[1]周　洁,吉　玲,王新华,等.细胞间黏附分子-1在支气管哮喘儿童支气管黏膜中的表达及其对支气管上皮细胞凋亡的影响[J].新乡医学院学报,2019,36(2):126-130.[doi:10.7683/xxyxyxb.2019.02.006]
　ZHOU Jie,JI Ling,WANG Xin-hua,et al.Expression of intercellular adhesion molecule-1 in bronchial mucosa of asthmatic children and its effect on apoptosis of bronchial epithelial cells[J].Journal of Xinxiang Medical University,2019,36(2):126-130.[doi:10.7683/xxyxyxb.2019.02.006]
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细胞间黏附分子-1在支气管哮喘儿童支气管黏膜中的表达及其对支气管上皮细胞凋亡的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
36
期数:
2019年2
页码:
126-130
栏目:
基础研究
出版日期:
2019-02-05

文章信息/Info

Title:
Expression of intercellular adhesion molecule-1 in bronchial mucosa of asthmatic children and its effect on apoptosis of bronchial epithelial cells
作者:
周　洁1; 吉　玲2; 王新华1; 周栩平1
（1.驻马店市中心医院新生儿科，河南　驻马店　463000；2.郑州大学第三附属医院新生儿科，河南　郑州　450000）
Author(s):
ZHOU Jie1; JI Ling2; WANG Xin-hua1; ZHOU Xu-ping1
（1.Department of Neonatology，the Central Hospital of Zhumadian City，Zhumadian 463000，Henan Province，China；2.Department of Neonatology，the Third Affiliated Hospital of Zhengzhou University，Zhengzhou 450000，Henan Province，China）
关键词:
哮喘; 细胞间黏附分子-1; 支气管上皮细胞; 细胞凋亡
Keywords:
asthma; intercellular adhesion molecule-1; bronchial epithelial cells; apoptosis
分类号:
R562.2+5
DOI:
10.7683/xxyxyxb.2019.02.006
文献标志码:
A
摘要:
目的　探讨细胞间黏附分子-1（ICAM-1）在支气管哮喘儿童支气管黏膜中的表达及其对支气管上皮细胞凋亡的影响。方法　收集驻马店市中心医院2014年5月至2017年12月收治的28例哮喘儿童的支气管黏膜组织标本，同时选取28例支气管扩张非哮喘儿童的支气管黏膜组织作为对照；采用实时荧光定量聚合酶链反应（qRT-PCR）和Western blot法检测支气管哮喘儿童支气管黏膜组织中ICAM-1 mRNA和蛋白表达。将正常培养的支气管上皮细胞16HBE分为对照组、感染组、阴性组和干扰组。对照组细胞正常培养，不做任何处理；感染组细胞感染呼吸道合胞病毒（RSV），但不进行转染；阴性组细胞转染siRNA-control后感染RSV；干扰组细胞转染ICAM-1 siRNA后感染RSV。采用qRT-PCR和Western blot法检测各组细胞中ICAM-1 mRNA和蛋白表达，流式细胞术检测细胞凋亡情况，酶联免疫吸附试验检测细胞培养基上清液中白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）水平，Western blot法检测细胞中活化的含半胱氨酸的天冬氨酸蛋白水解酶3（Cleaved Caspase-3）、核因子-κB p65（NF-κB p65）表达。结果　哮喘支气管黏膜组织中ICAM-1 mRNA和蛋白表达显著高于正常支气管黏膜组织（P<0.05）。感染组、阴性组、干扰组细胞中ICAM-1 mRNA和蛋白表达显著高于对照组（P<0.05）。干扰组细胞中ICAM-1 mRNA和蛋白表达显著低于感染组和阴性组（P<0.05）。感染组、阴性组、干扰组细胞凋亡率显著高于对照组（P<0.05）。干扰组细胞凋亡率显著低于感染组和阴性组（P<0.05）。感染组、阴性组、干扰组细胞培养基上清液中IL-1β、TNF-α水平显著高于对照组（P<0.05）。干扰组细胞培养基上清液中IL-1β、TNF-α水平显著低于感染组和阴性组（P<0.05）。感染组、阴性组、干扰组细胞中Cleaved Caspase-3、NF-κB p65蛋白水平显著高于对照组（P<0.05）。干扰组细胞中Cleaved Caspase-3、NF-κB p65蛋白水平显著低于感染组和阴性组（P<0.05）。结论　ICAM-1在哮喘儿童支气管黏膜组织中表达上调，抑制ICAM-1可以降低RSV诱导的支气管上皮细胞凋亡，降低支气管上皮细胞中Cleaved Caspase-3、NF-κB p65蛋白表达，减少细胞分泌炎性因子。
Abstract:
Objective　To investigate the expression of intercellular adhesion molecule-1（ICAM-1） in bronchial mucosa of asthmatic children and its effect on the apoptosis of bronchial epithelial cells.Methods　Twenty-eight bronchial mucosa of asthmatic children in the Central Hospital of Zhumadian City from May 2014 to December 2017，and twenty-eight bronchial mucosa of bronchiectasis without asthmatic children were selected as control.The expression of ICAM-1 mRNA and protein in bronchial mucosa of asthmatic children were detected by quantitative real-time polymerase chain reaction（qRT-PCR） and Western blot.The bronchial epithelial cells who was cultured normally were divided into control group，infection group，negative group and interference group.The cells in the control group were cultured normally without any treatment.The cells in the infection group were infected with respiratory syncytial virus（RSV），but they were not transfected.The cells in the negative group were transfected with siRNA-control and then they were infected with RSV.The cells in the interference group were transfected with ICAM-1 siRNA and then they were infected with RSV.The expressions of ICAM-1 mRNA and protein in each group were determined by qRT-PCR and Western blot,and the cell apoptosis was measured by flow cytometry.The interleukin （IL）- 1β and tumor necrosis factor（TNF）-α levels in supernatant of cell culture medium were detected by enzyme linked immunosorbent assay.The expressions of cleaved cysteinyl aspartate specific proteinase (Cleaved Caspase-3) and nuclear factor κB （NF-κB）p65 in cells were determined by Western blot.Results　The expressions of ICAM-1 mRNA and protein in bronchial mucosa of asthmatic patients were significantly higher than those in normal bronchial mucosa （P<0.05）.The expressions of ICAM-1 mRNA and protein in the infection group，negative group and interference group were significantly higher than those in the control group （P<0.05）.The expressions of ICAM-1 mRNA and protein in the interference group were significantly lower than those in the infection group and the negative group （P<0.05）.The cell apoptosis rate of the infection group，negative group and interference group was significantly higher than that of the control group （P<0.05）.The cell apoptosis rate in the interference group was significantly lower than that in the infection group and the negative group （P<0.05）.The levels of IL-1β and TNF-α in supernatant of cell culture medium in the infection group，the negative group and the interference group were significantly higher than those in the control group （P<0.05）.The levels of IL-1β and TNF-α in supernatant of cell culture medium in the interference group were significantly lower than those in the infection group and the negative group （P<0.05）.The levels of Cleaved Caspase-3 and NF-κB p65 protein in the infection group，the negative group and the interference group were significantly higher than those in the control group （P<0.05）.The levels of Cleaved Caspase-3 and NF-κB p65 protein in the interference group were significantly lower than those in the infection group and the negative group （P<0.05）.Conclusion　The ICAM-1 expression in bronchial mucosa of asthmatic children is up-regulated.Inhibiting ICAM-1 can reduce the level of apoptosis induced by RSV in bronchial epithelial cells，down-regulate the levels of Cleaved Caspase-3 and NF-κB p65 protein in bronchial epithelial cells,and reduce the secretion of inflammatory factors.

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更新日期/Last Update: 2019-02-05