Development of a high-throughput screening assay for cytoprotective agents in rotenone-induced cell death

被引：8

作者：

Yoon, Il Sang ^{[1
]}

Au, Qingyan ^{[1
]}

Barber, Jack R. ^{[1
]}

Ng, Shi Chung ^{[1
]}

Zhang, Bin ^{[1
]}

机构：

[1] CytRx Corp, Dept Biol, San Diego, CA 92109 USA

来源：

ANALYTICAL BIOCHEMISTRY | 2010年 / 407卷 / 02期

关键词：

Rotenone; High-throughput screening; High-content analysis; Small molecule; Parkinson's disease; Carnosic acid; PARKINSONS-DISEASE; IN-VITRO; ACTIVATION; PATHOGENESIS; APOPTOSIS; MODEL;

D O I：

10.1016/j.ab.2010.08.011

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Parkinson's disease (PD) is a neurodegenerative disease featured by selective loss of substantia nigra neurons. Rotenone administration in animals induces neurodegeneration accompanied by alpha-synuclein-positive Lewy body-like inclusions, recapturing typical histopathological features of PD. In an effort to screen for small-molecule agents to reverse rotenone-induced cytotoxicity, we developed and validated a sensitive and robust assay with neuroblastoma SK-N-SH cells. This assay was amenable to a high-throughput screening format with Z' factor of 0.56. Robotic screening of a bioactive compound library led to the identification of carnosic acid that can effectively protect cells from rotenone treatment. Using a high-content image-based assay and Western blot analysis, we demonstrated that carnosic acid protects cells from rotenone stress by significant induction of HSP70 expression. Therefore, the assay reported here can be used to identify novel cytoprotective agents for clinical therapeutics of PD. (C) 2010 Elsevier Inc. All rights reserved.

引用

页码：205 / 210

页数：6

共 29 条

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