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Journal of Southern Medical University ; (12): 1388-1393, 2006.
Article in Chinese | WPRIM | ID: wpr-334919

ABSTRACT

<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>


Subject(s)
Animals , Humans , Rats , Alzheimer Disease , Genetics , Metabolism , Pathology , Amyloid beta-Peptides , Toxicity , Animals, Newborn , Brain-Derived Neurotrophic Factor , Genetics , Physiology , Cell Line , Cell Survival , Genetics , Physiology , Cells, Cultured , Dependovirus , Genetics , Genetic Vectors , Hippocampus , Cell Biology , Metabolism , Immunohistochemistry , Microscopy, Electron, Scanning , Neurons , Metabolism , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Rats, Sprague-Dawley , Transfection
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