[Effects of brain-derived neurotrophic factor on H2O2-induced apoptosis of human arterial smooth muscle cell].
OBJECTIVE
To investigate the protective effects of brain-derived neurotrophic factor (BDNF) on hydrogen peroxide (H2O2) induced apoptosis of human arterial smooth muscle cells (SMC).


METHODS
The hBDNF cDNA was transfected into the SMC mediated by Lipofect- AMINE, and ectopic expression in SMC was driven by the regulatory elements of I type collagen gene(COLIA1). The cell proliferation was measured with a colorimetric assay based on the cleavage of the tetrazolium salt MTT. Microscopic analysis of cell apoptosis was performed with fluorescent stain of acridine orange (AO). Apoptosis rate of SMC was tested by flow cytometry (FCM). DNA pattern was examined by agarose gel electrophoresis.


RESULTS
Exposure of growing SMC to 100-800 mumol/L H2O2 for 24 h remarkably suppressed the cell proliferation. After treatment of the cells with 200 mumol/L H2O2 for 24 h, morphological and biochemical changes of classic apoptosis such as condensation of cytoplasm, fragmentation of the nuclear chromatin, DNA "ladder" pattern after electrophoresis were observed. Yet, the above-mentioned apoptosis changes were not observed and the apoptosis rate was reduced obviously in the BDNF-transfected SMC treated with the same concentration of H2O2.


CONCLUSIONS
H2O2 can induce apoptosis of SMC, but BDNF inhibits H2O2-induced SMC apoptosis markedly and has strong anti-apoptotic activity.
Chen
Jianye
J
Institute of Biochemistry and Molecular Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Liu
Ji
J
Wang
Ruohan
R
Liu
Zhimin
Z
Li
Ji
J
Chen
Junjie
J
D017209:Apoptosis / Q000187:drug effects*; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D006801:Humans; D006861:Hydrogen Peroxide; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D014162:Transfection
D019208:Brain-Derived Neurotrophic Factor; D006861:Hydrogen Peroxide