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Protective Effects of Fisetin Against 6-OHDA-Induced Apoptosis by Activation of PI3K-Akt Signaling in Human Neuroblastoma SH-SY5Y Cells

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Abstract

6-Hydroxydopamine (6-OHDA) induces the production of reactive oxygen species (ROS) that are associated with various neurodegenerative diseases such as Parkinson’s disease. 3,3′,4′,7-Tetrahydroxyflavone (fisetin), a plant flavonoid has a variety of physiological effects such as antioxidant activity. In this study, we investigated the molecular mechanism of the neuroprotective effects of fisetin against 6-OHDA-induced cell death in human neuroblastoma SH-SY5Y cells. 6-OHDA-mediated cell toxicity was reduced in a fisetin concentration-dependent manner. 6-OHDA-mediated elevation of the expression of the oxidative stress-related genes such as hemeoxygenase-1, NAD(P)H dehydrogenase quinone 1, NF-E2-related factor 2, and γ-glutamate-cysteine ligase modifier was suppressed by fisetin. Fisetin also lowered the ratio of the proapoptotic Bax protein and the antiapoptotic Bcl-2 protein in SH-SY5Y cells. Moreover, fisetin effectively suppressed 6-OHDA-mediated activation of caspase-3 and caspase-9, which leads to the cell death, while, 6-OHDA-induced caspase-3/7 activity was lowered. Furthermore, fisetin activated the PI3K-Akt signaling, which inhibits the caspase cascade, and fisetin-mediated inhibition of 6-OHDA-induced cell death was negated by the co-treatment with an Akt inhibitor. These results indicate that fisetin protects 6-OHDA-induced cell death by activating PI3K-Akt signaling in human neuronal SH-SY5Y cells. This is the first report that the PI3K-Akt signaling is involved in the fisetin-protected ROS-mediated neuronal cell death.

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Abbreviations

PD:

Parkinson’s disease

ROS:

Reactive oxygen species

6-OHDA:

6-Hydroxydopamine

fisetin:

3,3′,4′,7-Tetrahydroxyflavone

ERK:

Extracellular signal-regulated kinase

HRP:

Horseradish peroxidase

DCDFA:

2′,7′-Dichlorofluorescein diacetate

DMEM:

Dulbecco’s modified Eagle’s medium

LDH:

Lactate dehydrogenase

qPCR:

Quantitative PCR

HO-1:

Hemeoxygenase-1

NQO1:

NAD(P)H dehydrogenase quinone 1

NRF2:

NF-E2-related factor 2

GCLM:

γ-glutamate-cysteine ligase modifier

PI3K:

Phosphoinositide 3-kinase

Akti:

Akt inhibitor

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Author notes

  1. Yuta Morishige

    Present address: Research Institute of Tuberculosis, 3-1-24 Matsuyama, Kiyose, Tokyo, 204-8533, Japan

Authors and Affiliations

  1. Laboratory of Pathobiochemistry, Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka, 569-1094, Japan

    Ryoko Watanabe, Takumi Kurose & Ko Fujimori

  2. Laboratory of Biodefense and Regulation, Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka, 569-1094, Japan

    Yuta Morishige

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  1. Ryoko Watanabe
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  2. Takumi Kurose
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Correspondence to Ko Fujimori.

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Watanabe, R., Kurose, T., Morishige, Y. et al. Protective Effects of Fisetin Against 6-OHDA-Induced Apoptosis by Activation of PI3K-Akt Signaling in Human Neuroblastoma SH-SY5Y Cells. Neurochem Res 43, 488–499 (2018). https://doi.org/10.1007/s11064-017-2445-z

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  • DOI: https://doi.org/10.1007/s11064-017-2445-z

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