Interleukin-1β Increases Baseline Expression and Secretion of Interleukin-6 by Human Uveal Melanocytes In Vitro via the p38 MAPK/NF-κB Pathway
Interleukin-1β Increases Baseline Expression and Secretion of Interleukin-6 by Human Uveal Melanocytes In Vitro via the p38 MAPK/NF-κB Pathway
复制标题
DOI：
10.1167/iovs.10-6908
复制
发表时间：
2011-05-01
影响因子：
4.4
通讯作者：
Chan, Chi-Chao
中科院分区：
文献类型：
--
作者：
Hu, Dan-Ning;Chen, Min;Chan, Chi-Chao
目的。黑素细胞是葡萄膜的主要细胞成分。白细胞介素6(IL-6)是一种促炎细胞因子。研究IL-6在体外培养的人葡萄膜黑素细胞(Um)中的表达及IL-1β对其表达的调节作用，并探讨相关的信号转导途径。用酶联免疫吸附试验试剂盒检测IL-6蛋白表达，RT-PCR检测IL-6转录产物。用酶联免疫吸附试验检测核抽提物中的核转录因子-kappaB和p38MAPK、ERK、JNK的磷酸化水平。加入p38(SB203580)、ERK(UO1026)、JNK(SP600125)和核因子-kappaB(BAY11-7082)的抑制剂，观察它们的作用。结果：在无血清培养的UM条件培养液中检测到低水平的IL-6蛋白。血清的加入增加了IL-6的分泌。IL-1β(0.1-10 ng/mL)以剂量和时间依赖的方式使IL-6转录和蛋白水平增加6倍，同时核提取液中的核转录因子-kappaB和细胞裂解产物中的磷酸化p38MAPK显著增加。单独使用核因子-kappaB和p38抑制剂后，IL-6的表达完全消失。结论首次报道了UM细胞表达和分泌IL-6的情况。IL-1β可促进黑素细胞产生IL-6。这些数据表明，UMS可能在眼炎性和自身免疫性疾病的发病机制中发挥作用。(投资眼科VS科学。2011年；52：37673774)doi：10.1167/iovs.10-6908
PURPOSE. Melanocyte is the major cell component in the uvea. Interleukin (IL)-6 is a proinflammatory cytokine. The authors studied the expression of IL-6 in cultured human uveal melanocytes (UMs) and their modulation by IL-1 beta and explored the relevant signal pathways.METHODS. Conditioned media and cells were collected from UMs cultured in medium with and without serum and were stimulated by IL-1 beta. IL-6 protein and transcript were measured using an ELISA kit and RT-PCR, respectively. NF-kappa B in nuclear extracts and phosphorylated p38 MAPK, ERK, and JNK in cells cultured with and without IL-1 beta were measured by ELISA kits. Inhibitors of p38 (SB203580), ERK (UO1026), JNK (SP600125), and NF-kappa B (BAY11-7082) were added to the cultures to evaluate their effects.RESULTS. Low levels of IL-6 protein were detected in the conditioned medium in UMs cultured without serum. The addition of serum increased the secretion of IL-6. IL-1 beta (0.1-10 ng/mL) increased IL-6 transcript and protein levels in a dose-and time-dependent manner up to sixfold, accompanied by a significant increase of NF-kappa B in nuclear extracts and phosphorylated p38 MAPK in cell lysates. NF-kappa B and p38 inhibitors alone decreased, whereas a combination of these two inhibitors completely abolished the IL-1 beta-induced expression of IL-6.CONCLUSIONS. This is the first report on the expression and secretion of IL-6 by UMs. IL-1 beta augments IL-6 production from the melanocytes. The data suggest that UMs may play a role in the pathogenesis of ocular inflammatory and autoimmune diseases. (Invest Ophthalmol Vis Sci. 2011;52:3767-3774) DOI: 10.1167/iovs.10-6908