The effects of propofol on neurotoxicity induced by α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid in rat mixed cortical cultures

Abstract

Background

The pattern of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-mediated neurotoxicity (necrosis vs apoptosis) and the neuroprotective effect of propofol on AMPA-mediated neurotoxicity are still unclear.

Methods

Thirteen-day-old primary rat mixed cortical cultures were used. To measure the neuroprotective effect of propofol, AMPA (50µM), AMPA (50µM) plus propofol (0.1, 1, 25, 50µM), AMPA (50µM) plus DMSO, propofol (50µM) and DMSO were administered (n = 45). Seventy-two h later, surviving cells were counted using trypan blue staining and were converted to cell death rate (CDR). To measure the effect of propofol (50µM) on AMPA (50µM)-induced apoptosis, a triple stain was done. In a fixed field (×400), the number of neuronal cells stained by neuronal nuclei (NeuN) and Hoechst staining and apoptotic cells stained by terminal deoxynucleotidyl transferase mediated dUTP nick-end-labeling (TUNEL) assays were counted. Apoptotic cell rates (ACR) were also calculated. Statistical analyses were performed using one way-analysis of variance followed by Bonferroni's test. P < 0.05 was considered statistically significant.

Results

AMPA (50µM) stimulation demonstrated 49.3% CDR, and adding propofol 50µM decreased CDR to 29.4% (P < 0.05). In the TUNEL assay, cells with no drug treatment demonstrated 12.3% ACR and 50µM AMPA increased ACR to 28% (P < 0.05). Adding 50µM propofol to AMPA decreased the ACR to 20.1% (P < 0.05).

Conclusions

Propofol (50µM) had neuroprotective effects against AMPA (50µM)-induced cell death by reducing apoptosis.