Development of a high-throughput screening assay for cytoprotective agents in rotenone-induced cell death.
Parkinson's disease (PD) is a neurodegenerative disease featured by selective loss of substantia nigra neurons. Rotenone administration in animals induces neurodegeneration accompanied by α-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.
Yoon
Il Sang
IS
Department of Biology, CytRx Corporation, 3030 Bunker Hill Street, Suite 101, San Diego, CA 92109, USA.
Au
Qingyan
Q
Barber
Jack R
JR
Ng
Shi Chung
SC
Zhang
Bin
B
D045784:Abietanes / Q000737:chemistry / Q000494:pharmacology; D017209:Apoptosis; D015153:Blotting, Western; D045744:Cell Line, Tumor; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007306:Insecticides / Q000633:toxicity*; D018696:Neuroprotective Agents / Q000737:chemistry / Q000494:pharmacology*; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology; D012402:Rotenone / Q000633:toxicity*; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology
D045784:Abietanes; D018840:HSP70 Heat-Shock Proteins; D007306:Insecticides; D018696:Neuroprotective Agents; D010936:Plant Extracts; D054852:Small Molecule Libraries; D012402:Rotenone; C018381:salvin