RESUMO
Doxorubicin (Dox), an effective antineoplastic drug, was limited use for cardiotoxicity. Xinshuitong Capsule (XST), a patented herbal formula, showed desirable beneficial effects in the treatment of chronic heart failure (CHF) patients. However, the drug on Dox-induced cardiotoxicity remains unclear. Ninety male Sprague-Dawley rats were randomized into two groups: 15 rats were selected as the normal group and 75 rats were injected intraperitoneally with Dox to establish CHF rat models, the success ones were randomly divided into five groups: low XST (LXST), medium XST (MXST) or high XST (HXST) (4.9, 9.8, or 19.6 g/kg d) administrated intragastrically twice a day for 4 weeks, with the captopril-treated group and the model group as comparison. The model group showed the cardiac functions generally impaired, and CHF mortality rate higher (47%) than those in the XST-treated groups (averaged 24%, P < 0.05). Compared with XST-treated groups, myocardial remodeling, inflammation and desarcomerization, and higher water content more severe in the cardiac tissue in the model group (P < 0.05), which was associated with higher expressions of mRNA or protein levels of AQP1, 4 and 7. Dox-impaired cardiac functions, cardiac remodeling and myocardial edema could be dose-dependently reverted by XST treatment. XST could inhibit AQP1, 4 and 7 at mRNA levels or at protein levels, which was associated with the attenuation of myocardial edema and cardiac remodeling, decreasing the ventricular stiffness and improving the cardiac functions and rats' survival. AQPs is involved in cardiac edema composed one of the mechanisms of Dox-induced cardiotoxicity, XSTvia inhibition of AQPs relieved the Dox-induced side effects.
Assuntos
Aquaporinas/antagonistas & inibidores , Medicamentos de Ervas Chinesas/farmacologia , Edema Cardíaco/prevenção & controle , Insuficiência Cardíaca/prevenção & controle , Miocárdio/metabolismo , Administração Oral , Animais , Aquaporina 1/antagonistas & inibidores , Aquaporina 1/genética , Aquaporina 1/metabolismo , Aquaporina 4/antagonistas & inibidores , Aquaporina 4/genética , Aquaporina 4/metabolismo , Aquaporinas/genética , Aquaporinas/metabolismo , Água Corporal/metabolismo , Cápsulas , Cardiotoxicidade , Doença Crônica , Modelos Animais de Doenças , Doxorrubicina , Medicamentos de Ervas Chinesas/administração & dosagem , Edema Cardíaco/induzido quimicamente , Edema Cardíaco/metabolismo , Edema Cardíaco/patologia , Insuficiência Cardíaca/induzido quimicamente , Insuficiência Cardíaca/metabolismo , Insuficiência Cardíaca/patologia , Masculino , Miocárdio/patologia , Ratos Sprague-Dawley , Transdução de Sinais , Função Ventricular Esquerda/efeitos dos fármacos , Remodelação Ventricular/efeitos dos fármacosRESUMO
OBJECTIVE: Neuromyelitis optica (NMO) is a severe neurological demyelinating autoimmune disease that affects the optic nerves and spinal cord with no cure and no FDA-approved therapy. Research over the last decade revealed that the binding of NMO-IgG to the water channel protein astrocyte aquaporin 4 (AQP4) might be the primary cause of NMO pathogenesis. The purpose of this study was to identify potential blockers of NMO-IgG and AQP4 binding. METHODS: We developed a two-step screening platform consisting of a reporter cell-based high-throughput screen assay and a cell viability-based assay. Purified NMO-IgG from NMO patient serum and transfected Chinese hamster lung fibroblast V79 cells stably expressing human M23-AQP4 were used for primary screening of 40,000 small molecule fractions from 500 traditional Chinese herbs. RESULTS: Thirty-six positive fractions were identified, of which 3 active fractions (at 50 µg/ml) were found to be from the same Chinese traditional herb Mahonia japonica (Thunb.). A bioactivity-guided method based on a primary screening assay for blocking activity led to the isolation of an active single natural compound, isotetrandrine, from the 3 fractions. Our immunofluorescence staining results showed that isotetrandrine can block NMO-IgG binding to AQP4 without affecting the expression and function of AQP4. It can also inhibit NMO-IgG binding to astrocyte AQP4 in NMO patient sera and block NMO-IgG-dependent complement-mediated cytotoxicity with the IC50 at â¼3 µM. CONCLUSIONS: The present study developed a cell-based high-throughput screen to identify small molecule inhibitors for NMO-IgG and AQP4 binding, and suggests a potential therapeutic value of isotetrandrine in NMO.
Assuntos
Aquaporina 4/metabolismo , Astrócitos/metabolismo , Benzilisoquinolinas/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Imunoglobulina G/metabolismo , Neuromielite Óptica/metabolismo , Animais , Aquaporina 4/antagonistas & inibidores , Astrócitos/efeitos dos fármacos , Astrócitos/patologia , Benzilisoquinolinas/uso terapêutico , Células CHO , Células Cultivadas , Cricetinae , Cricetulus , Avaliação Pré-Clínica de Medicamentos/métodos , Medicamentos de Ervas Chinesas/uso terapêutico , Humanos , Imunossupressores/farmacologia , Imunossupressores/uso terapêutico , Camundongos , Camundongos Knockout , Neuromielite Óptica/tratamento farmacológico , Neuromielite Óptica/patologia , Ligação Proteica/efeitos dos fármacos , Ligação Proteica/fisiologiaRESUMO
Repetitive binge intoxication with ethanol (alcohol) in adult rats, mimicking chronic ethanol abuse in alcoholics, causes trauma-like brain edema and relatively selective neurodegeneration of hippocampal dentate granule cells and pyramidal neurons in the temporal cortex (especially entorhinal cortex). We have now modeled the aspects of this type of acquired brain damage in vitro with rat entorhino-hippocampal slice cultures of adult brain age (62 ± 3 days). When sequentially treated (four 16-h overnight exposures) with 100 mM ethanol, the slices display elevated levels of aquaporin-4 (AQP4) water channels accompanied by significant neurodegeneration. Increased AQP4 has been associated with neuroinflammatory responses including edema, pro-inflammatory cytokine elevations, arachidonic acid release, and oxidative stress. Co-treatment of ethanol-binged slice cultures with docosahexaenoic acid (DHA), an omega-3 fatty acid known to suppress brain damage from other insults, prevents both the AQP4 elevations and the neurodamage. Surmising that AQP4 augmentation is a causative neuroinflammatory component in this model, we are investigating several possibilities to explain the protective actions of the omega-3 fatty acid. Since the worldwide incidence of cognitive dysfunction and dementia from ethanol abuse and alcoholism is not inconsequential, DHA supplementation with chronic alcoholics could emerge to be a rational approach to potentially lessening brain disabilities.
Assuntos
Aquaporina 4/antagonistas & inibidores , Consumo Excessivo de Bebidas Alcoólicas/prevenção & controle , Ácidos Docosa-Hexaenoicos/uso terapêutico , Etanol/toxicidade , Hipocampo/efeitos dos fármacos , Degeneração Neural/prevenção & controle , Fatores Etários , Animais , Aquaporina 4/metabolismo , Consumo Excessivo de Bebidas Alcoólicas/metabolismo , Consumo Excessivo de Bebidas Alcoólicas/patologia , Ácidos Docosa-Hexaenoicos/farmacologia , Etanol/administração & dosagem , Hipocampo/metabolismo , Hipocampo/patologia , Masculino , Degeneração Neural/metabolismo , Degeneração Neural/patologia , Técnicas de Cultura de Órgãos , Ratos , Ratos Sprague-DawleyRESUMO
The potential of antiepileptic drugs (AEDs) to inhibit the water transport properties of aquaporin 4 (AQP4) was investigated using a combination of in silico and in vitro screening methods. Virtual docking studies on 14 AEDs indicated a range of docking energies that spanned approximately 40 kcal/mol, where the most stabilized energies were consistent with that of the previously identified AQP4 inhibitor acetazolamide. Nine AEDs and one bio-active metabolite were further investigated in a functional assay using AQP4 expressing Xenopus oocytes. Seven of the assayed compounds were found to inhibit AQP4 function, while three did not. A linear correlation was indicated between the in silico docking energies and the in vitro AQP4 inhibitory activity at 20 microM.