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Cellular and Molecular Neurobiology

, Volume 37, Issue 4, pp 655–664 | Cite as

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

  • Jie Yu
  • Jiacheng Zheng
  • Jiajia Lin
  • Linlu Jin
  • Rui Yu
  • Shinghung Mak
  • Shengquan Hu
  • Hongya Sun
  • Xiang Wu
  • Zaijun Zhang
  • Mingyuen Lee
  • Wahkeung Tsim
  • Wei Su
  • Wenhua Zhou
  • Wei Cui
  • Yifan Han
  • Qinwen Wang
Original Research

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.

Keywords

Indirubin-3-oxime H2O2 GSK3β PI3-K ERK 

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

Notes

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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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Jie Yu
    • 1
  • Jiacheng Zheng
    • 1
  • Jiajia Lin
    • 1
  • Linlu Jin
    • 2
  • Rui Yu
    • 1
  • Shinghung Mak
    • 3
  • Shengquan Hu
    • 3
  • Hongya Sun
    • 4
  • Xiang Wu
    • 1
  • Zaijun Zhang
    • 5
  • Mingyuen Lee
    • 6
  • Wahkeung Tsim
    • 7
  • Wei Su
    • 2
  • Wenhua Zhou
    • 1
  • Wei Cui
    • 1
  • Yifan Han
    • 3
  • Qinwen Wang
    • 1
  1. 1.Ningbo Key Laboratory of Behavioral Neuroscience, Zhejiang Provincial Key Laboratory of Pathophysiology, School of MedicineNingbo UniversityNingboChina
  2. 2.Ningbo Xiaoshi High SchoolNingboChina
  3. 3.Department of Applied Biology and Chemistry Technology, Institute of Modern Chinese MedicineThe Hong Kong Polytechnic UniversityHong Kong SARChina
  4. 4.The Affiliated Yinzhou Hospital, School of MedicineNingbo UniversityNingboChina
  5. 5.Institute of New Drug Research, Guangdong Province Key Laboratory of Pharmacodynamic, Constituents of Traditional Chinese Medicine and New Drug Research, College of PharmacyJinan UniversityGuangdongChina
  6. 6.State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical SciencesUniversity of MacauMacauChina
  7. 7.Division of Life Science and Center for Chinese MedicineThe Hong Kong University of Science and TechnologyHong KongChina

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