Protective effect of adeno-associated viral vector-mediated expression of human brain-derived neurotrophic factor in rat neurons against beta-amyloid-induced Alzheimer's disease in vitro.
- Author:
Zhao-hui LIU
1
;
Dong-liang MA
;
Hui JIN
;
Yan-bing MA
;
Hai-tao HU
Author Information
- Publication Type:Journal Article
- MeSH: Alzheimer Disease; genetics; metabolism; pathology; Amyloid beta-Peptides; toxicity; Animals; Animals, Newborn; Brain-Derived Neurotrophic Factor; biosynthesis; genetics; physiology; Cell Line; Cell Survival; drug effects; genetics; physiology; Cells, Cultured; Dependovirus; genetics; Genetic Vectors; Hippocampus; cytology; metabolism; Humans; Immunohistochemistry; Microscopy, Electron, Scanning; Neurons; drug effects; metabolism; ultrastructure; Proto-Oncogene Proteins c-bcl-2; metabolism; Rats; Rats, Sprague-Dawley; Transfection
- From: Journal of Southern Medical University 2006;26(10):1388-1393
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSUsing 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).
RESULTSThe 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.
CONCLUSIONhBDNF 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.
