Amburana cearensis seed extracts protect PC-12 cells against toxicity induced by glutamate

ABSTRACT Amburana cearensis (Allemão) A.C. Sm., Fabaceae, has been widely studied for its medicinal activities. Many neurodegenerative disorders are caused by oxidative stress, mitochondrial dysfunction, excitotoxicity induced by glutamate and ultimately cell death. This study describes the chemical profile of the ethanolic, hexane, dichloromethane, and ethyl acetate extracts obtained from seeds of A. cearensis. The objective of this study was to investigate the chemical profile of extracts obtained from seeds of A. cearensis, as well as their cytotoxicity and neuroprotective effects in cultures of neural PC12 cells. Metabolite profile was performed by GC–MS. PC12 cells were treated with increasing concentrations of the extracts (0.01–2000 µg/ml) and the cell viability was analyzed after 24 and 72 h using an MTT test. For the excitotoxicity assay, PC12 cells were pre-treated with glutamate (1 mM) for 6 h and treated with increasing concentrations (0.1–1000 µg/ml) of the extracts. The chromatographic analysis of the extracts detected various compounds with antioxidant properties, with the majority of peaks corresponding to the isoflavone coumarin. Only the hexane extract showed toxicity after 72 h exposure at the highest concentration (1000 µg/ml). By contrast, all extracts increased the cellular viability of PC12 cells against the toxicity caused by glutamate. Therefore, the extracts from the seeds of A. cearensis showed no toxicity and have neuroprotective potential against neuronal damage induced by glutamate, which may be related to their antioxidant properties.

Micropropagação de umburana de cheiro / Micropropagation of umburana de cheiro

Umburana-de-cheiro é uma espécie nativa da Caatinga, amplamente utilizada pela população local, devido o valor madeireiro, ornamental, forrageiro e, principalmente medicinal. Considerando o significativo risco de extinção e a inexistência de estudos relacionados ao cultivo e propagação, objetivou-se com este trabalho estabelecer um protocolo para micropropagação de Amburana cearensis. Para isso, avaliou-se a influência do regulador vegetal 6-benzilaminopurina (BAP) em cinco concentrações (0,0; 2,22; 4,44; 8,88 e 17,76µM) sobre a multiplicação in vitro de diferentes explantes (segmento nodal, ápice caulinar e segmento cotiledonar) e na etapa de enraizamento testou-se diferentes concentrações de ácido indol butírico (AIB) (0,0; 2,5; 5,0 e 10,0µM) na presença e ausência de 1,0g L-1 de carvão ativado. Maior número de brotos (2,36) foi obtido na presença de 4,44µM de BAP, a partir do segmento cotiledonar e maior porcentagem de enraizamento (92,0%) ocorreu na presença de 10,0µM de AIB na ausência de carvão ativado. Não existiram dificuldades para o estabelecimento do protocolo de micropropagação de A. cearensis.

Umburana de cheiro is a species native to the Caatinga, widely used by local people, because the value of timber, ornamental, forage and especially medicinal. Considering the significant risk of extinction and the ausence of studies related to the cultive and propagation, the aim of this work was to establish a protocol for micropropagation Amburana cearensis. For this, we evaluated the influence of plant growth regulator BAP at five different concentrations (0.0, 2.22, 4.44, 8.88 and 17.76mM) on the in vitro multiplication of different explants (nodal segments, apex cotyledon and stem segment) and the rooting stage we tested different concentrations of IBA (0.0, 2.5, 5.0 and 10.0mM) in the presence and absence of 1.0g L-1 of activated charcoal. Highest number of shoots (2.36) was obtained in 4.44mM of BAP from cotyledonary segment and higher rooting percentage (92.0%) ocurred in 10.0mM of IBA in the absence of activated charcoal. There were no difficulties to the establishment of micropropagation protocol A. cearensis.

Influence of process conditions on the physicochemical characteristics of cumaru (Amburana cearensis) powder produced by spray drying

The aim of the work was to study the spray-drying of ethanolic extract from Amburana cearensis (Allemão) A.C. Sm., Fabaceae, in order to obtain powders with better pharmacological and technological properties for herbal medicine. A 2³ fractional factorial statistical design was used to find adequate spray-drying operating conditions (inlet air temperature; feed flow rate and air flow rate) to produce A. cearensis powder with adequate concentration of active principles (amburoside and coumarin), low moisture content and high process yield. The HPLC analyses showed that the spray-drying powder of A. cearensis production did not cause alterations in the chromatographic profile when related to the fluid extract. The most significant factor that affected the amburoside concentration was air flow rate, while the concentration of coumarin, a thermolabile molecule, was influenced mainly by inlet air temperature. The moisture content of the spray-drying powder of A. cearensis varied from 3.72 to 5.85% (w/w), while the maximal process yield was 41.1% (w/w). The present study demonstrates for the first time the best operating conditions to produce A. cearensis extract powder that were adequate when related to the coumarin and amburoside concentrations and moisture content. However, additional studies are still needed to improve mainly it technological characteristics.

Avaliação da atividade antiedematogênica, antimicrobiana e mutagênica das sementes de Amburana cearensis (A. C. Smith) (Imburana-de-cheiro) / Assessment of the antiedematogenic, antimicrobial and mutagenic activity of Amburana cearensis seeds (A.C. Smith) (Imburana-de-cheiro)

Foram avaliados os efeitos antiinflamatório, antibacteriano e mutagênico do extrato aquoso das sementes de Amburana cearensis. A atividade antiinflamatória foi avaliada em modelo de edema de pata induzido por carragenina, utilizando o extrato em concentrações de 10 % e 20 % nos grupos experimentais; AAS 10 mg/kg (v.o) no grupo padrão e água destilada no grupo controle. A atividade antimicrobiana foi determinada através do método de diluição em Agar, utilizando concentrações de extrato de 10 %, 7,5 %, 5 %, 2,5 % e 1 % em cepas de Sthaphylococcus aureus ATCC 27853, Escherichia coli ATCC 25922 e Pseudomonas aeruginosas ATCC 25923) e a atividade mutagênica foi determinada pelo teste de Allium cepa, utilizando extrato em concentrações de 0,02 mg/mL, 0,1 mg/mL e 0,5 mg/mL. O extrato aquoso das sementes de Amburana cearensis nas concentrações de 10 % e 20 % apresentou efeito antiedematogênico, estatisticamente significativo a partir de duas horas após administração do flogógeno, e tal efeito persistiu até 24 horas após a indução da resposta inflamatória. Quanto à atividade antibacteriana, o extrato não apresentou ação contra as cepas bacterianas de Sthaphylococcus aureus, Escherichia coli e Pseudomonas aeruginosas nas concentrações testadas. A análise dos resultados do teste de Allium cepa evidenciou ação tóxica (em concentração de 0,5 mg/mL) e mutagênica (micronúcleo 0,1 mg/mL e aberrações cromossômicas 0,1 mg/mL e 0,5 mg/mL) do extrato de Amburana cearensis em células meristemáticas de Allium cepa. Tais resultados sugerem potencial aplicação terapêutica no tratamento da inflamação. Contudo, também demonstram a necessidade de estudar para comprovar a segurança na utilização dessa espécie.

The anti-inflammatory, antibacterial and mutagenic effects of the aqueous extract of Amburana cearensis seeds were evaluated. The anti-inflammatory activity was evaluated by a paw edema model induced by carrageenan, using the extract at 10 % and 20 % concentrations in the experimental groups: AAS 10 mg/kg (orally administrated) in the standard group and distilled water in the control group. The antimicrobial activity was determined by the agar dilution method, using extract concentrations of 10 %, 7.5 %, 5 %, 2.5 % and 1% in strains of Staphylococcus aureus ATCC 27853, Escherichia coli ATCC 25922 and Pseudomonas aeruginosas ATCC 25923), and the mutagenic activity was determined by the Allium cepa test using extract concentrations of 0,02 mg/mL, 0,1 mg/mL and 0,5 mg/mL. The aqueous extract of Amburana cearensis seeds at 10 % and 20 % concentrations had an statistically significant antiedematogenic effect two hours after administering the flogogen, and this effect persisted for up to 24 hours after inducing the inflammatory response. Regarding the antibacterial activity, the extract showed no action against the bacterial strains of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosas at the concentrations tested. The results of the Allium cepa test showed the toxic (at a concentration of 0,5 mg/mL) and mutagenicity (0.1 mg/mL micronucleus and 0.1 mg/mL and 0.5 mg/mL chromosomal aberrations) actions of the Amburana cearensis extract on meristematic cells of Allium cepa. These results suggest potential therapeutic applications to treat inflammations. However, they also show the need for further studies to demonstrate the safe use of this species.

Efficacy and safety of cumaru syrup as complementary therapy in mild persistent asthma: a double-blind, randomized, placebo-controlled study

Amburana cearensis is a medicinal plant known as "cumaru". It is used in Northeast Brazil in the treatment of respiratory diseases. This was a randomized, double-blind, placebo-controlled study, with the aim of evaluating the efficacy and safety of cumaru syrup as complementary therapy in mild persistent asthma. The study consisted of 3 phases, pre-treatment, treatment and post-treatment. The primary efficacy outcome was comparison of the changes reported by patients of the cumaru and placebo groups after treatment, using the "Asthma Quality of Life Questionnaire" (AQLQ). The secondary outcome was the effect of cumaru syrup on lung function based on spirometry. The results showed that in the cumaru group, the proportion of patients who had global improvement in asthma symptoms was significantly greater (61.90%, P=0.0009) than in the placebo group (9.52%). Only the spirometric parameters Forced Vital Capacity (FVC) and Forced Expiratory Volume in 1 second (FEV1) showed significant intergroup differences in post-treatment (P<0.05). The hematological and serum chemistry tests performed in the pre-treatment and post-treatment showed no statistically significant differences (P>0.05). Adverse events were reported by 3 patients (14.29%) in the cumaru group and 3 patients (14.29%) in the placebo group. All adverse events were considered non-serious and mild.

Amburana cearensis é uma planta medicinal conhecida como "cumaru". No Nordeste do Brasil é usada no tratamento de doenças respiratórias. Este é um estudo randomizado, duplo-cego e controlado por placebo, com o objetivo de avaliar a eficácia e segurança do xarope de cumaru como terapia complementar da asma persistente leve. O estudo consistiu de três fases, pré-tratamento, tratamento e pós-tratamento. A variável primária para determinação da eficácia foi a comparação das mudanças referidas pelos pacientes dos grupos cumaru e placebo após o tratamento, usando o "Questionário sobre Qualidade de Vida na Asma" (QQVA). A variável secundária foi o efeito do xarope de cumaru na função pulmonar baseado na espirometria. Os resultados mostraram que no grupo cumaru, a proporção de pacientes com melhora global dos sintomas da asma foi significativamente maior (61,90%, P=0.0009) que no grupo placebo (9,52%). Somente os parâmetros espirométricos, capacidade vital forçada (CVF) e volume expiratório forçado no primeiro segundo (VEF1), mostraram diferença intergrupo significtivas no pós-tratamento (P<0.05). Os testes hematológicos e do soro realizados no pré-tratamento e pós-tratamento não mostraram diferenças estatisticamente significativas (P>0.05). Eventos adversos foram reportados por 3 pacientes (14,29%) no grupo cumaru e 3 (14,29%) no grupo placebo. Todos os eventos adversos foram não sérios e leves.

Antinociceptive effect of the ethanolic extract of Amburana cearensis (Allemão) A.C. Sm., Fabaceae, in rodents / Efeito antinociceptivo do extrato etanólico de Amburana cearensis (Allemão) A.C. Sm., Fabaceae, em roedores

The ethanolic extract of the trunk bark of Amburana cearensis (EEA) was examined for its oral (p.o.) analgesic activity at the doses of 100, 200 and 400 mg/kg body weight. In the acetic acid-induced writhing test, the EEA (200 and 400 mg/kg, p.o.) reduced the number of writhing by 33.4 percent and 40.7 percent, respectively. Additionally, EEA (100, 200 and 400 mg/kg, p.o.) decreased by 77.5 percent, 79.7 and 91.3 percent, respectively, the paw liking time in the second phase of the formalin test. Therefore, EEA showed a dose-dependent analgesic effect in formalin test and was effective in reducing writhing in mice.

O extrato etanólico da entrecasca de A. cearensis (EEA) foi avaliado em modelos experimentais de nocicepção. No teste das contorções abdominais induzidas pelo ácido acético o EEA (200 e 400 mg/kg, v.o.) foi significativamente efetivo em inibir o estímulo álgico (33,4 por cento e 40,7 por cento), respectivamente, em relação aos animais do grupo controle. O EEA, em todas as doses, promoveu uma redução significativa do tempo de lambidas das patas na segunda fase do teste da formalina (77,5 por cento; 79,7 e 91,3 por cento). Os resultados sugerem uma ação antinociceptiva do EEA.