过氧化氢诱导的急性氧化应激对人晶状体上皮细胞形态、生存率及衰老标记蛋白30(SMP30)表达的影响

摘要

Objective To investigate the effects of acute oxidative stress induced by H2O2 on expression of senescence marker protein30 (SMP30) and morphology,survival rate of human lens epithelial cells (HLECs).Methods HLECs were treated with H2O2(0 μmol · L-1,100 μmol · L-1,200 μmol · L-1,300 μmol · L-1) for 24 hours,the acute oxidative stress models were established,the changes of cell morphology was observed,and MTT was used to analyze the cells state,the expressions of SMP30 were measured by Western blot.Results The cell density decreased,morphological changed and viability of cells significant decreased in 100 μmol · L-1 and 200 μmol ·L-1 treated group,the large and round cells appeared,the cell body stretched with unclear boundary.With the H2O2 concentration increased,the viability of cells were gradually decreased in treated group,there were statistical differences compared with 0 μmol · L-1 treated group (all P ＜ 0.05).The relative expression of SMP30 in control group and 100 μmol · L-1 and 200 μmol · L-1 treated group were 0.273 ±0.055,0.464 ± 0.058,0.442 ± 0.050,respectively.There were significant differences between 100 μmol · L-1,200 μmol · L-1 treated group and control group (all P ＜ 0.05),and there was no statistical difference between 100 μmol · L-1 and 200 μmol · L-1 treated group (P ＞ 0.05).Conclusion SMP30 is up-regulated in HLECs under acute oxidative stress induced by H2O2,the cell morphology is changed,the viability of cells is decreased,and SMP30 may be involved in the process of acute oxidative stress in HLECs.%目的 探讨过氧化氢诱导的急性氧化应激对人晶状体上皮细胞形态、生存率及衰老标记蛋白30(senescence marker protein30,SMP30)表达的影响.方法 采用不同浓度过氧化氢(0μmol·L-1、100 μmol·L-1、200 μmol·L-1、300 μmol·L-1)处理人晶状体上皮细胞24 h,建立不同浓度急性氧化应激模型,通过光镜观察、MTT分析细胞形态、生存率,Western blot检测SMP30蛋白的表达情况.结果 不同浓度过氧化氢处理细胞24 h后,100 μmol·L-1处理组和200 μmol·L-1处理组细胞密度、生长活力下降,细胞形态改变,开始出现变圆变大的细胞,胞体拉长,边界不清;300 μmol·L-1处理组细胞密度明显降低,细胞出现破碎,溃解.随着过氧化氢浓度的增高,细胞生存率逐渐下降,各处理组与对照组(Oμmol·L-)间相比,差异均有统计学意义(均为P ＜0.05).在对照组及100 μmol·L-1处理组、200 μmol·L-1处理组晶状体上皮细胞SMP30蛋白相对表达量分别为0.273±0.055、0.464±0.058、0.442±0.050,100 μmol·L-1处理组、200 μmol·L-处理组SMP30蛋白的表达均较对照组提高,差异均有统计学意义(均为P ＜0.05),100 μmol·L-1处理组和200 μmol·L-1处理组间差异无统计学意义(P＞0.05).结论 在急性氧化应激状态下人晶状体上皮细胞形态改变,生存率下降,SMP30蛋白表达上调,SMP30蛋白可能参与晶状体上皮细胞急性氧化应激损伤过程.