Yamada Takechiyo, Ogi Kazuhiro, Sakashita Masafumi, Kanno Masafumi, Kubo Seita, Ito Yumi, Imoto Yoshimasa, Tokunaga Takahiro, Okamoto Masayuki, Narita Norihiko, Fujieda Shigeharu
Department of Otorhinolaryngology, University of Fukui, Fukui 910-1193, Japan; Department of Otorhinolaryngology, Akita University Graduate School of Medicine, Akita 010-8543, Japan.
Department of Otorhinolaryngology, University of Fukui, Fukui 910-1193, Japan.
Auris Nasus Larynx. 2017 Aug;44(4):398-403. doi: 10.1016/j.anl.2016.10.007. Epub 2016 Nov 21.
Against recent reports concerning cytokine or chemokine in mouse or rat inner ear cells, it is almost unknown whether human inner ear cells would produce cytokine or chemokine. We have for the first time established the human inner-ear-derived fibroblasts from endolymphatic sac.
The expression levels of Toll-like receptors (TLRs) in human endolymphatic sac fibroblasts, and the effect on cytokine or chemokine production of the TLR ligands have been examined. To demonstrate the intracellular pathways involved in the regulation of cytokine-production, we used specific inhibitors of c-Jun N-terminal kinase (JNK), extracellular signal-related kinase (ERK), p38 mitogen-activated protein kinase (p38 MAPK)-signaling and N-acetyl-l-cysteine (NAC).
TLR 2, 3, 4 and 9 were highly expressed in human endolymphatic sac fibroblasts. The TLR 3 ligand, polyinosinic-polycytidylic acid (poly(I:C)) significantly enhanced the secretion of thymic stromal lymphopoietin (TSLP), B lymphocyte stimulator (BLyS), IFNγ-inducible protein 10 (IP-10), and macrophage inflammatory protein 1 alpha (MIP-1α) from the cells. The inhibitor of JNK strongly reduced the poly(I:C)-induced TSLP-production. The antioxidant drug, NAC also reduced the TSLP-production in fibroblasts stimulated with poly(I:C).
Our findings suggest human inner-ear-endolymphatic sac derived fibroblasts can produce the cytokine and chemokine in response to TLR ligands and play a certain role during the initiation of an immune response.
鉴于近期有关小鼠或大鼠内耳细胞中细胞因子或趋化因子的报道，人类内耳细胞是否会产生细胞因子或趋化因子几乎未知。我们首次从内淋巴囊建立了人内耳来源的成纤维细胞。
检测了人内淋巴囊成纤维细胞中Toll样受体（TLR）的表达水平，以及TLR配体对细胞因子或趋化因子产生的影响。为了证明参与细胞因子产生调节的细胞内途径，我们使用了c-Jun氨基末端激酶（JNK）、细胞外信号调节激酶（ERK）、p38丝裂原活化蛋白激酶（p38 MAPK）信号通路的特异性抑制剂和N-乙酰-L-半胱氨酸（NAC）。
TLR 2、3、4和9在人内淋巴囊成纤维细胞中高表达。TLR 3配体聚肌苷酸-聚胞苷酸（poly(I:C)）显著增强了细胞中胸腺基质淋巴细胞生成素（TSLP）、B淋巴细胞刺激因子（BLyS）、γ干扰素诱导蛋白10（IP-10）和巨噬细胞炎性蛋白1α（MIP-1α）的分泌。JNK抑制剂强烈降低了poly(I:C)诱导的TSLP产生。抗氧化药物NAC也降低了poly(I:C)刺激的成纤维细胞中TSLP的产生。
我们的研究结果表明，人内耳内淋巴囊来源的成纤维细胞可响应TLR配体产生细胞因子和趋化因子，并在免疫反应启动过程中发挥一定作用。
