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Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-334919

RESUMO

<p><b>OBJECTIVE</b>To achieve expression of human brain-derived neurotrophic factor (hBDNF) mediated by recombinant adeno-associated virus (rAAV) and explore the mechanism of its neuroprotective effects in rat neurons against beta-amyloid-induced Alzheimer's disease.</p><p><b>METHODS</b>Using molecular cloning technique, rAAV vector containing hBDNF gene (AAV-hBDNF) was constructed to transfect SD rat hippocampal neurons exposed to beta-amyloid treatment. The changes in cell apoptosis were observed by MTT assay and flow cytometry, and the expression of hBDNF and Bcl-2 protein were determined by immunocytochemical staining. Laser scanning confocal microscopy (LSCM) was used to observe the changes of [Ca(2+)](i).</p><p><b>RESULTS</b>The cultured rat hippocampal neurons were effectively transfected with AAV-hBDNF and expression of BDNF protein was obviously increased. hBNDF expression showed significant protective effects against beta-amyloid-induced neuronal damage, and the expression of Bcl-2 protein was increased significantly and the balance of [Ca(2+)](i) was maintained in BDNF-treated cells with beta-amyloid exposure.</p><p><b>CONCLUSION</b>hBDNF expression can effectively protect cultured rat hippocampal cells from beta-amyloid-induced apoptosis through inhibiting the intracellular calcium overload and increasing the expression of Bcl-2 protein.</p>


Assuntos
Animais , Humanos , Ratos , Doença de Alzheimer , Genética , Metabolismo , Patologia , Peptídeos beta-Amiloides , Toxicidade , Animais Recém-Nascidos , Fator Neurotrófico Derivado do Encéfalo , Genética , Fisiologia , Linhagem Celular , Sobrevivência Celular , Genética , Fisiologia , Células Cultivadas , Dependovirus , Genética , Vetores Genéticos , Hipocampo , Biologia Celular , Metabolismo , Imuno-Histoquímica , Microscopia Eletrônica de Varredura , Neurônios , Metabolismo , Proteínas Proto-Oncogênicas c-bcl-2 , Metabolismo , Ratos Sprague-Dawley , Transfecção
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