Cold-induced signal transduction mechanism in airway epithelial cells

李敏超; 何叶; Juliy M. Perelman; Victor P. Kolosov; 周向东

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Cold-induced signal transduction mechanism in airway epithelial cells

更新时间：2026-01-09

- Cold-induced signal transduction mechanism in airway epithelial cells
- Journal of Army Medical University Vol. 36, Issue 20, Pages: 2071-2076(2014)
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  CLC： R322.3;R329.26;R394.2
- Published：2014
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李敏超,何叶,Juliy M Perelman,Victor P Kolosov,周向东.人气道上皮细胞感知冷刺激的受体机制[J].陆军军医大学学报(原第三军医大学学报),2014,36(20):2071-2076.

　Li Minchao,He Ye,Juliy M. Perelman,Victor P. Kolosov,Zhou Xiangdong.Cold-induced signal transduction mechanism in airway epithelial cells[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(20):2071-2076.
李敏超,何叶,Juliy M Perelman,Victor P Kolosov,周向东.人气道上皮细胞感知冷刺激的受体机制[J].陆军军医大学学报(原第三军医大学学报),2014,36(20):2071-2076. DOI：

　Li Minchao,He Ye,Juliy M. Perelman,Victor P. Kolosov,Zhou Xiangdong.Cold-induced signal transduction mechanism in airway epithelial cells[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(20):2071-2076. DOI：

摘要

目的探讨温度感受器瞬时受体电位蛋白-8（transient receptor potential melastatin 8，TRPM8）在人气道上皮细胞感受冷刺激过程中发挥的作用及其机制。方法用薄荷醇及冷空气（18 ℃）刺激人气道上皮16HBE细胞：①以TRPM8 通道特异性拮抗剂BCTC、TRPM8 shRNA为干预手段，动态观察在薄荷醇、冷刺激作用下细胞内Ca

荧光强度变化来监测气道上皮细胞上TRPM8的功能；②以磷脂酰肌醇4，5-二磷酸（phosphatidylinositol 4，5-bisphosphate，PIP2）荧光分子探针PLCδ1-PH-GFP转染细胞，通过动态观察各组细胞内PLCδ1-PH-GFP从细胞膜至细胞质的荧光转位变化来检测PIP2的动态变化来研究冷刺激作用下TRPM8的信号通路。结果 ①细胞内相对钙离子浓度变化：薄荷醇组处理4 min后、冷刺激组处理6 min后分别为（5.80±0.34）、（5.02±0.36），明显高于对照组（1.04±0.12）、（1.03±0.08）（P0.05），1 mmol/L 薄荷醇组+BCTC组、1 mmol/L薄荷醇组+转染TRPM8 shRNA组最高值分别为（2.68±0.19）、（1.08±0.16），明显低于1 mmol/L 薄荷醇组，18 ℃+ BCTC组、18 ℃+转染TRPM8 shRNA组最高值分别为（1.77±0.22）、（1.02±0.18），明显低于18 ℃组（均P0.05）；②PLCδ1-PH-GFP从细胞膜至细胞质的转位变化（Fm/Fc值）：1 mmol/L薄荷醇组、冷刺激组最高值分别为（0.90±0.01）、（0.90±0.01），明显高于对照组最高值（1.01±0.02）（P0.05），细胞处理200 s 时1 mmol/L薄荷醇+BCTC15 μmol/L组、1 mmol/L薄荷醇组+转染TRPM8 shRNA组分别为（0.86±0.03）、（0.89±0.02），明显低于1 mmol/L薄荷醇组（0.36±0.06）（P0.05），细胞处理220 s时18 ℃+BCTC15 μmol/L组、18 ℃+转染TRPM8 shRNA组分别为（0.82±0.02）、（0.93±0.01），明显低于18 ℃组（0.40±0.06）（P0.05）。结论 TRPM8受体是人气道上皮细胞感受冷刺激的主要受体，并通过TRPM8→Ca

→ PLC→PIP2信号通路引起气道黏液高分泌等反应。

Abstract

Objective

To explore the role of transient receptor potential melastatin 8 (TRPM8) receptor and related cold-induced signal transduction mechanism in airway epithelial cells. Methods The 16 HBE human airway epithelial cells were treated with cold temperature (18 ℃) and menthol. By means of Ca

imaging

we explored the function of TRPM8 when the cells were pretreated with TRPM8 channel antagonist BCTC and exon-18-specific TRPM8 shRNA. Hydrolysis of phosphatidylinositol 4

5-bisphosphate (PIP2) were characterized by means of spatiotemporal dynamics of phospholipase C (PLC) δ1-pleckstrin homology-green fluorescent protein (δ1-PH-GFP). Results The concentration of intercellular calcium in the menthol and cold air treatment group (5.80±0.34

5.02±0.36) higher than that in the control group (0.94±0.12) (P0.05). The intracellular calcium concentration was in the menthol+BCTC and menthol+TRPM8 shRNA groups (2.68±0.19

1.08±0.16) than in the menthol treatment group (P0.05). The intracellular calcium concentration in the cold air + BCTC and cold air+TRPM8 shRNA groups (1.77±0.22

1.02±0.18) were lower than that in the cold air treatment group (P0.05). The translocation of PLC δ1-PH-GFP from the membrane to the cytosol (Fm/Fc) in the menthol treatment group and cold air treatment group (0.90±0.01

0.90±0.01) were higher than that in the control group (1.01±0.02) (P0.05). Fm/Fc was lower in the menthol+BCTC group and TRPM8 shRNA+menthol group (0.86±0.03

0.89±0.02) than in the menthol treatment group (P0.05)

and also lower in the cold air+BCTC group and cold air+TRPM8 shRNA group (0.82±0.02

0.93±0.01) than in the cold air treatment group (P0.05). Conclusion TRPM8 receptor is involved in the response to cold temperature in airway epithelial cells

and contributes to mucus hypersecretion through the TRPM8→Ca

→ PLC→PIP2 signaling pathway.

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