Rutin Inhibits Hydrogen Peroxide-induced Apoptosis Through PI3K/AKT Signaling Pathway In Human Lens Epithelial Cells

ObjectiveThere is very high blindness rate in cataract patients, and scholars are generally believed that the oxidative damage and apoptosis of lens epithelial cells(LEC) is the main mechanism of cataract formation. The studies of scholars prove that hydrogen peroxide(H2O2) induce human lens epithelial cells(HLEC) apoptosis by PI3K/AKT signaling pathway; The early experimental results of this topic prove the protective effect of rutin(Ru) on H2O2-induced HLEC apoptosis. Therefore, based on the above basis, we researched the relationship between PI3K/AKT signaling pathway and the protective effect of Ru.Methods(1) Normal SRA01/04 HLEC were cultured in two 96-well plates. The cells were incubated with 200 μmol/L H2O2 at different cultured times in one plate; In the other plate, The cells were incubated with 100 μmol/L Ru at different pretreatment times, then 200 μmol/L H2O2 cultivating 24 h. MTT determined the cell survival rates of two plates.(2) The experimental cells were divided into four groups(control group, H2O2 group, rutin group, LY294002 group). Western blot measured the expression of AKT and p-AKT, in order to verify that Ru inhibits H2O2-induced cell apoptosis via PI3K/AKT signaling pathway.(3) According to the above groups, flow cytometry(FCM) was used to measure each cell apoptosis rate.Results(1) H2O2 induced apoptosis of HLEC. And with the extension of time, the cell survival rate gradually decreased. When at 24 h, the result was closest to 50% inhibition rate(IC50), so we chose 24 h as H2O2 treatment time.(2) Ru protected HLEC from apoptosis induced by H2O2, and the result was the highest in 1 h pretreatment. After 1 h, the survival rates gradually decreased. So we chose 1 h as Ru pretreatment time.(3) In control group, H2O2, rutin and LY294002 group, H2O2 group had the highest cell apoptosis rate; Rutin group was lower than in H2O2 group(P < 0.01); LY294002 group was higher than in rutin group, but there was no statistically significant difference with H2O2 group(P > 0.05).(4) In control group, H2O2, rutin and LY294002 group, the relative expression of p- AKT decreased in H2O2 and LY294002 group(p < 0.01, vs control group), the relative expression of p- AKT rised in rutin group.ConclusionRutin inhibit H2O2-induced cell apoptosis and may be associated with PI3K/AKT signaling pathway.