Oxidative stress and premature senescence in corneal endothelium following penetrating keratoplasty in an animal model.
Oxidative stress and premature senescence in corneal endothelium following penetrating keratoplasty in an animal model.
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DOI：
10.1186/s12886-016-0192-6
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发表时间：
2016-02-02
影响因子：
2
通讯作者：
Chen P
中科院分区：
文献类型：
--
作者：
Zhao X;Wang Y;Wang Y;Li S;Chen P
本研究的目的是解决穿透性角膜移植术后过早衰老过程如何影响角膜内皮的问题，因为供体角膜内皮细胞的质量对于角膜移植的成功非常重要。使用正常风险原位小鼠角膜移植模型评估角膜内皮细胞衰老和诱导的氧化应激。衰老相关的β-半乳糖苷酶（SA-β-Gal）染色用于评估同种异体角膜移植物内皮的过早衰老。使用氧化应激和抗氧化防御 RT2-PCR 阵列和使用 H2O2 处理的体外实验模型来研究可能的机制。同种异体组小鼠角膜内皮有明显的SA-β-Gal阳性，同种异体组小鼠角膜内皮p16INK4a信息和蛋白水平较同基因组升高。通过PCR阵列，发现PKP后同种异体角膜移植物内皮细胞存在氧化-抗氧化失衡。使用 PKP 后同种异体角膜移植物的人内皮样本验证了小鼠模型的结果。我们还开发了一个体外实验模型，使用 H2O2 处理来模拟培养的人角膜内皮细胞 (HCEC) 的氧化应激状态，发现 HCEC 中 ROS 水平升高、CDK 抑制剂上调以及 ROS 介导的 p16INK4A 上调通过 ASK1-p38 MAPK 途径发生。我们的结果表明 PKP 后同种异体角膜移植物的内皮细胞中存在氧化应激和过早衰老。本文的在线版本 (doi:10.1186/s12886-016-0192-6) 包含补充材料，可供授权用户使用。
The purpose of this study was to address the question of how the premature senescence process may affect corneal endothelium after penetrating keratoplasty, because the quality of donor corneal endothelial cells is important for corneal transplant success. The cell senescence and induced oxidative stress in corneal endothelium were assessed using a normal-risk orthotopic mice corneal transplantation model. Senescence associated beta-galactosidase (SA-beta-Gal) staining was used to evaluate premature senescence in the endothelium of corneal allografts. Oxidative Stress and Antioxidant Defense RT2-PCR Arrays and in vitro experimental model using H2O2 treatment were used to investigate the possible mechanism. SA-beta-Gal positivity was observed obviously in mice corneal endothelium of allogenic group and the levels of p16INK4a message and protein increased in endothelium of allogenic group compared to syngenic group. By PCR array, an oxidant-antioxidant imbalance was found in the endothelium of corneal allograft after PKP. The results from mice model were validated using human endothelium samples of corneal allograft after PKP. We also developed an in vitro experimental model using H2O2 treatment to simulate a state of oxidative stress in cultured human corneal endothelial cells (HCECs) and found that elevated ROS levels, the up-regulation of CDK inhibitors and ROS-mediated p16INK4A up-regulation in HCECs occur via the ASK1-p38 MAPK pathway. Our results demonstrate the presence of oxidative stress and premature senescence in the endothelium of corneal allografts following PKP. The online version of this article (doi:10.1186/s12886-016-0192-6) contains supplementary material, which is available to authorized users.