Nitric oxide-induced cytotoxicity attenuation by thiopentone sodium but not pentobarbitone sodium in primary brain cultures.
1. We describe the effects of barbiturates on the neurotoxicity induced by nitric oxide (NO) on foetal rat cultured cortical and hippocampal neurones. Cessation of cerebral blood flow leads to an initiation of a neurotoxic cascade including NO and peroxynitrite. Barbiturates are often used to protect neurones against cerebrovascular disorders clinically. However, its neuroprotective mechanism remains unclear. 2. In the present experiment, we established a new in vitro model of brain injury mediated by NO with an NO-donor, 1-hydroxy-2-oxo-3-(3-aminopropyl)-3-isopropyl-1-triazene (NOC-5) on grid tissue culture wells. We also investigated the mechanisms of protection of CNS neurones from NO-induced neurotoxicity by thiopentone sodium, which contains a sulphydryl group (SH-) in the medium, and pentobarbitone sodium, which does not contain SH-. 3. Primary cultures of cortical and hippocampal neurones (prepared from 16-day gestational rat foetuses) were used after 13-14 days in culture. The cells were exposed to NOC-5 at the various concentrations for 24 h in the culture to evaluate a dose-dependent effect of NOC-5. 4. To evaluate the role of the barbiturates, neurones were exposed to 4, 40 and 400 microM of thiopentone sodium or pentobarbitone sodium with or without 30 microM NOC-5. In addition, superoxide dismutase (SOD) at 1000 u ml(-1) and 30 microM NOC-5 were co-administered for 24 h to evaluate the role of SOD. 5. Exposure to NOC-5 induced neural cell death in a dose-dependent manner in both cortical and hippocampal cultured neurones. Approximately 90% of the cultured neurones were killed by 100 microM NOC-5. 6. This NOC-5-induced neurotoxicity was significantly attenuated by high concentrations of thiopentone sodium (40 and 400 microM) as well as SOD, but not by pentobarbitone sodium. The survival rates of the cortical neurones and hippocampal neurones that were exposed to 30 microM NOC-5 were 11.2+/-4.2% and 37.2+/-3.0%, respectively, and in the presence of 400 microM thiopentone sodium, the survival rate increased to 65.3+/-3.5% in the cortical neurones and 74.6+/-2.2% in the hippocampal neurones. 7. These findings demonstrate that thiopentone sodium, which acts as a free radical scavenger, protects the CNS neurones against NO-mediated cytotoxicity in vitro. In conclusion, thiopentone sodium is one of the best of the currently available pharmacological agents for protection of neurones against intraoperative cerebral ischaemia.
Shibuta
S
S
Department of Anaesthesiology, Osaka University Medical School, Suita-city, Japan.
Kosaka
J
J
Mashimo
T
T
Fukuda
Y
Y
Yoshiya
I
I
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D016166:Free Radical Scavengers / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006993:Hypnotics and Sedatives / Q000494:pharmacology*; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D010424:Pentobarbital / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D013874:Thiopental / Q000494:pharmacology*; D014226:Triazenes / Q000633:toxicity
C114535:1-hydroxy-2-oxo-3-(3-aminopropyl)-3-isopropyl-1-triazene; D016166:Free Radical Scavengers; D006993:Hypnotics and Sedatives; D014226:Triazenes; D009569:Nitric Oxide; D010424:Pentobarbital; D013874:Thiopental