Interleukin-17A Induces Extracellular Matrix Protein Expression in Osteoblastic ROS17/2.8 Cells
Interleukin-17A Induces Extracellular Matrix Protein Expression in Osteoblastic ROS17/2.8 Cells
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DOI：
10.2485/jhtb.20.247
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发表时间：
2011-12-01
影响因子：
0.4
通讯作者：
Maeno, Masao
中科院分区：
文献类型：
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作者：
Kuwabara, Akiko;Tanabe, Natsuko;Maeno, Masao
白细胞介素（IL）-17在许多自身免疫性和炎症性疾病中起重要作用。我们最近证明了IL-17 A对破骨细胞生成的各种直接和间接作用。然而，IL-17 A对成骨细胞骨基质形成的影响尚不清楚。因此，我们检测了IL-17 A对成骨细胞ROS 17/2.8细胞中IL-17受体（IL-17 R）、细胞外基质蛋白（ECMP）和环氧合酶（考克斯）表达、前列腺素（PG）E-2产生、碱性磷酸酶（ALP）活性和矿化结节形成的影响。我们还使用考克斯-2活性的特异性抑制剂NS 398检测了PGE（2）对IL-17诱导的ECMP表达的间接作用。在存在或不存在50 ng/mL中和抗IL-17 A抗体或1 μ M NS 398的情况下，用0（对照）、1、10或100 ng/mL IL-17 A培养细胞。采用实时定量PCR和Western blotting检测IL-17 RA、IL-17 RC、I型胶原、骨唾液蛋白（BSP）、骨钙素、骨桥蛋白、考克斯-1和考克斯-2的表达。用ELISA法检测PGE（2）的产生。在IL-17 A存在下，IL-17 RA、I型胶原、BSP、骨钙素、骨桥蛋白和考克斯-2的表达以及PGE（2）的产生显著增加，而IL-17 RC和考克斯-1的表达、细胞增殖、ALP活性和矿化结节形成不受影响。抗IL-17抗体阻断IL-17 A诱导的ECMP表达。NS 398阻断IL-17 A诱导的PGE（2）产生，但不影响IL-17 A诱导的ECMP表达。这些结果表明，IL-17 A通过IL-17 RC和/或IL-17 A诱导的IL-17 RA在成骨细胞中刺激ECMP表达;然而，细胞的矿化结节形成不受IL-17 A添加的影响。此外，我们的结果表明IL-17 A对ECMP表达的刺激作用不依赖于IL-17 A诱导的PGE的间接作用（2）。
Interleukin (IL)-17 plays an important role in many autoimmune and inflammatory diseases. We recently demonstrated the various direct and indirect effects of IL-17A on osteoclastogenesis. However, the effect of IL-17A on bone matrix formation by osteoblasts is unclear. Therefore, we examined the effect of IL-17A on IL-17 receptor (IL-17R), extracellular matrix protein (ECMP), and cyclooxygenase (COX) expression; prostaglandin (PG) E-2 production; alkaline phosphatase (ALPase) activity; and mineralized nodule formation in osteoblastic ROS17/2.8 cells. We also examined the indirect effect of PGE(2) on IL-17-induced ECMP expression using NS398, a specific inhibitor of COX-2 activity. Cells were cultured with 0 (control), 1, 10, or 100 ng/mL IL-17A in the presence or absence of 50 ng/mL neutralizing anti-IL-17A antibodies or 1 mu M NS398. IL-17RA, IL-17RC, type I collagen, bone sialoprotein (BSP), osteocalcin, osteopontin, COX-1, and COX-2 expression was examined by real-time PCR and Western blotting. PGE(2) production was examined by ELISA. The expression of IL-17RA, type I collagen, BSP, osteocalcin, osteopontin, and COX-2, and PGE(2) production were increased significantly in the presence of IL-17A, whereas the expression of IL-17RC and COX-1, cell proliferation, ALPase activity, and mineralized nodule formation were unaffected. Anti-IL-17 antibodies blocked IL-17A-induced ECMP expression. NS398 blocked IL-17A-induced PGE(2) production, but it did not affect IL-17A-induced ECMP expression. These results suggest that IL-17A stimulates ECMP expression via IL-17RC and/or IL-17A-induced IL-17RA in osteoblasts; however, mineralized nodule formation by the cells was unaffected by the addition of IL-17A. Furthermore, our results indicate that the stimulatory effect of IL-17A on ECMP expression is independent of the indirect effect of IL-17A-induced PGE(2).