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目的探讨高迁移率族蛋白B1(HMGB1)参与调节支气管上皮细胞分泌哮喘相关炎症因子的机制。方法20只雌性BALB/c小鼠随机分对照组和哮喘组,采用卵清蛋白腹腔注射致敏联合雾化吸入激发建立哮喘小鼠模型。支气管上皮细胞16-HBE分组处理:空白对照组,HMGB1组(浓度分别为100、200、500ng/mL),500ng/mL HMGB1+AG490组,500ng/mL HMGB1+雷帕霉素组。HE染色观察小鼠气道结构;qRT-PCR和免疫组化法检测两组小鼠肺部组织HMGB1的表达;ELISA法测定两组小鼠支气管肺泡灌洗液(BALF)上清中HMGB1的含量和各组细胞上清液中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和白细胞介素-1α(IL-1α)水平。Western blot检测各组细胞中p-JAK2和pSTAT3的蛋白水平。结果哮喘组小鼠HMBG1的表达比正常小鼠明显升高。HMGB1升高细胞中TNF-α、IL-6和IL-1α水平,上调p-JAK2和p-STAT3水平,并且呈现浓度依赖性关系。AG490和雷帕霉素处理抑制细胞中p-JAK2和pSTAT3表达,同时降低TNF-α、IL-6和IL-1α水平。结论HMGB1参与调控哮喘慢性炎症反应,可能与活化JAK2/STAT3信号通路促进支气管上皮细胞分泌炎症因子有关。
Objective To investigate the mechanism of high mobility group box 1 (HMGB1) involved in the regulation of asthma-related inflammatory cytokines secretion in bronchial epithelial cells. Methods Twenty female BALB / c mice were randomly divided into control group and asthma group. The asthma model was established by intraperitoneal injection of ovalbumin combined with inhalation stimulation. Bronchial epithelial cells were treated with 16-HBE: blank control group, HMGB1 group (concentration of 100,200,500ng / mL, respectively), 500ng / mL HMGB1 + AG490 group and 500ng / mL HMGB1 + rapamycin group. The airway structure of mice was observed by HE staining. The expression of HMGB1 in the lung tissue of the two groups was detected by qRT-PCR and immunohistochemistry. The content of HMGB1 in the supernatant of bronchoalveolar lavage fluid (BALF) (TNF-α), interleukin-6 (IL-6) and interleukin-1α (IL-1α) in the supernatant of each group. Western blot was used to detect the protein levels of p-JAK2 and pSTAT3 in each group. Results The expression of HMBG1 in asthmatic mice was significantly higher than that in normal mice. HMGB1 increased the levels of TNF-α, IL-6 and IL-1α, increased the levels of p-JAK2 and p-STAT3 in a concentration-dependent manner. AG490 and rapamycin treatment inhibited the expression of p-JAK2 and pSTAT3 in cells while decreasing the levels of TNF-α, IL-6 and IL-1α. Conclusion HMGB1 is involved in the regulation of chronic inflammatory response in asthma, which may be related to the activation of JAK2 / STAT3 signaling pathway to promote bronchial epithelial cells to secrete inflammatory cytokines.