芦丁对氧化损伤环境下晶状体透明度的影响及其作用机制OA

Objective To investigate the effect of rutin on the transparency of rat lens models cultured in vitro under oxidative stress and its underlying mechanism.Methods In this study,the intact binocular lenses of 60 8-week-old SD rats were taken.After excluding the lenses with turbidity caused by trauma,10 lenses were selected for intervention with 50μmol·L-1 rutin as the rutin group,which was only used to observe the changes in the transparency of the lenses.The re-maining lenses were randomly divided into 3 groups for subsequent experiments:the control group,the hydrogen peroxide(H2O2)group,and the H2O2+rutin group.The opacity of the lenses in each group was observed under a dark-field micro-scope and then statistically analyzed using ImageJ software.The expressions of endoplasmic reticulum stress proteins like PERK,IRE1α,and ATF6 in lenses were detected by Western blot.The content of reactive oxygen species(ROS)in lenses was measured using immunofluorescence staining,while the content of calcium ions and malondialdehyde(MDA)in lenses was measured using the calcium ion and lipid oxidative damage detection kit.Results Microscopic observations revealed that the lens opacity time of rats in the rutin group was delayed to 9 days compared with the control group(7 days).After 5 days of culture in vitro,the lens opacity of rats in the H2O2 group(6.89±0.15)significantly increased compared with that in the control group(1.03±0.07)(P＜0.001);compared with the H2O2 group,the lens opacity of rats in the H2O2+rutin group(2.52±0.15)significantly decreased(P＜0.001).Immunofluorescence staining showed that the relative fluorescence intensity of ROS in rat lenses in the H2O2+rutin group(4.35±0.11)significantly decreased compared to the H2O2 group(8.53±0.44),and the difference was statistically significant(P＜0.001).Compared with the H2O2 group,the relative ex-pression levels of endoplasmic reticulum stress-related proteins PERK,IRE1a,and ATF6 in rat lenses in the H2O2+rutin group were significantly reduced,with all differences being statistically significant(all P＜0.001).The results from the cal-cium ion and lipid oxidative damage assays showed that the calcium ion content(0.09±0.01)mmol·L-1 and MDA con-tent(6.82±0.42)μmol·L-1 in rat lenses in the H2O2+rutin group were remarkably lower than those in the H2O2 group,which had a calcium ion content of(0.24±0.03)mmol·L-1 and an MDA content of(10.63±1.13)μmol·L-1,and the differences were both statistically significant(both P＜0.01).Conclusion Under the oxidative damage induced by H2O2,rutin protects the normal function of the endoplasmic reticulum in rat lenses by inhibiting endoplasmic reticulum stress,thereby delaying the occurrence of lens opacity.