Grewia tiliaefolia and its active compound vitexin regulate the expression of glutamate transporters and protect Neuro-2a cells from glutamate toxicity

Life Sci. 2018 Jun 15:203:233-241. doi: 10.1016/j.lfs.2018.04.047. Epub 2018 Apr 26.

Abstract

Aim: Glutamate is a major neurotransmitter involved in several brain functions and glutamate excitotoxicity is involved in Alzheimer's disease (AD). In the current study, the neuroprotective effect of the Indian medicinal plant Grewia tiliaefolia (GT) and its active component vitexin was evaluated in Neuro-2a cells against glutamate toxicity.

Materials and methods: Neuro-2a cells were exposed to glutamate to cause excitotoxicity and the neuroprotective effect of GT and vitexin were evaluated using biochemical studies (estimation of reactive oxygen species, reactive nitrogen species, protein carbonyl content, lipid peroxidation level, mitochondrial membrane potential and caspase-3 activity), molecular docking studies, gene expression and western blot analysis.

Key findings: Glutamate exposure to Neuro-2a cells induced oxidative stress, loss of membrane potential, suppressed the expression of antioxidant response genes (Nrf-2, HO-1, NQO-1), glutamate transporters (GLAST-1, GLT-1) and induced the expression of NMDAR, Calpain. However, pre-treatment of cells with GT/vitexin inhibited oxidative stress mediated damage by augmenting the expression of Nrf-2/HO-1 pathway, inducing the expression of glutamate transporters and downregulating Calpain, NMDAR. Molecular docking showed that vitexin effectively binds to NMDAR and GSK-3β and thereby can inhibit their activation. GT/vitexin also inhibited glutamate induced Bax expression.

Significance: Methanol extract of G. tiliaefolia and its active component vitexin can act in an antioxidant dependent mechanism as well as by regulating glutamate transporters in mitigating the toxicity exerted by glutamate in Neuro-2a cells. Our results conclude that GT/vitexin can act as potential drug leads for the therapeutic intervention of AD.

Keywords: Calpain; GLT-1; GSK-3β; Glutamate; NMDAR; Vitexin.

MeSH terms

  • Amino Acid Transport System X-AG / metabolism*
  • Animals
  • Apigenin / pharmacology*
  • Apoptosis / drug effects
  • Cell Proliferation / drug effects
  • Glutamic Acid / toxicity*
  • Grewia / chemistry*
  • Lipid Peroxidation / drug effects
  • Membrane Potential, Mitochondrial / drug effects
  • Mice
  • Molecular Docking Simulation
  • Neuroblastoma / chemically induced
  • Neuroblastoma / drug therapy
  • Neuroblastoma / metabolism
  • Neuroblastoma / pathology*
  • Neuroprotective Agents / pharmacology*
  • Oxidative Stress / drug effects
  • Reactive Oxygen Species / metabolism
  • Signal Transduction / drug effects
  • Tumor Cells, Cultured

Substances

  • Amino Acid Transport System X-AG
  • Neuroprotective Agents
  • Reactive Oxygen Species
  • Glutamic Acid
  • Apigenin
  • vitexin