The cytotoxicity effect of 7-hydroxy-3,4-dihydrocadalene from Heterotheca inuloides and semisynthetic cadalenes derivates towards breast cancer cells: involvement of oxidative stress-mediated apoptosis.

Background: Heterotheca inuloides, traditionally employed in Mexico, has demonstrated anticancer activities. Although it has been proven that the cytotoxic effect is attributed to cadinane-type sesquiterpenes such as 7-hydroxy-3,4-dihydrocadalene, the mechanism of action by which these agents act in tumor lines and their regulation remain unknown. This study was undertaken to investigate for first time the cytotoxic activity and mechanism of action of 7-hydroxy-3,4-dihydrocadalene and two semi-synthetic cadinanes derivatives towards breast cancer cells. Methods: Cell viability and proliferation were assayed by thiazolyl blue tetrazolium bromide (MTT) assay and Trypan blue dye exclusion assay. Cell migration measure was tested by wound-healing assay. Moreover, the reactive oxygen species (ROS) and lipid peroxidation generation were measured by 2',7'-dichlorofluorescein diacetate (DCFH-DA) assay and thiobarbituric acid reactive substance (TBARS) assay, respectively. Furthermore, expression of caspase-3, Bcl-2 and GAPDH were analyzed by western blot. Results: The results showed that 7-hydroxy-3,4-dihydrocadalene inhibited MCF7 cell viability in a concentration and time dependent manner. The cytotoxic potency of semisynthetic derivatives 7-(phenylcarbamate)-3,4-dihydrocadalene and 7-(phenylcarbamate)-cadalene was remarkably lower. Moreover, in silico studies showed that 7-hydroxy-3,4-dihydrocadalene, and not so the semi-synthetic derivatives, has optimal physical-chemical properties to lead a promising cytotoxic agent. Further examination on the action mechanism of 7-hydroxy-3,4-dihydrocadalene suggested that this natural product exerted cytotoxicity via oxidative stress as evidenced in a significantly increase of intracellular ROS levels and in an induction of lipid peroxidation. Furthermore, the compound increased caspase-3 and caspase-9 activities and slightly inhibited Bcl-2 levels. Interestingly, it also reduced mitochondrial ATP synthesis and induced mitochondrial uncoupling. Conclusion: Taken together, 7-hydroxy-3,4-dihydrocadalene is a promising cytotoxic compound against breast cancer via oxidative stress-induction.

Nutritional Value of Moringa oleifera Lam. Leaf Powder Extracts and Their Neuroprotective Effects via Antioxidative and Mitochondrial Regulation.

Age-related neurodegenerative disorders are an increasing public health problem. Oxidative stress is one of the major causes. Medicinal plant-based functional foods can be effective for these diseases. The aim of this work is to investigate the neuroprotective role of methanol extracts of Moringa oleifera leaf powder on antioxidant/oxidant imbalance and mitochondrial regulation in a H2O2-induced oxidative stress model in human neuroblastoma cells. On nutritional analysis, results showed that moringa contained 28.50% carbohydrates, 25.02% proteins, 10.42% fat, 11.83% dietary fiber, 1.108 mg ß-carotene, 326.4 µg/100 g vitamin B1 and 15.2 mg/100 g vitamin C. In-vitro assays revealed that moringa methanol extracts had more phenolic content and higher antioxidant activity than acetone extracts. Moreover, pretreatments with methanol extracts showed a protective effect against H2O2-induced oxidative damage through increasing cell viability and reducing free radicals. Furthermore, the extract decreased lipid peroxidation and enhanced glutathione levels and antioxidant enzyme activity. Finally, moringa also prevented mitochondrial dysfunction by regulating calcium levels and increasing mitochondrial membrane potential. The most active concentration was 25 µg/mL. In summary, the nutritional and functional properties of Moringa oleifera as a neuroprotective agent could be beneficial to protect against oxidative stress and provide necessary nutrients for a healthy diet.

Pharmacological Update Properties of Aloe Vera and its Major Active Constituents.

Aloe vera has been traditionally used to treat skin injuries (burns, cuts, insect bites, and eczemas) and digestive problems because its anti-inflammatory, antimicrobial, and wound healing properties. Research on this medicinal plant has been aimed at validating traditional uses and deepening the mechanism of action, identifying the compounds responsible for these activities. The most investigated active compounds are aloe-emodin, aloin, aloesin, emodin, and acemannan. Likewise, new actions have been investigated for Aloe vera and its active compounds. This review provides an overview of current pharmacological studies (in vitro, in vivo, and clinical trials), written in English during the last six years (2014-2019). In particular, new pharmacological data research has shown that most studies refer to anti-cancer action, skin and digestive protective activity, and antimicrobial properties. Most recent works are in vitro and in vivo. Clinical trials have been conducted just with Aloe vera, but not with isolated compounds; therefore, it would be interesting to study the clinical effect of relevant metabolites in different human conditions and pathologies. The promising results of these studies in basic research encourage a greater number of clinical trials to test the clinical application of Aloe vera and its main compounds, particularly on bone protection, cancer, and diabetes.

Current knowledge on Parmelia genus: Ecological interest, phytochemistry, biological activities and therapeutic potential.

Parmelia Acharius is one of the most representative genera within Parmeliaceae family which is the largest and the most widespread family of lichen-forming fungi. Parmelia lichens present a medium to large foliose thallus and they are distributed from the Artic to the Antartic continents, being more concentrated in temperate regions. According to its current description, the genus encompasses up to 41 different species and it is phylogenetically located within the Parmelioid clade (the largest group in the family). Interestingly, some of its species are among the most common epiphytic lichens in Europe such as Parmelia sulcata Taylor and Parmelia saxatilis (L.) Ach. The present work aims at providing a complete overview of the existing knowledge on the genus, from general concepts such as taxonomy and phylogeny, to their ecological relevance and biological interest for pharmaceutical uses. As reported, Parmelia lichens arise as valuable tools for biomonitoring environmental pollution due to their capacity to bioaccumulate metal elements and its response to acid rain. Moreover, they produce a wide array of specialized products/metabolites including depsides, depsidones, triterpenes and dibenzofurans, which have been suggested to exert promising pharmacological activities, mainly antimicrobial, antioxidant and cytotoxic activities. Herein, we discuss past and recent data regarding to the phytochemical characterization of more than 15 species. Even though the knowledge is still scarce in comparsion to other groups of organisms such as higher plants and other non-lichenized fungi. Reviewed works suggest that Parmelia lichens are worthy of further research for determining their actual possibilities as sources of bioactive compounds with potential therapeutic applications.

Antioxidant activity, neuroprotective properties and bioactive constituents analysis of varying polarity extracts from Eucalyptus globulus leaves.

Eucalyptus globulus is employed as herbal tea and therapeutical purposes. In this work, it is investigated for first time the neuroprotective activities, based on antioxidant properties, of varying polarity extracts (acetone, ethanol and methanol) from E. globulus leaves and elucidate their main bioactive constituents. Methanol and acetone extracts contained the highest phenolic compounds amount and chlorogenic acid was the major compound identified by UPLC-ESI-MS/MS. Moreover, the three tested extracts showed significant antioxidant properties, varying their potency depending on the in vitro technique used. Furthermore, E.globulus extracts were effective in ameliorating H2O2-induced oxidative stress by increasing cell viability, GSH levels and antioxidant enzymes activity and, by decreasing ROS production and lipid peroxidation levels in SH-SY5Y cells. Taken together, E.globulus leaves extracts could be used as raw material for food and pharmaceutical supplements for their high content in antioxidant compounds with health benefits properties against oxidative stress.

Evaluation of the adaptogenic potential exerted by ginsenosides Rb1 and Rg1 against oxidative stress-mediated neurotoxicity in an in vitro neuronal model.

BACKGROUND: Ginseng (Panax sp.) is a drug with multiple pharmacological actions that has been largely used in traditional medicines for the treatment of many health problems. In the therapy of neurodegenerative disorders, it has been employed due to its capacity to strengthen mental processes by enhancing cognitive performance and psychological function. Current work aimed at evaluating the adaptogenic potential of Rb1 and Rg1 against oxidative-stress mediated degeneration in a model of nervous cells. METHODS: Oxidative stress and mitochondrial dysfunction were achieved by exposing SH-SY5Y cells to the mitochondrial complex I inhibitor rotenone. The cytoprotective activity of pre-treatments with ginsenosides Rb1 and Rg1 against rotenone was assessed by determining biochemical markers regarding oxidative stress (ROS scavenging, glutathione and lipid peroxidation levels, SOD activity and Nrf2 activation) and apoptosis-related alterations (mitochondrial membrane potential, calcium levels, aconitase activity and pro/antiapoptotic proteins). Their capacity to cross the blood brain barrier was also estimated. RESULTS: At their optimal doses, ginsenosides Rb1 and Rg1 significantly ameliorated redox status within the cells; they reduced ROS and TBARS levels and improved the glutathione system, as well as they enhanced SOD activity and Nrf2 pathway activation. They protected neuronal cells against MMP loss, calcium homeostasis disruption and aconitase inhibition. Consequently, apoptotic cell death was attenuated by the pre-treatment with ginsenosides, as evidenced by the reduction in caspase-3 and Bax, and the increase in Bcl-2 expressions; also, lower levels of cytochrome C were found in the cytosol. Poor BBB permeation was demonstrated for both ginsenosides. CONCLUSIONS: In conclusion, ginsenosides Rb1 and Rg1 exhibit neuroprotective potential which is achieved, at least in part, via mitochondrial protection and the plausible involvement of Nrf2 pathway activation. Our results contribute to validate the traditional use of ginseng for cognitive-enhancing purposes and provide basis to encourage further research on the potential of ginsenosides in the treatment of neurodegenerative diseases.

Protective effects of lichen metabolites evernic and usnic acids against redox impairment-mediated cytotoxicity in central nervous system-like cells.

Lichens species produce unique secondary metabolites that attract increasing pharmacological interest, including their redox modulatory activities. Current work evaluated for the first time the in vitro cytoprotective properties, based on the antioxidant activities, of the Parmeliaceae lichens Evernia prunastri and Usnea ghattensis and the mechanism of action of their major phenolic constituents: the evernic and usnic acids, respectively. In two models of central nervous system-like cells (U373-MG and SH-SY5Y cell lines), exogenous H2O2 induced oxidative stress-mediated cytotoxicity. We first assessed their radical scavenging capacities (ORAC and DPPH tests) and the phenolic content of the extracts. At the optimal concentrations, pretreatments with evernic acid displayed significant protection against H2O2-induced cytotoxic damage in both models. It reversed the alterations in oxidative stress markers (including ROS generation, glutathione system and lipid peroxidation levels) and cellular apoptosis (caspase-3 activity). Such effects were in part mediated by a notable enhancement of the expression of intracellular phase-II antioxidant enzymes; a plausible involvement of the Nrf2 cytoprotective pathway is suggested. Usnic acid exerted similar effects, to some extent more moderate. Results suggest that lichen polyketides evernic and usnic acids merit further research as promising antioxidant candidates in the therapy of oxidative stress-related diseases, including the neurodegenerative disorders.

In vitro neuroprotective potential of lichen metabolite fumarprotocetraric acid via intracellular redox modulation.

The lichen-forming fungi Cetraria islandica has been largely used in folk medicines, and it has recently showed promising in vitro antioxidant effects in glial-like cells. Current work aimed at investigating the neuroprotective potential of its major isolated secondary metabolite: the depsidone fumarprotocetraric acid (FUM). H2O2 was used herein to induce oxidative stress (OS)-mediated cytotoxicity in two models of neurons and astrocytes cells (SH-SY5Y and U373-MG cell lines). We found that a pre-treatment with FUM significantly enhanced cell viability compared to H2O2-treated cells, and we selected the optimal concentrations in each model (1 and 25µg/ml, respectively) for assessing its cytoprotective mechanisms. FUM, which exerted effective peroxyl radical scavenging effect in the chemical oxygen radical antioxidant capacity (ORAC) assay, alleviated the alterations in OS markers provoked by H2O2. It attenuated intracellular ROS formation, lipid peroxidation and GSH depletion. At mitochondrial level, FUM prevented from the dissipation of mitochondrial membrane potential and the increase in mitochondrial calcium, implying a protective role against oxidative damage in mitochondrial membrane. Similarly, FUM pre-treatment diminished H2O2-induced apoptosis, as evidenced by the reduction in caspase-3 activity and expression; inmunoblot analysis also revealed a decrease in Bax and an increase in Bcl-2 proteins levels. Furthermore, FUM up-regulated the expression of the antioxidant enzymes catalase, superoxide dismutase-1, and hemeoxigenase-1. These findings and the activation of Nrf2 binding activity in nuclear extracts suggest a plausible involvement of Nrf2 signaling pathway in the cytoprotection by FUM. In conclusion, FUM emerges as a potential drug candidate in the therapy of OS-related diseases, such as the neurodegenerative disorders.

In vitro neuroprotective potential of the monoterpenes α-pinene and 1,8-cineole against H2O2-induced oxidative stress in PC12 cells.

Oxidative stress is involved in the pathogenesis of several neurodegenerative diseases such as Alzheimer's and Parkinson's diseases. Natural products are considered as therapeutically useful antioxidant agents against reactive oxygen species (ROS). We have evaluated the antioxidant and protective potential of the monoterpenes 1,8-cineole and α-pinene against H2O2-induced oxidative stress in PC12 (rat pheochromocytoma) cells. Pretreatment with these monoterpenes was found to attenuate the loss of cell viability and the changes in cell morphology. Moreover, they inhibited the intracellular ROS production and markedly enhanced the expression of antioxidant enzymes including catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR) and heme-oxygenase 1 (HO-1). In addition, they were able to decrease apoptosis as is evident from reduced capase-3 activity. The mechanisms of their antioxidant action appear to involve ROS scavenging and induction of the nuclear Nrf2 factor. This study demonstrates the potential beneficial therapeutic effect of these common monoterpenes on the oxidant/antioxidant balance in diseases of the nervous system.

Kaurane diterpenes as mitochondrial alterations preventive agents under experimental oxidative stress conditions.

CONTEXT: Foliol, linearol, and sidol are the most common diterpenes found in Sideritis L. spp. (Lamiaceae) with a wide range of demonstrated properties including anti-inflammatory, antioxidant, and anti-apoptotic effects. OBJECTIVE: For the first time, the present work was studied for the potential protective role of these kaurane-type diterpenes on mitochondrial oxidative stress induced by H2O2 in the human astrocytoma U373-MG cell line and in the rat adrenal pheochromocytoma PC12 cell line. MATERIALS AND METHODS: Mitochondrial protection was assayed at 5 and 10 µM concentrations for 24 h (for kaurane diterpenes) and H2O2 as oxidative stress inducer (0.1 mM for PC12 cells and 1 mM for U373-MG, for 30 min). ATP concentration was determined by high-performance liquid chromatography (HPLC), and changes in mitochondrial membrane potential, caspase-3 activity as well as in cytosolic and mitochondrial calcium levels were assessed by fluorometric techniques, by using specific fluorescent probes. RESULTS: Pretreatments for 24 h with linearol and sidol, prior to H2O2 exposure, acted as mitochondrial alterations preventive agents by increasing membrane potential (over 40-60% in PC12 cells and over 10-20% in U373-MG), restoring both cytosolic and mitochondrial calcium homeostasis (linearol at 10 µM caused a 3.5-fold decrease in cytosolic calcium concentration in PC12 cells), decreasing caspase-3 activity (over 1.25-1.5-fold for linearol and sidol) and avoiding ATP depletion (linearol increased over 20% ATP level in both cell types). CONCLUSION: Our results suggest that linearol and sidol could provide protective activity by targeting mitochondria in response to the deleterious changes induced by H2O2.

Neuroprotective activity and cytotoxic potential of two Parmeliaceae lichens: Identification of active compounds.

BACKGROUND: Lichens are symbiotic organisms capable of producing unique secondary metabolites, whose pharmacological activities are attracting much interest. PURPOSE: The present study aimed to investigate the in vitro neuroprotective effects and anticancer potential of methanol extracts of two Parmeliaceae lichens: Cetraria islandica and Vulpicida canadensis. The chemical composition of the two lichens was also determined. METHODS: Neuroprotective activity was studied with respect to the antioxidant properties of the extracts; radical scavenging tests (ORAC and DPPH assays) were performed and oxidative stress markers (intracellular ROS production, caspase-3 activity, MDA and glutathione levels) were assessed in a hydrogen peroxide-induced oxidative stress model in astrocytes. Cytotoxic activity was tested against human HepG2 (hepatocellular carcinoma) and MCF-7 (breast adenocarcinoma) cell lines. RESULTS: Cell viability studies identified a single concentration for each extract that was subsequently used to measure oxidative stress markers. Lichen extracts were able to reverse the oxidative damage caused by hydrogen peroxide, thus promoting astrocyte survival. Both lichen extracts also had anticancer activity in the cell lines, with IC50 values of 19.51-181.05 µg/ml. The extracts had a high total phenolic content, and the main constituents identified by HPLC were fumarprotocetraric acid in Cetraria islandica, and usnic, pinastric and vulpinic acids in Vulpicida canadensis. The biological activities of the lichen extracts can be attributed to these secondary metabolites. CONCLUSION: The lichen species studied are promising sources of natural compounds with neuroprotective activity and cytotoxic potential, and warrant further research.

Mitochondria-Targeted Protective Compounds in Parkinson's and Alzheimer's Diseases.

Mitochondria are cytoplasmic organelles that regulate both metabolic and apoptotic signaling pathways; their most highlighted functions include cellular energy generation in the form of adenosine triphosphate (ATP), regulation of cellular calcium homeostasis, balance between ROS production and detoxification, mediation of apoptosis cell death, and synthesis and metabolism of various key molecules. Consistent evidence suggests that mitochondrial failure is associated with early events in the pathogenesis of ageing-related neurodegenerative disorders including Parkinson's disease and Alzheimer's disease. Mitochondria-targeted protective compounds that prevent or minimize mitochondrial dysfunction constitute potential therapeutic strategies in the prevention and treatment of these central nervous system diseases. This paper provides an overview of the involvement of mitochondrial dysfunction in Parkinson's and Alzheimer's diseases, with particular attention to in vitro and in vivo studies on promising endogenous and exogenous mitochondria-targeted protective compounds.

Protective properties of Salvia lavandulifolia Vahl. essential oil against oxidative stress-induced neuronal injury.

Salvia lavandulifolia Vahl., known as "Spanish sage", has potential value in dementia for its sedative, antioxidant, anti-inflammatory and anticholinesterase properties. This work aimed to evaluate the in vitro neuroprotective activity of S. lavandulifolia essential oils, obtained from plants at different phenological stages (vegetative and flowering phases) and plants grown at different densities, against H2O2-induced oxidative stress in PC12 cells. The effect on cell viability and morphology, lipid peroxidation, GSH/GSSG ratio, intracellular ROS levels, antioxidant enzymes (CAT, SOD, GR, GPx, HO-1) and apoptotic enzymes was investigated. Comparing with H2O2-treated PC12 cells, pretreatments with essential oil samples attenuated morphological changes and loss of cell viability, decreased MDA levels and intracellular ROS production and increased GSH/GSSG ratio. Moreover, Spanish sage increased antioxidant status as evidenced in an increase of antioxidant enzyme activity and protein expression and inhibited caspase-3 activity. Furthermore, our results suggest that S. lavandulifolia essential oils are able to activate Nrf2 transcription factor. Collectively, the sample of essential oil obtained with the highest densities of planting and at flowering phase exerted the major neuroprotective activity. Our findings demonstrate that S. lavandulifolia essential oils may have therapeutic value for the prevention and treatment of neurodegenerative diseases associated with oxidative stress-induced neuronal injury.

Phenolic content, antioxidant and astroprotective response to oxidative stress of ethanolic extracts of Mentha longifolia from Sinai.

The aerial parts ofMentha longifolia L. are used as herbal remedies for curing different diseases through traditional Bedouin medicine. The antioxidant activity of the ethanolic extracts of M longifolia was investigated measuring peroxyl radical-scavenging activity by ORAC assay, with Trolox (a water-soluble analogue of α-tocopherol) employed as reference compound. In addition, the total content of phenolic compounds estimated by the Folin-Ciocalteau method and the identification of the polyphenols using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) have been performed. Furthermore, the effect of these extracts on cell viability and intracellular ROS production was assayed using the U373-MG human astrocytoma cell line in a H2O2-induced oxidative stress model. Results showed that the major type of polyphenols found were benzoic acids, cinnamic acids, flavones and flavanones. The total phenolic content was 37.7 mg gallic acid/g sample and the ORAC value was 1.355 .mol TE/mg sample. The data obtained in cellular assays demonstrated that these ethanolic extracts protected H2O2-induced astrocyte damage by increasing cell viability and inhibiting production of intracellular ROS. These results suggest that the investigated extracts obtained from the aerial parts of M longifolia have antioxidant potential related to their phenol content which have important beneficial health effects, especially in those disease associated with ROS.

Compositional analysis and in vitro protective activity against oxidative stress of essential oils from egyptian plants used in traditional medicine.

The Sinai desert in Egypt contains great variability in plants extensively used for traditional medicines such as Achillea fragrantissima, Chiliadenus montanus, Mentha longifolia and Haplophyllum tuberculatum. The essential oils extracted by hydrodistillation from the aerial parts have been analyzed. Subsequently, their potential protective activity against oxidative stress has been evaluated, employing H2O2 as oxidant inductor and astrocytes as the cell model. The chemical composition of the essential oils was analyzed by GC/MS. Most of the compounds identified in A. fragrantissima and M. longifolia samples were oxygenated monoterpene derivatives, whereas for H. tuberculatum they were monoterpenes hydrocarbons and oxygenated compounds, and for C. montanus oxygenated monoterpenes and oxygenated sesquiterpenes predominated. The in vitro evaluation of antioxidant properties, using ORAC assay, revealed that M. longifolia essential oil possessed the highest scavenging activity against peroxyl radicals, following by H. tuberculatum, A. fragrantissima and C. montanus. Under oxidative stress conditions, M. longifolia and H. tuberculatum essential oils were the only ones that protected human astrocytoma U373-MG cells against H2O2 damage. Both essential oils prevented cell death and inhibited ROS production caused by H2O2. M. longifolia essential oil was the most active, suggesting an interesting prevention role in those CNS disorders associated with oxidative stress.

Nrf2-dependent neuroprotective activity of diterpenoids isolated from Sideritis spp.

ETHNOPHARMACOLOGICAL RELEVANCE: The species of the genus Sideritis are extensively used in the Mediterranean area in folk medicine for their digestive, antimicrobial, anti-inflammatory and antioxidant properties, among others. Moreover, Sideritis species as tea infusions are popularly known for improving memory function and cognitive ability. Diterpenoids are one of the most abundant and important pharmacological interest of the classes of natural products presented in these medicinal plants. AIM OF THE STUDY: To determine for the first time the neuroprotective effects, based on their antioxidant properties, of the three isolated major diterpenoids andalusol, conchitriol and lagascatriol in an oxidative stress model. MATERIALS AND METHODS: H2O2 was used as oxidant inductor and rat adrenal pheochromocytoma PC12 cells as cellular model. Cell viability was measured using MTT and LDH assays, lipid peroxidation was determined by HPLC, GSH and GSSG levels assessed by fluorometric techniques, enzymatic activity and protein expression were determined by spectrofometric techniques and Western blot, respectively. RESULTS: Pretreatments with the three diterpenoids significantly attenuated H2O2-induced changes in mitochondrial integrity and activity (MTT assay), in cell membrane integrity (LDH assay) and in cell morphology. Moreover, these diterpenoids inhibited intracellular ROS production H2O2-induced, reduced lipid peroxidation and counteracted GSH/GSSG changes. Furthermore, both activities and protein expression of the endogenous antioxidant enzymes (CAT, SOD, GR, GPx and HO-1) were increased. The Nrf2 pathway was involved, at least in part, in the protective effects of these diterpenoids. CONCLUSION: These findings suggest that these natural compounds provide significant antioxidant effects in PC12 under for counteracting the oxidative damage H2O2-induced and their potential role as useful agents for the prevention of those oxidative stress-mediated dementia disorders. Andalusol was the most active compound among the studied diterpenoids.

Protective effects of Merlot red wine extract and its major polyphenols in PC12 cells under oxidative stress conditions.

UNLABELLED: The potential effect of the extracts from free-run and pressed Merlot red wine has been evaluated in PC12 cells under oxidative stress situation. Comparing both vinification process, pressed Merlot red wine extract possessed higher neuroprotective activity than the free run wine, possibly attributed to the major content in all global polyphenolic families. High performance liquid chromatography determination of individual polyphenols showed that the major compounds found in Merlot red wine extract were quercetin, catechin, epicatechin, tyrosol, gallic acid, and procyanidins. Pretreatments with these polyphenolic compounds (0.25 mM and 0.1 mM, 24 h) significantly increased cell viability of H(2)O(2) and Fenton reaction treated cells. Moreover, these polyphenols attenuated ROS production and decreased the Redox Index of glutathione (RI = GSSG/GSH + GSSG) in cells treated only with Fenton reaction. Furthermore, some polyphenols induced antioxidant enzymes activity and protein expression. Quercetin was the most active. These results support the beneficial effects of red wine extracts and some of its polyphenols under oxidative stress conditions. PRACTICAL APPLICATION: This research provides evidences of the preventive properties of wine extracts and its major polyphenols under oxidative stress conditions.

Effect of cooking and germination on phenolic composition and biological properties of dark beans (Phaseolus vulgaris L.).

Legumes are the basés diet in several countries. They hold a high nutritional value, but other properties related to human health are nowadays being studied. The aim of this work was to study the influence of processes (boiling or germination) on the phenolic composition of dark beans (Phaseolus vulgaris L. c.v. Tolosana) and their effect on their antioxidant, neuroprotective and anticancer ability. Phenolic composition of raw and processed dark beans was analysed by HPLC-PAD and HPLC-ESI/MS. The antioxidant activity was evaluated by ORAC. Astrocytes cultures (U-373) have been used to test their neuroprotective effect. Anticancer activities were evaluated on three different cell lines (renal adenocarcinoma (TK-10), breast adenocarcinoma (MCF-7) and melanoma (UACC-62)) by sulphorhodamine B method. Qualitative and quantitative differences in phenolic composition have been observed between raw and processed dark beans that influence the antioxidant activity, mainly for germinated samples which show a decrease of antioxidant capacity. Although every assayed extracts decreased reactive oxygen species release and exhibited cytotoxicity activities on cancer cell lines, raw beans proved to be the most active in neuroprotective and antitumoral effects; this sample is especially rich in phenolic compounds, mainly anthocyanins. This study further demonstrated that phenolic composition of dark beans is related with cooking process and so with their neuroprotective and anticancer activity; cooking of dark beans improves their digestion and absorption at intestinal level, while maintaining its protective ability on oxidative process at cellular level.

Diterpenoids isolated from Sideritis species protect astrocytes against oxidative stress via Nrf2.

Andalusol (1), conchitriol (2), and lagascatriol (3) are diterpenoids produced by Sideritis species. Hydrogen peroxide-induced oxidative stress in U373-MG cells was used as an in vitro model to evaluate the cytoprotective potential, based on antioxidant properties, of these isolated compounds and to investigate the underlying mechanisms. Experimental results revealed that pretreatment with compounds 1 and 3 restored H2O2-induced oxidative changes by increasing cell viability, attenuating morphological changes, inhibiting intracellular ROS production and lipid peroxidation, and enhancing the antioxidant defense system. Furthermore, nuclear factor E2-related factor 2 (Nrf2) signaling was involved in the protective mechanisms of 1-3. The present findings suggest that two of the compounds studied (1 and 3) might play a preventive role in neurodegenerative diseases associated with oxidative stress.

Neuroprotective properties of Spanish red wine and its isolated polyphenols on astrocytes.

The neuroprotective effect of Merlot red wine and its isolated polyphenols was evaluated in an oxidative stress model induced by Fenton reaction and hydrogen peroxide in the human astrocytoma U373 MG cell line. Compared with cells treated only with oxidative stress inductors, the pre-incubation with Merlot red wine for 24h caused a significant increase in cell viability for all concentrations assayed. The most abundant polyphenols found in Merlot red wine were the flavonoids catechin (37.8mg/l), epicatechin (52.3mg/l), quercetin (5.89mg/l) and procyanidins (15.2mg/l), the hydroxybenzoic acid gallic acid (16.7mg/l), and the phenolic alcohol tyrosol (31.4mg/l). The potential protective role of these polyphenols when isolated was then assessed in treated Fenton reaction U373 MG cells. Polyphenols decreased reactive oxygen species generation and increased the activity and the protein expression of the antioxidant enzymes catalase, superoxide dismutase, glutathione reductase and glutathione peroxidase. Of the polyphenols, quercetin and procyanidins showed the highest neuroprotective effect.