[Toll样受体2介导的信号分子高迁移率族蛋白1在小鼠哮喘中的作用机制]

[Mechanism of signal molecule high mobility group box protein 1 mediated by Toll-like receptor 2 in murine asthma].

作者信息

He Fang, Shen Qiying, Fang Lei, Jiang Xuqin, Wu Huimei, Liu Rongyu

机构信息

Department of Geriatric Pulmonary, First Affiliated Hospital, Anhui Medical University; Geriatric Institute of Anhui Province; Key Laboratory of Geriatric Molecule Medical of Anhui Province, Hefei 230022, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2014 Apr 29;94(16):1219-22.

PMID:24924884

Abstract

OBJECTIVE

To explore the role and mechanism of signal molecule high mobility group box protein 1 (HMGB1) mediated by Toll-like receptor 2 (TLR2) in a murine asthma model.

METHODS

Fourteen specific pathogen free (SPF) female C57 and TLR2(-/-) mice each were randomly divided into 4 groups of C57 control, C57 asthma, TLR2(-/-) control and TLR2(-/-) asthma (n = 7 each). The animals were sensitized and challenged with ovalbumin (OVA) for asthmatic modeling. The same amount of normal saline was used in the control group. The supernatant of bronchoalveolar lavage fluid (BALF) was collected for detecting the level of HMGB1 by enzyme-linked immunosorbent assay (ELISA). And the expression of HMGB1 in lung tissue was detected by Western blot and immunohistochemistry.

RESULTS

Asthmatic murine model was successfully established. The level of HMGB1 in the BALF of C57 asthma group was significantly higher than that in C57 control, TLR2(-/-) asthma and TLR2(-/-) control groups ((59.0 ± 13.9) vs (42.3 ± 1.6), (47.5 ± 2.3), (42.4 ± 1.4) ng/L; P = 0.001, 0.001, 0.037) . The results of immunohistochemistry showed that the marker of HMGB1 in lung tissue was less than those in the C57 control and TLR2(-/-) control groups. However, the C57 asthma and TLR2(-/-) asthma groups were obviously more and they were located in airway epithelium. Western blot showed that the expression of HMGB1 was significantly higher in C57 asthma group than that in the C57 control, TLR2(-/-) asthma and TLR2(-/-) control groups (0.92 ± 0.29 vs 0.18 ± 0.09, 0.31 ± 0.16, 0.21 ± 0.14; P = 0.007, 0.022, 0.009).

CONCLUSIONS

HMGB1 promotes the airway inflammation mediated by TLR2. And it may participate in the pathogenesis of asthma.

摘要

目的

探讨Toll样受体2（TLR2）介导的信号分子高迁移率族蛋白1（HMGB1）在小鼠哮喘模型中的作用及机制。

方法

将14只无特定病原体（SPF）雌性C57小鼠和14只TLR2基因敲除（TLR2(-/-)）小鼠分别随机分为C57对照组、C57哮喘组、TLR2(-/-)对照组和TLR2(-/-)哮喘组（每组n = 7）。用卵清蛋白（OVA）对动物进行致敏和激发以建立哮喘模型。对照组使用等量的生理盐水。收集支气管肺泡灌洗液（BALF）上清液，采用酶联免疫吸附测定（ELISA）法检测HMGB1水平。并通过蛋白质印迹法（Western blot）和免疫组织化学法检测肺组织中HMGB1的表达。

结果

成功建立哮喘小鼠模型。C57哮喘组BALF中HMGB1水平显著高于C57对照组、TLR2(-/-)哮喘组和TLR2(-/-)对照组（(59.0 ± 13.9) 对 (42.3 ± 1.6)、(47.5 ± 2.3)、(42.4 ± 1.4) ng/L；P = 0.001、0.001、0.037）。免疫组织化学结果显示，肺组织中HMGB1标记物少于C57对照组和TLR2(-/-)对照组。然而，C57哮喘组和TLR2(-/-)哮喘组明显更多，且位于气道上皮。蛋白质印迹法显示，C57哮喘组中HMGB1的表达显著高于C57对照组、TLR2(-/-)哮喘组和TLR2(-/-)对照组（0.92 ± 0.29对0.18 ± 0.09、0.31 ± 0.16、0.21 ± 0.14；P = 0.007、0.022、0.009）。