摘要
目的 观察并初步探讨地拉罗司(DFX)对人视网膜血管内皮细胞(HREC)脂质过氧化和铁死亡的影响.方法 细胞实验.将体外培养的HREC分为正常糖组(NG组)、高糖组(HG组)、正常糖+DFX组(NG+DFX组)、高糖+DFX组(HG+DFX组)、正常糖+DFX+柠檬酸铁胺(FAC)组(NG+DFX+FAC组)和高糖+DFX+FAC组(HG+DFX+FAC组).光学显微镜观察细胞形态;细胞计数试剂盒-8检测细胞增殖率;钙黄绿素乙酰甲酯试剂盒检测不稳定铁池(LIP)含量;酶联免疫吸附试验检测活性氧(ROS)、脂质过氧化物丙二醛(MDA)、谷胱甘肽(GSH)和氧化型GSH(GSSG)含量;免疫蛋白印迹法检测GSH过氧化物酶4(GPX4)和溶质载体家族7成员11(SLC7A11)的蛋白相对表达量.两组间比较采用双尾Student t检验;多组间比较采用单因素方差分析.结果 与HG组和HG+DFX+FAC组比较,HG+DFX组细胞增殖率、GSH含量及GPX4、SLC7A11蛋白相对表达量明显增高,差异均有统计学意义(F=150.70、21.02、26.09、52.62,P＜0.001);LIP、ROS、MDA、GSSG含量明显降低,差异均有统计学意义(F=807.20、16.94、31.62、19.21,P＜0.001).结论 高糖显著诱导HREC LIP含量升高、脂质过氧化和铁死亡;地拉罗司通过下调LIP水平从而抑制HREC脂质过氧化和铁死亡.
Abstract
Objective To observe and preliminarily explore the effects of Deferasirox(DFX)on lipid peroxidation and ferroptosis in human retinal endothelial cells(HREC).Methods A cell experimental study.Divided the in vitro cultured HREC into normal glucose(NG)group,high glucose(HG)group,NG+DFX group,HG+DFX group,NG+DFX+ferric ammonium citrate(FAC)group,and HG+DFX+FAC group.Light microscope was used to observe the morphology of the cells;cell proliferation was detected by Cell Counting Kit-8 assay,and Calcein-AM staining was used to detect the unstable iron pool(LIP)content;enzyme-linked immunosorbent assay reader was used to detect the reactive oxygen species(ROS),malondialdehyde(MDA),glutathione(GSH),and oxidized glutathione(GSSG);Western blot was used to detect the relative protein expression of Glutathione Peroxidase 4(GPX4)and Solute Carrier Family 7 Member 11(SLC7A11).Two-tailed Student t test was used for comparison between the two groups;one-way ANOVA was used for comparison between multiple groups.Results Compared with the HG group and the HG+DFX+FAC group,the cell proliferation rate and the contents of GSH and the relative protein expression of GPX4,and SLC7A11 in the HG+DFX group were significantly increased,and the differences were statistically significant(F=150.70,21.02,26.09,52.62;P＜0.001).The contents of LIP,ROS,MDA,and GSSG were significantly decreased,and the differences were statistically significant(F=807.20,16.94,31.62,19.21;P＜0.001).Conclusions High glucose significantly induces an increase in LIP,lipid peroxidation,and ferroptosis in HREC.Deferasirox inhibits lipid peroxidation and ferroptosis in HREC by downregulating LIP levels.
维普
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