Novel quantitative method for evaluating oxygen/glucose deprivation-induced injury of hippocampal slices
- Author:
Le-Qin YAN
1
;
Er-Qing WEI
;
Hai-Tao HU
;
Wei-Ping ZHANG
;
Meng-Ling WANG
;
Jian-Zong SHEN
Author Information
1. College of Medical Sciences, Zhejiang University, Hangzhou 310031, China.
- Publication Type:Journal Article
- From:
Journal of Zhejiang University. Medical sciences
2002;31(2):81-85
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE: To establish a simple, sensitive in vitro method to evaluate oxygen/glucose deprivation (OGD)-induced injury of brain hippocampal slices in rats. METHODS: Rat hippocampal slices were incubated in 2% 2, 3, 5-triphenyltetrazolium chloride (TTC) solution after oxygen/glucose deprivation. They were then soaked in a measured volume of ethanol and dimethylsulfoxide (50:50) to extract the TTC formazan Product which was then measured by spectrophotometry. OGD induced LDH release was simultaneously measured. RESULTS: Progressive prolongation of OGD induced hippocampal injury resulted in decreased formazan coloration as determined by spectrophotometry. There was a parallel increase in LDH release, thus a negative correlation in these two products was noted. (r=-0.933,P <0.01). The injury was attenuated in the brain slices pre-treated with nimodipine, dexamethasone, and ketamine, but not ONO-1078. CONCLUSION: Solvent extraction and spectrophotometric quantification of formazan represents an objective measurement of OGD-induced injury of rat hippocampal slices.
