飞燕草素对光诱导小鼠视网膜感光细胞661W铁过载的调控作用  

Regulatory effect of delphinidin on iron overload in light-induced retinal photoreceptor cells 661W of mice

作　　者：傅晓颖 邓晓敏 陈倩云 彭佳媛 杜旌畅 朱彦锋[1] 余小平[1] FU Xiaoying;DENG Xiaomin;CHEN Qianyun;PENG Jiayuan;DU Jingchang;ZHU Yanfeng;YU Xiaoping(Chengdu Medical College,Chengdu 610500,Sichuan Province,China;Department of Pathology,Leshan People’s Hospital,Leshan 614000,Sichuan Province,China)

机构地区：[1]成都医学院,四川省成都市610500 [2]乐山市人民医院病理科,四川省乐山市614000

出　　处：《眼科新进展》2021年第7期615-620,共6页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助项目(编号81773432,82073539)。

摘　　要：目的探究飞燕草素(Delphinidin)对光诱导661W细胞铁过载的调控作用及机制。方法将小鼠视网膜感光细胞661W分为Control组、Light组、Light+铁螯合剂(DFP)组、Light+Delphinidin组、Control+Delphinidin组。以(2000±200)lx白色荧光持续照射661W细胞48 h建立光损伤模型,采用CCK-8法筛选Delphinidin和DFP的最佳作用浓度。利用CCK-8法测定各组细胞活力,Hoechst-PI荧光观察各组细胞的细胞膜受损程度,流式细胞仪检测各组细胞铁含量及脂质过氧化物水平,Western blot检测各组细胞铁代谢相关蛋白表达水平。结果CCK-8法检测结果显示,5μmol·L^(-1) Delphinidin和20μmol·L^(-1) DFP为最佳作用浓度。与Control组相比,Light组661W细胞损伤严重,细胞活力下降为(56.69±1.48)%,铁含量增加,脂质过氧化物水平升高,差异均有统计学意义(均为P<0.05);与Light组相比,Light+DFP组和Light+Delphinidin组661W细胞损伤减轻,细胞活力分别升高为(62.85±0.46)%和(63.41±0.68)%,铁含量减少,脂质过氧化物水平降低,差异均有统计学意义(均为P<0.05)。Western blot检测结果显示,与Control组相比,Light组细胞二价金属离子转运体1(DMT1)、转铁蛋白受体1(TfR1)、铁蛋白轻链(FTL)、铁蛋白重链1(FTH1)表达水平均升高,膜铁转运蛋白(Fpn)表达水平降低,差异均有统计学意义(均为P<0.05);与Light组相比,Light+DFP组和Light+Delphinidin组细胞TfR1、Fpn蛋白表达水平均升高,DMT1、FTL和FTH1蛋白表达水平均降低,差异均有统计学意义(均为P<0.05)。结论Delphinidin可通过调节铁代谢相关蛋白表达,缓解光诱导的661W细胞铁过载,进而抑制脂质过氧化反应,减少视网膜氧化损伤。Objective To explore the regulatory effect and mechanism of Delphinidin on light-induced iron overload in 661W cells.Methods Mouse retinal photoreceptor cells 661W were divided into Control group,Light group,Light+iron chelator(DFP)group,Light+Delphinidin group,Control+Delphinidin group.The light damage model was established with(2000±200)lx white fluorescence for continuous irradiation of 661W cells at 48 h,and the optimal concentrations of Delphinidin and DFP were screened by CCK-8 methods.The cell viability of each group was measured by the CCK-8 methods,the cell membrane damage degree of each group was observed by Hoechst-PI fluorescence,the iron content and lipid peroxide level of each group were detected by flow cytometry and the expression levels of iron metabolism-related proteins in each group were analyzed by Western blot.Results The results of CCK-8 methods showed that 5μmol·L^(-1) Delphinidin and 20μmol·L^(-1) DFP were the optimal drug concentrations.Compared with the Control group,the 661W cells in the Light group were seriously damaged,cell viability decreased to(56.69±1.48)%,iron content increased,lipid peroxide level increased,and the differences were statistically significant(all P<0.05);compared with the Light group,cell damage in the Light+DFP group and the Light+Delphinidin group was alleviated,cell viability increased to(62.85±0.46)%and(63.41±0.68)%,respectively,iron content decreased,lipid peroxides level decreased,and the differences were statistically significant(all P<0.05).Western blot results showed that compared with the Control group,the expression levels of divalent metal ion transporter 1(DMT1),transferrin receptor 1(TfR1),ferritin light chain(FTL)and ferritin heavy chain1(FTH1)were increased in the Light group,while the expression level of ferroportin(Fpn)protein decreased,and the differences were all statistically significant(all P<0.05);compared with the Light group,the protein expression levels of TfR1 and Fpn were increased in the Light+DFP group and the Light+Delphinid

关 键 词：飞燕草素 光化学损伤 氧化应激 铁过载 

分 类 号：R774[医药卫生—眼科]