Correlation analysis of intestinal flora with lung function and airway inflammatory cells in children with acute bronchial asthma
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摘要: 目的 探讨肠道菌群与急性发作期支气管哮喘患儿肺功能和气道炎症的相关性。方法 选择2018年2月-2021年6月儿科病房收治的105例急性发作期支气管哮喘患儿为研究对象，根据哮喘急性发作严重程度分级，将患儿分为轻度组(36例)、中度组(42例)和重度组(27例)。另选择31例健康儿童为对照组。所有受试者均采集大便标本，荧光定量PCR检测肠道菌群相对丰度。比较各组肠道菌群状态，分析肠道菌群丰度与患儿肺功能和气道炎症细胞、炎症因子的关系。结果 重度组、中度组、轻度组乳酸杆菌、脆弱拟杆菌、毛螺菌、韦荣球菌、普拉梭菌和罗氏菌丰度低于对照组(P＜0.001)，且重度组低于中度组和轻度组(P＜0.05)；重度组、中度组、轻度组分节丝状菌、艰难梭菌丰度高于对照组(P＜0.001)，且重度组高于中度组和轻度组(P＜0.05)。重度组第1秒用力呼气容积(FEV1)、用力肺活量(FVC)、FEV1/FVC、最大呼气峰流量(PEF)占正常预计值的百分率(PEF%)低于中度组和轻度组(P＜0.05)，诱导痰细胞总数、嗜酸粒细胞比例、肿瘤坏死因子-α(TNF-α)、IL-6高于中度组和轻度组(P＜0.05)。支气管哮喘患儿乳酸杆菌、脆弱拟杆菌相对丰度与气道细胞总数、嗜酸粒细胞、TNF-α、IL-6呈负相关(r=-0.513、-0.493；-0.491、-0.401；-0.506、-0.477；-0.385、-0.401，均P＜0.05)，与FEV1、FVC、FEV1/FVC、PEF%呈正相关(r=0.477、0.395、0.362、0.509；0.394、0.305、0.293、0.411，均P＜0.05)；分节丝状菌、艰难梭菌相对丰度与气道细胞总数、嗜酸粒细胞、TNF-α、IL-6呈正相关(r=0.439、0.397；0.409、0.351；0.341、0.369；0.321、0.309，均P＜0.05)，与FEV1、FVC、FEV1/FVC、PEF%呈负相关(r=-0.413、-0.336、-0.274、-0.492；-0.369、-0.273、-0.391、-0.416，均P＜0.05)。结论 急性发作期支气管哮喘患儿肠道菌群紊乱与肺功能降低、气道炎症细胞增加、炎症因子水平增高有关。Abstract: Objective To investigate the relationship between intestinal flora and lung function and airway inflammation in children with acute bronchial asthma.Methods One hundred and five patients with acute bronchial asthma treated in the pediatric inpatient ward of our hospital from February 2018 to June 2021 were selected. According to the severity of acute asthma attack, the children were divided into mild group(36 cases), moderate group(42 cases) and severe group(27 cases). Another 31 healthy children were selected as the control group. Stool samples were collected from all subjects, and the relative abundance of intestinal flora was detected by fluorescence quantitative PCR. The intestinal flora status of each group was compared to analyze the relationship between intestinal flora abundance and lung function and airway inflammatory cells and inflammatory cytokines.Results The abundance ofLactobacillus,Bacteroides fragilis,Spirillum tricillum,Verronococcus,Clostridium difficileandrochein severe, moderate and mild groups was lower than that in control group(P< 0.001), and the abundance of severe group were lower than that in moderate and mild groups(P< 0.05). The abundance ofFilamentous fungiandClostridium difficilein severe group were higher than those in control group(P< 0.001), and the abundance of severe group were higher than that in moderate and mild groups(P< 0.05). The forced expiratory volume in the first second(FEV1), forced critical capacity(FVC), FEV1/FVC, and maximum peak expiratory flow(PEF)% in the severe group were lower than those in the moderate and mild groups(P< 0.05), while the total number of induced sputum cells and the proportion of eosinophil, the level of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) were higher than those in the moderate and mild groups(P< 0.05). The relative abundance ofLactobacillusandBacteroides fragiliswas negatively correlated with the total number of airway cells, eosinophils, TNF-α and IL-6(r=-0.513, -0.493; -0.491, -0.401; -0.506, -0.477; -0.385, -0.401;P< 0.05), were positively correlated with FEV1, FVC, FEV1/FVC and PEF%(r=0.477, 0.395, 0.362, 0.509; 0.394, 0.305, 0.293, 0.411,P< 0.05); the relative abundance ofFilamcinomaandClostridium difficilewere positively correlated with the total number of airway cells eosinophils, TNF-α and IL-6(r=0.439, 0.397; 0.409, 0.351; 0.341, 0.369; 0.321, 0.309,P< 0.05), and were negatively correlated with the FEV1, FVC, FEV1/FVC and PEF%(r=-0.413, -0.336, -0.274, -0.492; -0.369, -0.273, -0.391, -0.416,P< 0.05).Conclusion Intestinal flora disorder in children with acute bronchial asthma is associated with decreased lung function and increased airway inflammatory cells and inflammatory cytokines.
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Key words:
- intestinal flora /
- bronchial asthma /
- children /
- lung function /
- inflammatory cells /
- immune
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表 1 4组受试儿基线资料的比较
例，X±S，例(%) 组别 例数 年龄/岁 男/女 体重/kg 心率/(次·min-1) 血氧饱和度/% 用药情况 吸入型糖皮质激素 吸入型糖皮质激素联合白三烯 吸入型糖皮质激素联合茶碱类药物 轻度组 36 8.42±1.99 20/16 36.00±7.06 107.81±6.901) 94.44±4.391) 20(55.56) 10(27.78) 6(16.67) 中度组 42 7.79±2.16 27/15 35.10±8.96 110.36±6.311) 91.24±3.411) 23(54.76) 12(28.57) 7(16.67) 重度组 27 8.67±1.92 16/9 35.81±6.94 107.81±7.771) 88.07±2.121)2)3) 13(48.15) 10(37.04) 4(14.81) 对照组 31 8.42±2.23 18/13 36.61±6.80 103.48±6.96 98.81±1.03 F/χ2 1.186 0.825 0.242 5.908 64.406 0.745 P 0.318 0.844 0.867 0.001 ＜0.001 0.946 与对照组比较，1)P＜0.05；与轻度组比较，2)P＜0.05；与中度组比较，3)P＜0.05。 
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表 2 4组受试儿肠道菌群相对丰度的比较
X±S 组别 例数 乳酸杆菌 脆弱拟杆菌 毛螺菌 韦荣球菌 普拉梭菌 罗氏菌 轻度组 36 18.86±3.491) 21.56±4.071) 7.86±1.861) 9.42±1.851) 3.03±0.831) 5.03±1.071) 中度组 42 18.95±4.401) 21.93±4.021) 8.31±1.351) 10.19±2.111) 3.12±0.881) 5.19±0.981) 重度组 27 12.70±3.231)2)3) 14.81±3.641)2)3) 6.30±1.121)2)3) 7.30±1.701)2)3) 2.33±0.771)2)3) 3.52±1.001)2)3) 对照组 31 32.03±5.67 26.71±3.67 9.19±2.07 14.00±3.20 4.00±1.05 6.13±1.68 F 106.745 45.478 15.612 44.711 17.328 23.178 P ＜0.001 ＜0.001 ＜0.001 ＜0.001 ＜0.001 ＜0.001 组别 例数 梭状芽胞杆菌 分节丝状菌 艰难梭菌 大肠埃希菌 瘤胃球菌 肠球菌 轻度组 36 21.25±4.24 4.72±0.731) 2.03±0.691) 8.33±2.25 6.22±1.55 7.19±1.27 中度组 42 21.21±3.36 4.98±1.281) 2.19±0.731) 8.26±1.56 6.17±1.48 7.17±1.99 重度组 27 21.48±3.92 7.00±1.361)2)3) 3.30±0.661)2)3) 8.04±1.40 6.00±1.56 7.15±1.04 对照组 31 23.03±4.22 2.90±0.69 1.03±0.18 8.42±1.86 6.10±1.59 7.03±1.26 F 1.596 72.439 64.850 0.230 0.118 0.076 P 0.193 ＜0.001 ＜0.001 0.875 0.949 0.973 与对照组比较，1)P＜0.05；与轻度组比较，2)P＜0.05；与中度组比较，3)P＜0.05。
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表 3 3组支气管哮喘患儿气道炎症细胞、炎症因子和肺功能比较
X±S 组别 例数 细胞总数/(×108·L-1) 巨噬细胞/% 嗜酸粒细胞/% 中性粒细胞/% 淋巴细胞/% TNF-α/(ng·L-1) 轻度组 36 2.42±0.28 42.30±10.55 3.02±0.44 44.21±6.41 5.58±0.95 9.22±1.90 中度组 42 3.02±0.661) 40.44±9.64 5.28±1.251) 44.48±6.51 5.54±0.95 11.99±2.681) 重度组 27 3.49±0.821)2) 37.24±12.90 8.33±2.001)2) 44.85±6.17 5.65±0.73 19.13±6.091)2) F 24.329 1.685 126.898 0.077 0.123 57.815 P ＜0.001 0.191 ＜0.001 0.926 0.884 ＜0.001 组别 例数 IL-6/(pg·mL-1) FEV1/L FVC/L FEV1/FVC/% PEF% 轻度组 36 12.13±3.50 1.75±0.34 2.41±0.40 74.13±15.53 83.77±3.64 中度组 42 15.25±3.201) 1.44±0.271) 2.14±0.211) 67.72±14.411) 69.80±8.301) 重度组 27 17.21±3.301)2) 1.08±0.181)2) 1.76±0.281)2) 63.92±14.381)2) 42.08±4.361)2) F 18.952 44.925 35.203 3.911 366.723 P ＜0.001 ＜0.001 ＜0.001 ＜0.001 ＜0.001 与轻度组比较，1)P＜0.05；与中度组比较，2)P＜0.05。
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表 4 肠道菌群与肺功能和气道炎症细胞的相关性分析
指标 乳酸杆菌 脆弱拟杆菌 分节丝状菌 毛螺菌 r P r P r P r P 细胞总数 -0.513 ＜0.001 -0.491 ＜0.001 0.439 ＜0.001 -0.209 0.109 嗜酸粒细胞 -0.493 ＜0.001 -0.401 ＜0.001 0.397 0.001 -0.163 0.231 TNF-α -0.506 ＜0.001 -0.385 0.002 0.341 0.016 -0.102 0.469 IL-6 -0.477 ＜0.001 -0.401 ＜0.001 0.369 0.008 -0.154 0.239 FEV1 0.477 ＜0.001 0.394 0.004 -0.413 ＜0.001 0.158 0.295 FVC 0.395 0.003 0.305 0.020 -0.336 0.018 0.143 0.342 FEV1/FVC 0.362 0.010 0.293 0.022 -0.274 0.024 0.136 0.392 PEF% 0.509 ＜0.001 0.411 ＜0.001 -0.492 ＜0.001 0.128 0.432 指标 韦荣球菌 普拉梭菌 罗氏菌 艰难梭菌 r P r P r P r P 细胞总数 -0.138 0.386 -0.096 0.581 -0.162 0.237 0.409 ＜0.001 嗜酸粒细胞 -0.157 0.302 -0.105 0.536 -0.152 0.323 0.351 0.012 TNF-α -0.143 0.245 -0.107 0.415 -0.096 0.623 0.321 0.032 IL-6 -0.165 0.229 -0.137 0.269 -0.127 0.354 0.309 0.039 FEV1 0.201 0.129 0.133 0.418 0.203 0.115 -0.369 0.008 FVC 0.169 0.225 0.117 0.488 0.178 0.207 -0.273 0.030 FEV1/FVC 0.172 0.213 0.126 0.449 0.141 0.356 -0.391 0.006 PEF% 0.155 0.318 0.145 0.337 0.139 0.371 -0.416 ＜0.001
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