[关键词]
[摘要]
目的:研究锌对半乳糖诱导的人晶状体上皮细胞(human lens epithelial cells,HLEC)凋亡的影响。
方法:将培养的SRA01/04细胞系分为六组,对照组、半乳糖处理组、补锌组、补锌联合半乳糖处理组、缺锌组、缺锌联合半乳糖处理组。采用MTT法检测半乳糖对HLEC存活率的影响,荧光显微镜和流式细胞仪分别检测各实验分组细胞核形态和细胞凋亡率水平的变化。
结果:不同浓度半乳糖(0、25、50、75、100、125mmol/L)处理HLEC细胞后,细胞存活率分别为(100.0±5.4)%、(97.5±3.2)%、(91.3±5.3)%、(93.4±0.6)%、(86.6±1.4)%和(83.5±0.4)%,与未处理细胞相比,半乳糖浓度为100、125mmol/L时,细胞存活率显著下降,差异有统计学意义(P<0.05)。荧光显微镜结果显示对照组和补锌组的细胞核呈现均一的染色,半乳糖处理组、补锌联合半乳糖处理组、缺锌组和缺锌联合半乳糖处理组中一些细胞的细胞核收缩,表现出典型的细胞凋亡形态。各组的细胞凋亡率分别为(1.5±0.1)%、(7.1±0.2)%、(1.4±0.1)%、(4.4±0.2)%、(5.5±0.2)%和(15.8±0.3)%。与对照组相比,半乳糖处理组、补锌联合半乳糖处理组、缺锌组和缺锌联合半乳糖处理组的细胞凋亡率显著增加,差异有统计学意义(P < 0.05)。与半乳糖处理组相比,补锌联合半乳糖处理组的细胞凋亡率显著下降,差异有统计学意义(P<0.05),缺锌联合半乳糖处理组的细胞凋亡率则显著升高,差异有统计学意义(P<0.05)。
结论:补锌可以保护人晶状体上皮细胞抵抗由半乳糖引起的细胞凋亡,可能对白内障的形成有抑制作用。
[Key word]
[Abstract]
AIM:To study the effect of zinc on galactose-induced cell apoptosis in human lens epithelial cells(HLEC).
METHODS:HLEC cell line SRA01/04 cells were cultured in DMEM medium and divided into six groups: control group, galactose treatment group, zinc supplement group, zinc supplementation combined with galactose treatment group, zinc deficiency group, zinc deficiency combined with galactose treatment group. The cell viabilities were assayed by MTT, cell morphology and apoptosis were detected by fluorescence microscope and flow cytometry, respectively.
RESULTS:The cell viabilities induced by galactose(0, 25, 50, 75, 100, 125mmol/L)were(100.0±5.4)%,(97.5±3.2)%,(91.3±5.3)%,(93.4±0.6)%,(86.6±1.4)% and(83.5±0.4)%, respectively. When the concentration of galactose was 100 and 125mmol/L, cell viability was significantly decreased, compared with the untreated cells(P<0.05). Fluorescence microscopy results showed that the cell nucleus remained uniformly stained in control group and zinc supplement group. Nuclear shrinkage, a typical apoptotic morphology, was visible in some cells in the galactose treatment group, zinc supplementation combined with galactose treatment group, zinc deficiency group and zinc deficiency combined with galactose treatment group. The cell apoptosis in the six groups were(1.5±0.1)%,(7.1±0.2)%,(1.4±0.1)%,(4.4±0.2)%,(5.5±0.2)% and(15.8±0.3)%, respectively. The cell apoptosis were significant increased in galactose treatment group, zinc supplementation combined with galactose treatment group, zinc deficiency group and zinc deficiency combined with galactose treatment group, compared with the control group(P<0.05), and those of zinc supplementation combined with galactose treatment group were significant decreased, and zinc deficiency combined with galactose treatment group were increased(P<0.05), compared with the galactose treatment group.
CONCLUSION:Zinc supplementation protects human lens epithelial cells against apoptosis induced by galactose and may have an inhibition effect on cataract formation.
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[基金项目]
国家自然科学基金(No.21561006,21867007); 贵州省科学技术基金(No.黔科合LH字\〖2016\〗7372)