Brain Insulin Resistance in Alzheimer's disease: Targeting phosphoinositide 3-kinase (PI3K)/Akt/GSK-3? pathway
Author(s): Ansab Akhtar, Sangeeta Pilkhwal Sah
Abstract
Protein concentrations were determined by the biuret method. Cholinergic activity was evaluated by acetylcholinesterase assay to assess the cholinergic dysfunction which is one of the core pathologies of dementia and AD. Inflammatory cytokines like TNF-α, IL-6 was determined by ELISA method to evaluate the neuroinflammation which is aggravated by insulin resistance. C-reactive protein, a marker of neuroinflammation and neurodegeneration was also determined by ELISA. Mitochondrial dysfunction was evaluated estimating mitochondrial enzyme complex-I, II, III, IV depicting picture of viable and non-viable neuronal cells. Histopathology was done by H&E staining to find out apoptotic cells, neuroinflammation, and neurodegeneration. Molecular technique like RT-PCR for IRS-1, PI3-K, AKT, GSK 3-β and BDNF was performed for gene expression analysis