RESUMEN
Objective:To discuss the protective effect of gingerone on the hippocampal neuron HT22 cells after oxygen-glucose deprivation/reoxygenation(OGD/R),and to clarify the related mechanism.Methods:The HT22 cells were cultured,and the OGD/R cell injury model was established by setting the gradient of OGD/R time.The HT22 cells were divided into control group,OGD/R group,OGD/R+ 1 μmol·L-1 gingerone group,OGD/R + 10 μmol·L-1 gingerone group,OGD/R+100 μmol·L-1 gingerone group,and OGD/R+0.2%dimethyl sulfoxide(DMSO)group.The viability of the cells in various groups was detected by CCK-8 assay;the survival rates of the cells in various groups were calculated to determine the optimal drug concentration of gingerone.The cells were divided into control,OGD/R group,OGD/R+ gingerone,and OGD/R+gingerone+nuclear factor erythroid-2-related factor 2(Nrf2)inhibitor(ML385)groups.The cells in OGD/R + gingerone group were treated with gingerone for 4 h before OGD treatment for 8 h followed by reoxygenation for 8 h,and the cells in OGD/R+gingerone+ ML385 group were treated with 10 μmol·L-1 ML385 for 6 h before gingerone treatment.The viability of the cells in various groups was detected by CCK-8 assay;the expression levels of Nrf2,heme oxygenase-1(HO-1),B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)proteins in the cells in various groups were detected by Western blotting method;the activity of superoxide dismutase(SOD)and the level of malondialdehyde(MDA)in the cell culture supernatant in various groups were detected by enzyme-linked immunosorbent assay(ELISA)method.Results:Compared with control group,the survival rate of the HT22 cells was below 50%after treated with OGD for 8 h and reoxygenation for 8 h,so the HT22 cell OGD/R model was established by treated with OGD for 8 h and reoxygenation for 8 h.Compared with OGD/R group,the survival rates of the cells in OGD/R+different doses of gingerone groups were increased to various extents,and the survival rate of the cells in OGD/R+ 100 μmol·L-1 gingerone group was significantly increased(P<0.01);so 100 μmol·L-1 gingerone was used for the subsequent experiment.Compared with control group,the viability of the cells in OGD/R group was significantly decreased(P<0.01),and the expression levels of Nrf2,HO-1,and Bax proteins in the cells were significantly increased(P<0.01),while the expression level of Bcl-2 protein in the cells was significantly decreased(P<0.05),and the SOD activity in the cell culture supernatant was significantly decreased(P<0.01),and the level of MDA was significantly increased(P<0.01);compared with OGD/R group,the viability of the cells in OGD/R + gingerone group was significantly increased(P<0.01),and the expression levels of Nrf2,HO-1,and Bcl-2 proteins in the cells were significantly increased(P<0.05 or P<0.01),while the expression level of Bax protein in the cells was decreased(P<0.05),the SOD activity in the cell culture supernatant was significantly increased(P<0.01),and the level of MDA was significantly decreased(P<0.01);compared with OGD/R + gingerone group,the viability of the cells in OGD/R + gingerone + ML385 group was significantly decreased(P<0.01),and the expression levels of Nrf2,HO-1,and Bcl-2 proteins were significantly decreased(P<0.01),while the expression level of Bax protein in the cells was significantly increased(P<0.01),the SOD activity in the cell culture supernatant was significantly decreased(P<0.01),and the level of MDA was significantly increased(P<0.05).Conclusion:Gingerone alleviates the oxidative stress damage,and thereby plays an inhibiory effect on the apoptosis of the HT22 neurons by activating the Nrf2/HO-1 signaling pathway after OGD/R.