摘要
Abstract
Objective To explore the mechanism of pirenoxine inhibiting ferroptosis by regulating micro RNA-29b(miR-29b)to improve oxidative stress damage to lens epithelial cells in cataract.Methods Hydrogen peroxide(H2O2)induced human lens epithelial cell B3(HLE-B3)to establish oxidative damage model in vitro.HLE-B3 cells were divided into control group(normal culture),model group(200 μmol·L-1 H2O2),pirenoxine group(53.33 μg·mL-1 pirenoxine 300 μL+200 μmol·L-1 H2O2)and NC inhibitor group(transfected NC inhibitor+200 μmol·L-1 H2O2),miR-29b inhibitor group(transfected with miR-29b inhibitor+200 μmol·L-1 H2O2),pirenoxine combined with NC inhibitor group(transfected with NC inhibitor+53.33 μg·mL-1 pirenoxine 300 μL+200 μmol·L-1H2O2)and pirenoxine combined with miR-29b inhibitor group(transfected with miR-29b inhibitor+53.33μg·mL-1 pirenoxine 300 μL+200 μmol·L-1 H2O2).The relative expression level of miR-29b in each group was detected by reverse transcription real-time fluorescence quantitative polymerase chain reaction.The activity of superoxide dismutase(SOD),catalase(CAT);the level of malondialdehyde(MDA)in cells of each group were measured by enzyme-linked immunosorbent assay.The positive expression of Ferritin in cells of each group was detected by immunofluorescence.Ferrous ion(Fe2+)assay kit was used to detect the Fe2+level in each group.Results The relative expression levels of miR-29b in model group and pirenoxine group were 1.00±0.18 and 2.54±0.51,respectively.The SOD activities of control group,model group,pirenoxine group,pirenoxine combined with NC inhibitor group,pirenoxine combined with miR-29b inhibitor group were(37.29±7.45),(12.61±2.52),(26.51±5.31),(22.37±4.47),(16.48±3.29)U·mL-1,respectively;the CAT activities were(7.42±1.48),(1.33±0.27),(5.25±1.05),(4.72±0.94)and(2.19±0.43)U·mL-1,respectively;the MDA levels were(3.15±0.63),(10.69±2.13),(6.22±1.24),(6.54±1.31)and(8.92±1.77)mmol·mL-1,respectively;the ferritin relative fluorescence intensities were 1.00±0.16,5.37±1.07,2.45±0.49,2.77±0.55 and 4.12±0.82,respectively;the Fe2+levels were(1.41±0.28),(4.76±0.95),(3.11±0.62),(3.26±0.65)and(4.33±0.85)μmol·L-1,respectively.The relative expression level of miR-29b in the pirenoxine group showed statistically significant difference compared with that in the model group(P＜0.05).There were statistically significant differences between the model group and the control group,between the pirenoxine group and the model group,and between the pirenoxine combined with miR-29b inhibitor group and the pirenoxine combined with NC inhibitor group(all P＜0.05).However,there were no statistically significant differences between the pirenoxine combined with NC inhibitor group and the pirenoxine group(all P＞0.05).Conclusion Pirenoxine may inhibit ferroptosis by regulating miR-29b,thereby ameliorating oxidative stress damage to lens epithelial cells.关键词
吡诺克辛/白内障/铁死亡/晶状体/过氧化氢Key words
cataract/pirenoxine/ferroptosis/lens/hydrogen peroxide分类
医药卫生
