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Indirubin-3-Oxime Prevents H2O2-Induced Neuronal Apoptosis via Concurrently Inhibiting GSK3β and the ERK Pathway

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Abstract

Oxidative stress-induced neuronal apoptosis plays an important role in many neurodegenerative disorders. In this study, we have shown that indirubin-3-oxime, a derivative of indirubin originally designed for leukemia therapy, could prevent hydrogen peroxide (H2O2)-induced apoptosis in both SH-SY5Y cells and primary cerebellar granule neurons. H2O2 exposure led to the increased activities of glycogen synthase kinase 3β (GSK3β) and extracellular signal-regulated kinase (ERK) in SH-SY5Y cells. Indirubin-3-oxime treatment significantly reversed the altered activity of both the PI3-K/Akt/GSK3β cascade and the ERK pathway induced by H2O2. In addition, both GSK3β and mitogen-activated protein kinase inhibitors significantly prevented H2O2-induced neuronal apoptosis. Moreover, specific inhibitors of the phosphoinositide 3-kinase (PI3-K) abolished the neuroprotective effects of indirubin-3-oxime against H2O2-induced neuronal apoptosis. These results strongly suggest that indirubin-3-oxime prevents H2O2-induced apoptosis via concurrent inhibiting GSK3β and the ERK pathway in SH-SY5Y cells, providing support for the use of indirubin-3-oxime to treat neurodegenerative disorders caused or exacerbated by oxidative stress.

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Abbreviations

ANOAVA:

Analysis of variance

CGNs:

Cerebellar granule neurons

ERK:

Extracellular signal-regulated kinase

FBS:

Fetal bovine serum

FDA:

Fluorescein diacetate

GSK3β:

Glycogen synthase kinase 3β

H2O2 :

Hydrogen peroxide

LDH:

Lactate dehydrogenase

MAPK:

Mitogen-activated protein kinase

MTT:

3(4,5-Dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide

NO:

Nitric oxide

PBS:

Phosphate-buffered saline

PI:

Propidium iodide

PI3-K:

Phosphoinositide 3-kinase

ROS:

Reactive oxygen species

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Acknowledgments

This work was supported by the Natural Science Foundation of Zhejiang Province (LY15H310007), Research Grants Council of Hong Kong (561011 & 15101014), the Applied Research Project on Nonprofit Technology of Zhejiang Province (2016C37110), the National Natural Science Foundation of China (U1503223, 81202150, 81471398), the Ningbo International Science and Technology Cooperation Project (2014D10019), Ningbo municipal innovation team of life science and health (2015C110026), Ningbo Natural Science Foundation (2015A610219), the Scientific Research Foundation for the Returned Overseas Chinese Scholars, the State Education Ministry, and the K. C. Wong Magna Fund in Ningbo University.

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Authors and Affiliations

  1. Ningbo Key Laboratory of Behavioral Neuroscience, Zhejiang Provincial Key Laboratory of Pathophysiology, School of Medicine, Ningbo University, Ningbo, 315211, China

    Jie Yu, Jiacheng Zheng, Jiajia Lin, Rui Yu, Xiang Wu, Wenhua Zhou, Wei Cui & Qinwen Wang

  2. Ningbo Xiaoshi High School, Ningbo, 315010, China

    Linlu Jin & Wei Su

  3. Department of Applied Biology and Chemistry Technology, Institute of Modern Chinese Medicine, The Hong Kong Polytechnic University, Hong Kong SAR, China

    Shinghung Mak, Shengquan Hu & Yifan Han

  4. The Affiliated Yinzhou Hospital, School of Medicine, Ningbo University, Ningbo, 315211, China

    Hongya Sun

  5. Institute of New Drug Research, Guangdong Province Key Laboratory of Pharmacodynamic, Constituents of Traditional Chinese Medicine and New Drug Research, College of Pharmacy, Jinan University, Guangdong, China

    Zaijun Zhang

  6. State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macau, China

    Mingyuen Lee

  7. Division of Life Science and Center for Chinese Medicine, The Hong Kong University of Science and Technology, Hong Kong, China

    Wahkeung Tsim

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  1. Jie Yu
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  15. Wei Cui
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Correspondence to Wei Cui, Yifan Han or Qinwen Wang.

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Yu, J., Zheng, J., Lin, J. et al. Indirubin-3-Oxime Prevents H2O2-Induced Neuronal Apoptosis via Concurrently Inhibiting GSK3β and the ERK Pathway. Cell Mol Neurobiol 37, 655–664 (2017). https://doi.org/10.1007/s10571-016-0402-z

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