IL-17、IL-22和IL-10在支气管哮喘患儿外周血单个核细胞中的表达及意义

doi:10.3969/j.issn.1673-8640.2019.01.006

摘要/Abstract

摘要：

目的探讨白细胞介素（IL）-17、IL-22和IL-10在支气管哮喘患儿外周血单个核细胞中的表达及意义。方法选取75例哮喘患儿,根据病情分为发作组（41例）和缓解组（34例）,以体检健康儿童40名作为正常对照组。分离所有对象的外周血单个核细胞,分别采用实时定量聚合酶链反应（PCR）及免疫印迹法检测外周血单个核细胞中IL-17、IL-22和IL-10基因及蛋白的表达,同时检测所有对象的肺功能[第1秒用力呼气量（FEV1）、FEV1/用力肺活量（FVC）比值]。采用Pearson相关分析评估各项目间的相关性。结果发作组FEV1和FEV1/FVC比值均低于缓解组和正常对照组（P<0.05）,且缓解组低于正常对照组（P<0.05）。发作组外周血单个核细胞中IL-17、IL-22 mRNA和蛋白的相对表达量均高于缓解组和正常对照组（P<0.05）,且缓解组高于正常对照组（P<0.05）;而IL-10 mRNA和蛋白的相对表达量均低于缓解组和正常对照组（P<0.05）,且缓解组低于正常对照组（P<0.05）。哮喘患儿外周血单个核细胞中IL-17、IL-22 mRNA和蛋白的相对表达量与IL-10 mRNA和蛋白的相对表达量、FEV1、FEV1/FVC比值均呈负相关（P<0.05）,IL-10 mRNA和蛋白的相对表达量与FEV1、FEV1/FVC比值均呈正相关（P<0.05）。结论哮喘患儿外周血单个核细胞中IL-17、IL-22表达上调,而IL-10表达下调,且与患儿病情有关,提示辅助性T细胞17（Th17）/调节性T细胞（Treg）比例失调可能参与了哮喘发病。

关键词: 白细胞介素17, 白细胞介素22, 白细胞介素10, 支气管哮喘, 儿童

Abstract:

Objective To investigate the expressions of interleukin （IL）-17,IL-22 and IL-10 in peripheral blood mononuclear cells in children with bronchial asthma. Methods A total of 75 children with asthma were classified into exacerbation group （41 cases） and remission group （34 cases）. A total of 40 healthy children were enrolled as control group. The peripheral blood mononuclear cells were isolated. The expressions of IL-17,IL-22 and IL-10 genes in peripheral blood mononuclear cells were determined by real-time quantitation polymerase chain reaction （PCR）. The expressions of IL-17,IL-22 and IL-10 proteins in peripheral blood mononuclear cells were determined by western blotting. The forced expiratory volume in 1 second （FEV1） and the ratio of FEV1/forced vital capacity （FVC） were determined. The correlations among the variables were analyzed by Pearson correlation analysis. Results The FEV1 and FEV1/FVC in exacerbation group were lower than those in remission group and control group （P<0.05）,and those in remission group were lower than those in control group（P<0.05）. The relative expression levels of IL-17 mRNA and IL-22 mRNA in peripheral blood mononuclear cells in exacerbation group were higher than those in remission group and control group （P<0.05）,and those in remission group were higher than those in control group （P<0.05）. However, the relative expression level of IL-10 mRNA was lower than those in remission group and control group （P<0.05）,and that in remission group was lower than that in control group （P<0.05）. The relative expression levels of IL-17 mRNA, IL-17 protein and IL-22 mRNA, IL-22 protein in peripheral blood mononuclear cells in asthmatic children were negatively correlated with the relative expression levels of IL-10 mRNA and IL-10 protein,FEV1 and FEV1/FVC（P<0.05）,and the relative expression levels of IL-10 mRNA and IL- 10 protein were positively correlated with FEV1 and FEV1/FVC （P<0.05）. Conclusions The expressions of IL-17 and IL-22 in peripheral blood mononuclear cells of children with asthma are upregulated,while the expression of IL- 10 is downregulated,which are related with disease status. It prompts that T helper cell 17（Th17）/regulatory T cell （Treg） imbalance might be involved in the pathogenesis of asthma.

Key words: Interleukin 17, Interleukin 22, Interleukin 10, Bronchial asthma, Children

中图分类号:

R446.62

陆光辉, 褚志华, 周发为, 杨年安, 杨洁, 王姣平, 曾守逵. IL-17、IL-22和IL-10在支气管哮喘患儿外周血单个核细胞中的表达及意义[J]. 检验医学, 2019, 34(1): 28-32.

LU Guanghui, CHU Zhihua, ZHOU Fawei, YANG Nianan, YANG Jie, WANG Jiaoping, ZENG Shoukui. Expressions of IL-17,IL-22 and IL-10 in peripheral blood mononuclear cells in children with bronchial asthma[J]. Laboratory Medicine, 2019, 34(1): 28-32.

图/表 5

参考文献 12

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基因名称	引物序列（5'~3'）	长度（bp）
IL-17	上游：GACTCTCCACCGCAATGAA	205
	下游：ACACCCACCAGCATCTTCTC
IL-22	上游：ACAGCAAATCCAGTTCTCCAA	144
	下游：TCCAGAGGAATGTGCAAAAG
IL-10	上游：TGGGGGAGAACCTGAAGAC	178
	下游：CCTTGCTCTTGTTTTCACAGG
GAPDH	上游：TGCTGAGTATGTCGTGGAGTCT	494
	下游：AGAAGGGGCGGAGATGAT

基因名称	引物序列（5'~3'）	长度（bp）
IL-17	上游：GACTCTCCACCGCAATGAA	205
	下游：ACACCCACCAGCATCTTCTC
IL-22	上游：ACAGCAAATCCAGTTCTCCAA	144
	下游：TCCAGAGGAATGTGCAAAAG
IL-10	上游：TGGGGGAGAACCTGAAGAC	178
	下游：CCTTGCTCTTGTTTTCACAGG
GAPDH	上游：TGCTGAGTATGTCGTGGAGTCT	494
	下游：AGAAGGGGCGGAGATGAT

组别	例数	性别		年龄（岁）	FEV1（%）	FEV1/FVC比值（%）
组别	例数	男[例（%）]	女[例（%）]	年龄（岁）	FEV1（%）	FEV1/FVC比值（%）
发作组	41	23（56.1）	18（43.9）	6.2±1.9	73.7±9.5^*#	68.3±7.8^*#
缓解组	34	20（58.8）	14（41.2）	6.5±2.3	89.8±6.7^*	87.7±9.9^*
正常对照组	40	24（60.0）	16（40.0）	6.3±2.1	93.1±3.9	96.5±11.3
F值		0.133		1.275	15.382	24.183
P值		0.936		0.118	<0.001	<0.001

组别	例数	性别		年龄（岁）	FEV1（%）	FEV1/FVC比值（%）
组别	例数	男[例（%）]	女[例（%）]	年龄（岁）	FEV1（%）	FEV1/FVC比值（%）
发作组	41	23（56.1）	18（43.9）	6.2±1.9	73.7±9.5^*#	68.3±7.8^*#
缓解组	34	20（58.8）	14（41.2）	6.5±2.3	89.8±6.7^*	87.7±9.9^*
正常对照组	40	24（60.0）	16（40.0）	6.3±2.1	93.1±3.9	96.5±11.3
F值		0.133		1.275	15.382	24.183
P值		0.936		0.118	<0.001	<0.001

组别	例数	IL-17		IL-22		IL-10
组别	例数	mRNA	蛋白	mRNA	蛋白	mRNA	蛋白
发作组	41	2.53±0.10^*#	0.81±0.06^*#	3.36±0.15^*#	0.89±0.08^*#	3.05±0.16^*#	0.40±0.06^*#
缓解组	34	1.92±0.11^*	0.63±0.08^*	2.92±0.17^*	0.68±0.06^*	3.57±0.18^*	0.72±0.10^*
正常对照组	40	1.64±0.09	0.35±0.05	2.15±0.14	0.39±0.04	4.02±0.21	0.93±0.11
F值		39.541	52.939	48.273	67.184	37.195	30.999
P值		<0.001	<0.001	<0.001	<0.001	<0.001	<0.001