摘要
目的: 蛋白质去甲酰化是一种成熟的调节机制,调节染色质结构和动力学、细胞增殖和分化、应激反应和细胞凋亡。在脊椎动物的眼睛中,我们和其他人已经表明,在调节眼睛发育方面,sumoylation起着不可或缺的作用。在应激诱导和衰老过程中,眼组织逐渐失去正常状态,出现白内障、老年性黄斑变性等主要眼病。我们最近已经证明,SUMOYlation积极调节晶状体细胞的分化,这一过程是否与晶状体发病机制有关还有待研究。在这项研究中,我们已经证明,用葡萄糖氧化酶和紫外线照射处理的透明小鼠晶状体在体外发生白内障,并且与此过程相关的3种sumoylation酶的表达模式已经被发现有明显的改变。 方法: 实验采用4周龄的C57BL/6J小鼠。从眼睛中小心地取出镜片,在M199培养基(Sigma 3769)中培养至少12小时。实验选用透明镜片(无手术损伤)。将晶状体暴露于紫外线照射60分钟或用30μm/ml葡萄糖氧化酶(Go,MP Biomedicals,1673)治疗以诱导白内障形成。用qRT-PCR分析mRNA水平。蛋白质水平用蛋白质印迹分析法测定,并用图像J定量。 结果: 我们得到了以下结果:1)Go治疗和uva照射均能诱导体外培养小鼠晶状体白内障的形成;2)Go治疗后,5种sumoylation酶的mRNAs和蛋白质均显著下调;3)uva照射后,mRNAs和protein的表达模式发生变化。SAE1, UBA2 , UBC9 和 PIAS1的Ns是相反的,而mrnas要么显著上调(sae1、uba2和ubc9),要么轻微上调(pias1),所有4种sumoylation酶的蛋白质都下调;对于ranbp2,uva诱导的mRNA和蛋白质的变化与go治疗有关。 结论:在Go和UVA照射条件下,三种主要苏门答腊酶的mRNA和蛋白表达水平均发生了显著变化。我们的结果表明,苏门答腊酶表达模式的改变与氧化应激诱导的白内障发生有关。
关键词: SAE1
Current Molecular Medicine
Title:Altered Expression Patterns of the Sumoylation Enzymes E1, E2 and E3 Are Associated with Glucose Oxidase- and UVA-Induced Cataractogenesis
Volume: 18 Issue: 8
关键词: SAE1
摘要: Purpose: Protein sumoylation is a well established regulatory mechanism that regulates chromatin structure and dynamics, cell proliferation and differentiation, stress response and cell apoptosis. In the vertebrate eye, we and others have shown that sumoylation plays an indispensable role in regulating eye development. During stress induction and aging process, the ocular tissues gradually loss their normality and develop major ocular diseases such as cataract and aging-related macular degeneration. We have recently demonstrated that sumoylation actively regulates differentiation of lens cells, whether this process is implicated in lens pathogenesis remains to be investigated. In this study, we have demonstrated that transparent mouse lenses treated with glucose oxidase and UVA irradiation undergo in vitro cataract formation, and associated with this process, the expression patterns of the 3 sumoylation enzymes have been found significantly altered.
Methods: Four-week-old C57BL/6J mice were used in our experiment. Lenses were carefully excised from eyes and cultured in M199 medium (Sigma 3769) for at least 12 hours. Transparent lenses (without surgical damage) were selected for experimentation. The lenses were exposed to UVA for 60 min or treated with 30 mU/mL glucose oxidase (GO, MP Biomedicals, 1673) to induce cataract formation. The mRNA levels were analysed with qRT-PCR. The protein levels were determined with western blot analysis and quantitated with Image J.
Results: we have obtained the following results: 1) Both GO treatment and UVA irradiation can induce cataract formation in the in vitro cultured mouse lenses; 2) With GO treatment, the mRNAs and proteins for the 5 sumoylation enzymes were all significantly downregulated; 3) With UVA irradiation, the changes in the expression patterns of the mRNAs and proteins for the SAE1, UBA2 , UBC9 and PIAS1 were opposite, while the mRNAs were upregulated either significantly (for SAE1, UBA2 and UBC9) or slightly (PIAS1), the proteins for all 4 sumoylation enzymes were downregulated; For RanBP2, the UVA induced changes in both mRNA and protein are consist with the GO treatment.
Conclusion: Under GO and UVA irradiation conditions, the expression levels of both mRNA and protein for the three major sumoylation enzymes were significantly changed. Our results suggest that altered expression patterns of the sumoylation enzymes are associated with oxidative stressinduced cataractogenesis.
Export Options
About this article
Cite this article as:
Altered Expression Patterns of the Sumoylation Enzymes E1, E2 and E3 Are Associated with Glucose Oxidase- and UVA-Induced Cataractogenesis, Current Molecular Medicine 2018; 18 (8) . https://dx.doi.org/10.2174/1566524019666190111152324
DOI https://dx.doi.org/10.2174/1566524019666190111152324 |
Print ISSN 1566-5240 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5666 |
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers