Department of Ophthalmology, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA.
Invest Ophthalmol Vis Sci. 2011 Jul 1;52(7):4859-65. doi: 10.1167/iovs.10-7066.
Various ocular and orbital tissues differ in their manifestations of inflammation, although the reasons for this are unclear. Such differences may be due to behaviors exhibited by resident cell types, including fibroblasts. Fibroblasts mediate immune function and produce inflammatory mediators. Chronic stimulation of ocular fibroblasts can lead to prolonged inflammation and, in turn, to impaired vision and blindness. Interleukin (IL)-1β, which is produced by various cells during inflammation, is a potent activator of fibroblasts and inducer of the expression of inflammatory mediators. The hypothesis for this study was that that human fibroblasts derived from distinct ocular tissues differ in their responses to IL-1β and that variations in the IL-1 signaling pathway account for these differences.
Human fibroblasts were isolated from the lacrimal gland, cornea, and Tenon's capsule and treated with IL-1β in vitro. Cytokine and prostaglandin (PG)E(2) production were measured by ELISA and EIA. Cyclooxygenase (Cox)-2 expression was detected by Western blot. Components of the IL-1 signaling pathway were detected by flow cytometry, ELISA, Western blot, and immunofluorescence.
Cytokine and PGE(2) production and Cox-2 expression were greatest in corneal fibroblasts. VEGF production was greatest in Tenon's capsule fibroblasts. Variations in IL-1 receptor and receptor antagonist expression, IκBα degradation and p65 nuclear translocation, however, did not account for these differences, but overexpression of the NF-κB member RelB dampened Cox-2 expression in all three fibroblast types.
The results highlight the inherent differences between ocular fibroblast strains and provide crucial insight into novel, tissue-specific treatments for ocular inflammation and disease, such as RelB overexpression.
尽管原因尚不清楚，但各种眼部和眼眶组织在炎症表现上存在差异。这种差异可能是由于驻留细胞类型的行为所致，包括成纤维细胞。成纤维细胞介导免疫功能并产生炎症介质。眼成纤维细胞的慢性刺激可导致炎症持续存在，并进而导致视力受损和失明。白细胞介素 (IL)-1β是炎症过程中各种细胞产生的一种强效成纤维细胞激活物和炎症介质诱导物。本研究的假设是，源自不同眼组织的人成纤维细胞对 IL-1β的反应不同，而 IL-1 信号通路的变化解释了这些差异。
从泪腺、角膜和 Tenon 囊分离人成纤维细胞，并在体外用 IL-1β处理。通过 ELISA 和 EIA 测量细胞因子和前列腺素 (PG)E2 的产生。通过 Western blot 检测环氧化酶 (Cox)-2 的表达。通过流式细胞术、ELISA、Western blot 和免疫荧光检测 IL-1 信号通路的组成部分。
角膜成纤维细胞产生的细胞因子和 PGE2 以及 Cox-2 表达最多。Tenon 囊成纤维细胞产生的 VEGF 最多。然而，IL-1 受体和受体拮抗剂表达、IκBα 降解和 p65 核易位的变化并不能解释这些差异，但 NF-κB 成员 RelB 的过表达可抑制所有三种成纤维细胞类型的 Cox-2 表达。
这些结果突出了眼成纤维细胞株之间的固有差异，并为眼部炎症和疾病的新型组织特异性治疗提供了重要的见解，例如 RelB 过表达。
