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目的了解支气管哮喘患儿外周血中性粒细胞(PMN)功能状况及其与哮喘病情的关系。方法对2005—2008年广西医科大学第一附属医院收治的67例临床确诊为支气管哮喘的患儿和作为健康对照组的16名儿童进行哮喘症状评分、肺功能检查,将哮喘组分为轻-中度哮喘组(MA)和重度哮喘组(SA)。所有研究对象抽取静脉血分离外周血中性粒细胞,体外培养12h,收集培养上清液。采用荧光标记流式细胞仪检测中性粒细胞活化率及中性粒细胞吞噬功能,比色法检测中性粒细胞弹性蛋白酶(NE)活性,还原比色法检测上清液一氧化氮(NO)水平,酶联免疫吸附实验(ELISA)法测定上清白细胞介素-8(IL-8)水平。比较对照组与哮喘组以及轻-中度哮喘组与重度哮喘组反映中性粒细胞功能各指标的差异;比较哮喘急性发作期与缓解期外周血中性粒细胞功能各指标的差异。结果 (1)与对照组比较,哮喘组PMN活化率及PMN噬菌率高于对照组,差异有统计学意义(P<0.01);哮喘组上清液NE、NO、IL-8高于对照组,差异有统计学意义(P<0.01)。(2)重度哮喘组PMN活化率高于轻-中度哮喘组,PMN噬菌率低于轻-中度哮喘组,差异有统计学意义(P<0.01),上清NE、NO、IL-8高于轻-中度哮喘组,差异有统计学意义(P<0.01)。(3)急性发作期PMN活化率、上清液NE、NO、IL-8与缓解期比较差异无统计学意义(P>0.05),急性发作期患儿PMN噬菌率高于缓解期,差异有统计学意义(P<0.01)。结论哮喘患儿外周血存在中性粒细胞活化的证据,表现为吞噬、趋化、释放功能的改变。哮喘患儿外周血中性粒细胞活化与哮喘的病情程度有关,外周血中性粒细胞活化相关指标可用于判断哮喘病情。布地奈德吸入治疗对哮喘患儿外周血中性粒细胞功能可能无明显影响。
Objective To investigate the functional status of peripheral blood neutrophil (PMN) in children with bronchial asthma and its relationship with asthma. Methods Sixty-seven children with clinically diagnosed bronchial asthma and 16 children as healthy control group were enrolled in the First Affiliated Hospital of Guangxi Medical University from 2005 to 2008. Their asthma symptom scores and pulmonary function tests were performed. The asthma group was divided into mild- Moderate asthma group (MA) and severe asthma group (SA). All subjects were drawn venous blood from peripheral blood neutrophils, cultured in vitro 12h, the culture supernatant was collected. Neutrophil activation and neutrophil phagocytosis were detected by fluorescence-labeled flow cytometry. Neutrophil elastase (NE) activity was measured by colorimetric assay. Nitric oxide (NO) ), And the level of interleukin-8 (IL-8) in supernatant was determined by enzyme-linked immunosorbent assay (ELISA). The differences of neutrophil function indexes between the control group and the asthma group, the mild-moderate asthma group and the severe asthma group were compared. The differences of the peripheral blood neutrophil function indexes between the acute and remission stages of asthma were compared. Results (1) Compared with the control group, the PMN activation rate and PMN phagocytosis rate in the asthma group were significantly higher than those in the control group (P <0.01). The levels of NE, NO and IL-8 in the asthma group were higher than those in the control group Group, the difference was statistically significant (P <0.01). (2) The PMN activation rate in severe asthma group was higher than that in mild to moderate asthma group, the PMN phage rate was lower than that in mild to moderate asthma group (P <0.01), the levels of NE, NO and IL- 8 was higher than mild to moderate asthma group, the difference was statistically significant (P <0.01). (3) The PMN activation rate in acute exacerbation stage was not significantly different from that in remission stage (P> 0.05). The PMN phagocytosis rate in acute exacerbation period was higher than that in remission stage, and the difference was not significant There was statistical significance (P <0.01). Conclusion There is evidence of neutrophil activation in the peripheral blood of children with asthma, showing the changes of phagocytosis, chemotaxis and release. Peripheral blood neutrophil activation in children with asthma and the severity of asthma, peripheral blood neutrophil activation related indicators can be used to determine the condition of asthma. Inhaled budesonide may have no effect on peripheral blood neutrophil function in children with asthma.