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Effects of insulin-like growth factor-1 on okadaic acid-induced apoptosis in SH-SY5Y cells
Authors:Xing Changhong  Peng Ying  Chang Rui  Yin Yanling  Xie Zuoping
Affiliation:Department of Biological Science and Biotechnology, State Key Laboratory of Biomembrane and Membrane Biotechnology, Tsinghua University, Beijing 100084, China.
Abstract:The effects of insulin-like growth factor-1 (IGF-1) on the cytotoxicity and apoptosis induced by okadaic acid (OA) in SH-SY5Y cells were investigated. Cell viability was measured using the MTT (3-(4,5-dimethylthiazolyl-2)-2,-5-diphenyltetrazolium bromide) assay. Early and late apoptosis/necrosis were analyzed by flow cytometry using Annexin V and propidium iodide (PI) double-staining. Caspase-3 activation was detected by Western blot analysis. Preincubation with IGF-1 for 24 h prevented cytotoxicity induced by 40 nM OA given for 24 h, and the MTT value significantly increased. Incubation with 20 nM OA for 24 h caused a marked increase in the percentage of early apoptotic and late apoptotic/necrotic cells, which was not dependent on the activation of caspase-3. OA-induced apoptosis was significantly decreased by pretreatment with 10 ng/ml of IGF-1 for 24 h. The results supported the hypothesis that IGF-1 may be useful in the treatment of Alzheimer's disease.
Keywords:Insulin‐like growth factor‐1  Okadaic acid  Apoptosis  Caspase  Flow cytometry  Alzheimer's disease
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