ABSTRACT
BACKGROUND: Propofol and ketamine are have been known to have neuroprotective effects. However, the effect of combined therapy with these 2 drugs is not well known with in vitro model. This study was conducted to determine whether combined administration of propofol and ketamine could have additive effects in protecting cortical neurons from the oxygen-glucose deprivation (ischemia) - reoxygenation (reperfusion) injury. METHODS: Thirteen-day-old primary mixed cortical cultures were exposed to a 5-min combined oxygen-glucose deprivation (OGD, in vitro ischemia model), followed by 2 hr of reperfusion. Propofol (1, 10, 25, 50, 100micrometer) and ketamine (1, 2.5, 5, 10, 50micrometer) were added as alone or combination from the initiation of the OGD injury to the end of the reperfusion periods. The survived cells were counted using trypan-blue staining. The data were converted to the cell death rate. Statistical analysis was done by oneway-ANOVA tests and Bonferroni's test. P < 0.05 was considered as statistically significant. RESULTS: OGD-reperfusion demonstrated about a 70% cell death rate. 5-50micrometer of ketamine decreased the cell death rate compared with the no drug treated group (P < 0.05). 10-100micrometer of propofol decreased the cell death rate compared with the no drug treated group (P < 0.05). Combined administration of ketamine 2.5micrometer + propofol 50, 100micrometer, ketamine 10micrometer + propofol 100micrometer and propofol 1, 10micrometer + ketamine 5, 10micrometer decreased cell death rate compared with the same dosage of propofol or ketamine alone treated group (P < 0.05). CONCLUSIONS: Propofol or ketamine demonstrated neuroprotective effects. And, combined administration ofpropofol and ketamine demonstrated additive neuroprotective effects against OGD-reperfusion injury.
Subject(s)
Animals , Rats , Cell Death , Ischemia , Ketamine , Neurons , Neuroprotective Agents , Propofol , Reperfusion , Reperfusion InjuryABSTRACT
BACKGROUND: The pattern of alpha-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 (50micrometer), AMPA (50micrometer) plus propofol (0.1, 1, 25, 50micrometer), AMPA (50micrometer) plus DMSO, propofol (50micrometer) 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 (50micrometer) on AMPA (50micrometer)-induced apoptosis, a triple stain was done. In a fixed field (x400), 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 (50micrometer) stimulation demonstrated 49.3% CDR, and adding propofol 50micrometer decreased CDR to 29.4% (P < 0.05). In the TUNEL assay, cells with no drug treatment demonstrated 12.3% ACR and 50micrometer AMPA increased ACR to 28% (P < 0.05). Adding 50micrometer propofol to AMPA decreased the ACR to 20.1% (P < 0.05). CONCLUSIONS: Propofol (50micrometer) had neuroprotective effects against AMPA (50micrometer)-induced cell death by reducing apoptosis.
Subject(s)
Animals , Rats , alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid , Apoptosis , Cell Death , Deoxycytidine , Dimethyl Sulfoxide , Diminazene , DNA Nucleotidylexotransferase , In Situ Nick-End Labeling , Neurons , Neuroprotective Agents , Propofol , Trypan BlueABSTRACT
BACKGROUND: The pattern of alpha-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 (50micrometer), AMPA (50micrometer) plus propofol (0.1, 1, 25, 50micrometer), AMPA (50micrometer) plus DMSO, propofol (50micrometer) 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 (50micrometer) on AMPA (50micrometer)-induced apoptosis, a triple stain was done. In a fixed field (x400), 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 (50micrometer) stimulation demonstrated 49.3% CDR, and adding propofol 50micrometer decreased CDR to 29.4% (P < 0.05). In the TUNEL assay, cells with no drug treatment demonstrated 12.3% ACR and 50micrometer AMPA increased ACR to 28% (P < 0.05). Adding 50micrometer propofol to AMPA decreased the ACR to 20.1% (P < 0.05). CONCLUSIONS: Propofol (50micrometer) had neuroprotective effects against AMPA (50micrometer)-induced cell death by reducing apoptosis.