ABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: In Brazil, there are species of snakes that become involved in accidents and cause serious health problems to the inhabitants, highlighting the genus Bothrops for being responsible for approximately 90% of accidents reported annually. In the northern region of the country, this genus is responsible for the largest number of accidents, especially among rural dwellers. These populations invest in alternative treatments for with the purpose of improving the symptoms caused by snakebites. The species Mauritia flexuosa L. f., known as buriti, is traditionally used for the treatment of envenomation by snakes. AIM OF THE STUDY: This study aimed to evaluate the antiophidic potential of the oil of Mauritia flexuosa L. f. for Bothrops moojeni H. venom, confronting cultural and scientific knowledge. MATERIALS AND METHODS: The physicochemical properties were determined, and the components present in the oil, extracted from fruit pulp, were analyzed by Gas Chromatography Coupled with Mass Spectrometry. The in vitro inhibitory capacity of the oil for phospholipase, metalloprotease and serine protease activities was investigated. In the in vivo studies, male Swiss mice were used to evaluate the effect of oil on lethality and toxicity, and hemorrhagic, myotoxic and edematogenic activities were assessed. RESULTS: GCâMS analysis identification of 90.95% of the constituents of the oil, with the main components being 9-eicosenoic acid, (Z)- (34.54%), n-hexadecanoic acid (25.55%) and (E)-9-octadecenoic acid ethyl ester (12.43%). For the substrates, the outcomes indicate that the oil inhibited the activity of the main classes of toxins present in Bothrops moojeni H. venom (VBm) at the highest dose tested (0.5 µL), with inhibition of 84% for the hydrolysis of the selective substrate for serine protease and inhibition of 60% for the hydrolysis of substrates for PLA2 and metalloproteases. The antiophidic activity in vivo was evaluated with two concentrations of the oil: 1.5 mg, the dosage the population, diluted in mineral oil to a volume of 1 tablespoon and 15 mg, administered by gavage 30 min before poisoning and at time zero (concomitant to poisoning), and both concentrations administered by gavage in combination with topical use at time zero. The bleeding time in the group treated with oil at a concentration of 15 mg administered at time zero was significantly lower than that in the control group (p < 0.05). However, a greater inhibition of bleeding time was observed when local application was combined with the gavage treatment at both concentrations tested at time zero (p < 0.05). In the myotoxicity test, oil was efficient in reducing the myotoxic effects induced by the venom at the two concentrations tested, with gavage administration at time zero and gavage plus topical administration at time zero (p < 0.05). CONCLUSIONS: The data obtained show that the oil is safe to use at the concentrations studied and contains fatty acids that may collaborate for cellular-level repair of the injuries caused by Bm poisoning. The in vitro and in vivo experiments showed that oil inhibits the main proteolytic enzymes present in the venom and that it has important activities to control the local effects caused by bothropic venom.
Subject(s)
Bothrops , Crotalid Venoms , Snake Bites , Male , Animals , Mice , Snake Bites/drug therapy , Crotalid Venoms/toxicity , Gas Chromatography-Mass Spectrometry , Serine ProteasesABSTRACT
In Brazil, there are species of snakes that become involved in accidents and cause serious health problems to the inhabitants, highlighting the genus Bothrops for being responsible for approximately 90% of accidents reported annually. In the northern region of the country, this genus is responsible for the largest number of accidents, especially among rural dwellers. These populations invest in alternative treatments for with the purpose of improving the symptoms caused by snakebites. The species Mauritia flexuosa L. f., known as buriti, is traditionally used for the treatment of envenomation by snakes. Aim of the study This study aimed to evaluate the antiophidic potential of the oil of Mauritia flexuosa L. f. for Bothrops moojeni H. venom, confronting cultural and scientific knowledge. Materials and methods The physicochemical properties were determined, and the components present in the oil, extracted from fruit pulp, were analyzed by Gas Chromatography Coupled with Mass Spectrometry. The in vitro inhibitory capacity of the oil for phospholipase, metalloprotease and serine protease activities was investigated. In the in vivo studies, male Swiss mice were used to evaluate the effect of oil on lethality and toxicity, and hemorrhagic, myotoxic and edematogenic activities were assessed. Results GC‒MS analysis identification of 90.95% of the constituents of the oil, with the main components being 9-eicosenoic acid, (Z)- (34.54%), n-hexadecanoic acid (25.55%) and (E)-9-octadecenoic acid ethyl ester (12.43%). For the substrates, the outcomes indicate that the oil inhibited the activity of the main classes of toxins present in Bothrops moojeni H. venom (VBm) at the highest dose tested (0.5 μL), with inhibition of 84% for the hydrolysis of the selective substrate for serine protease and inhibition of 60% for the hydrolysis of substrates for PLA2 and metalloproteases. The antiophidic activity in vivo was evaluated with two concentrations of the oil: 1.5 mg, the dosage the population, diluted in mineral oil to a volume of 1 tablespoon and 15 mg, administered by gavage 30 min before poisoning and at time zero (concomitant to poisoning), and both concentrations administered by gavage in combination with topical use at time zero. The bleeding time in the group treated with oil at a concentration of 15 mg administered at time zero was significantly lower than that in the control group (p < 0.05). However, a greater inhibition of bleeding time was observed when local application was combined with the gavage treatment at both concentrations tested at time zero (p < 0.05). In the myotoxicity test, oil was efficient in reducing the myotoxic effects induced by the venom at the two concentrations tested, with gavage administration at time zero and gavage plus topical administration at time zero (p < 0.05). Conclusions The data obtained show that the oil is safe to use at the concentrations studied and contains fatty acids that may collaborate for cellular-level repair of the injuries caused by Bm poisoning. The in vitro and in vivo experiments showed that oil inhibits the main proteolytic enzymes present in the venom and that it has important activities to control the local effects caused by bothropic venom.
ABSTRACT
Introdução: A procura por novas alternativas terapêuticas, como as que utilizam as plantas medicinais, tem despertado grande interesse da comunidade científica na busca por tratamentos mais eficientes para as doenças, incluindo o câncer. Terminalia fagifolia Mart. é uma planta medicinal encontrada no Cerrado brasileiro, usada popularmente no tratamento de aftas e tumores. Objetivos: Avaliar a atividade citotóxica dos extratos etanólicos da casca e das folhas da Terminalia fagifolia em linhagens celulares NIH 3T3 e L929 e tumorais PC3 e B16F10. Métodos: Foi realizada a metodologia de determinação da viabilidade celular em ensaio com monocamada de células utilizando o ensaio MTS. As linhagens NIH 3T3, L929, PC3 e B16F10 foram expostas por 24 horas a diferentes concentrações dos extratos etanólicos da casca e folhas da Terminalia fagifolia. Resultados: Os resultados adquiridos mostraram que os extratos apresentaram viabilidade celular, sendo considerada de moderada a alta, para as células normais NIH 3T3 e L929 e citotoxicidade severa para as células tumorais PC3 e B16F10. Dessa forma, torna-se necessária a continuidade dos estudos com essa planta, pois os extratos da casca e das folhas apresentaram atividades antitumorais muito promissoras. Conclusões: Os extratos da casca e das folhas demonstraram viabilidade celular ≥ 50% nas linhagens celulares normais NIH 3T3 e L929 e demonstraram atividade citotóxica para as linhagens tumorais PC3 e B16F10, apresentando redução da viabilidade celular em torno de 60% e 70%, respectivamente. (AU)
Introduction: The search for new therapeutic alternatives, as the ones that use medicinal plants, has awaken a huge interest from the scientific community in seeking through more efficient treatments for diseases, including cancer. Terminalia fagifolia Mart. is a medicinal plant found in Brazilian "Cerrado", popularly used in aphthas and tumor treatment. Objectives: To evaluate the cytotoxic activity of ethanolic extracts of the bark and leaves of the Terminalia fagifolia in cell lines NIH 3T3 and L929 and tumor cells PC3 and B16F10. Methods: The determination methodology in cellular viability was held in an assay with cells monolayer's using the MTS assay. The NIH 3T3, L929, PC3 and B16F10 lines was exposed for 24 hours in differents ethanolic extracts concentrations. Results: The acquired results showed that the extracts had cellular variability is considered moderate to high for the normal cells NIH 3T3 and L929 and severe cytotoxicity to tumor cells PC3 and B16F10. This way, it is necessary to continue studying this plant, since both the bark and leaves extracts have great antitumor activity. Conclusion: The bark and leaves extracts showed cellular variability ≥ 50 % in normal cell lines NIH 3T3 and L929 and demonstrated cytotoxic activity for tumor cells PC3 and B16F10, presenting reduction of cell variability around 60% and 80%, respectively. (AU)
