Alpha-lipoic acid protects human dopaminergic neuronal cells against hydrogen peroxide-induced cell injury by inhibiting autophagy and apoptosis

서지정보

ㆍ발행기관 : 대한구강생물학회 ㆍ수록지정보 : International Journal of Oral Biology / 46권 / 1호
ㆍ저자명 : Kyeong-Rok Kang, Jae-Sung Kim, Tae-Hyeon Kim, Jeong-Yeon Seo, HyangI Lim, Jong-Hyun Park, Kwang Yeol Yang, Sun-Kyoung Yu, Heung-Joong Kim, Chun Sung Kim, Hong Sung Chun, Dong-Seol Lee, Joo-Cheol Park, Do Kyung Kim

목차

Introduction
Materials and Methods
1. Materials
2. Cell culture and cell treatments
3. Cytotoxicity assay
4. Cell Live/Dead assay
5. DAPI staining
6. Measurement of intracellular ROS
7. Immunoblotting
8. Haematoxylin and eosin staining
9. Immunocytochemistry
10. Statistical analysis
Results
1. ALA protects SH-SY5Y cells against H2O2-inducedcytotoxicity
2. ALA inhibited H2O2-induced apoptosis and ROSproduction in SH-SY5Y cells
3. ALA protects H2O2-induced cell death throughdown-regulating oxidative stress
4. ALA inhibits autophagy in H2O2-induced SH-SY5Ycell injury
Discussion
References

영어 초록

Alpha-lipoic acid (ALA) is a naturally occurring antioxidant and has been previously used to treat diabetes and cardiovascular disease. However, the autophagy effects of ALA against oxidative stress-induced dopaminergic neuronal cell injury remain unclear. The aim of this study was to investigate the role of ALA in autophagy and apoptosis against oxidative stress in the SH-SY5Y human dopaminergic neuronal cell line. We examined SH-SY5Y phenotypes using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay (cell viability/proliferation), 4′,6-diamidino-2-phenylindole dihydrochloride nuclear staining, Live/Dead cell assay, cellular reactive oxygen species (ROS) assay, immunoblotting, and immunocytochemistry. Our data showed ALA attenuated hydrogen peroxide (H2O2)-induced ROS generation and cell death. ALA effectively suppressed Bax up-regulation and Bcl-2 and BclxL down-regulation. Furthermore, ALA increased the expression of the antioxidant enzyme, heme oxygenase-1. Moreover, the expression of Beclin-1 and LC-3 autophagy biomarkers was decreased by ALA in our cell model. Combined, these data suggest ALA protects human dopaminergic neuronal cells against H2O2-induced cell injury by inhibiting autophagy and apoptosis.

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