氯化锌对紫外线损伤的人晶状体上皮细胞的保护作用  

Protective effects of zinc chloride with appropriate concentrations on human lens epithelial cell damages induced by UVB irradiation

作　　者：杜宇翔[1] 郭大东[2] 唐凯[1] 司俊康 沙芳[1] 毕宏生[2] 

机构地区：[1]山东中医药大学,山东省济南市250014 [2]山东中医药大学眼科研究所、山东省高校中西医结合眼病防治技术重点实验室,山东省济南市250002

出　　处：《眼科新进展》2014年第5期405-409,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:81072961);山东省自然科学基金资助(编号:ZR2010HM032)~~

摘　　要：目的研究适当浓度的ZnCl2对紫外线(ultraviolet B,UVB)损伤的人晶状体上皮细胞(human lens epithelial cell,HLEC)的保护作用及其作用机制。方法用MTT法检测空白对照组、单纯紫外照射组和2 mg·L-1、3 mg·L-1、4 mg·L-1ZnCl2预处理后UVB照射(实验组)HLEC B-3的生存率;实时细胞电子分析系统(RT-CES)动态监测ZnCl2(2mg·L-1、3 mg·L-1、4 mg·L-1)预处理对UVB照射后HLEC B-3细胞增殖的影响;DAPI染色法检测各组细胞核的变化;实时荧光定量PCR和酶联免疫吸附试验(ELISA)检测HLEC B-3中caspase-12 mRNA和蛋白质表达的变化。结果 MTT结果表明,单纯紫外照射组和ZnCl2(2 mg·L-1、3 mg·L-1、4 mg·L-1)预处理实验组24 h细胞生存率分别为34.47%±9.31%、65.91%±12.46%、64.30%±16.24%和46.36%±6.28%;48 h细胞生存率分别为8.31%±1.38%、53.82%±11.34%、59.25%±7.58%和36.57%±8.41%;72 h细胞生存率分别为6.75%±3.71%、48.29%±10.06%、56.53%±8.47%和39.57%±8.93%。RT-CES分析结果显示,ZnCl2(2 mg·L-1、3mg·L-1、4 mg·L-1)预处理可抑制UVB照射诱发的HLEC B-3的坏死和凋亡;DAPI染色结果表明,ZnCl2可减少UVB照射引起的HLEC B-3细胞核固缩和碎裂;RT-PCR和ELISA检测结果表明,ZnCl2可抑制UVB照射诱发的caspase-12 mRNA和蛋白质的表达升高。结论适当浓度的ZnCl2可抑制UVB照射后HLEC B-3中caspase-12的表达上调,进而对体外培养的HLEC B-3细胞UVB损伤起到保护作用。Objective To investigate the protective effects and its mechanism of zinc chloride （ ZnCl2 ） with appropriate concentrations on human lens epithelial cells （HLEC） B-3 damage induced by ultraviolet B （UVB） irradiations. Methods HLEC B- 3 were incubated with ZnC12 （ 2 mg· L- 1,3 mg · L-1, 4 mg · L- 1 ）, then cells were ex- posed to UVB and the survival rate of control group, UVB group and ZnCl2 （2 mg· L-1, 3 mg· L- 1,4 mg· L- 1 ） ＋ UVB groups were detected by MTT assay; The effects of ZnCl： on UVB-induced HLEC B-3 proliferation was dynamic detected using real-time cell electronic sensing （RT-CES） system; Meanwhile, 4＇, 5＇ -diamidino-2-phenylindole （ DA- PI） staining was performed to monitor the protective effects of ZnCl2 on UVB-induced HLEC B-3 cell nuclei; Further,the effects of ZnCl2 on UVB-induced caspase-12 protein and mRNA expression in HLEC B-3 were tested by both real-time quantitative PCR （ RT- PCR） and enzyme-linked inununosorbent assay （ELISA） technique. Results MTT as- say showed the survival rates of HLEC B-3 in UVB group and ZnC12 （2 mg · L-1 ,3 mg · L-1 ,4 mg · L-1） ＋UVB groups at24 hours were 34.47% ±9.31% ,65.91%± 12.46% , 54.30%± 16.24% and 46.36% ±6.28% ,respectively,which at 48 hours were 8.31% ±1.38% ,53.82% ± 11.34% ,59.25% ± 7.58% and 35.57% ± 8.41% ,respectively,which at 72 hours were 6.75% ±3.71% ,48.29% ±10.06% ,56.53% ± 8. 47% and 39.57% ± 8. 93% ,respectively. RT-CES results suggested that ZnCI： （2 mg· L-1, 3 mg · L-1, 4 mg · L-1 ） could decrease UVB-induced HLEC B-3 apoptosis and necrosis. DAPI results showed that ZnCl2 could decrease UVB-induced HLEC B-3 cell nuclear condensation and nuclear fragmentation. In the meantime, the expression levels of caspase-12 mRNA and protein were inhibited by ZnCl2 in HLEC B-3 under UVB irradiation. Conelusion Ap- propriate concentrations of ZnCl2 can efficiently decrease the expression of easpase-12 under UVB radiation and reduce the UVB-induced impairment on HLEC B-3. ZnC

关 键 词：锌离子 晶状体上皮细胞 紫外线 CASPASE-12 

分 类 号：R776.1[医药卫生—眼科]