细胞外信号调节激酶在调节哮喘大鼠模型淋巴细胞白细胞介素 13 表达中的作用。

Role of extracellular signal-regulated kinase in regulating expression of interleukin 13 in lymphocytes from an asthmatic rat model.

机构信息

Department of Respiratory Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.

出版信息

Chin Med J (Engl). 2010 Jul;123(13):1715-9.

PMID:20819635

Abstract

BACKGROUND

The extracellular signal-regulated kinase (ERK) is widely expressed in mammal cells and involved in airway proliferation and remodeling in asthma. In this study, we intend to explore the role of ERK in the expression of the Th2 cytokine, interleukin 13 (IL-13) in lymphocytes in asthma.

METHODS

Forty Sprague-Dawley rats were randomly divided into two groups: normal control and asthmatic groups. Peripheral blood lymphocytes were isolated and purified from the blood of each rat and divided into five groups: control, asthmatic lymphocytes, asthmatic cells stimulated with ERK activator epidermal growth factor (EGF), or with ERK inhibitor PD98059, or with EGF and PD98059 together. The expression of phosphorylated-ERK (p-ERK) was observed by immunocytochemical staining, the expression of ERK mRNA was determined by reverse transcriptase-PCR, IL-13 protein in supernatants was measured by ELISA.

RESULTS

(1) The ERK mRNA level and the percentage of cells with p-ERK in lymphocytes from asthmatic rats were significantly higher than those in normal controls, and were significantly increased by EGF administration. This effect of EGF was significantly inhibited by PD98059 pretreatment. (2) IL-13 protein in supernatants of asthmatic lymphocytes was higher than that produced by normal control lymphocytes, and was significantly increased by EGF treatment. This EGF effect was partly blocked by PD98059 pretreatment. (3) There was a significant positive correlation between the percentage of cells with p-ERK in peripheral blood lymphocytes and IL-13 protein in supernatants of lymphocytes from asthmatic rats.

CONCLUSIONS

In asthma the ERK expression and activation levels were increased, as was the protein level of IL-13. The ERK signaling pathway may be involved in the increased expression of the Th2 cytokine IL-13 in asthma.

摘要

背景

细胞外信号调节激酶（ERK）广泛存在于哺乳动物细胞中，参与哮喘气道增殖和重塑。本研究旨在探讨 ERK 在哮喘淋巴细胞中 Th2 细胞因子白细胞介素 13（IL-13）表达中的作用。

方法

40 只 Sprague-Dawley 大鼠随机分为正常对照组和哮喘组。从每只大鼠的血液中分离和纯化外周血淋巴细胞，并将其分为五组：对照组、哮喘淋巴细胞组、用 ERK 激活剂表皮生长因子（EGF）刺激的哮喘细胞组、用 ERK 抑制剂 PD98059 刺激的哮喘细胞组、用 EGF 和 PD98059 共同刺激的哮喘细胞组。通过免疫细胞化学染色观察磷酸化-ERK（p-ERK）的表达，通过逆转录聚合酶链反应（RT-PCR）测定 ERK mRNA 的表达，通过酶联免疫吸附试验（ELISA）测定上清液中 IL-13 蛋白的含量。

结果

（1）哮喘大鼠淋巴细胞中 ERK mRNA 水平和 p-ERK 阳性细胞比例明显高于正常对照组，EGF 给药后明显增加。PD98059 预处理可显著抑制 EGF 的这种作用。（2）哮喘淋巴细胞上清液中 IL-13 蛋白含量高于正常对照组，EGF 处理后明显增加。PD98059 预处理部分阻断了 EGF 的这种作用。（3）哮喘大鼠外周血淋巴细胞中 p-ERK 阳性细胞比例与淋巴细胞上清液中 IL-13 蛋白含量呈显著正相关。