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紫外线照射后人晶状体上皮细胞的凋亡机制。

Mechanisms of apoptosis on human lens epithelium after ultraviolet light exposure.

作者信息

Kim Seong-Taeck, Koh Jae-Woong

机构信息

Department of Ophthalmology, Chosun University School of Medicine, Gwangju, Korea.

出版信息

Korean J Ophthalmol. 2011 Jun;25(3):196-201. doi: 10.3341/kjo.2011.25.3.196. Epub 2011 May 24.

DOI:10.3341/kjo.2011.25.3.196
PMID:21655046
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3102824/
Abstract

PURPOSE

The purpose of this study is to understand the mechanism of apoptosis occurring on a cultured human lens epithelial cell line after exposure to ultraviolet (UV) light. We intended to confirm the presence of cellular toxicity and apoptosis and to reveal the roles of p53, caspase 3 and NOXA in these processes.

METHODS

Cells were irradiated with an ultraviolet lamp. Cellular toxicity was measured by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Hoechst staining and fluorescent anti-caspase 3 antibodies were used for apoptosis investigation. The quantities of p53, caspase 3, and NOXA were measured by Western blotting for to investigate the apoptosis pathway.

RESULTS

Cellular toxicity on the human lens epithelium markedly increased with time after UV exposure. On Hoechst staining, we found that apoptosis also remarkably increased after exposure to ultraviolet light, compared with a control group. In the immunochemical study using anti-caspase 3 antibodies, active caspase 3 significantly increased after exposure to ultraviolet light. On Western blotting, p53 decreased, while caspase 3 and NOXA increased.

CONCLUSIONS

Exposure of cultured human lens epithelial cell lines to ultraviolet light induces apoptosis, which promotes the expression of NOXA and caspase 3 increases without increasing p53. This may suggest that UV induced apoptosis is caused by a p53-independent pathway in human lens epithelial cells.

摘要

目的

本研究旨在了解人晶状体上皮细胞系在暴露于紫外线（UV）后发生凋亡的机制。我们旨在确认细胞毒性和凋亡的存在，并揭示p53、半胱天冬酶3（caspase 3）和NOXA在这些过程中的作用。

方法

用紫外线灯照射细胞。通过3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）法测定细胞毒性。使用Hoechst染色和荧光抗半胱天冬酶3抗体进行凋亡研究。通过蛋白质免疫印迹法测定p53、半胱天冬酶3和NOXA的量，以研究凋亡途径。

结果

紫外线暴露后人晶状体上皮细胞的细胞毒性随时间显著增加。在Hoechst染色中，我们发现与对照组相比，暴露于紫外线后凋亡也显著增加。在使用抗半胱天冬酶3抗体的免疫化学研究中，暴露于紫外线后活性半胱天冬酶3显著增加。在蛋白质免疫印迹法中，p53减少，而半胱天冬酶-3和NOXA增加。

结论

培养的人晶状体上皮细胞系暴露于紫外线可诱导凋亡，这促进了NOXA的表达，且半胱天冬酶3增加而p53不增加。这可能表明紫外线诱导的凋亡是由人晶状体上皮细胞中不依赖p53的途径引起的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/56d9/3102824/7b3067336b2e/kjo-25-196-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/56d9/3102824/d6b6fbd10c08/kjo-25-196-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/56d9/3102824/ff1e590dd1db/kjo-25-196-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/56d9/3102824/65d3321aa10a/kjo-25-196-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/56d9/3102824/7b3067336b2e/kjo-25-196-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/56d9/3102824/d6b6fbd10c08/kjo-25-196-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/56d9/3102824/ff1e590dd1db/kjo-25-196-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/56d9/3102824/65d3321aa10a/kjo-25-196-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/56d9/3102824/7b3067336b2e/kjo-25-196-g004.jpg

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