Protective Effect of Ginsenoside Rg3 on Lipopolysaccharide-Induced Neuronal-Galial Interaction Injury Model
Objective To investigate the protective effect of ginsenoside Rg3 on lipopolysaccharide-induced galial-neuronal interaction injury model.Methods Primary microglia cells and HT-22 cell lines were cultured,and the cells were divided into the control group(CON),the 100 ng/mL LPS group(LPS),the control inflammation model group(CM),the ginsenoside Rg3 group,and the inflammation model plus ginsenoside Rg3 treatment group(CM+Rg3).Ginsenoside Rg3 was administered in the ginsenoside Rg3 group and the CM+Rg3 group at the doses of 2.5,5,10,and 20 μmol/L.Galial-neuronal interaction modeling was performed in the CM group and the CM+Rg3 group.The purity of the primary microglia cells was assessed by immunofluorescence,the viability of the HT-22 cells in each group was assessed by CCK-8,and changes in the levels of inflammatory cytokines,including interleukin(IL)-1β,IL-6,tumor necrosis factor α(TNF-α),and IL-10,in the cell samples of each group were assessed with the ELISA kits.The level of cellular oxidative damage was measured with a ROS kit,and the expression of apoptosis-related proteins,including Bax and Bcl-2,was assessed by Western blot.The activity of Caspase-3 enzyme in each group was measured with a Caspase-3 enzyme activity kit.Results The purity of the microglia cultured reached over 95％and was suitable for subsequent experiments.After ginsenoside Rg3 stimulation at different doses,the survival rate of HT-22 was not much different from that of the CON group.The survival rate of neurons after 100 ng/mL LPS stimulation was 98％,indicating that Rg3 and LPS had no effect on the survival of neurons.Compared with that of the CON group,the survival rate of HT-22 cells in the CM group was significantly decreased(P＜0.01).Compared with the CM group,the CM+Rg3 group showed a significant increase in the viability of neurons(P＜0.0l),indicating that the glia-neuron interaction model was successfully constructed.When Rg3 dose was 10 μmol/L,the HT-22 cells in CM+Rg3 group showed the highest viability(P＜0.05).Hence,10 μmol/L Rg3 was selected for further experiments.After 100 ng/mL LPS stimulation,the concentrations of IL-1 β,IL-6,TNF-α,and IL-10 in HT-22 cells were not significantly different from those in the CON group.After 100 ng/mL LPS stimulation,the concentrations of IL-lβ and TNF-α in microglia were higher than those in the CON group(P＜0.05),but the concentrations of IL-1 β and TNF-α in the LPS+Rg3 group were lower than those in the LPS group(P＜0.05).The concentration of reactive oxygen species in the CM+Rg3 group was slightly higher than that in the CON group,and significantly lower than that in the CM group(P＜0.01).The expression of Bax in the CM+Rg3 group was higher than that in the CON group,and lower than that in the CM group.The expression of Bcl-2 was lower in the CON group,and higher than that in the CM group(P＜0.01).The Caspase-3 enzyme activity in the CM group was significantly higher than that in the CON group(P＜0.01).The Caspase-3 enzyme activity in the CM+Rg3 group was significantly lower than that in the CM group(P＜0.01).Conclusion Ginsenoside Rg3 may play a role in the alleviation of neuronal apoptosis by regulating glial cell damage,reducing the secretion of inflammatory factors,and inhibiting neuronal apoptosis.
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