East-West Medical Research Institute, Kyung Hee University, Seoul 130-702, Korea.
Mol Biol Rep. 2011 Jun;38(5):3207-12. doi: 10.1007/s11033-010-9993-5. Epub 2010 Feb 21.
15-deoxy-∆(12,14)-prostaglandin J(2) (15d-PGJ(2)) is an anti-inflammatory prostaglandin that plays a role in promoting the resolution of inflammation. We investigated the effects of 15d-PGJ(2) on the production of IL-8 and on the expression of Toll-like receptors (TLRs) 2 in human primary keratinocytes stimulated with lipopolysaccharide (LPS). Cell proliferation was analyzed using the MTT assay, TLR2 and -4 mRNA expression was detected by RT-PCR, and IL-8 production and NF-κB p65 activities were determined by ELISA. LPS and 15d-PGJ(2) did not influence the proliferation rate at low concentrations (0.5 and 2.0 μM) in keratinocytes, and showed toxicity at high concentrations (5.0 μM). LPS, compared with control, induced the expression of TLR2 mRNA, increased IL-8 production, and enhanced NF-κB activity. 15d-PGJ(2) decreased TLR2 mRNA, increased IL-8 production, and suppressed NF-κB activity. Costimulation with LPS and 15d-PGJ(2), compared with LPS stimulation alone, decreased TLR2 mRNA (1.8-fold), increased IL-8 production (1.8-fold at 0.5 μM and 3.7-fold at 2.0 μM), and inhibited NF-κB activity (3.3-fold at 0.5 μM and 5.1-fold at 2.0 μM). TLR4 mRNA was not expressed in primary keratinocytes. These results suggest that 15d-PGJ(2) suppresses TLR2 expression and that it up-regulates the production of IL-8 by inhibiting the NF-κB signaling pathway in primary keratinocytes. Thus, 15d-PGJ(2) can have both anti- and pro-inflammatory effects, and 15d-PGJ(2)-mediated IL-8 up-regulation is related to the mitogen-activated protein kinase (MAPK) and NF-κB signaling pathways.
15-脱氧-Δ(12,14)-前列腺素 J2(15d-PGJ2)是一种具有抗炎作用的前列腺素，可促进炎症消退。本研究旨在探讨 15d-PGJ2 对脂多糖(LPS)刺激的人原代角质形成细胞产生白细胞介素-8(IL-8)和 Toll 样受体(TLR)2 表达的影响。采用 MTT 法检测细胞增殖，RT-PCR 检测 TLR2 和 TLR4mRNA 表达，ELISA 检测 IL-8 产生和 NF-κB p65 活性。低浓度(0.5 和 2.0 μM)时 LPS 和 15d-PGJ2 对角质形成细胞增殖率无影响，高浓度(5.0 μM)时具有细胞毒性。与对照组相比，LPS 诱导 TLR2mRNA 表达增加，IL-8 产生增加，NF-κB 活性增强。15d-PGJ2 降低 TLR2mRNA 表达，增加 IL-8 产生，抑制 NF-κB 活性。与 LPS 单独刺激相比，LPS 和 15d-PGJ2 共同刺激时 TLR2mRNA 减少(1.8 倍)，IL-8 产生增加(0.5 μM 时增加 1.8 倍，2.0 μM 时增加 3.7 倍)，NF-κB 活性抑制(0.5 μM 时抑制 3.3 倍，2.0 μM 时抑制 5.1 倍)。原代角质形成细胞不表达 TLR4mRNA。上述结果提示 15d-PGJ2 抑制 TLR2 表达，通过抑制 NF-κB 信号通路而上调原代角质形成细胞 IL-8 的产生。因此，15d-PGJ2 具有抗炎和促炎双重作用，15d-PGJ2 介导的 IL-8 上调与丝裂原活化蛋白激酶(MAPK)和 NF-κB 信号通路有关。
