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Molecular Neurobiology

, Volume 53, Issue 10, pp 6716–6729 | Cite as

Activation of AKT1/GSK-3β/β-Catenin–TRIM11/Survivin Pathway by Novel GSK-3β Inhibitor Promotes Neuron Cell Survival: Study in Differentiated SH-SY5Y Cells in OGD Model

  • B. S. Darshit
  • M. RamanathanEmail author
Article

Abstract

The objective of this study is to elucidate the effect of a new glycogen synthase kinase-3β (GSK-3β) inhibitor in RA differentiated SH-SY5Y cells in oxygen and glucose deprivation (OGD) model. The pathway involved in GSK-3β signaling during OGD was measured to elucidate the mechanism of action. The differentiation of SH-SY5Y into mature neuronal cells was done with retinoic acid. During differentiation, upregulation of the growth-associated protein 43 (GAP43), neurogenin1 (NGN1), neuronal differentiation 2 (NeuroD2), and tripartite motif containing 11 (TRIM11) genes were observed. Twelve hours of optimal OGD exposure resulted in the alteration of GSK-3β functions of the neuron cells. Of the five molecules selected for this study, molecule G3 showed better effect in the initial phase of the study. Hence, G3 (0.5, 1, and 5 μM) was selected for further study in the OGD model. The standard GSK-3β inhibitor, AR-A014418 (1 μM), was used for comparison. Molecules were pretreated (30 min) and cotreated during OGD exposure. GSK-3β inhibitors showed antiapoptotic activity as evidenced by reduced caspase-3 enzyme activity and increased survivin transcription, as well as improved membrane integrity, evidenced by LDH assay. The inhibitor molecules also up-regulated survival AKT1/GSK-3β/β-catenin pathway and stabilized β-catenin. Inhibition of GSK-3β maintained neuronal survival by upregulating GAP43, Ngn1, and NeuroD2 gene transcription. Further GSK-3β inhibition reduced the TRIM11 gene transcription. In conclusion, both inhibitors have been found to control apoptosis and maintain neuronal functioning and this effect might have been mediated through AKT1/GSK-3β/β-catenin–TRIM11/survivin pathway.

Keywords

Glycogen synthase kinase-3β (GSK-3ββ-Catenin Apoptosis Neuroprotection Ischemia Purine 

Notes

Acknowledgments

This project was funded by the Medicine Division, Department of Biotechnology (DBT), New Delhi, India (reference no. BT/PR14062/MED/30/357/2010). The author would like to thank Dr. Sivaram Hariharan for his contribution in correcting the grammar of this manuscript.

Compliance with Ethical Standards

Conflict of Interest

No conflict of interest

Supplementary material

12035_2015_9598_MOESM1_ESM.docx (74 kb)
Figure S1 (DOCX 73 kb)
12035_2015_9598_MOESM2_ESM.docx (15 kb)
Table S1 (DOCX 15 kb)
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Figure S2 (DOCX 2109 kb)
12035_2015_9598_MOESM4_ESM.docx (15 kb)
Table S2 (DOCX 15 kb)

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of PharmacologyPSG College of PharmacyCoimbatoreIndia

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