ABSTRACT
The fungus Xylaria sp. KYJ-15 was isolated from Illigera celebica. Based on the one strain many compounds (OSMAC) strategy, the strain was fermented on potato and rice solid media, respectively. As a result, two novel steroids, xylarsteroids A (1) and B (2), which are the first examples of C28-steroid with an unusual β- and γ-lactone ring, respectively, along with two new dihydroisocoumarin glycosides, xylarglycosides A (3) and B (4), were identified. Their structures were elucidated by spectroscopic methods, X-ray diffraction and electronic circular dichroism (ECD) experiments. All isolated compounds were evaluated for cytotoxicity, DPPH radical scavenging activity, acetylcholinesterase inhibitory and antimicrobial effect. Compound 1 exhibited potent AChE inhibitory activity with an IC50 value of 2.61 ± 0.05 μmol·L-1. The β-lactone ring unit of 1 is critical for its AChE inhibitory activity. The finding was further confirmed through exploring the interaction of 1 with AChE by molecular docking. In addition, both compounds 1 and 2 exhibited obvious antibacterial activity against Bacillus subtilis with a minimum inhibitory concentration (MIC) of 2 μg·mL-1. Compounds 3 and 4 exhibited antibacterial activities against Staphylococcus aureus with MICs of 4 and 2 μg·mL-1, respectively, which also exhibited DPPH radical scavenging activity comparable to the positive control with IC50 values of 9.2 ± 0.03 and 13.3 ± 0.01 μmol·L-1, respectively.
Subject(s)
Humans , Acetylcholinesterase , Molecular Docking Simulation , Anti-Bacterial Agents , Glycosides , Lactones , PainABSTRACT
Chemical constituents in soft coral Sarcophyton glaucum were separated and purified by various chromatographic methods. Based on the spectral data, physicochemical properties, and comparison with the data reported in the literature, nine cembranoids, including a new cembranoid named sefsarcophinolide(1) together with eight known cembranoids, namely(+)-isosarcophine(2), sarcomilitatin D(3), sarcophytonolide J(4),(1S,3E,7E,13S)-11,12-epoxycembra-3,7,15-triene-13-ol(5), sarcophytonin B(6),(-)-eunicenone(7), lobophytin B(8), and arbolide C(9), were identified. As revealed by biological activity experiment results, compounds 2-6 had weak acetylcholinesterase inhibitory activity, and compound 5 displayed weak cytotoxicity against K562 tumor cell line.
Subject(s)
Animals , Anthozoa , Acetylcholinesterase , Cell Line, TumorABSTRACT
Acetylcholinesterase (AChE) activity levels can be used as an indicator for AChE inhibition due to pesticide poisoning in bird species. We assessed the comparative brain cholinesterase (AChE) activity level of five bird species inhabiting pesticide exposed croplands and Protected Area i.e. Deva Vatala National Park (DVNP), Bhimber by using a spectrophotometric method. AChE activity levels ranged from 56.3 to 85.9 µmol/min/g of brain tissue of birds representing DVNP. However, AChE activity levels ranged from 27.6 to 79.9 µmol/min/g of brain tissue of birds representing croplands. AChE activity levels observed in Jungle babbler, Common babbler, and Red-vented bulbul showed significant differences (P 0.05). Maximum inhibition was recorded in Jungle babbler (53%) followed by Common babbler (35%), Red-vented bulbul (18%), White wagtail (15%), and Black drongo (7%). The brain cholinesterase inhibition levels under-protected ecosystems (DVNP, Bhimber) and agricultural landscape suggest insecticidal contamination and its impact on avifauna diversity. The study also emphasizes on the importance of pesticide-free zones to protect the biodiversity of birds.
Os níveis de atividade da acetilcolinesterase (AChE) podem ser usados como um indicador para a inibição da AChE devido ao envenenamento por pesticidas em espécies de aves. Avaliamos o nível de atividade comparativa da colinesterase cerebral (AChE) de cinco espécies de aves que habitam áreas cultivadas expostas a pesticidas e Área Protegida, ou seja, Deva Vatala National Park (DVNP), Bhimber, usando um método espectrofotométrico. Os níveis de atividade da AChE variaram de 56,3 a 85,9 µmol / min / g de tecido cerebral de aves representando DVNP. No entanto, os níveis de atividade da AChE variaram de 27,6 a 79,9 µmol / min / g de tecido cerebral de aves representando áreas de cultivo. Os níveis de atividade de AChE observados no tagarela da selva, tagarela comum e bulbul vermelho exalado mostraram diferenças significativas (P 0,05). A inibição máxima foi registrada no tagarela da selva (53%), seguido pelo tagarela comum (35%), bulbul vermelho (18%), alvéola branca (15%) e drongo preto (7%). Os níveis de inibição da colinesterase cerebral nos ecossistemas subprotegidos (DVNP, Bhimber) e na paisagem agrícola sugerem contaminação por inseticida e seu impacto na diversidade da avifauna. O estudo também enfatiza a importância das zonas livres de pesticidas para proteger a biodiversidade das aves.
Subject(s)
Animals , Acetylcholinesterase/deficiency , Bird Diseases/diagnosis , Bird Diseases/chemically induced , Pesticides/poisoningABSTRACT
Abstract Acetylcholinesterase (AChE) activity levels can be used as an indicator for AChE inhibition due to pesticide poisoning in bird species. We assessed the comparative brain cholinesterase (AChE) activity level of five bird species inhabiting pesticide exposed croplands and Protected Area i.e. Deva Vatala National Park (DVNP), Bhimber by using a spectrophotometric method. AChE activity levels ranged from 56.3 to 85.9 µmol/min/g of brain tissue of birds representing DVNP. However, AChE activity levels ranged from 27.6 to 79.9 µmol/min/g of brain tissue of birds representing croplands. AChE activity levels observed in Jungle babbler, Common babbler, and Red-vented bulbul showed significant differences (P < 0.05) at two sites. However, White wagtail and Black drongo demonstrated non-significant differences (P > 0.05). Maximum inhibition was recorded in Jungle babbler (53%) followed by Common babbler (35%), Red-vented bulbul (18%), White wagtail (15%), and Black drongo (7%). The brain cholinesterase inhibition levels under-protected ecosystems (DVNP, Bhimber) and agricultural landscape suggest insecticidal contamination and its impact on avifauna diversity. The study also emphasizes on the importance of pesticide-free zones to protect the biodiversity of birds.
Resumo Os níveis de atividade da acetilcolinesterase (AChE) podem ser usados como um indicador para a inibição da AChE devido ao envenenamento por pesticidas em espécies de aves. Avaliamos o nível de atividade comparativa da colinesterase cerebral (AChE) de cinco espécies de aves que habitam áreas cultivadas expostas a pesticidas e Área Protegida, ou seja, Deva Vatala National Park (DVNP), Bhimber, usando um método espectrofotométrico. Os níveis de atividade da AChE variaram de 56,3 a 85,9 µmol / min / g de tecido cerebral de aves representando DVNP. No entanto, os níveis de atividade da AChE variaram de 27,6 a 79,9 µmol / min / g de tecido cerebral de aves representando áreas de cultivo. Os níveis de atividade de AChE observados no tagarela da selva, tagarela comum e bulbul vermelho exalado mostraram diferenças significativas (P < 0,05) em dois locais. No entanto, alvéola branca e drongo preto demonstraram diferenças não significativas (P > 0,05). A inibição máxima foi registrada no tagarela da selva (53%), seguido pelo tagarela comum (35%), bulbul vermelho (18%), alvéola branca (15%) e drongo preto (7%). Os níveis de inibição da colinesterase cerebral nos ecossistemas subprotegidos (DVNP, Bhimber) e na paisagem agrícola sugerem contaminação por inseticida e seu impacto na diversidade da avifauna. O estudo também enfatiza a importância das zonas livres de pesticidas para proteger a biodiversidade das aves.
Subject(s)
Animals , Pesticides/toxicity , Pakistan , Acetylcholinesterase , Birds , Cholinesterase Inhibitors/toxicity , Ecosystem , Crops, AgriculturalABSTRACT
Objetivo: descrever a atividade de inibição da enzima acetilcolinesterase (AChE), por meio de ativos extraídos de alcaloides naturais. Metodologia: este estudo se configura como uma revisão sistemática da literatura, no período de janeiro de 2015 a setembro de 2021, nas bases de dados PUBMED, LILACS e SCIENCE DIRECT, com os descritores Acetylcholinesterase; Alzheimer; Alkaloids. As informações obtidas foram tabuladas para avaliação dos alcaloides inibidores da acetilcolinesterase. Resultados: de 563 artigos encontrados, 17 foram utilizados. Dois deles relataram a atividade de alcaloides inibidores da AChE por meio de ensaios clínicos, enquanto os demais a realizaram por testes in vitro. De 160 substâncias estudadas, 48 apresentaram atividade anticolinesterásica, as quais foram avaliadas de acordo com a sua concentração inibitória média (IC50). Discussão: a eficiência dos alcaloides como inibidores da AChE, provavelmente está relacionada com sua carga positiva no pH do organismo e sua boa biodisponibilidade, tendo como consequência uma atividade duradoura in vivo, em comparação com os medicamentos sintéticos. Conclusão: no presente estudo, foi possível observar uma grande diversidade de substâncias alcalóidicas antiAChE. Contudo, torna-se necessária a realização de mais ensaios in vivo e in vitro para a constatação efetiva da atividade dessas moléculas.
Objective: describe the activity of the enzyme Acetylcholinesterase (AChE) through natural actives extracted from alkaloids. Methodology: this study is a systematic literature review, from January 2015 to September 2021, in the PUBMED, LILACS, and SCIENCE DIRECT databases, with the descriptors Acetylcholinesterase; Alzheimer's; Alkaloids. The information obtained was tabulated for the evaluation of Acetylcholinesterase inhibitor alkaloids. Results: of 563 articles found, 17 were used. Two of them reported the activity of AChE-inhibiting Alkaloids through clinical trials, while the others performed it through in vitro tests. Of 160 substances studied, 48 showed anticholinesterase activity, which was evaluated according to their mean inhibitory concentration (IC50). Discussion: the efficiency of Alkaloids as AChE inhibitors is probably related to their positive charge on the body's pH and their good bioavailability, resulting in a long-lasting activity in vivo compared to synthetic drugs. Conclusion: in the present study, it was possible to observe a great diversity of antiAChE alkaloid substances. However, it is necessary to carry out more in vivo and in vitro tests to verify the effective activity of these molecules.
Subject(s)
Alkaloids , Alzheimer Disease , Acetylcholinesterase , Therapeutics , Cholinesterases , Neurodegenerative Diseases , PubMed , Alkalies , Synthetic DrugsABSTRACT
Abstract: Comparative study GC - FID /M S of essential oils of fruits, leaves and roots of the endemic plant Angelica pancicii Vandas ex Velen. revealed a significant difference in their chemical composition. The enantiomeric purity of the main component in the fruit oil (+) - ß - phellandrene was a lso confirmed. In addition, imperatorin, isoimperatorin, oxypeucedanin, oxypeucedanin hydrate, angeloylpangelin and umbelliprenin were isolated from the fruit hexane extract. The content of these coumarins in the hexane extracts from different plant parts was further determined by HPLC. The essential oils and hexane extracts were assessed for their antioxidant potential and inhibitory effect towards ï¡ - amylase and acetylcholinesterase enzymes. The fruit and leaf essential oils (> 80%) as well as the fruit he xane extract (> 62%) significantly inhibited acetylcholinesterase enzyme. Distinguish free radical scavenging properties were detected for the leaf (Inh. 95.0 ± 2.2 %) and the root (Inh. 66.0 ± 2.4 %) extracts.
Resumen: Estudio comparativo GC - FID / MS de aceites esenciales de frutas, hojas y raíces de la planta endémica Angelica pancicii Vandas ex Velen revelaron una dife rencia significativa en su composición química. También se confirmó la pureza enantiomérica del componente principal del aceite de fruta (+) - ß - felandreno. Además, se aislaron imperatorina, isoimperatorina, oxipeucedanina, hidrato de oxipeucedanina, angeloi lpangelina y umbeliprenina del extracto de hexano del fruto. El contenido de estas cumarinas en los extractos de hexano de diferentes partes de la planta se determinó adicionalmente mediante HPLC. Los aceites esenciales y extractos de hexano se evaluaron p or su potencial antioxidante efecto inhibidor de las enzimas - ï¡ - amilasa y acetilcolinesterasa. Los aceites esenciales de frutas y hojas (> 80%), así como el extracto de hexano de frutas (> 62%) inhibieron significativamente la enzima acetilcolinesterasa. Se detectaron propiedades de captación de radicales libres diferenciadas para los extractos de hoja (Inh. 95,0 ± 2,2%) y de raíz (Inh. 66,0 ± 2,4%).
Subject(s)
Acetylcholinesterase/chemistry , Angelica/chemistry , alpha-Amylases/chemistry , Oils, Volatile/chemistry , Cholinesterase Inhibitors/chemistry , Plant Leaves/chemistry , AntioxidantsABSTRACT
Abstract: This article describes the chemical composition, physical properties and acetylcholinesterase (A ChE) and butyrylcholinesterase (BuChE) activity of stem - distilled essential oil (E O ) from Bursera graveolens wood chips, Burseraceae. The plant material was acquired in Quimis (Bosque de Sancán), city of Jipijapa in the province of Manabí, coastal region o f Ecuador. Thirty - six components were identified by CG - MS, which represented 98.54% of the volatile oil. The main components were limonene (68.52%) and mentofuran (20.37%). The hydrocarbon monoterpenes constituted the most abundant fractions. The average y ield of the E O was 1.26%. Regarding the physical properties of E O , the following values were obtained: relative density (1,029 g/mL), refractive index (1,477) and specific rotation (+4,567). The E O presented IC 50 inhibition values of 47.2 and 51.9 µg/mL fo r the enzymes AChE and BuChE, respectively.
Resumen: Este artículo describe la composición química, propiedades físicas y actividad acetilcolinesterasa (AChE) y butirilcolinesterasa (BuChE) del aceite esencial (AE) destilado a vapor de astillas de madera de Bursera graveolens , Burseraceae. La materia vegetal fue adquirida en Quimis (Bosque de Sancán), ciudad de Jipijapa en la provincia de Manabí, región costera d e Ecuador. Treinta y seis componentes fueron identificados por CG - MS, que representaron al 98.54 % del aceite volátil. Los componentes principales fueron limoneno (68.52%) y mentofurano (20.37%). Los monoterpenos hidrocarburos constituyeron las fracciones m ás abundantes. El rendimiento medio del AE fue de 1.26%. Con respecto a las propiedades físicas del AE se obtuvo los siguientes valores, densidad relativa (1.029 g/mL), índice de refracción (1.477) y rotación específica (+4.567). El AE presentó valores de inhibición IC 50 de 47.2 y 51.9 µg/mL para las enzimas AChE y BuChE, respectivamente.
Subject(s)
Oils, Volatile/chemistry , Bursera/metabolism , Bursera/chemistry , Acetylcholinesterase/pharmacology , Acetylcholinesterase/chemistry , Butyrylcholinesterase/pharmacology , Butyrylcholinesterase/chemistry , Oils, Volatile/pharmacology , EcuadorABSTRACT
Abstract The ß-carboline-1,3,5-triazine hydrochlorides 8-13 were evaluated in vitro against acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). The analysed compounds were selective to BuChE, with IC50 values in the range from 1.0-18.8 µM being obtained. The N-{2-[(4,6-dihydrazinyl-1,3,5-triazin-2-yl)amino]ethyl}-1-phenyl-ß-carboline-3-carboxamide (12) was the most potent compound and kinetic studies indicate that it acts as a competitive inhibitor of BuChE. Molecular docking studies show that 12 strongly interacts with the residues of His438 (residue of the catalytic triad) and Trp82 (residue of catalytic anionic site), confirming that this compound competes with the same binding site of the butyrylthiocholine
Subject(s)
Triazines/adverse effects , In Vitro Techniques/methods , Pain , Acetylcholinesterase/pharmacology , Butyrylcholinesterase/pharmacology , Butyrylthiocholine/adverse effects , Carbolines/agonists , Cholinesterase Inhibitors/administration & dosage , Molecular Docking Simulation/instrumentationABSTRACT
Abstract The purpose of this study was to investigate the relationship between the acetylcholinesterase (AChE) inhibitory and antigenotoxic effect with the neuroprotective activity of Glaucium corniculatum methanol and water extracts rich in rutin and quercetin flavonoids. Neuroprotective activity in terms of cell survival and development against oxidative damage was measured by MTT assay and microscopic analysis in H2O2-induced NGF-differentiated PC12 (dPC12) cells. QRT-PCR and western blot hybridization method was employed for the determination of AChE inhibition of the extracts in the same cell model, and the genotoxic and antigenotoxic effects were identified with Comet assay with human lymphocytes. H2O2-induced vitality loss in dPC12 cells was inhibited in pre-treated cells with these plant extracts. Moreover, extracts stimulated neurite formation and prevented the oxidative stress-induced reduction in neurite growth. In general, it was determined that G. corniculatum methanol extract containing higher amounts of rutin and quercetin was more effective than water extract in terms of AChE inhibitory, antigenotoxic and also neuroprotective effect. In this study, it was shown for the first time that both AChE inhibitory and antigenotoxic effects of G. corniculatum may be effective in neuroprotection and it's protective and therapeutic effects against neurodegeneration may be related to the flavonoid content.
Subject(s)
Acetylcholinesterase/adverse effects , Plant Extracts/agonists , Papaveraceae/classification , Neuroprotection , Pain/classification , Flavonoids/pharmacology , Blotting, Western , Neuroprotective AgentsABSTRACT
Abstract Papaveraceae is one of the prominent alkaloid-containing families, and plants of the genus Glaucium (Papaveraceae) are known for their bioactive alkaloids. Glaucium species have been used in traditional medicine in Turkey as an analgesic, narcotic, sedative, and antitussive. In this study, it was planned to evaluate the inhibitory activity of an alkaloidal extract of Glaucium corniculatum subsp. refractum on acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) and prolyl oligopeptidase (POP), as well as exploring the chemical profile of the plant by using Gas Chromatography-Mass Spectrometry (GC-MS). The AChE, BuChE and POP inhibition activities of the alkaloidal extract of G. corniculatum subsp. refractum were determined spectrophotometrically. A rapid GC-MS method was used to identify alkaloids that could be responsible for these inhibition activities. In total, eleven alkaloids were identified in the alkaloid extract of the plant by GC-MS. Allocyptopine (52.92%) and protopine (25.38%) were found as the major constituents. The alkaloidal extract of G. corniculatum subsp. refractum showed potent AChE inhibitory activity (IC50:1.25 µg/mL) and BuChE inhibitory activity (IC50: 7.02 µg/mL). The extract also showed a remarkable inhibitory effect on POP with an IC50 value of 123.69 µg/mL. This study presents the first GC-MS investigation and POP inhibitory activity of G. corniculatum subsp. refractum.
Subject(s)
Acetylcholinesterase/adverse effects , Butyrylcholinesterase/adverse effects , Papaveraceae/metabolism , Plant Extracts/agonists , Alkaloids/analysis , Gas Chromatography-Mass Spectrometry/methods , Medicine, TraditionalABSTRACT
Marine fungi are important members of the marine microbiome, which have been paid growing attention by scientists in recent years. The secondary metabolites of marine fungi have been reported to contain rich and diverse compounds with novel structures (Chen et al., 2019). Aspergillus terreus, the higher level marine fungus of the Aspergillus genus (family of Trichocomaceae, order of Eurotiales, class of Eurotiomycetes, phylum of Ascomycota), is widely distributed in both sea and land. In our previous study, the coral-derived A. terreus strain C23-3 exhibited potential in producing other biologically active (with antioxidant, acetylcholinesterase inhibition, and anti-inflammatory activity) compounds like arylbutyrolactones, territrems, and isoflavones, and high sensitivity to the chemical regulation of secondary metabolism (Yang et al., 2019, 2020; Nie et al., 2020; Ma et al., 2021). Moreover, we have isolated two different benzaldehydes, including a benzaldehyde with a novel structure, from A. terreus C23-3 which was derived from Pectinia paeonia of Xuwen, Zhanjiang City, Guangdong Province, China.
Subject(s)
Animals , Mice , Acetylcholinesterase/metabolism , Anthozoa/microbiology , Anti-Inflammatory Agents/pharmacology , Aspergillus/chemistry , Benzaldehydes/pharmacology , Signal TransductionABSTRACT
OBJECTIVES@#To evaluate the effect of echinacoside (ECH) on cognitive dysfunction in post cerebral stroke model rats.@*METHODS@#The post stroke cognitive impairment rat model was created by occlusion of the transient middle cerebral artery (MCAO). The rats were randomly divided into 3 groups by a random number table: the sham group (sham operation), the MCAO group (received operation for focal cerebral ischemia), and the ECH group (received operation for focal cerebral ischemia and ECH 50 mg/kg per day), with 6 rats in each group. The infarct volume and spatial learning were evaluated by triphenyl tetrazolium chloride staining and Morris water maze. The expression of α7nAChR in the hippocampus was detected by immunohistochemistry. The contents of acetylcholine (ACh), malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), activities of choline acetyltransferase (ChAT), acetylcholinesterase (AChE), and catalase (CAT) were evaluated by enzyme linked immunosorbent assay. The neural apoptosis and autophagy were determined by TUNEL staining and LC3 staining, respectively.@*RESULTS@#ECH significantly lessened the brain infarct volume and ameliorated neurological deficit in infarct volume and water content (both P<0.01). Compared with MCAO rats, administration of ECH revealed shorter escape latency and long retention time at 7, 14 and 28 days (all P<0.01), increased the α7nAChR protein expression, ACh content, and ChAT activity, and decreased AChE activity in MCAO rats (all P<0.01). ECH significantly decreased MDA content and increased the GSH content, SOD, and CAT activities compared with MCAO rats (all P<0.05). ECH suppressed neuronal apoptosis by reducing TUNEL-positive cells and also enhanced autophagy in MCAO rats (all P<0.01).@*CONCLUSION@#ECH treatment helped improve cognitive impairment by attenuating neurological damage and enhancing autophagy in MCAO rats.
Subject(s)
Animals , Rats , Acetylcholinesterase , Autophagy , Brain Ischemia/metabolism , Cerebral Infarction , Cognitive Dysfunction/drug therapy , Glutathione/metabolism , Glycosides , Infarction, Middle Cerebral Artery/drug therapy , Neuroprotective Agents/therapeutic use , Rats, Sprague-Dawley , Reperfusion Injury/drug therapy , Stroke/drug therapy , Superoxide Dismutase/metabolism , alpha7 Nicotinic Acetylcholine ReceptorABSTRACT
Abstract In this sense the objective of assessing the levels of pesticide poisoning in rural farmers of San Sebastian and take AL, using acetylcholinesterase enzymes Erythrocyte and plasma as biological indicator of intoxication. This is a prospective, transversal and descriptive variables analyzed were: year whose, sex, age group, education, location, condition, route of exposure. The data were acquired by collecting blood samples and socio demographic information of farmers. Was put as the determining factor the type of conventional and organic farming, and periods of drought and rainy. 56 volunteers were analyzed. The analyses were performed in the automatic biochemical Analyzer Cobas Integra 400 plus®. According to the results of the analyses, it was the largest number of individuals with reduced values of cholinesterase, specifically the Group of conventional farming, the period of greatest change index was in the rainy season, where the activity of AChE, expressive values presented in the city of São Sebastião, with 80% result of intoxicated, in the city of the foot - take, 21.73% over the same period. On analysis of the AChP, São Sebastião has obtained the highest number of contaminated with 18.75% and 30% respectively, in the District Take Foot stood between 10 and 21.73% of reduced levels of cholinesterase. This sets the organic system of cultivation, as the best alternative for prevention of future diseases, in addition to bringing quality of life for rural workers, as well as for consumers.
Resumo Nesse sentido objetivou-se avaliar os níveis de intoxicação por agrotóxicos em agricultores rurais de São Sebastião e Pé Leve - AL, utilizando as enzimas acetilcolinesterase Eritrocitária e Plasmática, como indicador biológico de intoxicação. Trata - se de um estudo prospectivo, transversal e descritivo cujas variáveis analisadas foram: ano, sexo, faixa etária, escolaridade, local, circunstância, via de exposição. Os dados foram adquiridos através da coleta das amostras de sangue e das informações sócio demográficas dos agricultores. Foi posto como fator determinante o tipo de cultivo convencional e orgânico, e os períodos de estiagem e chuvoso. Foram analisados 56 voluntários. As análises foram realizadas no analisador automático de bioquímica Cobas Integra® 400 plus. De acordo com os resultados das análises, percebeu-se maior número de indivíduos com valores diminuídos da colinesterase, especificamente no grupo de cultivo convencional, o período de maior índice de alteração foi na época chuvosa, onde a atividade da AChE, apresentou valores expressivos na cidade de São Sebastião, com resultado de 80% de intoxicados, na cidade do Pé - Leve, obteve-se 21,73% no mesmo período. Na análise da AChP, São Sebastião obteve maior número de contaminados com 18,75% e 30% respectivamente, já no Distrito Pé Leve ficou entre 10 e 21,73% de níveis diminuídos da colinesterase. Isso define o sistema orgânico de cultivo, como a melhor alternativa para prevenção de futuras patologias, além de trazer qualidade de vida para os trabalhadores rurais, como também para os consumidores.
Subject(s)
Humans , Pesticides , Occupational Exposure/analysis , Acetylcholinesterase , Quality of Life , Prospective Studies , FarmersABSTRACT
This study aimed to evaluate the anticholinesterase activities of extracts and fractions of Ocotea daphnifolia in vitro and characterize its constituents. The effects of hexane, ethyl acetate, and ethanolic extracts on acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activity were determined with a spectrophotometry assay. All extracts inhibited cholinesterase activity, and the ethanolic extract (2 mg/mL) exhibited the highest inhibition of both enzymes (99.7% for BuChE and 82.4% for AChE). The ethanolic extract was fractionated by column chromatography resulting in 14 fractions that were also screened for their anticholinesterase effects. Fraction 9 (2 mg/mL) showed the highest activity, inhibiting AChE and BuChE by 71.8% and 90.2%, respectively. This fraction was analyzed by high-performance liquid chromatography high-resolution mass spectrometry which allowed the characterization of seven glycosylated flavonoids (containing kaempferol and quercetin nucleus) and one alkaloid (reticuline). In order to better understand the enzyme-inhibitor interaction of the reticuline toward cholinesterase, molecular modeling studies were performed. Reticuline targeted the catalytic activity site of the enzymes. Ocotea daphnifolia exhibits a dual cholinesterase inhibitory activity and displays the same pattern of intermolecular interactions as described in the literature. The alkaloid reticuline can be considered as an important bioactive constituent of this plant.
Subject(s)
In Vitro Techniques/instrumentation , Cholinesterase Inhibitors/analysis , Lauraceae/classification , Ocotea/adverse effects , Molecular Docking Simulation/instrumentation , Plants, Medicinal/anatomy & histology , Acetylcholinesterase/adverse effects , Spectrophotometry/instrumentation , Flavonoids , Butyrylcholinesterase/adverse effects , AlkaloidsABSTRACT
Hippeastrum puniceum is a species that belongs to the Amaryllidaceae family. A particular characteristic of this family is the consistent and very specific presence of isoquinoline alkaloids, which have demonstrated a wide range of biological activities such as antioxidant, antiviral, antifungal, antiparasitic, and acetylcholinesterase inhibitory activity, among others. In the present work, fifteen alkaloids were identified from the bulbs of Hippeastrum puniceum (Lam.) Kuntz using a GC-MS approach. The alkaloids 9-O-demethyllycoramine, 9-demethyl-2α-hydroxyhomolycorine, lycorine and tazettine were isolated through chromatographic techniques. The typical Amaryllidaceae alkaloids lycorine and tazettine, along with the crude and ethyl acetate extract from bulbs of the species were evaluated for their inhibitory potential on α-amylase, α-glucosidase, tyrosinase and acetylcholinesterase activity. Although no significant inhibition activity was observed against α-amylase, α-glucosidase and tyrosinase from the tested samples, the crude and ethyl acetate extracts showed remarkable acetylcholinesterase inhibitory activity. The biological activity results that correlated to the alkaloid chemical profile by GC-MS are discussed herein. Therefore, this study contributed to the knowledge of the chemical and biological properties of Hippeastrum puniceum (Lam.) and can subsidize future studies of this species
Subject(s)
Amaryllidaceae Alkaloids/analysis , Amaryllidaceae/classification , Acetylcholinesterase/adverse effects , Cholinesterase Inhibitors/pharmacology , Acetates/agonists , Antioxidants/pharmacologyABSTRACT
A guanitoxina (GNT) é uma neurotoxina produzida por algumas cepas de cianobactérias dos gêneros Dolichospermum e Sphaerospermopsis>. A GNT é o único organofosforado natural, capaz de causar a morte de animais selvagens e domésticos devido à inibição irreversível da acetilcolinesterase. Apesar de sua alta toxicidade, o diagnóstico da GNT em amostras biológicas ainda é um grande desafio. A dificuldade para sua detecção está diretamente ligada à sua instabilidade em altas temperaturas e pH alcalino, tornando difícil seu monitoramento em corpos d'água. Por isso, esta pesquisa objetivou estudar a estabilidade e biodisponibilidade da GNT em amostras aquosas, com intuito de obter mais informações sobre a natureza química e biológica dessa potente neurotoxina. Para realizar este estudo, a cepa ITEP-24 (S. torques-reginae) produtora de GNT foi cultivada em laboratório sob condições controladas, para obter biomassa para os experimentos de extração, semi-isolamento, estabilidade, ensaio in vitro e identificação por LC-MS/MS. Primeiramente foram realizados testes de extração da GNT partir de células liofilizadas da cepa ITEP-24 utilizando água, metanol e etanol em pH ácido. Depois utilizou-se dois métodos de extração em fase sólida (SPE) com cartuchos preenchidos com fases estacionarias C18 em fase reversa e sílica gel em fase normal, com objetivo de avaliar qual método de SPE seria melhor para extrair e concentrar a GNT. Nós também testamos métodos para lisar as células com sondas de ultrassom, misturador e centrifugação. Além dos métodos de extração, nós avaliamos a estabilidade da toxina em diferentes temperaturas, para isso a biomassa seca contendo a GNT ficou condicionada a 4 °C, 23 °C, -20 °C, -80 °C durantes seis meses, e análises de identificação foram realizadas dentro período de 150 dias em uma sequência de 30 dias. A estabilidade da toxina foi analisada também a partir de extrações em soluções com diferentes valores de pH (1,5; 3,0; 5,0; 7,0; 8,5; 10,5) e temperatura (23 ºC e 37 ºC). Depois, analisou-se a biodisponibilidade da GNT em células frescas da linhagem ITEP-24 através de teste de dissolução in vitro. O objetivo deste teste foi avaliar a liberação da toxina intracelular em meio simulado do conteúdo gástrica e intestinal com e sem enzimas digestivas para compreender e estimar a disponibilidade da GNT in vivo. Os resultados de todos experimentos descritos neste estudo, foram obtidos a partir de análises por cromatografia líquida de interação hidrofílica (HILIC) acoplado ao espectrômetro de massas do tipo triplo quadrupolo LC-QqQ-MS/MS utilizando as transições 253>58, 253>159 e 159>58 [M+H]+ utilizando coluna com fase estacionária zwitteriônica (ZIC). A identificação da GNT foi realizada também por cromatografia líquida acoplada ao espectrômetro de massas de alta resolução (LC-HR-QTOF-MS) com coluna Luna C18, Hydro-RP C18 e ZIC-HILIC. Dos protocolos de extração testados, a combinação de metanol/água (70:30 v/v) com ácido acético (0.3%) extraiu maior quantidade relativa da GNT a partir de células frescas e liofilizadas da cepa ITEP-24 e a concentração da toxina foi maior em amostras de células frescas. Em relação aos métodos de lise celular, as extrações realizadas em sonda de ultrassom com banho-maria e centrifugação por 1h foram estatisticamente significantes para liberar a toxina intracelular. Não houve diferença significativa entre os testes de SPE, no entanto, a semipurificação da toxina foi melhor com cartucho preenchido com sílica gel em fase normal e adaptação desse método em coluna aberta permitiu obter uma fração enriquecida com GNT. A GNT mostrou ser mais estável em pH ácido, sendo o pH 3,0 o melhor para manter e extrair a toxina em amostras aquosas e a toxina intracelular presente em células secas podem degradar em temperatura de 23 °C por um período de 150 dias mesmo em solução com pH 3,0. Durante os testes de extração e purificação foi observado também a degradação da toxina em processos de secagem e ressuspensão. As análises realizadas no LC-HR-QTOF-MS com diferentes métodos cromatográficos possibilitou a identificação da GNT, porém o método realizado com coluna ZIC-HILIC mostrou melhor resolução cromatográfica dos picos relativos m/z e tempo de retenção de toxina. Os resultados obtidos nos testes de dissolução in vitro mostraram que a GNT fica mais disponível no simulado gástrico com e sem a enzima pepsina, mas também pode ser absorvida no intestino. Portanto, o teste de dissolução in vitro pode ser uma ferramenta útil para a avaliação de risco de cianotoxinas in vivo, devido ao seu potencial de monitorar qualitativa e quantitativamente substâncias dissolvidas em fluidos gastrointestinais. Os resultados apresentados neste estudo fornecem informações valiosas para uma melhor compreensão da estabilidade e biodisponibilidade do GNT. Além disso, os métodos apresentados neste estudo podem ser úteis para diversas aplicações projetadas para identificar a toxina em amostras ambientais, bem como orientações para procedimentos de purificação da GNT
Guanitoxin (GNT) is a neurotoxin produced by some strains of cyanobacteria of the genus Dolichospermum and Sphaerospermopsis. GNT is the only natural organophosphate, capable of causing the death of animals from wild and domestic animals due to irreversible inhibition of acetylcholinesterase. Despite its high toxicity, the diagnosis of GNT in biological samples is still a significant challenge. The difficulty in its detection is directly linked to its instability at high temperatures and alkaline pH, making it difficult to monitor in bodies of water. Therefore, this research aimed to study the stability and bioavailability of GNT in aqueous samples to provide more information about the chemical and biological nature of this molecule. The strain ITEP-24 (S. torques-reginae) producing GNT was grown in the laboratory under controlled conditions to obtain biomass for the extraction, semi-isolation, stability, in vitro tests, and toxin identification by LC-MS/MS. Firstly, tests were carried out to extract GNT from lyophilized cells strain ITEP-24 using water, methanol, and ethanol at acidic pH and, two SPE methods in cartridges with stationary phases of C18 reverse phase and normal phase gel silica, to evaluate which would be better to extract and concentrate the GNT. We also tested different methods of cell lysis, such as ultrasound probes, mixers, and centrifugation. In addition to the extraction methods, the stability of the toxin was evaluated at different temperatures, for this, the dry biomass containing the toxin was conditioned at 4 °C, 23 °C, -20 °C, -80 °C for 150 days and analysis of the identification of the GNT was carried out within that period in a sequence of 30 days. The toxin stability was also analyzed from extractions in solutions with different pH values (1.5; 3.0; 5.0; 7.0; 8.5; 10.5) and temperature (23 ºC and 37 ºC). In addition, we performed dissolution tests with fresh cells of the ITEP-24 strain to evaluate the bioavailability of GNT in simulated gastric and intestinal fluids with and without digestive enzymes to understand and estimate the availability of GNT in vivo. The results of all experiments described in this study were obtained from analyzes by hydrophilic interaction liquid chromatography (HILIC) coupled to the LC-QqQ-MS/MS triple quadrupole mass spectrometer using the transitions m/z 253> 58, m/z 253> 159 and m/z 159> 58 [M + H]+ using a column with the zwitterionic stationary phase (ZIC). Liquid chromatography coupled to the high-resolution mass spectrometer (LC-HR-QTOF-MS) with Luna column C18, Hydro-RP C18, and ZIC-HILIC carried out the identification of the GNT. From the extraction protocols tested, the combination of methanol/water (70:30 v/v) with acetic acid (0.3%) extracted a greater relative amount of GNT from fresh and lyophilized ITEP-24 cells, and the concentration of the toxin is higher previously fresh. Concerning cellular methods, the ultrasound probe with a water bath and centrifugation for 1h ware statistically significant to release the intracellular toxin. There was no significant difference between the SPE tests. However, the semi-purification of the toxin was better with a cartridge filled with gel silica in the normal phase and adaptation of this method in an open column allowed to obtain a fraction enriched with GNT. GNT was more stable at acid pH, with pH 3.0 being the best to maintain and the intracellular toxin present in dry cells can degrade at a temperature at 23 °C for 150 days even in pH 3.0 solution. The toxin can also hydrolyze in the drying and resuspension processes. The analyzes carried out in LC-HR-QTOF-MS with different chromatographic methods made it possible to identify the GNT itself, however, the ZIC-HILIC column method showed excellent chromatographic resolution of the relative m/z peaks and toxin retention time. The results obtained in the in vitro dissolution tests showed that GNT is more available in the gastric simulation with and without the enzyme pepsin, but it can also be absorbed in the intestine. Thus, in vitro dissolution tests can be used as a useful tool for the risk assessment of cyanotoxins in vivo due to their potential to qualitatively and quantitatively monitor substances dissolved in gastrointestinal fluids. The results presented in this study provide valuable information for a better understanding of the stability and bioavailability of GNT. Besides, the methods presented in this study can be useful for various applications designed to identify the toxin in environmental samples, as well as guidance on procedures for purifying GNT
Subject(s)
Acetylcholinesterase/adverse effects , Mass Spectrometry/methods , Diagnosis , Methods , Organophosphorus Compounds/antagonists & inhibitors , In Vitro Techniques/methods , Chromatography, Liquid/methods , Cyanobacteria/metabolism , Solid Phase Extraction/instrumentation , Hydrophobic and Hydrophilic Interactions , Hydrogen-Ion ConcentrationABSTRACT
Abstract This study reports the action of essential oils (EO) from five plants on the activity of native and recombinant acetylcholinesterases (AChE) from Rhipicephalus microplus. Enzyme activity of native susceptible AChE extract (S.AChE), native resistant AChE extract (R.AChE), and recombinant enzyme (rBmAChE1) was determined. An acetylcholinesterase inhibition test was used to verify the effect of the EO on enzyme activity. EO from Eucalyptus globulus, Citrus aurantifolia, Citrus aurantium var.dulcis inhibited the activity of S.AChE and R.AChE. Oils from the two Citrus species inhibited S.AChE and R.AChE in a similar way while showing greater inhibition on R.AChE. The oil from E. globulus inhibited native AChE, but no difference was observed between the S.AChE and R.AChE; however, 71% inhibition for the rBmAChE1 was recorded. Mentha piperita oil also inhibited S.AChE and R.AChE, but there was significant inhibition at the highest concentration tested. Cymbopogon winterianus oil did not inhibit AChE. Further studies are warranted with the oils from the two Citrus species that inhibited R.AChE because of the problem with R. microplus resistant to organophosphates, which target AChE. C. winterianus oil can be used against R. microplus populations that are resistant to organophosphates because its acaricidal properties act by mechanism(s) other than AChE inhibition.
Resumo Este estudo relata a ação de óleos essenciais de cinco plantas na atividade de acetilcolinesterases (AChE) nativas e recombinantes de Rhipicephalus microplus. A atividade enzimática do extrato de acetilcolinesterase nativa suscetível (S.AChE) e resistente (R.AChE) e da enzima recombinante (rBmAChE1) foi determinada. Um teste de inibição da AChE foi utilizado, para verificar o efeito dos óleos essenciais sobre a atividade enzimática. Óleos essenciais de Eucalyptus globulus, Citrus aurantifolia, Citrus aurantium var. dulcis inibiram a atividade de S.AChE e R.AChE. Os óleos das duas espécies de Citrus inibiram S.AChE e R.AChE de maneira semelhante, mas mostraram maior inibição sobre R.AChE. O óleo de E. globulus inibiu a AChE nativa, mas sem diferença entre a S.AChE e a R.AChE; no entanto, 71% de inibição para rBmAChE1 foi observada. O óleo de Mentha piperita também inibiu S.AChE e R.AChE, mas houve inibição significativa apenas nas concentrações mais altas testadas. O óleo de Cymbopogon winterianus não inibiu a AChE. Estudos adicionais são necessários com os óleos das duas espécies de Citrus que inibiram a R.AchE, devido ao problema de R. microplus resistente aos organofosforados ter como alvo AChE. O óleo de C. winterianus pode ser usado contra populações de R. microplus, que são resistentes a organofosforados, porque suas propriedades acaricidas agem por mecanismos diferentes.
Subject(s)
Animals , Oils, Volatile/pharmacology , Cholinesterase Inhibitors/pharmacology , Cymbopogon , Rhipicephalus/enzymology , Acaricides/pharmacology , Acetylcholinesterase , LarvaABSTRACT
Afr. J. Biomed. Res. Vol. 24 (May, 2021); 257- 263 Research Article Protective Effect of Alkaloid-rich Extract of Brimstone Tree (Morinda lucida) on Neurotoxicity in the Fruit-fly (Drosophila melanogaster) Model Nwanna E.E. Functional Food, Nutraceutical and Phytomedicine unit Department of Biochemistry, Federal University of Technology Akure PMB 704, Nigeria ABSTRACT Brimstone plant is one of the medicinal plants found in Nigeria used in fore-lore medicine with little scientific information on its alkaloid constituents. This study was aimed at investigating the effect of alkaloid-rich compounds from the plant in manganese- induced (MgCl2) neurotoxicity in the fruit fly. In addition, alkaloid compounds will be characterized using gas chromatograph coupled with flame ionization detector (GC-FID). Alkaloid-rich extract was prepared by solvent extraction method, fruit flies were pre-treated with the extract (0.5 1.0mg/ml) in a fortified diet before induction with MgCl2. The survival rate and negative geotaxis were observed. Thereafter, the activity of acetylcholinesterase (AChE) enzyme, antioxidantive potentials in in -vivo reactive oxygen species (ROS) thiobarbituric acid reactive species (TBARS), total thiol content, nitric oxide (NO*), hydroxyl oxide (OH*) scavenging ability, ferric reducing antioxidant property (FRAP) and 2, 2'-azino-bis (3-ethylbenzthiazoline-6- sulphonic acid) (ABTs*) radical scavenging ability were carried out on the flies' homogenate. Results obtained revealed that the alkaloid-rich extract regulated the neuro-protective activity of AchE, reduced the reactive oxygen species level in the induced flies with an increased in antioxidantive potential, higher survival rate and increases in the life span of the flies with 50% reduction in the mortality rate. The GC-MS showed a total of (1.55/100mg) of different plant-derived alkaloids such as dicentrine, atropine, aporphine. These alkaloids-rich compounds were found to have anti-oxidative, anti-nociceptive, anti-inflammatory and anti- cholinergic activities. In conclusion, this study suggests that alkaloids from brimstone plant could be the reason for the observed biological activities for the prevention of neuronal related complications.
Subject(s)
Humans , Sulfur , Neurotoxicity Syndromes , Acetylcholinesterase , Drosophila Proteins , AlkaloidsABSTRACT
The chemical constituents from the roots of Aconitum kongboense were studied. Twenty-five diterpenoid alkaloids were isolated from the 95% methanol extract of the roots of A. kongboense by silica gel, reverse-phase silica gel and basic alumina column chromatography. They included a new aconitine-type diterpenoid alkaloid, named as kongboensenine(1), and twenty-four known ones(2-25), i.e., acotarine F(2), acotarine G(3), 14-acetyltalatisamine(4), talatisamine(5), indaconitine(6), yunaconitine(7), chasmanine(8), 6-epi-foresticine(9), homochasmanine(10), 8-deacetyl-yunaconitine(11), chasmaconitine(12), ajaconine(13), franchetine(14), ezochasmanine(15), crassicautine(16), 14-O-deacylcrassicausine(17), genicunine A(18), falconeridine(19), sachaconitine(20), liljestrandisine(21), 8-methyl-14-acetyltalatisamine(22), kongboendine(23), 14-benzoylchasmanine(24) and pseudaconine(25). Their structures were elucidated by common spectroscopic methods including high-resolution electrospray ionization mass spectrometry(HR-ESI-MS) and nuclear magnetic resonance(NMR) techniques. Compounds 2-4, 10, 13, 15-19 and 21-22 were isolated from this plant for the first time. Experimental results showed that all compounds did not have a significant inhibitory activity against acetylcholinesterase(AChE).
Subject(s)
Acetylcholinesterase , Aconitum/metabolism , Alkaloids , Diterpenes , Magnetic Resonance Spectroscopy , Molecular Structure , Plant Roots/metabolismABSTRACT
The biocompatibility of nanomaterials has attracted much attention. Graphene oxide (GO) is a nanomaterial widely used in biomedicine, but its toxicity can not be ignored. In this study, the effect of GO on the blood system (the hemolysis rate, the fragility of erythrocyte, and acetylcholinesterase activity) was systematically investigated. The results showed that the hemolysis rate of erythrocytes was lower than 8% when the GO concentration was below 100 μg/mL (P5 μm (LGO) increased the activity of acetylcholinesterase by 42.67% (P<0.05). Then molecular dynamics simulation was used to study how GO interacted with acetylcholinesterase and increased its activity. The results showed that GO was attached to the cell membrane, thus may provide an electronegative environment that helps the hydrolysate to detach from the active sites more quickly so as to enhance the activity of acetylcholinesterase.