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Abstract：
OBJECTIVE: Corneal endothelial cells (CECs) are extremely vulnerable to injury. In this study, the role and mechanism of action of the long non-coding RNA HOXA11-AS during corneal endothelial injury (CEI) were evaluated in vivo and in vitro.
METHODS: Scratch wounds were made to induce CEI in the corneal endothelium of rats and mice. Homeobox A11 (HOXA11)-AS expression was determined at different time points using quantitative real-time PCR. Human CECs with HOXA11-AS overexpression or downregulation were examined for survival, ferroptosis, and migration. Bioinformatics and dual-luciferase reporter assays were used to investigate the correlation between HOXA11-AS and microRNA (miR)-155.
RESULTS: HOXA11-AS expression was reduced in the corneal endothelium in a time-dependent manner. Scratch wounds triggered high rates of ferroptosis and migration in CECs and impaired cell proliferation. HOXA11-AS overexpression partially attenuated the scratch wound-induced changes in proliferation, ferroptosis, and migration, whereas silencing HOXA11-AS had the opposite effects. Moreover, HOXA11-AS served as a competing endogenous RNA of miR-155. Levels of miR-155 were upregulated in the corneal endothelium following the scratch injury, and this upregulation abolished the effect of HOXA11-AS overexpression on the behavior of CECs after injury; miR-155 inhibition counteracted the effect of HOXA11-AS silencing.
CONCLUSIONS: HOXA11-AS exerts protective effects against CEI by sponging miR-155, suggesting that these loci are treatment targets for corneal endothelial disorders.
METHODS: Scratch wounds were made to induce CEI in the corneal endothelium of rats and mice. Homeobox A11 (HOXA11)-AS expression was determined at different time points using quantitative real-time PCR. Human CECs with HOXA11-AS overexpression or downregulation were examined for survival, ferroptosis, and migration. Bioinformatics and dual-luciferase reporter assays were used to investigate the correlation between HOXA11-AS and microRNA (miR)-155.
RESULTS: HOXA11-AS expression was reduced in the corneal endothelium in a time-dependent manner. Scratch wounds triggered high rates of ferroptosis and migration in CECs and impaired cell proliferation. HOXA11-AS overexpression partially attenuated the scratch wound-induced changes in proliferation, ferroptosis, and migration, whereas silencing HOXA11-AS had the opposite effects. Moreover, HOXA11-AS served as a competing endogenous RNA of miR-155. Levels of miR-155 were upregulated in the corneal endothelium following the scratch injury, and this upregulation abolished the effect of HOXA11-AS overexpression on the behavior of CECs after injury; miR-155 inhibition counteracted the effect of HOXA11-AS silencing.
CONCLUSIONS: HOXA11-AS exerts protective effects against CEI by sponging miR-155, suggesting that these loci are treatment targets for corneal endothelial disorders.
摘要：
目的：角膜内皮细胞（CECs）极易受到损伤。在这项研究中,在体内和体外评估了长链非编码RNAHOXA11-AS在角膜内皮损伤（CEI）中的作用和作用机制。
方法:进行划痕以在大鼠和小鼠的角膜内皮中诱导CEI。使用定量实时PCR在不同时间点测定同源盒A11(HOXA11)-AS表达。检查具有HOXA11-AS过表达或下调的人CEC的存活率，铁性凋亡,和移民。使用生物信息学和双荧光素酶报告基因检测来研究HOXA11-AS与microRNA(miR)-155之间的相关性。
结果：HOXA11-AS在角膜内皮中以时间依赖性方式表达降低。划伤引发了CECs中的高比例铁凋亡和迁移以及细胞增殖受损。HOXA11-AS过表达部分减弱了划痕伤口诱导的增殖变化，铁性凋亡,和移民,而沉默HOXA11-AS则有相反的作用。此外，HOXA11-AS充当miR-155的竞争性内源性RNA。在划痕损伤后，miR-155的水平在角膜内皮中上调，这种上调消除了HOXA11-AS过表达对损伤后CEC行为的影响；miR-155抑制抵消了HOXA11-AS沉默的影响。
结论：HOXA11-AS通过形成miR-155发挥对CEI的保护作用，提示这些位点是角膜内皮疾病的治疗靶点。
方法:进行划痕以在大鼠和小鼠的角膜内皮中诱导CEI。使用定量实时PCR在不同时间点测定同源盒A11(HOXA11)-AS表达。检查具有HOXA11-AS过表达或下调的人CEC的存活率，铁性凋亡,和移民。使用生物信息学和双荧光素酶报告基因检测来研究HOXA11-AS与microRNA(miR)-155之间的相关性。
结果：HOXA11-AS在角膜内皮中以时间依赖性方式表达降低。划伤引发了CECs中的高比例铁凋亡和迁移以及细胞增殖受损。HOXA11-AS过表达部分减弱了划痕伤口诱导的增殖变化，铁性凋亡,和移民,而沉默HOXA11-AS则有相反的作用。此外，HOXA11-AS充当miR-155的竞争性内源性RNA。在划痕损伤后，miR-155的水平在角膜内皮中上调，这种上调消除了HOXA11-AS过表达对损伤后CEC行为的影响；miR-155抑制抵消了HOXA11-AS沉默的影响。
结论：HOXA11-AS通过形成miR-155发挥对CEI的保护作用，提示这些位点是角膜内皮疾病的治疗靶点。
