[1]王道光,刘冬梅,郭大东,等.氧化锌抑制紫外线B诱导的人晶状体上皮细胞Ca-ATP酶1表达[J].眼科新进展,2017,37(11):1022-1026.[doi:10.13389/j.cnki.rao.2017.0259]
　WANG Dao-Guang,LIU Dong-Mei,GUO Da-Dong,et al.Zinc oxide nanoparticles inhibits Ca-ATPase1 expression induced by UVB irradiation in human lens epithelial cells[J].Recent Advances in Ophthalmology,2017,37(11):1022-1026.[doi:10.13389/j.cnki.rao.2017.0259]
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氧化锌抑制紫外线B诱导的人晶状体上皮细胞Ca2+-ATP酶1表达/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
37卷
期数:
2017年11期
页码:
1022-1026
栏目:
实验研究
出版日期:
2017-11-05

文章信息/Info

Title:
Zinc oxide nanoparticles inhibits Ca2+-ATPase1 expression induced by UVB irradiation in human lens epithelial cells
作者:
王道光; 刘冬梅; 郭大东; 毕宏生; 吴秋欣
250002　山东省济南市，山东中医药大学附属眼科医院（王道光，刘冬梅，毕宏生，吴秋欣）；250002　山东省济南市，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（郭大东）
Author(s):
WANG Dao-Guang; LIU Dong-Mei; GUO Da-Dong; BI Hong-Sheng; WU Qiu-Xin
Affiliated Eye Hospital of Shandong University of Traditional Chinese Medicine（WANG Dao-Guang，LIU Dong-Mei，BI Hong-Sheng，WU Qiu-Xin），Jinan 250002，Shandong
关键词:
氧化锌; 紫外线B; 钙稳态; 细胞质膜Ca2+-ATP酶1; 人晶状体上皮细胞
Keywords:
zinc oxide; ultraviolet B; calcium homeostasis; plasma membrane calcium ATPase1; human lens epithelial cells
分类号:
R776
DOI:
10.13389/j.cnki.rao.2017.0259
文献标志码:
A
摘要:
目的　研究纳米材料氧化锌在有/无紫外线B（ultraviolet B，UVB）照射下对人晶状体上皮细胞系B-3（human lens epithelial cell B-3，HLEB-3）细胞质膜Ca2+-ATP酶1（plasma membrane calcium ATPase1，PMCA1）表达的影响。方法　HLEB-3细胞进行培养和传代，在有/无UVB照射情况下，加入不同浓度的氧化锌，利用DAPI染色细胞核；Annexin V/PI染色检测细胞凋亡；Fluo-3/AM染色检测细胞内Ca2+浓度的变化；实时荧光定量PCR检测PMCA1 mRNA表达水平；Western blot方法检测PMCA1蛋白表达水平。结果　DAPI细胞核染色结果显示，氧化锌以浓度依赖方式使HLEB-3细胞产生典型的细胞凋亡反应；UVB照射可增加氧化锌对HLEB-3细胞的毒性效应；在UVB照射下，经氧化锌（2.5 μg·mL-1、5.0 μg·mL-1、10.0 μg·mL-1）处理的HLEB-3细胞凋亡及细胞内Ca2+浓度增加（均为P<0.05），细胞内Ca2+水平分别从（156.34±4.59）nmol·L-1增加到（173.88±7.17）nmol·L-1、（289.02±9.09）nmol·L-1、（488.36±48.16）nmol·L-1，差异均有统计学意义（均为P<0.05）；在无UVB照射下，2.5 μg·mL-1、5.0 μg·mL-1、10.0 μg·mL-1氧化锌组细胞PMCA1 mRNA表达分别是0 μg·mL-1氧化锌组的0.75倍、0.57倍和0.41倍，差异均有统计学意义（均为P＜0.05）；而在有UVB照射下，各浓度氧化锌组细胞PMCA1 mRNA表达分别是0 μg·mL-1氧化锌组的0.64倍、0.24倍和 0.09倍，差异均有统计学意义（均为P＜0.05）。结论　氧化锌和UVB照射通过Ca2+信号转导通路对HLEB-3细胞有协同抑制作用，提示氧化锌在有UVB照射的情况下对后发性白内障的治疗有巨大潜在作用。
Abstract:
Objective　To investigate the effect of ZnO nanoparticles on the expressions of plasma membrane calcium ATPase1 (PMCA1) of human lens epithelial cell B-3(HLEB-3) at both mRNA and protein levels in the presence and absence of ultraviolet B (UVB) irradiation.Methods　HLEB-3 was cultured in RPMI 1640 medium，and the cytotoxic effect of different concentrations of ZnO (0 μg·mL-1，2.5 μg·mL-1，5.0 μg·mL-1，10.0 μg·mL-1) on HLEB-3 was investigated in the presence and absence of UVB irradiation.DAPI staining was used to monitor the effect of ZnO on HLECB-3 nuclei，and cell apoptosis was evaluated using annexin V-FITC/PI staining in the presence and absence of UVB irradiation．In addition，the intracellular calcium ion (Ca2+) levels were assayed using Fluo-3/AM staining，and the expression levels of both PMCA1 mRNA and protein within HLEB-3 were detected by real-time PCR and Western blot，respectively.Results　DAPI staining showed that the ZnO-treated HLEB-3 displayed a concentration-dependent apoptosis，and UVB irradiation could further aggravate the cytotoxic effect of ZnO on HLEB-3.In addition，in the presence of UVB irradiation，concentration gradient of ZnO (2.5 μg·mL-1，5.0 μg·mL-1，10.0 μg·mL-1) increased the intracellular calcium ion levels ［from (156.34±4.59)nmol·L-1 to (173.88±7.17) nmol·L-1，(289.02±9.09)nmol·L-1，(488.36±48.16)nmol·L-1，respectively］ and upregulated HLEB-3 apoptosis，with statistical difference (all P<0.05).Moreover，the expression level of PMCA1 in the 2.5 μg·mL-1，5.0 μg·mL-1，10.0 μg·mL-1 ZnO-treated epithelial cells was accordingly 0.75，0.57 and 0.41 as much as that in the 0 μg·mL-1 ZnO-treated cells in the absence of UVB irradiation (all P<0.05)，and was accordingly 0.64，0.24 and 0.09 in the present of UVB irradiation，with significant difference (all P<0.05).Conclusion　Both ZnO nanoparticle and UVB irradiation can exert cosuppression effect on HLEB-3 via calcium-mediated signaling pathway，indicating it has great potential for the treatment of posterior capsular opacification with UVB irradiation.

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更新日期/Last Update: 2017-11-01