Abstract: ObjectiveTo observe the protective effect of tetrahydroxystilbene glucoside (TSG) on rats' hippocampal neuronal damage induced by glutamate (Glu) in the culture.MethodsHippocampus was isolated from newborn SD rats and dispersedly cultured in the medium for 9 days. Neurons were incubated with TSG (5—100μmol/L) for 24h, the cells were washed twice with Lock's solution without Mg²⁺,then Glu 500 μmol/L was added. Thirty min later, the reaction was terminated by washing the monolayer cells twice with the Lock's solution and then cultures were kept at 37℃ for 24h. Cell viability was measured by MTT method and cell membrane damage was determined by LDH leakage; with Fluo-3/AM as an intracellular calcium indicator and added into the bathing medium, fluorescent intensity of intracellular free calcium were observed through laser scanning confocal microscopy (LSCM).ResultsAfter the treatment with 5—100μmol/L TSG for 24h, the decrease of cell viability and the increase of LDH leakage caused by Glu was obviously resisted dose dependently. TSG inhibited increase of Ca²⁺ in cytoplasm, compared with model group.ConclusionTSG can significantly promote the cell viability and reduce the cell membrane damage in Glu treating hippocampal neurons. The neuroprotective activities of TGS is mediated by inhibiting Ca²⁺ overload in cytoplasm.

Key words: tetrahydroxystilbene glucoside, glutamate (Glu), hippocampal neuron, laser scanning confocal microscopy