机械压力敏感性通道蛋白表达水平与COPD患者气道重塑的关联性分析

doi:10.13481/j.1671-587x.20160217

摘要/Abstract

摘要：

目的: 探讨机械压力敏感性瞬时受体电位C1蛋白(TRPC1)在慢性阻塞性肺疾病(COPD)患者支气管上皮细胞中的表达情况,阐述TRRC1蛋白参与COPD气道重塑的分子机制。方法: 选择欲行纤维支气管镜检查的患者64例作为非手术组,根据COPD诊治指南分为轻中度COPD患者(COPD组)40例和无气道慢性炎症性疾病患者(对照组)24例。分别检测第1秒用力呼气容积/用力肺活量(FEV₁/FVC)和其实测值占预计值百分比(FEV₁%pred);在纤维支气管镜引导下得到肺泡灌洗液(BALF),采用ELISA法测定BALF中TRPC1、基质金属蛋白酶-9(MMP-9)和转化生长因子-β₁(TGF-β₁)的表达水平;分析BALF中TRPC1表达量与FEV₁%pred、FEV₁/FVC、MMP-9及TGF-β₁表达水平的相关性。选取行肺叶切除术患者的肺组织标本17例作为手术组,其中COPD组8例和对照组(无慢性气道炎症性疾病)9例。高分辨CT下测量同一部位支气管重塑指标,测定壁厚与外径比率(TDR%)、气道壁面积占总横截面积比(WA%),对比气道结构变化。免疫组织化学法检测TRPC1蛋白在人支气管上皮细胞中的分布特点,采用Imagepro-plus 6.0软件比较COPD组与对照组气道上皮细胞中TRPC1蛋白的表达差异,Western blotting法测定TRPC1表达水平。分析气道支气管上皮细胞中TRPC1表达水平与TDR%和WA%的相关性。结果: 非手术组患者肺功能,与对照组比较,COPD组患者FEV1%pred和FEV1/FVC均下降(P<0.05),TRPC1、MMP-9和TGF-β₁表达水平升高(P<0.05)。偏相关分析,COPD组TRPC1表达水平与FEV₁%pred(r=-0.34,P=0.002)和FEV₁/FVC(r=-0.38,P=0.004)呈负相关关系,与MMP-9(r=0.36,P=0.004)和TGF-β₁(r=0.61,P=0.002)表达水平呈正相关关系。手术组组织标本中TRPC1蛋白主要分布于支气管上皮细胞腔内侧,表达于柱状细胞或杯状细胞胞浆内;其中COPD组累积光密度(IOD)/面积(area)比值高于对照组(P<0.05);Western blotting法检测,COPD组支气管黏膜上皮内TRPC1蛋白表达水平高于对照组(P<0.05)。偏相关分析,COPD组TRPC1表达水平与TDR%(r=0.59,P=0.002)和WA%(r=0.60,P=0.002)呈正相关关系。结论: COPD患者支气管黏膜上皮细胞中TRPC1被激活并高水平表达,并与肺功能受损程度、气道重塑相关因子表达水平呈正相关关系,说明感知压力的TRPC1通道可促进COPD的发生发展。

关键词: 慢性阻塞性肺疾病, 瞬时受体电位C1蛋白, 气道重塑

Abstract:

Objective: To explore the expression of pressure-sensitive transient receptor potential channel (TRPC1) in bronchial epithelium cells of the patients with chronic obstructive pulmonary disease(COPD), and to explain the molecular mechanism about the participation of TRPC1 in airway remodeling.Methods: Sixty-four patients who needed fiberoptic bronchoscope for diagnosis were selected.All the patients were used as non-operation group and divided into COPD group(40 cases of mild to moderate grade) and control group(24 cases without chronic airway inflammatory disease) according to Respiratory Disease Guideline.The level of the forced expiatory volume/forced vital capacity (FEV₁/FVC) in the first second and forced expiatory volume in the first second of the expected value (FEV₁% pred) were detected by lung function test.The expression levels of TRPC1, matrix metalloproteinase(MMP-9) and transforming growth factor-β₁(TGF-β₁) in bronchoalveolar liquid(BALF) were measured by ELISA.The relevance among expression level of TRPC1 and levels of FEV₁/FVC, FEV₁% pred, MMP-9, TGF-β1 was analyzed.Seventeen lung samples from the patients underwent pulmonary lobectomy were selected. All the patients were used as operation group and divided into COPD group(8 cases of mild to moderate grade) and control group(9 cases without chronic airway inflammatory disease). Then indictator relevance to airway remodeling, such as thickness-diameter ratio(TDR%), percentage of wall thickness(WA%) were measured.The immunohistochemistry experiment was used to detect the distribution characteristics of TRPC1 protein in human bronchial epithelial cells.The Imagepro-plus 6.0 software was used to analyze the expression difference of TRPC1 protein between COPD and control groups.Western blotting method was used to detect the expression level of TRPC1.The correlation between the expression level of TRPC1 and TDR%, WA% was analyzed.Results: The lung function detection results showed that in non-operation group, the levels of FEV₁% pred and FEV₁/FVC in COPD group were decreased compared with control group(P<0.05).The expression levels of TRPC1, MMP-9 and TGF-β1 in COPD group were increased compared with control group(P<0.05). The expression level of TRPC1 was negatively correlated with FEV₁%pred(r=-0.34, P=0.002) and FEV1/FVC(r=-0.38, P=0.004).The expression level of TRPC1 was positively correlated with MMP-9(r=0.36, P=0.004) and TGF-β₁ (r=0.61, P=0.002).The immunohistochemistry results in operation group displayed that the TRPC1 protein was mainly distributed in the epithelial columnar cell nucleus and the cell membrane near lumen within the bronchial epithelium.The ratio of integrated optical density(IOD) to area within the bronchial epithelium in COPD group was significantly higher than that in control group(P<0.05).The expression level of TRPC1 protein detected by Western blotting method in COPD group was higher than that in control group(P<0.05).The expression level of TRPC1 was positively correlated with TDR%(r=0.59, P=0.002) and WA%(r=0.60, P=0.002).Conclusion: The TRPC1 channel in human bronchial epithelial cells of COPD patients is actived and up-regulated.The expression level of TRPC1 channel is positively correlated with impared extent of lung function and the expression levels of cytokines relevant to airway remodeling.All above imply that pressure-sensitive TRPC1 channel could promote the development of airway remodeling.

Key words: chronic obstructive pulmonary disease, transient receptor potential canonical 1, airway remodeling

中图分类号:

R562

李娜, 何叶, 李敏超. 机械压力敏感性通道蛋白表达水平与COPD患者气道重塑的关联性分析[J]. 吉林大学学报(医学版), 2016, 42(02): 277-282.

LI Na, HE Ye, LI Minchao. Analysis on correlation between expression level of pressure-sensitive channel protein and airway remodeling in COPD patients[J]. Journal of Jilin University Medicine Edition, 2016, 42(02): 277-282.

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