TRPA1/M8 agonists upregulate ciliary beating through the pannexin-1 channel in the human nasal mucosa,Molecular Biology Reports

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TRPA1/M8 agonists upregulate ciliary beating through the pannexin-1 channel in the human nasal mucosa
Molecular Biology Reports ( IF 2.6 ) Pub Date : 2022-12-21 , DOI: 10.1007/s11033-022-08201-7
Thi Nga Nguyen _{1,

2} , Yuma Koga ₁ , Tetsuro Wakasugi ₁ , Takuro Kitamura ₁ , Hideaki Suzuki ₁

Affiliation

Background

Nasal breathing is important for maintaining physiological respiration. However, airflow in the nasal cavity has an inherent cooling effect and may suppress ciliary beating, an essential frontline defense in the airway. Nasal airflow is thought to be perceived by thermoreceptors for cool temperatures. We herein investigated the effect of the activation of thermosensitive transient receptor potentials (TRPs) for cool/cold temperatures on ciliary beating to search for a compensatory mechanism.

Methods

Inferior turbinates were collected from patients with chronic hypertrophic rhinitis. Ex vivo ciliary beat frequency (CBF) and ATP release were measured using a high-speed digital video camera and by luciferin-luciferase assay, respectively. Intracellular Ca²⁺ ([Ca²⁺]_i) imaging of isolated ciliated cells was performed using Fluo-8. The nasal mucosae were also subjected to fluorescence immunohistochemistry and real-time RT-PCR for TRPA1/TRPM8.

Results

CBF was significantly increased by adding either cinnamaldehyde (TRPA1 agonist) or l-menthol (TRPM8 agonist). This increase was inhibited by pannexin-1 blockers, carbenoxolone and probenecid. Cinnamaldehyde and l-menthol also increased the ATP release from the nasal mucosa and [Ca²⁺]_i of isolated ciliated cells. Immunohistochemistry detected TRPA1 and TRPM8 on the epithelial surface including the cilia and in the submucosal nasal glands. Existence of these receptors were confirmed at the transcriptional level by real-time RT-PCR.

Conclusions

These results indicate the stimulatory effect of the activation of TRPA1/TRPM8 on ciliary beating in the nasal mucosa, which would be advantageous to maintain airway mucosal defense against the fall of temperature under normal nasal breathing. This stimulatory effect is likely to be mediated by pannexin-1.

中文翻译：

TRPA1/M8 激动剂通过人鼻粘膜中的 pannexin-1 通道上调纤毛跳动

背景

鼻呼吸对于维持生理呼吸很重要。然而，鼻腔内的气流具有固有的冷却作用，并可能抑制纤毛的跳动，而纤毛是气道中重要的前线防御。鼻气流被认为是由温度感受器感知到凉爽的温度。我们在此研究了冷/冷温度下热敏瞬时受体电位（TRP）激活对纤毛跳动的影响，以寻找补偿机制。

方法

下鼻甲是从慢性肥厚性鼻炎患者身上采集的。分别使用高速数码摄像机和荧光素-荧光素酶测定来测量离体纤毛跳动频率（CBF）和ATP释放。使用Fluo-8 对分离的纤毛细胞进行细胞内Ca ²⁺ ([Ca ²⁺ ] _i ) 成像。鼻粘膜还对 TRPA1/TRPM8 进行荧光免疫组织化学和实时 RT-PCR 检测。

结果

添加肉桂醛（TRPA1 激动剂）或 l-薄荷醇（TRPM8 激动剂）可显着增加 CBF。这种增加被 pannexin-1 阻滞剂、carbenoxolone 和丙磺舒抑制。肉桂醛和l-薄荷醇还增加了鼻粘膜的ATP 释放和分离的纤毛细胞的[Ca ²⁺ ] _i 。免疫组织化学检测到上皮表面（包括纤毛）和粘膜下鼻腺中的 TRPA1 和 TRPM8。通过实时 RT-PCR 在转录水平上证实了这些受体的存在。