To study the effects of Ramulus et Uncus Uncariae (REUU) on oxygen free radical-mediated damage by hydrogen peroxide (H-2O-2) on cultured spinal sensory neurons, in vitro assays such as MTT assay, NR assay, neurofilament enzymeimmuno assay (EIA), sulforhodamine B (SRB) assay, assay for lactate dehydrogenase (LDH) activity and assay for lipid peroxidation were used in cultured spinal dorsal root ganglion neurons derived from mice. Spinal dorsal root ganglion neurons were cultured in media containing various concentrations of H-2O-2 for 5 hours, after which the neurotoxic effect of H-2O-2 was measured by in vitro assay. The protective effect of the herb extract, Ramulus et Uncus Uncariae (REUU) against H2O2-induced neurotoxicity was also examined. The results are as follows.
1. In NR assay and MTT assay, H-2O-2 sigruficantly decreased the cell viability of cultured mouse spinal dorsal root ganglion neurons according to exposure concentration in these cultures. An additional time course study was done on these cultures.
2. Cultured spinal dorsal root ganglion neurons which were exposed to various concentrations of H-2O-2 showed a quantitative decrease of neuronal cells by EIA and of total protein by sulforhodamine B (SRB) assay, while they showed an increase of both lipid peroxidation and LDH activity.
3. The effect of Ramulus et Uncus Uncariae (REUU) on H-2O-2 induced neurotoxicity showed a quantitative increase in both neurofilament and total protein, but showed a decrease of lipid peroxidation and LDH activity.
These results suggest that H-2O-2 has a neurotoxic effect on cultured spinal dorsal root ganglion neurons from mice and that the herb extract, Ramulus et Uncus Uncariae (REUU), was very effective in protecting H-2O-2 induced neurotoxicity by decreasing lipid peroxidation and LDH activity.
“This work was supported by the Brain Korea 21 Project.”
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