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

• Published in: Molecular neurobiology Vol. 53; no. 10; pp. 6716 - 6729
• Main Authors: Darshit, B. S., Ramanathan, M.
• Format: Journal Article
• Language: English
• Published: New York Springer US 01-12-2016
• Subjects: Apoptosis - drug effects; beta Catenin - metabolism; Biomarkers - metabolism; Biomedical and Life Sciences; Biomedicine; Caspase 3 - metabolism; Cell Biology; Cell Differentiation - drug effects; Cell Line, Tumor; Cell Survival - drug effects; Down-Regulation - drug effects; Gene Expression Regulation, Neoplastic - drug effects; Glucose; Glycogen Synthase Kinase 3 beta - metabolism; Humans; Inhibitor of Apoptosis Proteins - metabolism; L-Lactate Dehydrogenase - metabolism; Models, Biological; Neurobiology; Neurology; Neurons - drug effects; Neurons - metabolism; Neurons - pathology; Neuroprotection - drug effects; Neurosciences; Oxygen; Protein Kinase Inhibitors - pharmacology; Proto-Oncogene Proteins c-akt - metabolism; Signal Transduction - drug effects; Time Factors; Tripartite Motif Proteins - metabolism; Ubiquitin-Protein Ligases - metabolism; Up-Regulation - drug effects; Neuroprotection; Ischemia; Glycogen synthase kinase-3; ); GSK-3; Purine; Catenin; Apoptosis; β-Catenin; Glycogen synthase kinase-3β (GSK-3β)
• ISSN: 0893-7648; 1559-1182
• DOI: 10.1007/s12035-015-9598-z

The objective of this study is to elucidate the effect of a new glycogen synthase kinase-3
β
(GSK-3
β
) inhibitor in RA differentiated SH-SY5Y cells in oxygen and glucose deprivation (OGD) model. The pathway involved in GSK-3
β
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-3
β
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-3
β
inhibitor, AR-A014418 (1 μM), was used for comparison. Molecules were pretreated (30 min) and cotreated during OGD exposure. GSK-3
β
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-3
β
/
β
-catenin pathway and stabilized
β
-catenin. Inhibition of GSK-3
β
maintained neuronal survival by upregulating GAP43, Ngn1, and NeuroD2 gene transcription. Further GSK-3
β
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-3
β
/
β
-catenin–TRIM11/survivin pathway.