锌对半乳糖诱导的人晶状体上皮细胞凋亡的影响
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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。
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( 0,25,50,75,100,125 mmol/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 125 mmol/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 galactosetreatment 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.
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( 0,25,50,75,100,125 mmol/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 125 mmol/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 galactosetreatment 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.
引文
1 Verhoeven VJM,Wong KT,Buitendijk GHS,et al.Visual consequences of refractive errors in the general population.Ophthalmology 2015;122(1):101-109
2 Chou CF,Cotch MF,Vitale S,et al.Age-related eye diseases and visual impairment among U.S.adults.Am J Prev Med 2013;45(1):29-35
3娄尚,袁兆康.我国老年性白内障流行病学的调查研究.南昌大学学报(医学版)2012;52(6):98-101
4 Resnikoff S,Pascolini D,Etya'ale D,et al.Global data on visual impairment in the year 2002.Bull World Health Organ 2004;82(11):844-851
5 Wu DM,Lu J,Zheng YL,et al.Purple sweet potato color repairs Dgalactoseinduced spatial learning and memory impairment by regulating the expression of synaptic proteins.Neurobiol Learn Mem 2008;90(1):19-27
6 Devi VG,Rooban BN,Sasikala V,et al.Isorhamnetin-3-glucoside alleviates oxidative stress and opacification in selenite cataract in vitro.Toxicol In Vitro 2010;24(6):1662-1669
7 Agarwal R,Lezhitsa I.Effects of magnesium taurate on the onset and progression of D-galactose-induced experimental cataract:In vivo and in vitro evaluation.Exp Eye Res 2013;110:35-43
8 Dai J,Liu H,Zhou J,et al.Selenoprotein R protects human lens epithelial cells against D-galactose-induced apoptosis by regulating oxidative stress and endoplasmic reticulum stress.Int J Mol Sci 2016;17(2):231
9 Behndig A,Karlsson K,Reaume AG,et al.In vitro photochemical cataract in mice lacking copper-zinc superoxide dismutase.Free Radic Biol Med 2001;31(6):738-744
10贾义,宋萍萍,周静,等.锌对3-吗啉-斯得酮亚胺(SIN-1)诱导的人晶状体上皮细胞Cu/Zn-SOD表达及活性的影响.眼科新进展2017;37(1):24-27
11 Long AC,Colitz CM,Bomser JA.Apoptotic and necrotic mechanisms of stress-induced human lens epithelial cell death.Soc Exp Biol Med2004;229(10):1072-1080
12 Li WC,Kuszak JR,Dunn K,et al.Lens epithelial cell apoptosis appears to be a common cellular basis for non-congenital cataract development in humans and animals.J Chem Biol 1995;130(1):169-181
13 Manikandan R,Arumugam M.Anticataractogenic effect of hesperidin in galactose-induced cataractogenesis in Wistar rats.World J Ophthalmol2016;6(1):1-9
14 Iezhitsa I,Agarwal R,Saad SDB,et al.Mechanism of the anticataract effect of liposomal MgT in galactose-fed rats.Mol Vis 2016;22:734-747
15 Dai J,Zhou J,Liu H,et al.Selenite and ebselen supplementation attenuates D-galactose-induced oxidative stress and increases expression of SELR and SEP15 in rat lens.J Biol Inorg Chem 2016;21(8):1037-1046
16 Kumar SD,Vijaya M,Samy RP,et al.Zinc supplementation prevents cardiomyocyte apoptosis and congenital heart defects in embryos of diabetic mice.Free Radic Biol Med 2012;53(8):1595-1606
17 Prasad AS,Beck FWJ,Bao B,et al.Zinc supplementation decreases incidence of infections in the elderly:effect of zinc on generation of cytokines and oxidative stress.Am J Clin Nutr 2007;85(3):837-844
18 Du Y,Guo D,Wu Q,et al.Zinc chloride inhibits human lens epithelial cell migration and proliferation involved in TGF-β1 and TNF-αsignaling pathways in HLE B-3 cells.Biol Trace Elem Res 2014;159(1-3):425-433
19 Grahn BH,Paterson PG,Gottschall-Pass KT,et al.Zinc and the eye.J Am Coll Nutr 2001;20(2):106-118
20 Shukla N,Moitra JK,Trivedi RC.Determination of lead,zinc,potassium,calcium,copper and sodium in human cataract lenses.Sci Total Environ 1996;181(2):161-165
2 Chou CF,Cotch MF,Vitale S,et al.Age-related eye diseases and visual impairment among U.S.adults.Am J Prev Med 2013;45(1):29-35
3娄尚,袁兆康.我国老年性白内障流行病学的调查研究.南昌大学学报(医学版)2012;52(6):98-101
4 Resnikoff S,Pascolini D,Etya'ale D,et al.Global data on visual impairment in the year 2002.Bull World Health Organ 2004;82(11):844-851
5 Wu DM,Lu J,Zheng YL,et al.Purple sweet potato color repairs Dgalactoseinduced spatial learning and memory impairment by regulating the expression of synaptic proteins.Neurobiol Learn Mem 2008;90(1):19-27
6 Devi VG,Rooban BN,Sasikala V,et al.Isorhamnetin-3-glucoside alleviates oxidative stress and opacification in selenite cataract in vitro.Toxicol In Vitro 2010;24(6):1662-1669
7 Agarwal R,Lezhitsa I.Effects of magnesium taurate on the onset and progression of D-galactose-induced experimental cataract:In vivo and in vitro evaluation.Exp Eye Res 2013;110:35-43
8 Dai J,Liu H,Zhou J,et al.Selenoprotein R protects human lens epithelial cells against D-galactose-induced apoptosis by regulating oxidative stress and endoplasmic reticulum stress.Int J Mol Sci 2016;17(2):231
9 Behndig A,Karlsson K,Reaume AG,et al.In vitro photochemical cataract in mice lacking copper-zinc superoxide dismutase.Free Radic Biol Med 2001;31(6):738-744
10贾义,宋萍萍,周静,等.锌对3-吗啉-斯得酮亚胺(SIN-1)诱导的人晶状体上皮细胞Cu/Zn-SOD表达及活性的影响.眼科新进展2017;37(1):24-27
11 Long AC,Colitz CM,Bomser JA.Apoptotic and necrotic mechanisms of stress-induced human lens epithelial cell death.Soc Exp Biol Med2004;229(10):1072-1080
12 Li WC,Kuszak JR,Dunn K,et al.Lens epithelial cell apoptosis appears to be a common cellular basis for non-congenital cataract development in humans and animals.J Chem Biol 1995;130(1):169-181
13 Manikandan R,Arumugam M.Anticataractogenic effect of hesperidin in galactose-induced cataractogenesis in Wistar rats.World J Ophthalmol2016;6(1):1-9
14 Iezhitsa I,Agarwal R,Saad SDB,et al.Mechanism of the anticataract effect of liposomal MgT in galactose-fed rats.Mol Vis 2016;22:734-747
15 Dai J,Zhou J,Liu H,et al.Selenite and ebselen supplementation attenuates D-galactose-induced oxidative stress and increases expression of SELR and SEP15 in rat lens.J Biol Inorg Chem 2016;21(8):1037-1046
16 Kumar SD,Vijaya M,Samy RP,et al.Zinc supplementation prevents cardiomyocyte apoptosis and congenital heart defects in embryos of diabetic mice.Free Radic Biol Med 2012;53(8):1595-1606
17 Prasad AS,Beck FWJ,Bao B,et al.Zinc supplementation decreases incidence of infections in the elderly:effect of zinc on generation of cytokines and oxidative stress.Am J Clin Nutr 2007;85(3):837-844
18 Du Y,Guo D,Wu Q,et al.Zinc chloride inhibits human lens epithelial cell migration and proliferation involved in TGF-β1 and TNF-αsignaling pathways in HLE B-3 cells.Biol Trace Elem Res 2014;159(1-3):425-433
19 Grahn BH,Paterson PG,Gottschall-Pass KT,et al.Zinc and the eye.J Am Coll Nutr 2001;20(2):106-118
20 Shukla N,Moitra JK,Trivedi RC.Determination of lead,zinc,potassium,calcium,copper and sodium in human cataract lenses.Sci Total Environ 1996;181(2):161-165