Division of Nephrology, Department of Medicine, Showa University School of Medicine, Tokyo, Japan.
Am J Physiol Renal Physiol. 2010 Mar;298(3):F779-87. doi: 10.1152/ajprenal.00198.2009. Epub 2009 Dec 30.
We investigated the role of IL-17 family members IL-17A and IL-17F in the induction of chemokines in mouse cultured mesangial cells (SV40 MES 13 cells). We evaluated the expression of the chemokines monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-2 (MIP-2) by ELISA and real-time RT-PCR (Q-PCR). Activation of MAPK was assessed by immunoblotting. IL-17RA and IL-17RC were inhibited by small interfering RNA (siRNA). We found that IL-17A or IL-17F stimulation of mesangial cells led to both a dose- and time-dependent increase in MCP-1 and MIP-2 release. This effect was dependent on mRNA transcription and protein translation. Both also enhanced TNF-alpha- and IL-1beta-mediated MCP-1 and MIP-2 release in the cells. Additionally, we observed that IL-17A and IL-17F induced MAPK (p38 MAPK, ERK1/2, and JNK) activation and that pharmacological inhibitors of p38 MAPK (SB203580) and ERK1/2 (U0126), but not JNK (SP600125), blocked the IL-17A/IL-17F-mediated MCP-1 and MIP-2 release. Mesangial cells expressed IL-17RA and IL-17RC, and the IL-17A-mediated MCP-1 and MIP-2 release was significantly blocked by soluble IL-17RA. Furthermore, inhibition of either IL-17RA or IL-17RC expression via siRNA led to significant reduction of IL-17A/IL-17F-stimulated chemokine production. We conclude that IL-17A and IL-17F induce the production of chemokines MCP-1 and MIP-2 via MAPK pathways (p38 MAPK and ERK1/2), as well as mRNA transcription and protein translation and have synergistic effects with TNF-alpha and IL-1beta in cultured mesangial cells.
我们研究了白细胞介素-17 家族成员白细胞介素-17A 和白细胞介素-17F 在诱导小鼠培养系膜细胞（SV40 MES 13 细胞）趋化因子表达中的作用。我们通过 ELISA 和实时 RT-PCR（Q-PCR）评估趋化因子单核细胞趋化蛋白-1（MCP-1）和巨噬细胞炎症蛋白-2（MIP-2）的表达。通过免疫印迹评估 MAPK 的激活。通过小干扰 RNA（siRNA）抑制 IL-17RA 和 IL-17RC。我们发现白细胞介素-17A 或白细胞介素-17F 刺激系膜细胞导致 MCP-1 和 MIP-2 释放呈剂量和时间依赖性增加。这种作用依赖于 mRNA 转录和蛋白质翻译。两者还增强了细胞中 TNF-α和 IL-1β介导的 MCP-1 和 MIP-2 释放。此外，我们观察到白细胞介素-17A 和白细胞介素-17F 诱导 MAPK（p38 MAPK、ERK1/2 和 JNK）激活，并且 p38 MAPK（SB203580）和 ERK1/2（U0126）的药理学抑制剂，但不是 JNK（SP600125），阻断了白细胞介素-17A/白细胞介素-17F 介导的 MCP-1 和 MIP-2 释放。系膜细胞表达 IL-17RA 和 IL-17RC，可溶性 IL-17RA 显著阻断白细胞介素-17A 介导的 MCP-1 和 MIP-2 释放。此外，通过 siRNA 抑制 IL-17RA 或 IL-17RC 表达导致白细胞介素-17A/白细胞介素-17F 刺激的趋化因子产生显著减少。我们得出结论，白细胞介素-17A 和白细胞介素-17F 通过 MAPK 途径（p38 MAPK 和 ERK1/2）以及 mRNA 转录和蛋白质翻译诱导趋化因子 MCP-1 和 MIP-2 的产生，并在培养的系膜细胞中与 TNF-α和 IL-1β具有协同作用。
Zotero 插件