儿童支气管哮喘患者血清NETs和IL-4水平表达与发作期中医证型的相关性分析

目的　探究支气管哮喘发作期患儿血清中性粒细胞胞外诱捕网（NETs）、白介素-4（IL-4）表达水平及其与中医证型的相关性。方法　选取2017年1月~2020年1月湖南中医药高等专科学校附属第一医院收治的支气管哮喘急性发作期患儿98例作为观察组，其中热喘证37例（热喘证组）、寒喘证28例（寒喘证组）、虚喘证33例（虚喘证组）。另选取同期在该院体检的25例健康儿童作为健康对照组。使用PicoGreen荧光染料及单管型多功能检测仪检测血清游离DNA（cf-DNA）/NETs水平，采用酶联免疫吸附法（ELISA）检测血清IL-4表达水平，采用Pearson法分析支气管哮喘发作期患儿血清cf-DNA/NETs和IL-4水平与中性粒细胞、淋巴细胞、单核细胞及肺功能指标第1秒用力呼气量（FEV1）占预计值百分比（FEV1%）、FEV1/用力肺活量（FVC）水平相关性。结果　健康对照组、虚喘证组、寒喘证组、热喘证组患儿血清cf-DNA/NETs和IL-4水平及外周血中性粒细胞、单核细胞水平依次升高，差异均有统计学意义（F=895.500，69.590，63.250和50.590，均P＜0.05），淋巴细胞水平及肺功能指标FEV1%，FEV1/FVC水平均依次降低，差异具有统计学意义（F=71.410，75.790，均P＜0.05）。支气管哮喘发作期患儿血清cf-DNA/NETs与IL-4水平呈正相关（r=0.431，P=0.000）；血清cf-DNA/NETs水平与中性粒细胞、单核细胞水平呈正相关（r=0.541，0.401，均P＜0.05），与淋巴细胞、FEV1%和FEV1/FVC水平呈负相关（r=-0.456，-0.462和-0.503，均P＜0.05）；血清IL-4水平与中性粒细胞、单核细胞水平呈正相关（r=0.461，0.405，均P＜0.05），与淋巴细胞、FEV1%和FEV1/FVC水平呈负相关（r=-0.438，-0.479和-0.534，均P＜0.05）。结论　 支气管哮喘发作期患儿血清cf-DNA/NETs，IL-4水平明显升高，且热喘证患儿血清中二者水平最高，其次为寒喘证和虚喘证，二者水平变化可能与不同中医证型存在一定相关性。

Correlation Analysis of Serum NETs,IL-4 Expression and TCM Syndromes in Children with Bronchial Asthma During Onset

Objective　To explore the expression levels of neutrophil extracellular traps (NETs) and interleukin-4 (IL-4) in serum of children with bronchial asthma attack and their correlation with TCM syndrome types. Methods　A total of 98 children with acute exacerbation of bronchial asthma in the First Affiliated Hospital of Hunan College of Traditional Chinese Medicine from January 2017 to January 2020 were selected as the observation group, including 37 cases of heat asthma syndrome (heat asthma syndrome group), 28 cases of cold asthma syndrome (cold asthma syndrome group), and 33 cases of deficiency asthma syndrome (deficiency asthma syndrome group). In addition, 25 healthy children who had physical examination in the same hospital during the same period were selected as the healthy control group. The level of serum free DNA (cf-DNA)/NETs was measured by PicoGreen fluorescent dye and single tube multifunctional detector, the expression level of IL-4 in serum was detected by enzyme-linked immunosorbent assay (ELISA), Pearson method was used to analyze the correlation between serum cf-DNA/NETs, IL-4 levels and neutrophils, lymphocytes, monocytes and pulmonary function indexes FEV1%, FEV1/FVC levels. Results　The serum levels of cf-DNA/NETs, IL-4 and neutrophil and monocytes in the healthy control group, the deficiency asthma syndrome group, the cold asthma syndrome group and the heat asthma syndrome group were significantly increased in turn, the difference were statistically significant (F=895.500, 69.590, 63.250 and 50.590, all P < 0.05), the levels of lymphocyte and lung function index FEV1 and FEV1/FVC were significantly decreased in turn, the difference were statistically significant(F=71.410, 75.790, all P < 0.05). The serum cf-DNA/NETs in children with asthma had a positive correlation with IL-4 level (r = 0.431, P = 0.000). The levels of serum cf-DNA/NETs were positively correlated with the levels of neutrophils and monocytes (r=0.541, 0.401, all P < 0.05), and negatively correlated with lymphocyte, FEV1 and FEV1/FVC levels (r=-0.456, -0.462 and -0.503, all P < 0.05). The levels of serum IL-4 were positively correlated with the levels of neutrophils and monocytes (r=0.461, 0.405, all P < 0.05), and negatively correlated with lymphocyte, FEV1 and FEV1/FVC levels (r=-0.438, -0.479 and -0.534, all P < 0.05). Conclusion　The serum levels of cf-DNA/NETs and IL-4 in children with bronchial asthma attack were significantly increased. The serum levels of the two were the highest in children with heat asthma syndrome, followed by cold asthma syndrome and deficiency asthma syndrome. The changes of the two levels may be related to different TCM syndromes.