J Korean Neurol Assoc > Volume 21(4); 2003 > Article

Journal of the Korean Neurological Association 2003;21(4): 401-407.

PC12 세포에서 산화스트레스에 의한 베타-아밀로이드 펩티드의 세포독성

이재규 , 김찬석* 김태정 김병학 김성용 하정상* 김정희 김재룡

영남대학교 의과대학 생화학?분자생물학교실, 신경과학교실*

"Involvements of Oxidative Stress in β-amyloid Peptide-induced Cytotoxicity in PC12 Cells"

Jae-Kyu Lee

"Department of Biochemistry and Molecular Biology, Department of Neurology* College of Medicine, Yeungnam University"

Abstract

"Background: Alzheimer s disease is a neurodegenerative disorder characterized by the extracellular deposition of β- amyloid peptide(Aβ) in the brain, presumed to play a pathogenic role. However, the precise molecular mechanisms of its neurotoxicity are not fully understood.
Methods: Aβ-mediated cytotoxicity in neuronal cell lines (PC12, SH-SY5Y, IMR32, and U87) was measured by an MTT assay. NF-κB activation by Aβwas examined by a luciferase assay and apoptosis induced by Aβwas measured by cytoplasmic DNA fragmentations.
Results: Aβcytotoxicity in the tested cell lines was more prominent in the absence of serum than in the presence of serum in culture media. PC12 cells showed the highest sensitivity to Aβcytotoxicity among the cell lines. The Aβ(25-35) cytotoxicity in PC12 cells was increased in a dose-dependent manner. For convincing oxidative stress involved in Aβcytotoxicity, antioxidants such as DTT, GSH, vitamin C, or NAC were pretreated. GSH protected PC12 cells from Aβcytotoxicity, but DTT or NAC did not. Aβ (25-35) treatment to PC12 cells increased the NF-κB activity in a dose-dependent manner. Cytoplasmic DNA fragmentations, one of the apoptotic indicators, were increased at lower concentrations of Aβ(25-35) from 0.01 to 0.1 μM, however, dose-dependent increments of DNA fragmentations were not observed at higher concentrations from 1 to 10 μM.
Conclusions: From these results, Aβ-induced cytotoxicity in PC12 cells might be mediated by oxidative stress. Key Words: Alzheimer disease, Amyloid beta-protein, Oxidative stress, PC12 cells"