Effects of olfactory ensheathing cells on hydrogen peroxide-induced apoptosis in cultured dorsal root ganglion neurons.
- Author:
Xiao-dong YU
1
;
Zhuo-jing LUO
;
Lin ZHANG
;
Kai GONG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Apoptosis; drug effects; Cell Survival; drug effects; Cells, Cultured; Coculture Techniques; Ganglia, Spinal; cytology; drug effects; Hydrogen Peroxide; toxicity; Male; Olfactory Bulb; cytology; physiology; Rats; Rats, Sprague-Dawley; Time Factors
- From: Chinese Medical Journal 2007;120(16):1438-1443
- CountryChina
- Language:English
-
Abstract:
BACKGROUNDOlfactory ensheathing cells (OECs) can promote many kinds of neuron growth and axonal extension. The aim of the study was to investigate the effects of co-culturing with OECs on neuron apoptosis in vitro.
METHODSApoptosis was induced by treatment of cultured dorsal root ganglion neurons with 1 mmol/L hydrogen peroxide (H(2)O(2)). Cells were randomly arranged into the following treatment groups. In group 1, OECs at different density (10(4)/ml to 8 x 10(5)/ml) were added immediately after H(2)O(2) treatment and cells were co-cultured for 24 hours. In group 2, OECs were added at different time points (0, 4, 8, 12 and 24 hours) after H(2)O(2) treatment. Apoptotic cell death was determined by Hoechst 33258 staining and flow cytometry (FCM). Cell viability was determined by using methyl thiazoleterazolium (MTT) assays.
RESULTSThe results showed in the Hoechest 33258 staining, FCM and MTT that OECs have both the density-dependent protection and time-dependent protection on neuron apoptosis. The apoptosis decreased and the dorsal root ganglion neuron viability increased, when the density of OECs was increased in co-culture groups. But further increasing OEC density above 2 x 10(5)/ml (i.e. 8 x 10(5)/ml) failed to exert additional protection. As the interval between adding H(2)O(2) and adding OECs was increased, the amounts of apoptosis cells were also increased. When OECs were added 24 hours after H(2)O(2), no significant protection was observed.
CONCLUSIONThese results indicated that OECs could protect dorsal root ganglion neurons from apoptosis induced by H(2)O(2) in a density- and time-dependent manner.