[1]杜宇翔,郭大东,唐凯,等.氯化锌对紫外线损伤的人晶状体上皮细胞的保护作用[J].眼科新进展,2014,34(5):405-409.[doi:10.13389/j.cnki.rao.2014.0112]
　DU Yu-Xiang,GUO Da-Dong,TANG Kai,et al.Protective effects of zinc chloride with appropriate concentrations on human lens epithelial cell damages induced by UVB irradiation[J].Recent Advances in Ophthalmology,2014,34(5):405-409.[doi:10.13389/j.cnki.rao.2014.0112]
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氯化锌对紫外线损伤的人晶状体上皮细胞的保护作用
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
34卷
期数:
2014年5期
页码:
405-409
栏目:
实验研究
出版日期:
2014-05-05

文章信息/Info

Title:
Protective effects of zinc chloride with appropriate concentrations on human lens epithelial cell damages induced by UVB irradiation
作者:
杜宇翔; 郭大东; 唐凯; 司俊康; 沙芳; 毕宏生
２５００１４　山东省济南市， 山东中医药大学
Author(s):
DU Yu-Xiang;  GUO Da-Dong;  TANG Kai;  SI Jun-Kang;  SHA Fang;  BI Hong-Sheng
关键词:
锌离子; 晶状体上皮细胞; 紫外线; ｃａｓｐａｓｅ-１２
Keywords:
zinc ion ; lens epithelial cell;  ultraviolet B ;  caspase-12
DOI:
10.13389/j.cnki.rao.2014.0112
文献标志码:
A
摘要:
目的　研究适当浓度的ＺｎＣｌ２对紫外线（ｕｌｔｒａｖｉｏｌｅｔＢ，ＵＶＢ）损伤的人晶状体上皮细胞（ｈｕｍａｎｌｅｎｓｅｐｉｔｈｅｌｉａｌｃｅｌｌ，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２ｍｇ?Ｌ－１、３ｍｇ?Ｌ－１、４ｍｇ?Ｌ－１ＺｎＣｌ２预处理后ＵＶＢ照射（实验组）ＨＬＥＣＢ-３的生存率；实时细胞电子分析系统（ＲＴ-ＣＥＳ）动态监测ＺｎＣｌ２（２ ｍｇ?Ｌ－１、３ｍｇ?Ｌ－１、４ｍｇ?Ｌ－１）预处理对ＵＶＢ照射后ＨＬＥＣＢ-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ＨＬＥＣＢ-３中ｃａｓｐａｓｅ-１２ｍＲＮＡ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（２ｍｇ?Ｌ－１、３ｍｇ? Ｌ－１、４ｍｇ? Ｌ－１）预处理实验组２４ｈ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％；４８ｈ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％；７２ｈ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（２ｍｇ?Ｌ－１、３ｍｇ?Ｌ－１、４ｍｇ?Ｌ－１）预处理可抑制ＵＶＢ照射诱发的ＨＬＥＣＢ-３的坏死和凋亡；ＤＡＰＩ染色结果表明，ＺｎＣｌ２可减少ＵＶＢ照射引起的ＨＬＥＣＢ-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-１２ｍＲＮＡ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ＨＬＥＣＢ-３细胞ＵＶＢ损伤起到保护作用。
Abstract:
Objective To investigate the protective effects and its mechanism of zinc chloride ( ZriC12 ) 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 . 1-1 ,3 mg . 1-1 ,4 mg . L-’) . then cells were exposed to UVB and the survival rate of control group ,UVB group and ZnC12 (2 mg . L-l , 3 mg . L",4 mg ’ L-’) + UVB groups were detected by MTT assay; The effects of ZnCI, on UVB-induced HLEC B-3 proliferation was dynamic detected using real-time cell electronic sensing ( RT-CES) system ; Meanwhile ,4 ’ ,6 ’ -diamidino-2-phenylindole ( DA-PI) stairung was performed to monitor the protective effects of ZnC12 0n UVB-induced HLEC B-3 cell nuclei; Further, the effects of ZnCI, 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 immunosorbent assay ( ELISA) technique. Results MTT as-say showed the survival rates of HLEC B-3 in UVB group and ZnC12 (2 mg . 1’] ,3 mg . 1’1 ,4 mg . L") + UVB groups at 24 hours were 34. 47% +9. 31% .65. 91% + 12. 46% . 64. 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 36. 57% + 8. 41% ,respectively , which at 72 hours were 6. 75% + 3. 71% .48. 2g% + 10. 06% ,56. 53% + 8. 47% and 39. 57% +8. 93% ,respectively. RT-CES results suggested that ZnC12 (2 mg ’ L-’,3 mg ’ L",4 mg . L ’l ) could decrease UVB-induced HLEC B-3 apoptosis and necrosis. DAPI results showed that ZnCI, 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 ZnCI, in HLEC B-3 under UVB irradiation. Conclusion Appropriate concentrations of ZnCL can efficiently decrease the expression of caspase-12 under UVB radiation and reduce the UVB-induced impairment on HLEC B-3. ZnC12 plays a protective role in HLEC B-3 under UVB irradiation.

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备注/Memo

备注/Memo:
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）
更新日期/Last Update: 2014-05-04