Effects of Inflammatory Cytokines on Human Fetal RPE in vitro

炎症细胞因子对体外人胎儿RPE的影响

• 批准号: 7594092
• 负责人: sheldon s miller
• 金额: $ 69.23万
• 依托单位: NATIONAL EYE INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7594092
• 关键词: Acute; Affect; Apical; Back; Bathing; CCL2 gene; CCL7 gene; CCL8 gene; CXC Chemokines; CXCL10 gene; CXCL11 gene; Cells; Chronic Phase; Complex; Disease; Eye; Fluids and Secretions; Homeostasis; Human; IL8 gene; Immune system; In Vitro; Inflammation; Inflammatory; Interferon Type II; Liquid substance; Maintenance; Mediator of activation protein; Pathogenesis; Physiological; Play; Posterior Uveitis; Property; Retina; Role; Side; Small Inducible Cytokine A14; Structure of retinal pigment epithelium; Testing; Tumor Necrosis Factor-alpha; absorption; beta-Chemokines; chemokine; cytokine; disease natural history; fetal; human TNF protein; monolayer; receptor; research study; response

The retinal pigment epithelium (RPE) plays an important role in maintenance of homeostasis in the back of the eye by transporting fluid and secreting a variety of molecules including cytokines/chemokines. In ocular inflammation, affected by the elevated pro-inflammatory cytokines, the physiologic properties of RPE such as fluid transport and chemokine secretion may change resulting in exacerbation of the diseases. To investigate this question, we stimulated the cultured human fetal RPE with a mixture of TNF-alpha, IFN-gamma and IL-1beta. Apical addition of the mixture produced no changes in Jv. In contrast, basal addition increased net fluid absorption by 8.6 1 2.9 ul7cm-27hr-1 (n=3). Jv increased by 5.5 1 1.3 ul7cm-27hr-1 (n=3) after addition to both sides. Stimulation with the mixture, profoundly increased the secretion of the tested 12 chemokines and 3 cytokines. For the majority of these molecules, stimulation from the apical side was much more effective than that from the basal side and the secretion from the apical side was greater by far than that from the basal side. The highly affected chemokines were four CC chemokines (MCP-1, RANTES, MCP-2, MCP-3) and four CXC chemokines (GRO-alpha, IL-8, IP-10, I-TAC). In 3 independent experiments (n=3), for CC chemokines, apical stimulation increased secretion on apical side from 6.9 + 1.2 ng/well to 106.2 + 15.1 ng/well for MCP-1, from 0 to 10.5 + 0.6 ng/well for RANTES, from 69.1 + 1.2 pg/well to 17.1 + 0.7 ng/well for MCP-2, from 6.5 + 1.4 pg/well to 11.2 + 0.7 ng/well for MCP-3. For CXC chemokines, apical stimulation increased secretion on apical side from 15.7 + 3.2 pg/well to 36.2 + 0.3 ng/well for GRO-alpha, from 140.6 + 9.6 pg/well to 55.4 + 9.6 ng/well for IL-8, from 5.1 + 0.3 pg/well to 119.9 + 10.8 ng/well for IP-10, from 9.5 + 2.1 pg/well to 44.6 + 1.0 ng/well for I-TAC. The pro-inflammatory-cytokine-induced activation of JV and the predominant increase of secretion of most chemokine/cytokines to the apical bath suggest a critical role of the RPE in the pathogenesis of acute and chronic phases of ocular inflammatory diseases.

视网膜色素上皮（RPE）通过运输液体和分泌包括细胞因子/趋化因子在内的多种分子，在维持眼后部的体内平衡中发挥着重要作用。在眼部炎症中，受促炎细胞因子升高的影响，RPE 的生理特性（例如液体运输和趋化因子分泌）可能会发生变化，导致疾病恶化。为了研究这个问题，我们用 TNF-α、IFN-γ 和 IL-1β 的混合物刺激培养的人类胎儿 RPE。 顶部添加混合物不会导致 Jv 发生变化。相反，基础添加使净液体吸收增加了 8.6 1 2.9 ul7cm-27hr-1 (n=3)。两侧添加后Jv增加了5.5×1×1.3ul7cm-27hr-1(n=3)。混合物的刺激显着增加了所测试的 12 种趋化因子和 3 种细胞因子的分泌。对于大多数这些分子，来自顶端侧的刺激比来自基底侧的刺激有效得多，并且来自顶端侧的分泌远远大于来自基底侧的分泌。受影响最大的趋化因子是四种 CC 趋化因子（MCP-1、RANTES、MCP-2、MCP-3）和四种 CXC 趋化因子（GRO-alpha、IL-8、IP-10、I-TAC）。在 3 个独立实验 (n=3) 中，对于 CC 趋化因子，顶端刺激增加了顶端侧的分泌，MCP-1 从 6.9 + 1.2 ng/孔增加到 106.2 + 15.1 ng/孔，RANTES 从 0 增加到 10.5 + 0.6 ng/孔，RANTES 从 69.1 + 1.2 pg/孔增加到 17.1 + 0.7 MCP-2 ng/孔，从 6.5 + 1.4 pg/孔至 11.2 + MCP-3 为 0.7 纳克/孔。对于CXC趋化因子，顶端刺激增加了顶端侧的分泌，GRO-α从15.7 + 3.2 pg/孔增加到36.2 + 0.3 ng/孔，IL-8从140.6 + 9.6 pg/孔增加到55.4 + 9.6 ng/孔，IL-8从5.1 + 0.3 pg/孔增加到119.9 + IP-10 为 10.8 ng/孔，从 9.5 + 2.1 pg/孔到 I-TAC 为 44.6 + 1.0 ng/孔。 促炎细胞因子诱导的 JV 激活和大多数趋化因子/细胞因子分泌到顶浴的显着增加表明 RPE 在眼部炎症疾病急性和慢性阶段的发病机制中发挥着关键作用。