RESUMEN
Natural products have played a dominant role in the discovery of lead compounds for the development of drugs aimed at the treatment of human diseases. This electrospray ionization-ion mobility spectrometry-mass spectrometry (ESI-IMS-MS)-based study demonstrates that dietary antioxidants, isolated components from the stigmas of saffron (Crocus sativus L.) may be effective in inhibiting Aß fibrillogenesis, a neuropathological hallmark of Alzheimer's Disease (AD). This study reveals a substantial alteration in the monomer/oligomer distribution of Aß1-40, concomitant with re-direction of fibril formation, induced by the natural product interaction. These alterations on the Aß1-40 aggregation pathway are most prominent for trans-crocin-4 (TC4). Use of ESI-IMS-MS, electron microscopy alongside Thioflavin-T kinetics, and the interpretation of 3-dimensional Driftscope plots indicate a correlation of these monomer/oligomer distribution changes with alterations to Aß1-40 amyloid formation. The latter could prove instrumental in the development of novel aggregation inhibitors for the prevention, or treatment of AD.
Asunto(s)
Enfermedad de Alzheimer/tratamiento farmacológico , Antioxidantes/farmacología , Crocus/química , Extractos Vegetales/farmacología , Agregación Patológica de Proteínas/tratamiento farmacológico , Enfermedad de Alzheimer/patología , Péptidos beta-Amiloides/metabolismo , Péptidos beta-Amiloides/ultraestructura , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Antioxidantes/uso terapéutico , Carotenoides/farmacología , Humanos , Microscopía Electrónica de Transmisión , Fragmentos de Péptidos/metabolismo , Fragmentos de Péptidos/ultraestructura , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/uso terapéutico , Agregación Patológica de Proteínas/patología , Espectrometría de Masa por Ionización de Electrospray , Espectrometría de Masas en TándemRESUMEN
There is currently no effective treatment to prevent the progress of Alzheimer's disease (AD). The traditional Chinese herbs Dengzhan Shengmai (DZSM) capsules and their active component scutellarin possess multiple effects and are clinically used for the treatment of cerebrovascular diseases. Scutellarin has been reported to affect Aß aggregation. However, the effects of DZSM capsules on AD remain unknown. Through in vivo experiments, our study proved that the alleviating effects of DZSM capsules on cognitive deficits of AD mice were due to the role of scutellarin, which up-regulated low toxic amyloid plaques and down-regulated highly toxic soluble Aß42 and Aß40 levels in cortex. In vitro, we confirmed scutellarin's role in accelerating transforming Aß42 monomers into high-molecular-mass aggregates by biochemical assays, which supported the results observed in drug-treated APP/PS1 mice. In detail, the 1:10 ratio of scutellarin/Aß42 mixtures promoted production of large ß-sheet-rich fibrils whereas the 1:1 ratio promoted production of protofibrils. In addition, the binding between scutellarin and Aß monomers was quantified by microscale thermophoresis test and the apparent dissociation constant (Kd) was 1284.4⯱â¯238.8⯵M. What's more, binding regions between scutellarin and Aß fibrils were predicted by computational docking models and scutellarin might bind parallel to the long axis of Aß42 fibrils targeting hydrophobic grooves at residues 35-36 or 39. In conclusion, DZSM capsules protected against cognitive defects of AD through scutellarin-mediated acceleration of Aß aggregation into fibrils or protofibrils and reduction of soluble Aß oligomers, thus suggesting potential clinical applications of DZSM capsules and scutellarin in the treatment of AD.
Asunto(s)
Péptidos beta-Amiloides/metabolismo , Apigenina/uso terapéutico , Disfunción Cognitiva/tratamiento farmacológico , Disfunción Cognitiva/prevención & control , Medicamentos Herbarios Chinos/uso terapéutico , Glucuronatos/uso terapéutico , Presenilina-1/metabolismo , Agregado de Proteínas , Multimerización de Proteína , Péptidos beta-Amiloides/química , Péptidos beta-Amiloides/ultraestructura , Animales , Apigenina/química , Apigenina/farmacología , Medicamentos Herbarios Chinos/farmacología , Glucuronatos/química , Glucuronatos/farmacología , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Masculino , Ratones Endogámicos C57BL , Ratones Transgénicos , Peso Molecular , Placa Amiloide/tratamiento farmacológico , Placa Amiloide/patología , Placa Amiloide/ultraestructura , SolubilidadRESUMEN
2-(piperazin-1-yl)N-(1H-pyrazolo[3,4-b]pyridin-3-yl)acetamides are described as a new class of selective and potent acetylcholinesterase (AChE) inhibitors and amyloid ß aggregation inhibitors. Formation of synthesized compounds (P1P9) was justified via H1 NMR, C13 NMR, mass spectra and single crystal X-Ray diffraction study. All compounds were evaluated for their acetylcholinesterase and butyrylcholinesterase inhibitory activity, inhibition of self-mediated Aß aggregation and Cu(II)-mediated Aß aggregation. Also, docking study carried out was in concordance with in vitro results. The most potent molecule amongst the derivatives exhibited excellent anti-AChE activity (IC50â¯=â¯4.8â¯nM). Kinetic study of P3 suggested it to be a mixed type inhibitor. In vitro study revealed that all the compounds are capable of inhibiting self-induced ß-amyloid (Aß) aggregation with the highest inhibition percentage to be 81.65%. Potency of P1 and P3 to inhibit self-induced Aß1-42 aggregation was ascertained by TEM analysis. Compounds were also evaluated for their Aß disaggregation, antioxidation, metal-chelation activity.
Asunto(s)
Enfermedad de Alzheimer/tratamiento farmacológico , Pirazoles/química , Pirazoles/uso terapéutico , Piridinas/química , Piridinas/uso terapéutico , Acetilcolinesterasa/efectos de los fármacos , Péptidos beta-Amiloides/antagonistas & inhibidores , Péptidos beta-Amiloides/ultraestructura , Antioxidantes/farmacología , Hidrolasas de Éster Carboxílico/efectos de los fármacos , Quelantes/uso terapéutico , Inhibidores de la Colinesterasa/uso terapéutico , Cobre/química , Cristalografía por Rayos X , Evaluación Preclínica de Medicamentos , Humanos , Concentración 50 Inhibidora , Microscopía Electrónica de Transmisión , Simulación del Acoplamiento Molecular , Pirazoles/síntesis química , Piridinas/síntesis química , Espectrometría de FluorescenciaRESUMEN
Aggregation of amyloid-ß (Aß) and tau plays a crucial role in the onset and progression of Alzheimer's disease (AD). Therefore, the inhibition of Aß and tau aggregation may represent a potential therapeutic target for AD. Herein, we designed and synthesized both Aß and tau dual aggregation inhibitors based on the structure of curcumin and developed the novel curcumin derivative PE859. In this study, we investigated the inhibitory activity of PE859 on Aß aggregationin vitro and the therapeutic effects of PE859 on cognitive dysfunction via dual inhibition of Aß and tau aggregation in vivo. PE859 inhibited Aß aggregation in vitro and protected cultured cells from Aß-induced cytotoxicity. Furthermore, PE859 ameliorated cognitive dysfunction and reduced the amount of aggregated Aß and tau in brains of senescence-accelerated mouse prone 8 (SAMP8). These results warrant consideration of PE859 as a candidate drug for AD.
Asunto(s)
Péptidos beta-Amiloides/metabolismo , Trastornos del Conocimiento/tratamiento farmacológico , Trastornos del Conocimiento/metabolismo , Indoles/uso terapéutico , Agregado de Proteínas/efectos de los fármacos , Pirazoles/uso terapéutico , Proteínas tau/metabolismo , Envejecimiento/genética , Péptidos beta-Amiloides/ultraestructura , Animales , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Encéfalo/ultraestructura , Línea Celular Tumoral , Trastornos del Conocimiento/genética , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Humanos , L-Lactato Deshidrogenasa/metabolismo , Aprendizaje por Laberinto/efectos de los fármacos , Ratones , Ratones Transgénicos , Microscopía Electrónica de Transmisión , Actividad Motora/efectos de los fármacos , Neuroblastoma/patología , Tecnicas de Microbalanza del Cristal de Cuarzo , Factores de Tiempo , Proteínas tau/ultraestructuraRESUMEN
The Aß complexes of some redox-active species, such as Cu, cause oxidative stress and induce severe toxicity by generating reactive oxygen species (ROS). Thus, Cu chelation therapy should be considered as a valuable strategy for the treatment of Alzheimer's disease (AD). However, more attention should be paid to the specific chelating ability of these chelating agents. Herein, a tripeptide GGH was used to selectively chelate the Cu(2+) in Aß-Cu complex in the presence of other metal ions (e.g., K(+), Ca(2+), Ni(2+), Mg(2+), and Zn(2+)) as shown by isothermal titration calorimetry results. GGH decreased the level of HO(â¢) radicals by preventing the formation of intermediate Cu(I) ion. Thus, the Cu species completely lost its catalytic activity at a superequimolar GGH/Cu(II) ratio (4:1) as observed by UV-visible spectroscopy, coumarin-3-carboxylic acid fluorescence, and BCA assay. Moreover, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay indicates that GGH increased PC-12 cell viability from 36% to 63%, and neurotoxicity partly triggered by ROS decreased. These results indicate potential development of peptide chelation therapy for AD treatment.
Asunto(s)
Péptidos beta-Amiloides/metabolismo , Quelantes/farmacología , Cobre/metabolismo , Oligopéptidos/farmacología , Oxidación-Reducción/efectos de los fármacos , Células PC12/efectos de los fármacos , Péptidos beta-Amiloides/antagonistas & inhibidores , Péptidos beta-Amiloides/ultraestructura , Animales , Diferenciación Celular/efectos de los fármacos , Quelantes/química , Cobre/química , Cobre/farmacología , Relación Dosis-Respuesta a Droga , Metales/metabolismo , Metales/farmacología , Microscopía Electrónica de Transmisión , Neuronas/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Células PC12/ultraestructura , Unión Proteica/efectos de los fármacos , Ratas , Especies Reactivas de Oxígeno/metabolismo , Factores de TiempoRESUMEN
There is a crucial need to develop new effective drugs for Alzheimer's disease (AD) as the currently available AD treatments provide only momentary and incomplete symptomatic relief. Amongst natural products, curcumin, a major constituent of turmeric, has been intensively investigated for its neuroprotective effect against ß-amyloid (Aß)-induced toxicity in cultured neuronal cells. The ability of curcumin to attach to Aß peptide and prevent its accumulation is attributed to its three structural characteristics such as the presence of two aromatic end groups and their co-planarity, the length and rigidity of the linker region and the substitution conformation of these aromatics. However, curcumin failed to reach adequate brain levels after oral absorption in AD clinical trials due to its low water solubility and poor oral bioavailability. A number of new curcumin analogs that mimic the active site of the compound along with analogs that mimic the curcumin anti-amyloid effect combined with anticholinesterase effect have been developed to enhance the bioavailability, pharmacokinetics, water solubility, stability at physiological conditions and delivery of curcumin. In this article, we have summarized all reported synthetic analogs of curcumin showing effects on ß-amyloid and discussed their potential as therapeutic and diagnostic agents for AD.
Asunto(s)
Enfermedad de Alzheimer/tratamiento farmacológico , Péptidos beta-Amiloides/metabolismo , Encéfalo/efectos de los fármacos , Curcumina/análogos & derivados , Curcumina/farmacología , Fármacos Neuroprotectores/química , Fármacos Neuroprotectores/farmacología , Enfermedad de Alzheimer/diagnóstico , Enfermedad de Alzheimer/metabolismo , Enfermedad de Alzheimer/patología , Péptidos beta-Amiloides/ultraestructura , Animales , Encéfalo/metabolismo , Encéfalo/patología , Curcumina/farmacocinética , Humanos , Fármacos Neuroprotectores/farmacocinética , Agregado de Proteínas/efectos de los fármacos , Agregación Patológica de Proteínas/diagnóstico , Agregación Patológica de Proteínas/tratamiento farmacológico , Agregación Patológica de Proteínas/metabolismo , Agregación Patológica de Proteínas/patologíaRESUMEN
Alzheimer's disease is a neurodegenerative disease with no known cure and few effective treatment options. The principal neurotoxic agent is an oligomeric form of the amyloid-ß peptide and one of the treatment options currently being studied is the inhibition of amyloid aggregation. In this work, we test a novel pseudopeptidic aggregation inhibitor designated as SG1. SG1 has been designed to bind at the amyloid-ß self-recognition site and prevent amyloid-ß from misfolding into ß sheet. We used atomic force spectroscopy, a nanoscale measurement technique, to quantify the binding forces between two single amyloid peptide molecules. For the first time, we demonstrate that single molecule atomic force spectroscopy can be used to assess the effectiveness of amyloid aggregation inhibitors by measuring the experimental yield of binding and can potentially be used as a screening technique for quick testing of efficacy of inhibitor drugs for amyloid aggregation.
Asunto(s)
Péptidos beta-Amiloides/antagonistas & inhibidores , Evaluación Preclínica de Medicamentos , Microscopía de Fuerza Atómica , Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/metabolismo , Péptidos beta-Amiloides/ultraestructura , Técnicas Biosensibles/métodos , Evaluación Preclínica de Medicamentos/métodos , Humanos , Microscopía de Fuerza Atómica/métodos , Simulación de Dinámica MolecularRESUMEN
Alzheimer's disease (AD) is characterized by pathological aggregation of ß-amyloid peptides and MAP-Tau protein. ß-Amyloid (Aß) is a peptide responsible for extracellular Alzheimer's plaque formation. Intracellular MAP-Tau aggregates appear as a result of hyperphosphorylation of this cytoskeletal protein. Small, oligomeric forms of Aß are intermediate products that appear before the amyloid plaques are formed. These forms are believed to be most neurotoxic. Dendrimers are highly branched polymers, which may find an application in regulation of amyloid fibril formation. Several biophysical and biochemical methods, like circular dichroism (CD), fluorescence intensity of thioflavin T and thioflavin S, transmission electron microscopy, spectrofluorimetry (measuring quenching of intrinsic peptide fluorescence) and MTT-cytotoxicity assay, were applied to characterize interactions of cationic phosphorus-containing dendrimers of generation 3 and generation 4 (CPDG3, CPDG4) with the fragment of amyloid peptide (Aß(1-28)) and MAP-Tau protein. We have demonstrated that CPDs are able to affect ß-amyloid and MAP-Tau aggregation processes. A neuro-2a cell line (N2a) was used to test cytotoxicity of formed fibrils and intermediate products during the Aß(1-28) aggregation. It has been shown that CPDs might have a beneficial effect by reducing the system toxicity. Presented results suggest that phosphorus dendrimers may be used in the future as agents regulating the fibrilization processes in Alzheimer's disease.
Asunto(s)
Péptidos beta-Amiloides/química , Dendrímeros/química , Fósforo/química , Proteínas tau/química , Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/ultraestructura , Animales , Línea Celular Tumoral , Dicroismo Circular , Humanos , Ratones , Microscopía Electrónica de Transmisión , Tirosina/químicaRESUMEN
Alzheimer's disease (AD) is a common form of neurodegenerative disease. Mounting evidence suggests that metal ions play a key role in the aggregation of amyloid ß peptide (Aß), which acts as a factor or cofactor in the etiopathogenesis of AD. Therefore, inhibition of Aß aggregation emerges as a potential approach for the treatment of AD. We have found that baicalin can interact with copper directly and inhibits Aß1-42 aggregation. In addition, baicalin protects SH-SY5Y cells from oxidative injuries induced by Aß1-42 aggregation through decreasing H(2)O(2) production that is normally formed as a deleterious by-product of beta amyloid aggregation and the formation of plaques. Taken together, these data indicate that baicalin may be a potential agent to inhibit Aß aggregation and thereby delay, mitigate or modify the progression of neurodegenerative diseases such as AD.
Asunto(s)
Péptidos beta-Amiloides/metabolismo , Antioxidantes/farmacología , Flavanonas/farmacología , Peróxido de Hidrógeno/metabolismo , Estrés Oxidativo/efectos de los fármacos , Fragmentos de Péptidos/metabolismo , Péptidos beta-Amiloides/farmacología , Péptidos beta-Amiloides/ultraestructura , Línea Celular Tumoral , Cobre/metabolismo , Relación Dosis-Respuesta a Droga , Interacciones Farmacológicas , Humanos , Neuroblastoma/patología , Fragmentos de Péptidos/farmacología , Fragmentos de Péptidos/ultraestructura , Fenol/farmacología , Extractos Vegetales/farmacología , Scutellaria baicalensis , Factores de TiempoRESUMEN
Full-length amyloid beta peptides (Abeta(1-40/42)) form neuritic amyloid plaques in Alzheimer's disease (AD) patients and are implicated in AD pathology. However, recent transgenic animal models cast doubt on their direct role in AD pathology. Nonamyloidogenic truncated amyloid-beta fragments (Abeta(11-42) and Abeta(17-42)) are also found in amyloid plaques of AD and in the preamyloid lesions of Down syndrome, a model system for early-onset AD study. Very little is known about the structure and activity of these smaller peptides, although they could be the primary AD and Down syndrome pathological agents. Using complementary techniques of molecular dynamics simulations, atomic force microscopy, channel conductance measurements, calcium imaging, neuritic degeneration, and cell death assays, we show that nonamyloidogenic Abeta(9-42) and Abeta(17-42) peptides form ion channels with loosely attached subunits and elicit single-channel conductances. The subunits appear mobile, suggesting insertion of small oligomers, followed by dynamic channel assembly and dissociation. These channels allow calcium uptake in amyloid precursor protein-deficient cells. The channel mediated calcium uptake induces neurite degeneration in human cortical neurons. Channel conductance, calcium uptake, and neurite degeneration are selectively inhibited by zinc, a blocker of amyloid ion channel activity. Thus, truncated Abeta fragments could account for undefined roles played by full length Abetas and provide a unique mechanism of AD and Down syndrome pathologies. The toxicity of nonamyloidogenic peptides via an ion channel mechanism necessitates a reevaluation of the current therapeutic approaches targeting the nonamyloidogenic pathway as avenue for AD treatment.
Asunto(s)
Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/metabolismo , Síndrome de Down/metabolismo , Fragmentos de Péptidos/metabolismo , Enfermedad de Alzheimer/patología , Péptidos beta-Amiloides/ultraestructura , Calcio/metabolismo , Muerte Celular , Síndrome de Down/patología , Humanos , Microscopía de Fuerza Atómica , Modelos Moleculares , Fragmentos de Péptidos/ultraestructura , Estructura Terciaria de ProteínaRESUMEN
As reported by some authors, clioquinol (CQ), a 8-hydroxyquinoline derivative, has produced very encouraging results in the treatment of Alzheimer's disease (AD). Its biological effects are most likely ascribed to complexation of specific metal ions, such as copper (II) and zinc (II), critically associated with beta-amyloid (A beta) aggregation/fibrillogenesis and degeneration processes in the brain. The present study was aimed at assessing the in vitro effects of CQ on the aggregation/fibrillogenesis properties of human A beta either alone or complexed with Cu(2+) and Zn(2+). Surprisingly, our data indicated that CQ promoted rather than inhibited the formation of A beta fibrillar aggregates when added metal ions were present. To understand whether the latter effects were related to the peptide amino acid sequence, we also investigated the aggregational profile of rat A beta, which differs from the human homologous for three amino acidic substitutions. Such a sequence alteration drastically reduced the tendency of the peptide to undergo spontaneous aggregation/fibrillization. In the presence of CQ and metals, however, also rat A beta showed a strong propensity to generate fibrillar aggregates. In agreement with the pro-aggregation effects observed in solution, studies with neuroblastoma cells demonstrated an impairment of cell functioning only in the presence of CQ + A beta-metals. Based on the present findings, the literature data on the potential effectiveness of CQ-based chelation therapy in AD should be re-interpreted.
Asunto(s)
Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/efectos de los fármacos , Clioquinol/farmacología , Metales/toxicidad , Placa Amiloide/efectos de los fármacos , Enfermedad de Alzheimer/fisiopatología , Enfermedad de Alzheimer/terapia , Sustitución de Aminoácidos/efectos de los fármacos , Sustitución de Aminoácidos/fisiología , Péptidos beta-Amiloides/metabolismo , Péptidos beta-Amiloides/ultraestructura , Animales , Línea Celular Tumoral , Terapia por Quelación/efectos adversos , Terapia por Quelación/métodos , Cobre/toxicidad , Sinergismo Farmacológico , Humanos , Metales/química , Neurotoxinas/toxicidad , Placa Amiloide/metabolismo , Placa Amiloide/ultraestructura , Ratas , Especificidad de la Especie , Zinc/toxicidadRESUMEN
AIM: To compare the effects of salvianolic acid B (Sal B) and Ginkgo biloba extract EGb 761 on beta-amyloid peptide (beta-AP) fibril formation and cytotoxicity to PC12 cells. METHODS: The inhibitory effects of Sal B and EGb 761 on beta-AP1-40 fibril formation were determined by using fluorescence analysis with Thioflavin T (ThT) and electron microscopic image. beta-AP25-35 was aged by incubating at 37 degrees C for 7 d, then the protein was incubated with PC12 cells. The protective effects of Sal B and EGb 761 against cytotoxicity induced by aged beta-AP25-35 in PC12 cells were evaluated by MTT reduction assay and flow cytometric analysis. beta-AP25-35-induced accumulation of intracellular reactive oxygen species (ROS) was determined by fluorescence analysis. RESULTS: Both Sal B and EGb 761 inhibited the formation of amyloid fibrils, protected PC12 cells from beta-AP25-35-induced cytotoxicity, and decreased ROS accumulation caused by beta-AP25-35. The effective doses of Sal B were far lower than those of EGb 761. CONCLUSION: Sal B was much more efficient than EGb 761 in inhibiting beta-AP aggregation and in protecting PC12 cells from beta-AP-induced cytotoxicity.
Asunto(s)
Péptidos beta-Amiloides/toxicidad , Benzofuranos/farmacología , Fármacos Neuroprotectores/farmacología , Fragmentos de Péptidos/toxicidad , Extractos Vegetales/farmacología , Péptidos beta-Amiloides/química , Péptidos beta-Amiloides/ultraestructura , Animales , Apoptosis/efectos de los fármacos , Benzofuranos/aislamiento & purificación , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Medicamentos Herbarios Chinos/aislamiento & purificación , Medicamentos Herbarios Chinos/farmacología , Citometría de Flujo , Ginkgo biloba/química , Líquido Intracelular/efectos de los fármacos , Líquido Intracelular/metabolismo , Microscopía Electrónica , Fármacos Neuroprotectores/aislamiento & purificación , Células PC12 , Fragmentos de Péptidos/química , Fragmentos de Péptidos/ultraestructura , Extractos Vegetales/aislamiento & purificación , Plantas Medicinales/química , Ratas , Especies Reactivas de Oxígeno/metabolismo , Salvia miltiorrhiza/químicaRESUMEN
Fibrillar amyloid beta-protein (Abeta) is the principal component of amyloid plaques in the brains of patients with Alzheimer's disease. We have studied the effect of walnut extract on Abeta fibrillization by Thioflavin T fluorescence spectroscopy and electron microscopy. The walnut extract not only inhibited Abeta fibril formation in a concentration and time- dependent manner but it was also able to defibrillize Abeta preformed fibrils. Over 90% inhibition of Abeta fibrillization was observed with 5 microl of methanolic extract of walnut (MEOW) both after 2 and 3 days of incubation. The maximum defibrillization (91.6%) was observed when preformed Abeta fibrils were incubated with 10 microl of MEOW for 2 days. These results suggest that walnuts may reduce the risk or delay the onset of Alzheimer's disease by maintaining Abeta in the soluble form. Further studies showed that anti-amyloidogenic compound in walnut is an organic compound of molecular weight less than 10 kDa, which is neither a lipid nor a protein. Chloroform extract of walnut had no effect on Abeta fibrillization while MEOW and its 10 kDa filtrate inhibited Abeta fibrillization equally. It is proposed that polyphenolic compounds (such as flavonoids) present in walnuts may be responsible for its anti-amyloidogenic activity.
Asunto(s)
Péptidos beta-Amiloides/metabolismo , Amiloide/antagonistas & inhibidores , Juglans/química , Extractos Vegetales/farmacología , Péptidos beta-Amiloides/ultraestructura , Benzotiazoles , Relación Dosis-Respuesta a Droga , Colorantes Fluorescentes , Metanol , Microscopía Electrónica , Extractos Vegetales/administración & dosificación , Solventes , Espectrometría de Fluorescencia , Tiazoles , Factores de TiempoRESUMEN
OBJECTIVE: To observe inhibiting effect of ethanol extract from Achyranthes bidentata (Nx-E) on A beta 42 on (beta-amyloid protein 42) aggregation and fibril formation. METHODS: A beta 42 and Nx-E were incubated in 37 degrees C, 5%, CO2 incubator first, fibril formation was monitored by thioflavine-T fluorescence and was observed with transmission electron microscopy. RESULTS: At 7 days, 1 microgram.microL-1 A beta 42 fluorescence intensity was significantly increased, and much amyloid-like fibril was observed. A beta 42 co-incubated with 15 and 30 micrograms.microL-1 Nx-E for 7 days, their fluorescence intensities were remarkably reduced, few fibrils but an amorphous material were often observed. CONCLUSIONS: After 1 microgram.microL-1 A beta 42 was incubated alone up to 7 days, A beta 42 aggregated and amyloid-like fibril formed. 15 and 30 micrograms.microL-1 Nx-E can prevent A beta 42 from polymerizing to some extent.
Asunto(s)
Achyranthes/química , Péptidos beta-Amiloides/metabolismo , Medicamentos Herbarios Chinos/farmacología , Fragmentos de Péptidos/metabolismo , Plantas Medicinales/química , Péptidos beta-Amiloides/ultraestructura , Medicamentos Herbarios Chinos/aislamiento & purificación , Etanol/química , Fragmentos de Péptidos/ultraestructura , Desnaturalización Proteica , Pliegue de ProteínaRESUMEN
A series of benzofuran derivatives have been identified as inhibitors of fibril formation in the beta-amyloid peptide. The activity of these compounds has been assessed by a novel fibril-formation-specific immunoassay and for their effects on the production of a biologically active fibril product. The inhibition afforded by the compounds seems to be associated with their binding to beta-amyloid, as identified by scintillation proximity binding assay. Binding assays and NMR studies also indicate that the inhibition is associated with self-aggregation of the compounds. There is a close correlation between the activity of the benzofurans as inhibitors of fibril formation and their ability to bind to beta-amyloid. Non-benzofuran inhibitors of the fibril formation process do not seem to bind to the same site on the beta-amyloid molecule as the benzofurans. Thus a specific recognition site might exist for benzofurans on beta-amyloid, binding to which seems to interfere with the ability of the peptide to form fibrils.