Ginsenoside Rg1 inhibits H2O2-induced apoptosis and autophagy in human periodontal ligament cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Our previous studies have confirmed that H2O2 and ginsenoside Rg1 can cause changes in reactive oxygen species levels in human periodontal ligament cells,but the correlation of reactive oxygen species with apoptosis and autophagy remains unclear. OBJECTIVE:To explore the effect and mechanism of ginsenoside Rg1 on H2O2-induced apoptosis and autophagy of human periodontal ligament cells. METHODS:Human periodontal ligament cells were divided into control group,H2O2 group and H2O2+Rg1 group.Ginsenoside Rg1(50 μmol/L)was pre-incubated for 24 hours and H2O2 was treated for 2 hours(500 μmol/L).CCK-8 assay was used to detect the proliferation ability of the cells.A fluorescent probe DCFH-DA was used to detect the reactive oxygen species level of the cells.qRT-PCR and western blot assay were used to detect heme oxygenase 1,apoptosis-related factor Caspase-3,Bax,anti-apoptotic factor Bcl-2,autophagy Beclin-1,P62,LC3,pathway-related factors phosphatidylinositol-3-kinase(PI3K),protein kinase B(AKT),mammalian target of rapamycin(mTOR)mRNA and protein expression. RESULTS AND CONCLUSION:(1)Compared with the control group,the proliferation activity of human periodontal ligament cells in the H2O2 group decreased.Compared with the H2O2 group,the proliferation activity of human periodontal ligament cells increased in the H2O2+Rg1 group.(2)Compared with the control group,the expression of reactive oxygen species and heme oxygenase-1 increased in the H2O2 group.Compared with the H2O2 group,the expression of reactive oxygen species and heme oxygenase-1 decreased in the H2O2+Rg1 group.(3)Compared with the control group,the expression of Caspase-3,Bax,Beclin-1 and LC3 increased,while the expression of Bcl-2 and P62 decreased in human periodontal ligament cells of the H2O2 group.Compared with the H2O2 group,the expressions of Caspase-3,Bax,Beclin-1 and LC3 decreased,and the expressions of Bcl-2 and P62 increased in the H2O2+Rg1 group.(4)Compared with the control group,the expressions of PI3K,AKT and mTOR decreased in human periodontal ligament cells of the H2O2 group.Compared with the H2O2 group,the expressions of PI3K,AKT and mTOR increased in human periodontal ligament cells of the H2O2+Rg1 group.(5)These results suggest that ginsenoside Rg1 can inhibit H2O2-induced apoptosis and autophagy in human periodontal ligament cells by reducing the content of reactive oxygen species and down-regulating the related factor expression of the PI3K/AKT/mTOR pathway.