miR-26a/miR-146a/miR-31 对哮喘小鼠和哮喘儿童气道炎症的影响。
Effects of miR-26a/miR-146a/miR-31 on airway inflammation of asthma mice and asthma children.
机构信息
Department of Pathophysiology, Binzhou Medical University, Yantai, China.
出版信息
Eur Rev Med Pharmacol Sci. 2019 Jun;23(12):5432-5440. doi: 10.26355/eurrev_201906_18212.
OBJECTIVE
This study detected the expressions of microRNA-26a (miR-26a), miR-146a and miR-31 in lung tissues and BALF (bronchoalveolar lavage fluid) of asthma mice and children. Besides, cytokine levels of interleukin-5 (IL-5), IL-8, IL-12 and tumor necrosis factor-α (TNF-α) were detected as well. We aim to provide an experimental basis for clinical treatment of asthma.
PATIENTS AND METHODS
Forty female BALB/c mice were randomly assigned into control group and asthma group, respectively. Mice in asthma group (n=20) were immunized by intraperitoneal injection of OVA (ovalbumin) and provoked by atomization inhalation of OVA from the 15th day for 10 days. Mice in control group (n=20) were immunized and provoked with isodose saline during the same period. At the 26th day, mice were sacrificed for collecting lung tissues and BALF. Besides, we enrolled 17 cases of asthma children and 13 cases of children with airway foreign body as controls. BALF of each subject was collected. Total cellular score and differential counting in BALF were recorded. Expression levels of miR-26a, miR-146a, and miR-31 were detected by reverse transcription-polymerase chain reaction (RT-PCR). Levels of IL-5, IL-8, IL-12, and TNF-α were detected by enzyme-linked immunosorbent assay (ELISA).
RESULTS
The total cellular score in BALF of asthma mice and asthma children was higher than that of controls (p<0.05). Percentages of eosinophils, neutrophils, and lymphocytes in BALF of asthma mice and asthma children were higher than those of controls, whereas the percentage of macrophages was lower (p<0.05). Levels of IL-5, IL-8, IL-12, and TNF-α in lung tissues of asthma mice were markedly elevated compared with those of controls (p<0.05). Similarly, levels of IL-5, IL-8, IL-12, and TNF-α were higher in BALF of asthma children than controls (p<0.05). RT-PCR data showed higher mRNA levels of miR-26a, miR-146a, and miR-31 in lung tissues of asthma mice than controls (p<0.05). The mRNA levels of miR-26a, miR-146a, and miR-31 in BALF of asthma children were highly expressed compared with those of controls as well (p<0.05).
CONCLUSIONS
MiR-26a, miR-146a, and miR-31 are involved in asthma progression mainly through regulating inflammatory factors and cells.
目的
本研究检测了哮喘小鼠和儿童的肺组织和支气管肺泡灌洗液（BALF）中 microRNA-26a（miR-26a）、miR-146a 和 miR-31 的表达，并检测了白细胞介素-5（IL-5）、IL-8、IL-12 和肿瘤坏死因子-α（TNF-α）等细胞因子的水平。旨在为哮喘的临床治疗提供实验依据。
患者和方法
40 只雌性 BALB/c 小鼠随机分为对照组和哮喘组，每组 20 只。哮喘组小鼠（n=20）通过腹腔注射卵清蛋白（OVA）免疫，第 15 天起用 OVA 雾化吸入激发 10 天。对照组小鼠（n=20）同期用等剂量生理盐水免疫和激发。第 26 天处死小鼠取肺组织和 BALF。同时纳入 17 例哮喘儿童和 13 例气道异物儿童作为对照组，采集 BALF。记录总细胞评分和 BALF 中的差异计数。采用逆转录聚合酶链反应（RT-PCR）检测 miR-26a、miR-146a 和 miR-31 的表达水平。采用酶联免疫吸附试验（ELISA）检测 IL-5、IL-8、IL-12 和 TNF-α 的水平。
结果
哮喘小鼠和哮喘儿童的 BALF 总细胞评分均高于对照组（p<0.05）。哮喘小鼠和哮喘儿童 BALF 中的嗜酸性粒细胞、中性粒细胞和淋巴细胞百分比均高于对照组，而巨噬细胞百分比较低（p<0.05）。哮喘小鼠肺组织中 IL-5、IL-8、IL-12 和 TNF-α 水平明显高于对照组（p<0.05）。同样，哮喘儿童 BALF 中的 IL-5、IL-8、IL-12 和 TNF-α 水平也高于对照组（p<0.05）。RT-PCR 数据显示，哮喘小鼠肺组织中 miR-26a、miR-146a 和 miR-31 的 mRNA 水平均高于对照组（p<0.05）。哮喘儿童 BALF 中 miR-26a、miR-146a 和 miR-31 的 mRNA 水平也明显高于对照组（p<0.05）。
结论
miR-26a、miR-146a 和 miR-31 主要通过调节炎症因子和细胞参与哮喘的发生发展。
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