哮喘患儿外周血单个核细胞中NLRP3炎症小体及血清中IL-1β和IL-18表达变化及其意义

doi:10.13481/j.1671-587x.20190121

摘要/Abstract

摘要： 目的：观察哮喘患儿外周血单个核细胞中NOD样受体热蛋白结构域相关蛋白3（NLRP3）炎症小体表达水平及血清中下游因子白细胞介素1β（IL-1β）和白细胞介素18（IL-18）水平变化，探讨其对患儿病情评估的意义。方法：176例哮喘患儿根据临床表现分为急性发作期组（n=91）、慢性持续期组（n=49）和缓解期组（n=36），同期从门诊体检中心选取健康儿童60名作为对照组，采用肺功能仪检查各组研究对象肺功能。采用实时荧光定量PCR法检测各组研究对象外周血单个核细胞中NLRP3、含有CARD结构域的凋亡相关斑点样蛋白（ASC）和含半胱氨酸的天冬氨酸蛋白水解酶1（Caspase-1） mRNA表达水平，采用酶联免疫吸附（ELISA）实验检测各组研究对象血清中IL-1β和IL-18水平。结果：与对照组比较，急性发作期组、慢性持续期组和缓解期组哮喘患儿第1秒用力呼气容积占预计值百分比（FEV1%）和第1秒用力呼气容积所占肺活量比值（FEV1/FVC）均降低，且急性发作期组<慢性持续期组<缓解期组，组间比较差异有统计学意义（P<0.05）；哮喘患儿外周血单个核细胞中NLRP3、ASC和Caspase-1 mRNA表达水平及血清中IL-1β和IL-18水平均高于对照组（P<0.01）；急性发作期组患儿外周血单个核细胞中NLRP3、ASC和Caspase-1 mRNA表达水平及血清中IL-1β和IL-18水平高于慢性持续期组和缓解期组（P<0.05），且慢性持续期组患儿高于缓解期组（P<0.05）。Pearson相关分析，哮喘患儿外周血单个核细胞中NLRP3 mRNA表达水平与ASC、Caspase-1 mRNA表达水平和血清中IL-1β、IL-18水平均呈正相关关系（P<0.05），而与FEV1%和FEV1/FVC呈负相关关系（P<0.05）；ASC mRNA表达水平与Caspase-1 mRNA表达水平和血清中IL-1β、IL-18水平呈正相关（P<0.05），而与FEV1%和FEV1/FVC呈负相关关系（P<0.05）；Caspase-1 mRNA表达水平与血清中IL-1β和IL-18水平呈正相关关系（P<0.05），而与FEV1%和FEV1/FVC呈负相关关系（P<0.05）；血清中IL-1β水平与FEV1%和FEV1/FVC呈负相关关系（P<0.05）；IL-18水平与FEV1%和FEV1/FVC呈负相关关系（P<0.05）。结论：哮喘患儿外周血NLRP3炎症小体及下游因子IL-1β和IL-18水平升高，且与哮喘患儿的临床分期有关，NLRP3炎症小体通路可能促进了儿童哮喘的发病过程。

关键词: 支气管哮喘, NOD样受体热蛋白结构域相关蛋白3, 炎症小体, 白细胞介素1β, 白细胞介素18

Abstract: Objective: To investigate the changes of expressions of NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome in the peripheral blood mononuclear cells and downstream factors interleukin-1β (IL-1β) and interleukin-18 (IL-18) in serum in the children with asthma, and to explore their significances on assessing the condition of the children.Methods: A total of 176 cases of children with asthma were divided into acute exacerbation group (n=91), chronic persistent group (n=49) and clinical remission group (n=36) according to the clinical manifestation. During the same period, 60 healthy children were selected from the outpatient physical examination center as control group.The pulmonary function of children was checked with lung function instrument. The expression levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) and cysteinyl aspartate-specific proteinase-1 (Caspase-1) mRNA in peripheral blood mononuclear cells of the subjects in various groups were detected by using real-time quantitative PCR. The serum levels of IL-1β and IL-18 of the subjects in various groups were detected by using enzyme-linked immunosorbent assay (ELISA).Results: Compared with control group, the forced expiratory volume in 1 second percentage of predicted value (FEV1%) and fixed ratio of forced expiratory volume in the first second/forced vital capacity (FEV1/FVC) of the children in acute exacerbation, chronic persistent and clinical remission groups were decreased (P<0.05);acute exacerbation groupP<0.05).The levels of NLRP3, ASC and Caspase-1 mRNA in the peripheral blood mononuclear cells and serum levels of IL-1β and IL-18 in the children with asthma were higher than those in control group (P<0.01).The expression levels of NLRP3, ASC and Caspase-1 mRNA in the peripheral blood mononuclear cells of the children in acute exacerbation group were higher than those in chronic persistent and clinical remission groups (P<0.05), and the expression levels of NLRP3,ASC and Caspase-1 mRNA in chronic persistent group were higher than those in clinical remission group (P<0.05).Pearson correlation analysis showed that the expression level of NLRP3 mRNA in the peripheral blood mononuclear cells of asthmatic children was positively correlated with the expression levels of ASC, Caspase-1 mRNA and the serum levels of IL-1β and IL-18 (P<0.05), while it was negatively correlated with FEV1% and FEV1/FVC; the expression level of ASC mRNA was positively correlated with the expression level of Caspase-1 mRNA and the serum levels of IL-1β and IL-18 (P<0.05), while it was negatively correlated with FEV1% and FEV1/FVC (P<0.05); the expression level of Caspase-1 mRNA was positively correlated with the expression level of Caspase-1 mRNA and the serum levels of IL-1β and IL-18 (P<0.05), while it was negatively correlated with FEV1% and FEV1/FVC (P<0.05); the serum level of IL-1β was negatively correlated with FEV1% and FEV1/FVC (P<0.05), and the serum level of IL-18 was negatively correlated with FEV1% and FEV1/FVC (P<0.05).Conclusion: The expression levels of NLRP3 inflammasome and the downstream factor IL-1β and IL-18 in peripheral blood of the children with asthma are increased, and they are related to the clinical stage of the children with asthma. NLRP3 inflammasome pathway might promote the pathogenesis of asthma in the children.

Key words: children, bronchial asthma, NOD-like receptor family pyrin domain containing 3, inflammasome, interleukin-1β, interleukin-18

中图分类号:

R725.6

胡博, 王小稳, 曹建晖, 孙晓敏, 崔亚杰, 石聪聪. 哮喘患儿外周血单个核细胞中NLRP3炎症小体及血清中IL-1β和IL-18表达变化及其意义[J]. 吉林大学学报(医学版), 2019, 45(01): 111-116.

HU Bo, WANG Xiaowen, CAO Jianhui, SUN Xiaomin, CUI Yajie, SHI Congcong. Changes of expressions of NLRP3 inflammasome in peripheral blood mononuclear cells and IL-1β and IL-18 in serum in children with asthma and their significances[J]. Journal of Jilin University(Medicine Edition), 2019, 45(01): 111-116.

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