Hida Richard Yudi, Takano Yoji, Okada Naoko, Dogru Murat, Satake Yoshiyuki, Fukagawa Kazumi, Fujishima Hiroshi
Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan.
Curr Eye Res. 2008 Jan;33(1):19-22. doi: 10.1080/02713680701817366.
Eotaxin, a CC-chemokine with selective chemotactic effects for eosinophils, has been reported to play an important role in allergic conjunctival diseases. We previously reported that eotaxin is produced by conjunctival fibroblasts and keratocytes on stimulation with Th2 cytokines. Tranilast is known to have anti-allergic properties. In this study, we examined the inhibitory effect of tranilast, an anti-allergic drug, on eotaxin-1 production from cultured conjunctival fibroblasts.
Conjunctival fibroblasts obtained from normal patients were cultured in DMEM/F12 medium. On the fifth passage, the cells were transferred to a 96-well plate and, after starvation for 24 hr, TNF-alpha, IL-4, and tranilast or dexamethasone were added. After 48 hr, the concentrations of eotaxin-1 in the supernatants were measured by ELISA, and the cells were tested for eotaxin-1 expression by real-time PCR.
Eotaxin-1 production was observed on simultaneous stimulation with TNF-alpha and IL-4. This production was inhibited by both tranilast and dexamethasone. Inhibition of eotaxin-1 expression was also observed by real-time PCR.
Eotaxin-1 production from conjunctival fibroblasts was inhibited by both tranilast and dexamethasone. These results suggest that the anti-allergic effect of tranilast may be partly due to the inhibition of eotaxin-1 production from conjunctival fibroblasts.
嗜酸性粒细胞趋化因子是一种对嗜酸性粒细胞具有选择性趋化作用的CC趋化因子，据报道在过敏性结膜疾病中起重要作用。我们之前报道过，结膜成纤维细胞和角膜细胞在受到Th2细胞因子刺激时会产生嗜酸性粒细胞趋化因子。曲尼司特已知具有抗过敏特性。在本研究中，我们检测了抗过敏药物曲尼司特对培养的结膜成纤维细胞产生嗜酸性粒细胞趋化因子-1的抑制作用。
从正常患者获取的结膜成纤维细胞在DMEM/F12培养基中培养。在传代至第五代时，将细胞转移至96孔板中，饥饿24小时后，加入肿瘤坏死因子-α、白细胞介素-4以及曲尼司特或地塞米松。48小时后，通过酶联免疫吸附测定法测量上清液中嗜酸性粒细胞趋化因子-1的浓度，并通过实时聚合酶链反应检测细胞中嗜酸性粒细胞趋化因子-1的表达。
在肿瘤坏死因子-α和白细胞介素-4同时刺激下可观察到嗜酸性粒细胞趋化因子-1的产生。曲尼司特和地塞米松均可抑制这种产生。通过实时聚合酶链反应也观察到嗜酸性粒细胞趋化因子-1表达受到抑制。
曲尼司特和地塞米松均可抑制结膜成纤维细胞产生嗜酸性粒细胞趋化因子-1。这些结果表明，曲尼司特的抗过敏作用可能部分归因于其对结膜成纤维细胞产生嗜酸性粒细胞趋化因子-1的抑制作用。
