TRPC1是人气道上皮细胞感受压力的受体

基础医学与临床 ›› 2015, Vol. 35 ›› Issue (5): 626-631.

TRPC1是人气道上皮细胞感受压力的受体

何叶¹,李娜²,黎力³,李敏超²

1. 重庆医科大学附二院
2. 重庆医科大学附属第二医院
3. 重庆医科大学附属第二医院胸心外科

收稿日期:2014-11-13 修回日期:2015-03-16 出版日期:2015-05-05 发布日期:2015-04-28
通讯作者: 李敏超 E-mail:liminc66@yahoo.com.cn
基金资助:
气道内高压诱导气道重塑的信号链接机制

TRPC1 is an important stretch-activated cation receptor in airway epithelial cells

Received:2014-11-13 Revised:2015-03-16 Online:2015-05-05 Published:2015-04-28
Supported by:
signaling menchanisms of airway remodeling induced by airway high pressure

摘要/Abstract

摘要： 目的探讨机械敏感瞬时受体电位蛋白-1（TRPC1）在人气道上皮细胞感受压力过程中的作用。方法 1）收集人肺癌患者肺癌旁5 cm以外的无病变组织，根据临床表现分为正常组、慢性阻塞性肺病组（COPD）和哮喘组，通过细胞免疫组织化学、Western-blot及实时荧光定量聚合酶链反应（qRT-PCR）检测人气道上皮细胞内TRPC1表达水平。2）TRPC1 siRNA、NC siRNA转染16HBE细胞，细胞免疫荧光技术检测基因沉默效率。3）将16HBE细胞分为对照组、单纯刺激组、刺激+TRPC1 siRNA转染组，以细胞牵张刺激系统给予刺激，激光共聚焦观察细胞内钙离子浓度变化。结果 1）COPD组、哮喘组TRPC1蛋白表达及转录水平明显高于对照组（均P<0.05）2） TRPC1 siRNA转染组细胞内TRPC1荧光强度明显低于对照组（均 P<0.05），NC siRNA组较正常组无明显差别。3）单纯刺激组细胞内钙离子荧光强度显著高于对照组（P<0.05），而刺激+TRPC1 siRNA转染组钙离子荧光强度与对照组无明显差异。结论 TRPC1是人气道上皮细胞内重要的压力敏感受体。

关键词: 气道压力, 气道上皮细胞, 压力敏感离子通道, TRPC1

Abstract: Objective To explore the role of transient receptor potential Canonical 1 in the pressure-sensing mechanism in airway epithelial cells. Methods Collect human lung tissue, and divided into control group, COPD group and asthma group. The level of TRPC1 protein and mRNA in human bronchial epithelial was detected by immunohistochemistry, western blot and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). The level of TRPC1 protein was detected by immunofluorescence after 16HBE cells were transfected by TRPC1 siRNA、NC siRNA. Then 16 HBE human airway epithelial cells were divided into control group, stretch group, stretch+TRPC1 siRNA group, by means of Ca2+ imaging, we explore the activation of TRPC1 in 16 HBE after they were stimulated with cyclic stretch for 6 minutes. Results The level of TRPC1 protein and mRNA was significant higher in COPD and asthma group compared with control group ( P<0.05). The level of TRPC1 protein in TRPC1 siRNA group was lower than control group( P<0.05), NC siRNA group is similar with control group. The concentration of intercellular calcium in stretch group was higher than control group ( P<0.05), but stretch+TRPC1 siRNA group was similar with control group. Conclusion TRPC1 protein plays an important role in the response to stress stimulation in airway epithelial cells.

Key words: airway pressure, airway epithelial cell, pressure sensitive cation channel, TRPC1

中图分类号:

R562.2

何叶李娜黎力李敏超. TRPC1是人气道上皮细胞感受压力的受体[J]. 基础医学与临床, 2015, 35(5): 626-631.