and LDH tests and caspase 3-like activity assay. The highest concentration, 2.0 mg ml(-1), decreased the viability of both cell lines, however, the cancerous melanoma cells were slightly susceptible to the effects. The observed cytotoxicity was not due to the caspase 3 activity. In conclusion, the toxicity might be explained by the pro-oxidative activity of components within the extract against proteins and/or DNA but it is not related to caspase 3-dependent apoptosis within cells.

Iron(III) reducing and antiradical activities of three Sideritis from Turkey.

CONTEXT: Sideritis species (Lamiaceae) are widely used as herbal tea and have been used in folk medicine for their anti-inflammatory, anti-rheumatic, digestive, and antimicrobial activities in Turkey. Sideritis dichotoma Huter., Sideritis erythrantha Boiss. var. cedrotorum, and Sideritis vuralii H. Duman et Baser are available as commercial products in Turkey. OBJECTIVE: The antiradical activities of the various solvent extracts of Sideritis species are investigated here for the first time. MATERIALS AND METHODS: Plant samples were sequentially extracted with n-hexane, dichloromethane, methanol, and aqueous methanol (50%, v/v) in Soxhlet apparatus. The extracts of Camellia sinensis (L.) Kuntze (Theaceae) were also prepared for use as a positive control. Total phenolics, iron(III) reductive effects, and 1,1-diphenyl-2-picrylhydrazyl (DPPH•) radical scavenging activities of the all extracts were measured colorimetrically. RESULTS: The aqueous MeOH and MeOH extracts contained the highest amount of total phenols, whereas the n-hexane extract contained the lowest amounts. The polar extracts of C. sinensis showed higher antiradical activity and also iron(III) reductive effects than the Sideritis species; however, the non-polar extracts of Sideritis species were found to be more active than those from C. sinensis in the iron(III) reductive assay and in the DPPH(•) assay as well. But none of the extracts was found to be as active as with positive controls, viz., ascorbic acid, butylated hydroxyanisole (BHA), and Trolox. DISCUSSION AND CONCLUSION: These results can be shown to have antioxidant activities of these Sideritis species and support the ethnopharmacological use of these Sideritis plants.

Salvia officinalis L.: composition and antioxidant-related activities of a crude extract and selected sub-fractions.

The composition and antioxidant properties of a methanol: acetic acid (99:1, v/v) soluble crude extract isolated from S. officinalis L. leaves through maceration and selected fractions isolated thereof are presented in this study. The total phenol content was estimated as gallic acid equivalents, whilst qualitative-quantitative phenolic content was determined using high performance liquid chromatography with photodiode array detection. Antioxidant evaluation consisted of ferric reductive capacity and 1,1-diphenyl-2-picrylhydrazyl and hydroxyl free radical scavenging determinations. The crude extract contained hydroxybenzoic acids, hydroxycinnamic acids, flavonoids and diterpenoids, whilst caffeic acid, carnosic acid, luteolin, luteolin-7-O-glucoside and rosmarinic acid were identified from their chromatographic and spectral characteristics and quantified from their respective calibration curves. The crude extract and sub-fractions demonstrated varying degrees of efficacy in the antioxidant-related assays used, except the n-hexane fraction, which was unable to reduce iron(III) at reasonable concentrations. Although the positive controls, ascorbic acid, BHA and BHT, were more potent than the S. officinalis samples, two fractions were significantly (p < 0.05) more potent iron(III) reducing agents than pycnogenol, a proanthocyanidin-rich commercial preparation.

Ocimum basilicum L.: phenolic profile and antioxidant-related activity.

Ocimum basilicum L. leaf material was extracted by maceration with (80:20:1 v/v/v) methanol: water: acetic acid to produce a crude extract (CE), which was further fractionated by liquid-liquid extraction to isolate light petroleum (PE), ethyl acetate (EtOAc), n-butanol (n-BuOH) and H2O-soluble sub-fractions. The total phenol and flavonoid contents of the resulting samples were estimated using colorimetric-based methods, and their iron(III) reductive and free radical scavenging activities were determined in a battery of in vitro assays. The CE and sub-fractions contained phenolic compounds and flavonoids. The samples, except for PE, gave a positive result for the presence of flavones and flavonols; however, flavanones only appeared to be present in the CE. In iron(III) reduction, CE and n-BuOH were the most potent followed by EtOAc and H2O (statistically indistinguishable, p > 0.05). However, in the ferric reducing antioxidant power assay, H2O was the most potent followed by CE and EtOAc (statistically indistinguishable, p > 0.05) and n-BuOH and PE. In 1,1-diphenyl-2-picrylhydrazyl scavenging, all the samples, except PE, were effective against this reactive nitrogen species, with CE, EtOAc and n-BuOH being the most potent (statistically indistinguishable, p > 0.05). In alkylperoxyl scavenging, all the samples, except for PE, were effective against this reactive oxygen species (ROS). In superoxide anion scavenging, all the samples were capable of scavenging this ROS with CE being the most effective, followed by n-BuOH and H2O (statistically indistinguishable, p > 0.05) and EtOAc and PE. Similarly, in hydroxyl scavenging, all the samples were capable of scavenging this ROS with CE and n-BuOH being the most effective (statistically indistinguishable, p > 0.05) followed by EtOAc and H2O (statistically indistinguishable, p > 0.05) and PE.

Phenolic profile and antioxidant evaluation of Mentha x piperita L. (peppermint) extracts.

Seven extracts were prepared from Mentha x piperita (peppermint) leaves in sequence using a Soxhlet apparatus, viz. (40-60 degrees) light petroleum (PE), dichloromethane (CH2Cl2), acetonitrile (ACN), ethyl acetate (EtOAc), methanol (MeOH), n-butanol and water (H2O) extracts. The phenolic and flavonoid content of each extract were estimated using spectrophotometric methods whilst a qualitative-quantitative analysis was made by reverse-phase high performance liquid chromatography coupled with photodiode array detection (HPLC-PDA). Each extract was assessed in a battery of six antioxidant-related assays so as to determine their iron(III) reductive, iron(II) chelating and free radical scavenging abilities. The MeOH-soluble extract contained the greatest content of total phenols and flavonoids based upon the Folin-Ciocalteu and 2-aminoethyl diphenylborate reagent data and HPLC-PDA analysis. Based upon the chromatographic and UV-spectral data, the leaves principally contained the cinnamic acid caffeic acid, the depside rosmarinic acid and flavonoids (flavones and flavanones). Eriocitrin (383.3 +/- 2.2 mg/g extract) and rosmarinic acid (381.2 +/- 1.9 mg/g extract) were the most abundant components identified within the leaves, whilst naringenin-7-O-glucoside (0.8 +/- 0.01 mg/g extract) was the least abundant component identified being found only in the EtOAc-soluble extract. The EtOAc, ACN and H2O-soluble extracts demonstrated the most potent iron(III) reductive and 1,1'-diphenyl-2-picrylhydrayl, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) and hydroxyl free radical scavenging properties; however, the H2O and CH2Cl2-soluble extracts were the most potent extracts in the beta-carotene-linoleic acid bleaching inhibition assay. In terms of iron(II) chelation--an important antioxidant property--the PE, MeOH and H2O extracts demonstrated moderate iron(II) chelating activity.

Chemical and antioxidant evaluation of Indian gooseberry (Emblica officinalis Gaertn., syn. Phyllanthus emblica L.) supplements.

Indian gooseberry (Emblica officinalis Gaertn.) (Euphorbiaceae) has a distinguished history in Ayurveda medicine and is ascribed a number of medicinal properties and as a dietary supplement, its use is increasing in Western countries. It is thought that its beneficial properties are a function of its antioxidant potency. The study investigated the chemistry and antioxidant properties of four commercial E. officinalis fruit extracts in order to determine if there are any qualitative-quantitative differences. All extracts produced positive responses in the total phenol, total flavonoid and total tannin assays. The presence of predominantly (poly)phenolic analytes, e.g. ellagic and gallic acids and corilagin, was confirmed by RP-HPLC coupled with photodiode array detection. Despite ascorbic acid being a major constituent of E. officinalis fruits, the furanolactone could not be identified in one of the samples. The extracts demonstrated varying degrees of antioxidative efficacy. The extract designated IG-3 was consistently amongst the most effective extracts in the iron(III) reduction and 1,1-diphenyl-2-picrylhydrazyl and superoxide anion radical scavenging assays while the extract designated IG-1 demonstrated the best hydroxyl radical scavenging activity. All extracts appeared to be incapable of chelating iron(II) at realistic concentrations.

Acetylcholinesterase inhibitory guided fractionation of Melissa officinalis L.

The plant Melissa officinalis L. has been used traditionally in the treatment of cognitive dysfunction. Based on its traditional medicinal use, it was assessed for its clinical efficacy in mild to moderate Alzheimer's patients. The plant was effective in the management of the disease. Therefore, based on this result, a similar plant extract was prepared in order to be screened for bioactivities which are relevant in Alzheimer's disease therapy. The extract was recently screened for antioxidant activity and it showed a wide range of antioxidant properties. Another important bioactivity is acetylcholinesterase inhibition, which the extract was screened for in the current investigation. The extract was capable of inhibiting the enzyme in a time and dose-dependent manner. Activity of the extract at 10 min was estimated as 1.72+/-0.16 microg equivalents of physostigmine/mg of the extract. Acetylcholinesterase inhibitory guided fractionation of the extract was then carried out. Most of the fractions showed inhibitory activity and were more potent than the extract. The contents of the most potent fraction were identified as cis- and trans-rosmarinic acid isomers and a rosmarinic acid derivative using LC-DAD-ESI-MS and NMR methods.

Chemical composition and in vitro antioxidant evaluation of commercial water-soluble willow herb (Epilobium angustifolium L.) extracts.

Five commercially available water-soluble extracts prepared from the aerial parts of Epilobium angustifolium L. (Onagraceae) were screened for antioxidant-related properties in a battery of six in vitro assays. Total phenol content and qualitative-quantitative analyses were also carried out. The extracts demonstrated varying degrees of efficacy in each screen. Two extracts, denoted as nonfermented and Tver, were the most effective toward reducing iron(III), scavenging 1,1-diphenyl-2-picrylhydrazyl free radicals, inhibiting hydroxyl radical-catalyzed bovine brain-derived phospholipid degradation, and non-site- and site-specific hydroxyl radical-catalyzed 2-deoxy-D-ribose degradation. The activity profile of the extracts changed, however, when their iron(II) chelating ability was assessed. The nonfermented and Tver extracts were not as effective iron(II) chelators as the extract denoted as Lotos. All the extracts contained Folin-Ciocalteu-reactive substances, which was confirmed by the presence of predominantly polar phenolic analytes (i.e., hydroxylated benzoic acid derivatives and flavonoids).

Screening of free radical scavenging compounds in water extracts of Mentha samples using a postcolumn derivatization method.

An on-line high-performance liquid chromatography-1,1-diphenyl-2-picrylhydrazyl (HPLC-DPPH*) method has been improved for the detection of polar and nonpolar radical scavenging compounds in complex plant extracts. Nine water extracts were prepared from different Mentha species, varieties, hybrids, and cultivars. After the components within each extract had been separated by reverse phase chromatography using 10-100% methanol with 2% acetic acid as a mobile phase, analytes within the eluent capable of scavenging a citric acid-sodium citrate-buffered methanol 1,1-diphenyl-2-picrylhydrazyl solution were detected by postcolumn derivatization at 517 nm. The HPLC-DPPH* on-line method was applied to the qualitative and quantitative analysis of Mentha extracts. There was a strong correlation between the scavenging (negative) peak area and the concentration of the radical scavenging reference substances used. The minimum detectable concentration (microg/mL) of the antioxidant compounds was determined. Caffeic acid, eriocitrin (eriodictyol-7-O-rutinoside), luteolin-7-O-glucoside, and rosmarinic acid were identified as the dominant radical scavengers in these extracts by this method.

Antioxidant properties of aqueous extracts from selected lamiaceae species grown in Turkey.

Water-soluble extracts from black thyme (Thymbra spicata L.), savory (Satureja cuneifolia Ten.), Spanish oregano (Coridothymus capitatus (L.) Reichb. f.), sweet marjoram (Majorana hortensis Moench), Syrian oregano (Origanum syriacum L.), Toka oregano (Origanum minutiflorum O. Schwarz et P. H. Davis), and Turkish oregano (Origanum onites L.) were screened for antioxidant properties in a battery of six in vitro assays. Total phenol content and qualitative-quantitative compositional analyses were also carried out. The extracts demonstrated varying degrees of efficacy in each screen. The savory extract was the most effective at reducing iron(III), scavenging 1,1-diphenyl-2-picrylhydrazyl radicals, inhibiting ascorbate-iron(III)-catalyzed hydroxyl radical-mediated brain phospholipid peroxidation, and site-specific hydroxyl radical-mediated 2-deoxy-d-ribose degradation. The Syrian oregano extract was the most effective chelator of iron(II), while Spanish and Turkish oregano extracts were the most effective inhibitors of nonsite-specific hydroxyl radical-mediated 2-deoxy-d-ribose degradation. All the extracts contained Folin-Ciocalteu reagent-reactive substances, which was confirmed by the presence of polar phenolic analytes (i.e., hydroxybenzoates, hydroxycinnamates, and flavonoids).

Antioxidant properties and composition of aqueous extracts from Mentha species, hybrids, varieties, and cultivars.

Water-soluble extracts from the Mentha species M. aquatica L. and M. haplocalyx Briq., the hybrids M. x dalmatica L. and M. x verticillata L., the varieties M. arvensis var. japanensis [M. arvensis L. var. piperascens Holmes ex Christ] and M. spicata L. var. crispa Benth, and M. x piperita L. "Frantsila", M. "Morocco", and M. "Native Wilmet" cultivars were screened for potential antioxidative properties. These properties included iron(III) reduction, iron(II) chelation, 1,1-diphenyl-2-picrylhydrazyl radical scavenging, and the ability to inhibit iron(III)-ascorbate-catalyzed hydroxyl radical-mediated brain phospholipid peroxidation. Total phenol content and qualitative and quantitative compositional analyses of each extract were also made. The extracts demonstrated varying degrees of efficacy in each assay, with the M. x piperita "Frantsila" extract being better than the other extracts, except for ferrous iron chelation. With the exception of iron chelation, it appeared that the level of activity identified was strongly associated with the phenolic content.