Hata Kazunori, Andoh Akira, Shimada Mitsue, Fujino Sanae, Bamba Shigeki, Araki Yoshio, Okuno Takafumi, Fujiyama Yoshihide, Bamba Tadao
Department of Internal Medicine, Shiga University of Medical Science, Seta-Tsukinowa, Otsu 520-2192, Japan.
Am J Physiol Gastrointest Liver Physiol. 2002 Jun;282(6):G1035-44. doi: 10.1152/ajpgi.00494.2001.
Colonic subepithelial myofibroblasts (SEMFs) may play a role in the modulation of mucosal inflammatory responses. We investigated the effects of interleukin (IL)-17 on IL-6 and chemokine [IL-8 and monocyte chemoattractant protein (MCP)-1] secretion in colonic SEMFs. Cytokine expression was determined by ELISA and Northern blotting. Nuclear factor kappa B (NF-kappaB) DNA-binding activity was evaluated by electrophortetic gel mobility shift assay (EMSA). The activation of mitogen-activated protein kinase (MAPK) was assessed by immunoblotting. IL-6, IL-8, and MCP-1 secretions were rapidly induced by IL-17. IL-17 induced NF-kappaB activation within 45 min after stimulation. A blockade of NF-kappaB activation markedly reduced these responses. MAPK inhibitors (SB-203580, PD-98059, and U-0126) significantly reduced the IL-17-induced IL-6 and chemokine secretion. The combination of either IL-17 + IL-1beta or IL-17 + tumor necrosis factor (TNF)-alpha enhanced cytokine secretion; in particular, the effects of IL-17 + TNF-alpha on IL-6 secretion were much stronger than the other responses. This was dependent on the enhancement of IL-6 mRNA stability. In conclusion, human SEMFs secreted IL-6, IL-8, and MCP-1 in response to IL-17. These responses might play an important role in the pathogenesis of gut inflammation.
结肠上皮下肌成纤维细胞(SEMFs)可能在调节黏膜炎症反应中发挥作用。我们研究了白细胞介素(IL)-17对结肠SEMFs中IL-6和趋化因子[IL-8和单核细胞趋化蛋白(MCP)-1]分泌的影响。通过酶联免疫吸附测定(ELISA)和Northern印迹法测定细胞因子表达。通过电泳凝胶迁移率变动分析(EMSA)评估核因子κB(NF-κB)的DNA结合活性。通过免疫印迹法评估丝裂原活化蛋白激酶(MAPK)的激活情况。IL-17可迅速诱导IL-6、IL-8和MCP-1的分泌。IL-17在刺激后45分钟内诱导NF-κB激活。阻断NF-κB激活可显著降低这些反应。MAPK抑制剂(SB-203580、PD-98059和U-0126)可显著降低IL-17诱导的IL-6和趋化因子分泌。IL-17 + IL-1β或IL-17 +肿瘤坏死因子(TNF)-α的组合可增强细胞因子分泌;特别是,IL-17 + TNF-α对IL-6分泌的作用比其他反应要强得多。这取决于IL-6 mRNA稳定性的增强。总之,人SEMFs对IL-17作出反应分泌IL-6、IL-8和MCP-1。这些反应可能在肠道炎症的发病机制中起重要作用。