Doheny Eye Institute, Department of Ophthalmology, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA.
Invest Ophthalmol Vis Sci. 2012 Jun 28;53(7):4197-204. doi: 10.1167/iovs.12-9465.
Recent discovery of microRNAs and their negative gene regulation have provided new understanding in the pathogenesis of inflammatory diseases. This study demonstrated microRNA expression profiling and their likely role in sympathetic ophthalmia, using formalin-fixed, paraffin embedded samples.
Two groups of four enucleated globes (total eight globes) from patients with clinical and histopathological diagnosis of SO (experimental samples) and one group of four age-matched, noninflamed enucleated globes (control samples) were used. Human genome-wide microRNA PCR array was performed and results were subjected to bioinformatics calculation and P values stringency tests. The targets were searched using the recently published and periodically updated miRWalk software. Quantitative real-time PCR and immunohistochemical staining were performed to confirm the validated targets in the mRNA and in the protein levels, respectively.
No microRNA was significantly upregulated in SO, but 27 microRNAs were significantly downregulated. Among these, four microRNAs (hsa-miR-1, hsa-let-7e, hsa-miR-9, and hsa-miR-182) were known to be associated with the inflammatory signaling pathway. Only hsa-miR-9 has the validated targets, tumor necrosis factor-α, and nuclear factor kappa B1, which have been previously shown to be associated with mitochondrial oxidative stress-mediated photoreceptor apoptosis in eyes with SO.
Identification of altered levels of microRNAs by microRNA expression profiling may yield new insights into the pathogenesis of SO by disclosing specific microRNA signatures. In the future these may be targeted by synthetic microRNA mimic-based therapeutic strategies.
最近发现的 microRNAs 及其对基因的负向调控作用，为炎症性疾病的发病机制提供了新的认识。本研究通过福尔马林固定、石蜡包埋的样本，展示了 microRNA 表达谱及其在交感性眼炎中的可能作用。
使用两组（每组 4 只眼球，共 8 只眼球）临床和组织病理学诊断为交感性眼炎的眼（实验组）和一组（4 只眼球）年龄匹配、无炎症的眼（对照组）进行研究。进行了人类全基因组 microRNA PCR 阵列分析，并对结果进行了生物信息学计算和 P 值严格检验。使用最近发布并定期更新的 miRWalk 软件搜索靶标。进行定量实时 PCR 和免疫组织化学染色，分别在 mRNA 和蛋白质水平上验证验证的靶标。
交感性眼炎中没有 microRNA 明显上调，但有 27 个 microRNA 明显下调。其中，有 4 个 microRNAs（hsa-miR-1、hsa-let-7e、hsa-miR-9 和 hsa-miR-182）与炎症信号通路有关。只有 hsa-miR-9 有已验证的靶标，即肿瘤坏死因子-α和核因子 kappa B1，它们以前被证明与交感性眼炎眼中的线粒体氧化应激介导的光感受器凋亡有关。
通过 microRNA 表达谱鉴定 microRNA 水平的变化，可能通过揭示特定的 microRNA 特征，为交感性眼炎的发病机制提供新的见解。在未来，这些可能会成为基于合成 microRNA 模拟的治疗策略的靶点。
