Boldo Research Articles

Boldine Activates Intrinsic Apoptotic Pathway in DU-145 Androgen-Independent Prostate Cancer Cell Line

Prostate cancer is one of the most common forms of cancer in men and continues to be a problem in the developed world. The treatment approaches for androgen-independent prostate cancer are unsatisfactory and the survival of those patients remains poor. Thus, there is a strong demand to develop novel therapeutic agents to treat and prevent this advanced malignancy. The present study evaluated the effect of boldine (2,9-dihydroxy-1,10-dimethoxy-aporphine), an aporphine alkaloid occurs abundantly in the leaves of Boldo (Peumus boldus Molina), on growth and cell death of DU-145 androgen-independent prostate cancer cell line. The cell viability was measured by MTT test and LDH release was used to quantify necrosis cell death. Genomic DNA, caspase-3 activity, expression of cleaved caspase-9, Hsp70, Bcl-2 and Bax proteins were analyzed in order to study the apoptotic process. The results showed that boldine was able to reduce cell viability in the range of 60-240 mM concentrations, and suggest this aporphine alkaloid induces cell death by intrinsic apoptotic pathway that probably involves the down-regulation of heat shock protein 70 (Hsp70). In fact, an increase of caspase-3 enzyme activity and Bax protein expression, in conjunction with the more pronounced decrease in Bcl-2 occurred in DU-145 cells treated with boldine at 60-120 mM concentrations. In addition, caspase-9 was shown to be observably activated. Moreover, boldine such as quercetin, a well-known Hsp70 protein inhibitor, induced a reduction of Hsp70 expression. The hypothesis of apoptosis induction in our experimental conditions was reinforced by a high DNA fragmentation at 60-120 µM concentrations, not correlated to LDH release. The present findings, starting point for further investigation, suggest that boldine structure might be used to design novel derivatives for the developing of potential new drugs for advanced prostate cancer therapy.

Boldine Ameliorates Estrogen Deficiency-Induced Bone Loss via Inhibiting Bone Resorption.

Osteoporosis is an enormous health problem caused by the imbalance between bone resorption and bone formation. The current therapeutic strategies for osteoporosis still have some limitations. Boldine, an alkaloid isolated from Peumus boldus, has been shown to have antioxidant and anti-inflammatory effects in vivo. For the first time, we discover that boldine has a protective effect for the estrogen deficiency-induced bone loss in mice. According to the Micro-CT and histomorphometry assays, boldine conducts this protective effect through inhibiting bone resorption without affecting bone formation in vivo. Moreover, we showed that boldine can inhibit receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast formation via impairing the AKT signaling pathways, while SC79 (an AKT agonist) partially rescue this effect. In conclusion, our results suggest that boldine can prevent estrogen deficiency-induced osteoporosis by inhibiting osteoclastogenesis. Thus, boldine may be served as a novel therapeutic agent for anti-osteoporotic therapy.

Improved drug carriage and protective potential against Cisplatin-induced toxicity using Boldine-loaded PLGA nanoparticles

BackgroundCisplatin is a widely-used potent anti-cancer drug having severe side-effects precluding its sustained use. ObjectivesPoly (lactide-co-glycolide) (PLGA)-nanoparticles loaded Boldine, an antioxidant ingredient of ethanolic extract of Boldo plant (Peumus boldus) was tested in cancer mice model, Mus musculus to examine if it could reduce unwanted Cisplatin-induced toxicity in normal tissue. Material and methodsNano-encapsulation of Boldine was done by following the standardized solvent displacement method. Physico-chemical characterization of PLGA-encapsulated nano-Boldine (NBol) was accomplished through analyses of various spectroscopic techniques. Status of major antioxidant enzymes, functional markers, and lipid peroxidation (LPO) was also determined in certain tissue and serum samples. Percentage of cells undergoing cytotoxic death, Reactive oxygen species (ROS) accumulation and mitochondrial functioning were analyzed in both normal and cancer mice. Nanoscale changes in chromatin organization were assessed by Transmission electron microscopy (TEM). mRNA and protein expressions of Top II, Bax, Bcl-2, Cyt c, caspase 3 were studied by RT-PCR, immunoblot and immunofluorescence. ResultsNBol had faster mobility, site-specific action and ability of sustained particle release. NBol readily entered cells, prevented Cisplatin to intercalate with dsDNA resulting in reduction of chromatin condensation, with corresponding changes in ROS levels, mitochondrial functioning and antioxidant enzyme activities, leading to reduction in Deoxyribose nucleic acid (DNA) damage and cytotoxic cell death. Expression pattern of apoptotic genes like Top II, p53, Bax, Bcl-2, cytochrome c and caspase-3 suggested greater cytoprotective potentials of NBol in normal tissues. ConclusionsCompared to Boldine (Bol), NBol had better ability of drug carriage and protective potentials (29.00% approximately) against Cisplatin-induced toxicity. Combinational therapeutic use of PLGA-NBol can reduce unwanted Cisplatin-induced cellular toxicity facilitating use of Cisplatin.

Immobilization of Aspergillus ficuum tannase in calcium alginate beads and its application in the treatment of boldo (Peumus boldus) tea

Tannase (tannin acyl hydrolase, E.C. 3.1.1.20) is an enzyme that catalyzes the hydrolysis of ester and depside linkages in hydrolysable tannins such as tannic acid, releasing gallic acid and glucose. It has several commercial applications in food industry, among which are gallic acid production, reduction of tannin content in fruit juices, and preparation of instantaneous tea. In this study we immobilized Aspergillus ficuum tannase in calcium alginate beads and then used it to treat boldo (Peumus boldus) tea. Such a technique allowed entrapping tannase with a 75% efficiency and appreciably increasing its thermal and pH stability compared with the free enzyme. Storage stability and reuse of the immobilized enzyme were very promising, in that about 60% of starting enzyme activity was retained after bead storage for 90 days at 4 °C or after six cycles of use. Boldo tea treatment with immobilized tannase for 120 min at 40 °C led to 31 and 60% removals of tannins and epigallocatechin gallate, an increase of about two orders of magnitude in gallic acid content, 56 and 109% increases in total flavonoids and epigallocatechin contents, a 42.8% increase in antioxidant activity and significant enhancements of tea color, clarity and pH.

Telomerase Inhibition by a New Synthetic Derivative of the Aporphine Alkaloid Boldine.

Telomerase, the enzyme responsible for cell immortality, is an important target in anti-cancer drug discovery. Boldine, an abundant aporphine alkaloid of Peumus boldus, is known to inhibit telomerase at non-toxic concentrations. Cytotoxicity of N-benzylsecoboldine hydrochloride (BSB), a synthetic derivative of boldine, was determined using the MTT method in MCF7 and MDA-MB231 cells. Aliquots of cell lysates were incubated with various concentrations of BSB in qTRAP (quantitative telomere repeat amplification protocol)-ligand experiments before substrate elongation by telomerase or amplification by hot-start Taq polymerase. The crystal structure of TERT, the catalytic subunit of telomerase from Tribolium castaneum, was used for docking and molecular dynamics analysis. The qTRAP-ligand data gave an IC50 value of about 0.17 ± 0.1 µM for BSB, roughly 400 times stronger than boldine, while the LD50 in the cytotoxicity assays were 12.5 and 21.88 µM, respectively, in cells treated for 48 h. Although both compounds interacted well with the active site, MD analysis suggests a second binding site with which BSB interacts via two hydrogen bonds, much more strongly than boldine. Theoretical analyses also evaluated the IC50 for BSB as submicromolar. BSB, with greater hydrophobicity and flexibility than boldine, represents a promising structure to inhibit telomerase at non-toxic concentrations.

Evaluation on the effects of 0.1% Peumus boldus leaf and Spiraea ulmaria plant extract combination on bacterial colonization in canine atopic dermatitis: A preliminary randomized, placebo controlled, double-blinded study

Defective skin barrier characterize canine atopic dermatitis (AD). Pyoderma is the most common complication. Herbal compounds have been suggested as alternatives to control bacterial colonization for their effect on natural antimicrobial peptides (AMPs). This study evaluated the effects of 0.1% Peumus boldus leaf and Spiraea ulmaria plant extract combination on clinical signs, bacterial colonization and AMPs secretion in atopic dogs compared to placebo.Twenty privately-owned atopic dogs were randomly divided in 2 groups (treatment: n = 10; placebo: n = 10) and their abdomen was sprayed every 24 h for 4 weeks. Total and inguinal clinical scores (CADESI-03), manual bacterial count, and skin washes for AMPs (cBD3-like and cCath) were performed on days 0, 14 and 28. AMPs were detected using in-house, previously-validated, canine-specific ELISAs. Data were statistically analyzed and a p < 0.05 was considered significant.Clinical scores and AMPs secretion did not differ significantly between the two groups at any time point. A significant reduction of the clinical scores was seen in the placebo group at 14 and 28 days (p < 0.04). On days 14 and 28, a reduction in the bacterial count was seen in the treated group compared with placebo (p < 0.009 and p = 0.04, respectively). Compared to baseline, a reduction in Staphylococcus spp. was seen in the treated group after 14 days of treatment (p < 0.03).These results show the efficacy of this plant extract combination against bacterial colonization, suggesting its potential usefulness in preventing bacterial infection in atopic dogs. The influence of this compound on AMPs secretion or other mechanisms should be further evaluated.

Microencapsulation of Peumus boldus essential oil and its impact on peanut seed quality preservation

Argentinean peanut provides a significant source of food worldwide, therefore is essential to preserve the quality of seeds during the storage period in order to ensure the yield of this crop. In this study in situ effect of a formulation based on microencapsulated boldo (Peumus boldus) essential oil (EO) was evaluated on in-pod stored peanut (Arachis hipogaea) to preserve the seed quality. Statistically significant (p < 0.05) fungitoxic effects were observed at the end of the storage period. Seed damages caused by insects was very low during the 5 months, however, reductions of seed germinations were produced by the formulation. Low levels of residual boldo oil were recorded at the end of the assay. The application of boldo oil microcapsules is useful to preserve stored peanuts avoiding its deterioration, but not when it is intended for seed, but for another purposes as by-product elaboration.

Variation of the alkaloid content of Peumus boldus (boldo)

Eighteen alkaloids were detected in the bark, leaves, wood and roots of Peumus boldus, including traces of secoboldine, N-methylsecoboldine (boldine methine), glaucine and norreticuline, not reported previously as constituents of this species. Using appropriate standards, we quantified thirteen of them by UHPLC-MS/MS. Boldine was dominant in the bark, and laurolitsine in wood and roots. The alkaloid composition of the leaves, determined for 130 individually identified trees, classified by age and sex, was highly variable, where N-methyllaurotetanine, laurotetanine, coclaurine and in some cases isocorydine predominated, but not boldine.

The potential use of Mentha x piperita L., Peumus boldus Mol. and Baccharis trimera Iless. extracts as functional food ingredients

We studied the comparative antioxidant and anti-glycation activities of Mentha x piperita L., Baccharis trimera Iless. and Peumus boldus Mol, in order to evaluate their potential interest as ingredients in functional foods. The total content of polyphenol compounds was determined by Folin-Ciocalteau assay as their antioxidant and anti-glycation capacities, using ABTS and ORAC for the first one and a model with methylglyoxal and bovine serum albumin for the latter. Then, paraoxonase 1 (PON 1) arylesterase activity was measured as well as apolipoprotein A-1 (ApoA-1) structure by SDS-PAGE, in the presence of an oxidative agent and the herbal extracts. Finally, the same procedure was applied to high density lipoprotein (HDL) particles using a Lipoprint kit. Results show that herbal extracts have a considerable amount of total polyphenols, thus a high antioxidant activity and a considerable anti-glycation activity. Furthermore, these extracts restore PON 1 activity as well as the original configuration of apoA-1 and the distribution of HDL subclasses, favouring anti-atherogenic particles. These herbal extracts are interesting targets to use as ingredients in functional foods.

ESTUDOS PRÉ-CLÍNICOS E CLÍNICOS DE ESPÉCIES VEGETAIS SELECIONADAS DE PAÍSES PERTENCENTES AO MERCOSUL E ASPECTOS TOXICOLÓGICOS

As plantas medicinais são utilizadas para a prevenção e tratamento de doenças, sendo usada por cerca de 80% da população para o cuidado primário à saúde. A rica biodiversidade do Brasil e países vizinhos oferece um potencial único e incomparável para a descoberta e desenvolvimento de agentes bioativos. O objetivo desse trabalho foi fazer uma revisão sobre estudos pré-clínicos e clínicos de espécies vegetais selecionadas dos países do Mercosul e aspectos toxicológicos. Com base em trabalho publicado anteriormente pelo autor, foram selecionadas as espécies com maior número de registro na Anvisa no Brasil (Aesculus hippocastanum - Castanha da Índia e Mikania glomerata- Guaco); as mais utilizadas como medicamentos fitoterápicos simples na Argentina (Cynara scolymus L. - Alcachofra e Solanum dulcamara - Dulcamara); as plantas medicinais mais vendidas no Paraguai (Peumus boldus Mol. - Boldo e Borago officinalis - Borragem); as mais consumidas no norte do Uruguai (Aloe vera – Aloe e Passiflora edulis - Maracujá) e da Venezuela a Hedera helix (Hera) por ser muito popular no país seu consumo. Foram realizadas pesquisas nas bases de dados Google Acadêmico, PubMed e Science Direct no mês de abril de 2016, sobre ensaios pré-clínicos e clínicos dessas espécies vegetais, em português e inglês, sendo selecionados os artigos mais relevantes, com uso de animais (ratos, coelhos ou camundongos), linhagens celulares ou bacterianas, ou estudos clínicos. Em relação aos estudos pré-clínicos e clínicos, Aloe e Alcachofra, foram as que apresentaram maior número de publicações.

In vitro Modulating Activity of aqueous extracts from American Plants on Chlorpyrifos-induced toxicity on Murine Splenocytes

Chlorpyrifos is an highly toxic pesticide, which can induce immunotoxicity with deleterious effects on health worldwide. On the other hand, American plants can provide derivatives with protective and immunostimulating activity. Thus, plant potential against chlorpyrifos should be assayed. To identify bioactive aqueous extracts from Lantana grisebachii (LG), Aspidosperma quebracho-blanco (AQ), Peumus boldus (PB), and Ilex paraguariensis (IP), against chlorpyrifos-induced toxicity on female Balb/c splenocytes. Material and method: Splenocytes were treated in vitro for 72 hours with 0-35 µg/mL of chlorpyrifos, 0-100 µg/mL of each extract (LG, AQ, PB, IP), and 0-5 µg/mL of concanavalin A. Then, cellular viability and death (resazurin-based and propidium iodide stainings), hydroperoxides, lipoperoxides (xylenol orange-based assay), ?-glutamyl transpeptidase activity (Szasz method) were measured and analyzed statistically. Chlorpyrifos reduced splenocyte viability in a dose-dependent manner, which was counteracted by AQ and IP, which was less active in concanavalin A-responsive cells (p<0.05). Chlorpyrifos toxicity involved ?-glutamyltranspeptidase induction with a consequent peroxide reduction, whereas AQ and mainly IP antagonized these responses (p<0.05). The extracts of Ilex paraguariensis and Aspidosperma quebracho-blanco protected splenocytes in vitro against chlorpyrifos. This effect depended on cellular type, given that concanavalin A-responsive cells were more susceptible to this toxic.

Peumus boldus attenuates copper-induced toxicity in Drosophila melanogaster

Peumus boldus (P. boldus) is a medicinal plant popularly used in the treatment of gastrointestinal disorders. P. boldus aqueous extract is rich in phenolic compounds and alkaloids that possess antiinflammatory and antioxidant effects. In the present study, the potential protective effect of P. boldus against Cu2+-induced toxicity was investigated. Adult Drosophila melanogaster were exposed to Cu2+ (1mM and 3mM) and/or P. boldus aqueous extract (5mg/mL) in the food during 4days. Cu2+-fed flies had impairment in the negative geotaxis performance (i.e. motor climbing capability) as well as a higher incidence of mortality when compared to the control group. P. boldus co-treatment afforded protection against the Cu2+-induced toxicity. Acetylcholinesterase (AChE) and glutathione S-transferase (GST) activity decreased significantly in D. melanogaster after Cu2+ exposure. P. boldus co-exposure for 4days restored enzyme activities to control levels. In addition, Cu2+ exposure caused a significant increase in the mRNA levels of antioxidant enzymes, superoxide dismutase (Sod1), catalase (Cat), thioredoxin reductase (TrxR1) and nuclear factor erythroid 2–related factor 2 (Nrf2), as well as increased the mRNA levels of acetylcholinesterase (Ace). The expression of P-type ATPase (Atp7A) and copper uptake protein 1 (Ctr1A) mRNAs were up-regulated in D. melanogaster exposed to Cu2+. The co-treatment with P. boldus blunted Cu2+-induced up-regulation of Atp7A and down-regulated Ctr1A mRNA expression. These findings suggest that P. boldus extracts reduce Cu2+-induced toxicity but not Cu2+ absorption in D. melanogaster. Consequently, P. boldus can be a potential therapeutical alternative for modulating Cu2+-associated toxicity.

Hospedabilidade de plantas ornamentais e medicinais ao nematoide das galhas (Meloidogyne incognita) raça 2

Em razão do aumento na produção de plantas ornamentais e medicinais no estado do Ceará, da importância agrícola do gênero Meloidogyne e da escassez de dados sobre a hospedabilidade desse patógeno nessas espécies, objetivou-se avaliar a susceptibilidade de 30 espécies, sendo 20 ornamentais (Antirrhimum majus), (Gazania ringens), (Carthamus tinctorius), (Bryophyllum cayicinum), (Ceasalpinia pulcherrima), (Thumbergia alata), (Petunia hibryda), (Exacum affine) (Catharanthus roseus), (Opuntia sp.), (Sansevieria trifasciata), (Asparagus densiflorus), (Hibiscus mutabilis-roreus), (Impatiens balsamiana), (Celosia spicata), (Antirrhimum sp.), (Dianthus chinensis), (Zinnia elegans), (Tagetes patula), (Capsicum annuum) e 10 medicinais (Peumus boldus), (Ocimum gratissimum), (Mentha arvensis), (Mentha x Vilosa), (Plectranthus amboinicus), (Ocimum bassilicum), (Rosmarinus officinalis), (Cymbopogon citratus), (Lippia alba), (Cymbopogon winterianus). Realizou-se o ensaio em casa de vegetação, do Setor de Fitossanidade, Departamento de Fitotecnia, da Universidade Federal do Ceará. A inoculação foi realizada com 4.000 ovos/J2 por vaso. A avaliação das plantas deu-se aos 60 dias após a inoculação. Avaliou-se a reação das plantas, mensurando-se: número de galhas e de ovos, índice de massa de ovos, fator de reprodução e redução do fator de reprodução. A partir dos resultados, classificou-se a reação das plantas ao nematoide por meio de cinco critérios. Verificou-se que das plantas ornamentais apenas a espécie T. patula não apresentou galhas em suas raízes e das medicinais, as espécies M. vilosa, L. Alba, C. citratus, C. winterianus e P. boldus também não apresentaram galhas. Assim, conclui-se que as plantas ornamentais são potenciais introdutoras de nematoide das galhas em áreas indenes e que as medicinais, podem ser utilizadas em programas de rotação de culturas em função da pouca susceptibilidade ao nematoide das galhas.

Antioxidant and antibacterial activity of different extracts from herbs obtained by maceration or supercritical technology

Supercritical fluid extraction with carbon dioxide has been demonstrated to be an attractive method to apply to different processes involved in the food industry due to several advantages when it is compared with conventional organic solvents.Supercritical CO2 extracts from rosemary (Rosmarinus officinalis), boldo (Peumus boldus), cedron (Aloysia citrodora), congorosa (Maytenus ilicifolia), yerba mate (Ilex paraguariensis) and pitanga (Eugenia uniflora) were obtained at different extraction conditions; their antioxidant effect was studied and compared with that obtained by traditional solvent maceration methods and with that shown by different common synthetic antioxidants. Additionally, antibacterial activity of extracts from the different herbs obtained was determined against different microorganisms.Supercritical extracts from these herbs were efficient in stabilizing sunflower oil; thus, they could represent a valuable natural alternative to synthetic antioxidants. Herbal extracts prove to be a source of compounds with antibacterial activity, with promising applications in the control of microbial deterioration of food.

Redox Potential of Pomegranate (Punica granatum) and Boldo (Peumus boldus) Included in the Diets of New Zealand White Rabbits

Redox Potential of Pomegranate (Punica granatum) and Boldo (Peumus boldus) Included in the Diets of New Zealand White Rabbits

Evaluation of Boldine Activity against Intracellular Amastigotes of Leishmania amazonensis.

Leishmaniasis is a neglected and endemic disease that affects poorest population mainly in developing countries. A lack of adequate and definitive chemotherapeutic agents to fight against this infection has led to the investigation of numerous compounds. The aim of this study was to investigate in vitro activity of boldine against Leishmania amazonensis murine cell infection. Boldine ((S)-2,9-dihydroxy-1,10-dimethoxy-aporphine) is an aporphine alkaloid found abundantly in the leaves/bark of boldo (Peumus boldus Molina), a widely distributed tree native to Chile. The in vitro system consisted of murine macrophage infection with amastigotes of L. amazonensis treated with different concentrations from 50 to 600 μg/ml of boldine for 24 hr. Intracellular parasite destruction was assessed by morphological examination and boldine cytotoxicity to macrophages was tested by the MTT viability assay. When cells were treated with 100 μg/ml of boldine the reduction of parasite infection was 81% compared with untreated cultures cells. Interestingly, boldine-treatment caused a concentration-dependent decrease of macrophage infection that culminated with 96% of reduction when cells were submitted to 600 μg/ml of boldine. Cell cultures exposed to 100 μg/ml of boldine and 300 μg/ml of Glucantime® during 24 hr showed a significant reduction of 50% in parasitized cells compared with cell cultures exposed just to Glucantime®. The study showed that treatment with boldine produces a better effect than treatment with the reference antimonial drug, glucantime, in L. amazonensis infected macrophage. Our results suggest that boldine is a potentially useful agent for the treatment of leishmaniasis.

Computer-Aided 13C NMR Chemical Profiling of Crude Natural Extracts without Fractionation.

A computer-aided, 13C NMR-based dereplication method is presented for the chemical profiling of natural extracts without any fractionation. An algorithm was developed in order to compare the 13C NMR chemical shifts obtained from a single routine spectrum with a set of predicted NMR data stored in a natural metabolite database. The algorithm evaluates the quality of the matching between experimental and predicted data by calculating a score function and returns the list of metabolites that are most likely to be present in the studied extract. The proof of principle of the method is demonstrated on a crude alkaloid extract obtained from the leaves of Peumus boldus, resulting in the identification of eight alkaloids, including isocorydine, rogersine, boldine, reticuline, coclaurine, laurotetanine, N-methylcoclaurine, and norisocorydine, as well as three monoterpenes, namely, p-cymene, eucalyptol, and α-terpinene. The results were compared to those obtained with other methods, either involving a fractionation step before the chemical profiling process or using mass spectrometry detection in the infusion mode or coupled to gas chromatography.

Nutritional Composition and Polyphenolic Content of Edible Peumus boldus Mol. Fruits.

Peumus boldus is an endemic Chilean tree whose edible fruit lack a history of scientific research. The aim of this study was to determine its nutritional composition (moisture, ash, crude fiber, proteins, carbohydrates and lipids) and polyphenolic content (total phenolic content, HPLC-DAD, HPLC-MS/MS analysis) at different stages of development. The results show that the fruit decreased in ash, crude fiber and total phenolic content throughout its development, while its carbohydrate and protein content increased. An increase in phenolic molecules such as catechin and epicatechin was also observed. This is the first report on the nutritional composition and phenolic content of Peumus boldus Mol. fruits.

Neuroinflammatory response to experimental stroke is inhibited by boldine.

Inflammation plays a pivotal role in the development of ischemic brain damage. Astrocyte activation promotes the production of several proinflammatory mediators, such as TNF-α and iNOS. Eventually, neuronal death occurs, leading to the development of motor and memory deficits in patients. Boldine is the main alkaloid in the leaves and bark of the Peumus boldus Molina, and has anti-inflammatory and antioxidant properties. The aim of this work was to investigate the neuroprotective effect of boldine on neuroinflammation and memory deficits induced by permanent middle cerebral artery occlusion (pMCAO) in mice. Thirty minutes before pMCAO and during the next 5 days, animals received vehicle (0.025 µmol/l HCl) or boldine (8, 16 and 25 mg/kg, intraperitoneally). The extension of the infarct area, neurological scores, and myeloperoxidase activity were evaluated 24 h after pMCAO. Locomotor activity, working, and aversive memory were evaluated 72 h after pMCAO, object recognition memory was tested 96 h after pMCAO, and spatial memory was tested 120 h after pMCAO. Cresyl violet, Fluoro-Jade C staining, and immunohistochemical for GFAP, TNF-α, and iNOS were also carried out. The treatment with boldine significantly decreased the infarct area, improved the neurological scores, and increased cell viability. The vertical exploratory activity and aversive, spatial, object recognition, and working memory deficits induced by pMCAO were prevented by boldine. Moreover, myeloperoxidase activity and GFAP, TNF-α, and iNOS immunoreactivity were decreased significantly by boldine. Although various mechanisms such as its antioxidant activity should be considered, these results suggest that the neuroprotective effect of boldine might be related in part to its anti-inflammatory properties.

Essential Oils from &amp;lt;i&amp;gt;Mentha piperita, Cymbopogon citratus, Rosmarinus officinalis, Peumus boldus&amp;lt;/i&amp;gt; and &amp;lt;i&amp;gt;Foeniculum vulgare&amp;lt;/i&amp;gt;: Inhibition of Phospholipase A2 and Cytotoxicity to Human Erythrocytes

The essential oils from Mentha piperita, Cymbopogon citratus, Rosmarinus officinalis, Peumus boldus and Foeniculum vulgare were extracted by hydrodistillation and characterized and quantified by GC-MS and GC-DIC. The oils induced hemolysis with all the doses evaluated (0.6 to 1.8 μL), and the diameters of the halos varied between 9 and 15 mm. Pre-incubation of P. boldus oil with Bothrops jararacussu venom resulted in potentiation of venom-induced hemolysis (30%) (proteases and phospholipases A2). The essential oil from M. piperita (0.6 μL) inhibited venom-induced hemolysis by 45%, whereas 0.6 μL of R. officinalis oil increased the hemolysis by 20%. For the essential oil from F. vulgare, 100% inhibition of activity (0.6 and 1.2 μL) was observed. The application of C. citratus oil induced hemolysis with all the volumes evaluated. Phospholipase activity induced by the venom was only inhibited (10%) with the 0.6 μL volume of R. officinalis oil. The oils from M. piperita and F. vulgare (1.8 μL) and C. citratus oil (0.6 μL) potentiated the phospholipase activity. The results highlight the need for a broad characterization and regulation of the use of natural products, because they can have therapeutic or toxic actions.