External stimuli such as cigarette smoke and house dust mite are often involved in the development and exacerbation of asthma. These risk factors could activate or sensitize transient receptor potential channel ankyrin 1 (TRPA1), which are primarily expressed in neuronal structures but also in non-neuronal cells such as fibroblasts. However, the role of non-neuronal TRPA1 in the pathophysiology of airway diseases including asthma remains unclear. We investigated TRPA1 expression on human fibroblast cells and whether inflammatory mediators could modulate its function. This study utilized human lung fibroblast cell lines, Medical Research Council cell strain 5 (MRC-5) and HF19 cells frequently used on experimental studies regarding allergic and respiratory disorders. The human lung fibroblasts were stimulated with house dust mite (Der p1) or tumor necrosis factor alpha (TNF-α) for 24 h, and we quantified TRPA1 mRNA and protein by qRT-PCR and western blot analysis, respectively. TRPA1 mRNA expressions were upregulated after TNF-α treatment. Calcium imaging analysis revealed that TNF-α treatment apparently sensitized TRPA1-mediated calcium influx by TRPA1 agonist allyl isothiocyanate (AITC) and the selective TRPA1 channel blocker HC-030031 effectively reduced the calcium response. Lastly, TRPA1 activation was not only involved in increased IL-8 cytokine release, but also in upregulating gene expression of matrix metalloprotease 9 (MMP9) in the human lung fibroblasts treated with TNF-α Together, these results indicate that presence of inflammatory mediators such as TNF-α could upregulate the non-neuronal expression of TRPA1 on fibroblasts which may aggravate further the release of inflammatory cytokines observed in human airway diseases.

中文翻译：

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炎症刺激使 TRPA1 通道敏感以增加人肺成纤维细胞中细胞因子的释放

外部刺激如香烟烟雾和屋尘螨通常与哮喘的发生和恶化有关。这些风险因素可以激活或敏化瞬时受体电位通道锚蛋白 1 (TRPA1)，其主要在神经元结构中表达，但也在非神经元细胞（如成纤维细胞）中表达。然而，非神经元 TRPA1 在气道疾病（包括哮喘）的病理生理学中的作用仍不清楚。我们研究了人成纤维细胞上的 TRPA1 表达以及炎症介质是否可以调节其功能。本研究利用人肺成纤维细胞系、医学研究委员会细胞株 5 (MRC-5) 和 HF19 细胞，这些细胞经常用于有关过敏和呼吸系统疾病的实验研究。用屋尘螨（Der p1）或肿瘤坏死因子α（TNF-α）刺激人肺成纤维细胞24小时，我们分别通过qRT-PCR和蛋白质印迹分析定量TRPA1 mRNA和蛋白质。TNF-α处理后TRPA1 mRNA表达上调。钙成像分析显示，TNF-α 治疗明显使 TRPA1 激动剂异硫氰酸烯丙酯 (AITC) 和选择性 TRPA1 通道阻滞剂 HC-030031 介导的钙流入敏感，有效降低了钙反应。最后，TRPA1 激活不仅涉及增加 IL-8 细胞因子的释放，而且还上调用 TNF-α 处理的人肺成纤维细胞中基质金属蛋白酶 9（MMP9）的基因表达。