The role of epidermal growth factor receptor (EGFR) in mucin MUC5AC gene expression in airways

表皮生长因子受体（EGFR）在气道粘蛋白MUC5AC基因表达中的作用

• 批准号: 14570567
• 负责人: TAKEYAMA Kiyoshi
• 金额: $ 1.86万
• 依托单位: Tokyo Women's Medical University, School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570567/
• 关键词: bronchial asthma; goblet cell; airway remodeling; epidermal growth factor receptor; apoptosis; MUC5AC; mucin; airway hyperresponsiveness; ムチイン; EGF受容体(EGFR); 気道抵抗; 気道上皮

We determined the role of EGFR in pathogenesis of asthma both in vivo and in vitro. Brown Norway rats were sensitized (day 0, day 7) and challenged (days 14, 15 and 16) with ovalbumin (OVA). Morphological analysis (Alcian blue/PAS staining, immunohistochemistry for MUC5AC and EGFR), measurement of airway hyperreactivity (penH) and cell analysis (BAL) were performed on days 14 (before OVA challenge), 17, 24, 31 and day 38. On day 17, OVA-sensitized and challenged rats showed a marked increase in both MUC5AC and EGFR immunoreactivity in airway epithelium. Without additional OVA challenge, spontaneous resolution of both inflammatory cell infiltration and goblet cell hyperplasia were observed simultaneously with the reduction of EGFR expression on day 38. In contrast, AHR persisted for three weeks. Intratracheal instillation of the selective EGFR tyrosine kinase inhibitor AG1478 induced a significant reduction in all established asthma phenotypes, which was accompanied by apoptosis of airway epithelial cells. In in vitro studies, incubation of NCI-H292 cells with TGFα for 24h increased Bcl-2, MUC5AC and mucous glycoconjugates production, effects that were inhibited by pretreatment of AG1478. AG1478 also induced apoptosis in mucous producing cells, but a MEK inhibitor PD98059 had no effect on apoptosis. Z-VAD-fmk, a caspase inhibitor reduced AG1478-induced apoptosis. From these results, we concluded that the expression and activation of EGFR may play a crucial role in the development of asthma phenotypes and that a tyrosine kinase inhibitor of EGFR may be a useful therapeutic candidate for the restoration of airway remodeling in patients with asthma.

我们在体内和体外确定EGFR在哮喘发病机制中的作用。用卵清蛋白（OVA）致敏Brown Norway大鼠（第0天、第7天）并激发（第14、15和16天）。在第14天（在OVA攻击之前）、第17天、第24天、第31天和第38天进行形态学分析（阿辛蓝/PAS染色、MUC 5AC和EGFR的免疫组织化学）、气道高反应性测量（penH）和细胞分析（BAL）。在第17天，OVA致敏和激发的大鼠显示气道上皮中MUC 5AC和EGFR免疫反应性显著增加。在没有额外的OVA激发的情况下，在第38天观察到炎性细胞浸润和杯状细胞增生的自发消退以及EGFR表达的降低。相反，AHR持续了三周。气管内滴注选择性EGFR酪氨酸激酶抑制剂AG 1478可显著降低所有已确定的哮喘表型，并伴有气道上皮细胞凋亡。在体外研究中，将NCI-H292细胞与TGFα孵育24小时可增加Bcl-2、MUC 5AC和粘液糖缀合物的产生，这些作用可被AG 1478预处理抑制。AG 1478也诱导粘液产生细胞的凋亡，但MEK抑制剂PD 98059对凋亡没有影响。半胱天冬酶抑制剂Z-VAD-fastrin可减少AG 1478诱导的细胞凋亡。从这些结果中，我们得出结论，EGFR的表达和激活可能在哮喘表型的发展中起着至关重要的作用，EGFR的酪氨酸激酶抑制剂可能是一个有用的治疗候选人，用于恢复哮喘患者的气道重塑。

项目成果

Takeyama K et al.: "Effect of macrolide antibiotics on MUC5AC production in airway epithelial cells in vitro."Eur Respir J. 20. 90S (2002)

Takeyama K 等人：“大环内酯类抗生素对体外气道上皮细胞中 MUC5AC 产生的影响。”Eur Respir J. 20. 90S (2002)

Takeyama K et al.: "Restoration from goblet cell hyperplasia to normal epithelial phenotype in ovalbumin-sensltized rats."Am J Respir Crit Care Med. 167. A203 (2003)

Takeyama K 等人：“卵清蛋白致敏大鼠从杯状细胞增生恢复到正常上皮表型。”Am J Respir Crit Care Med。

武山廉: "気管支喘息における過分泌"内科. 90. 704-707 (2002)

Ren Takeyama：“支气管哮喘的分泌过多”《内科》90. 704-707 (2002)。

Tamaoki J, Takeyama K, et al.: "A Th2 cytokine inhibitor for airway inflammation in mild asthma."J Allergy Clin Immunol. 111. 197-198 (2003)

Tamaoki J、Takeyama K 等人：“一种用于治疗轻度哮喘气道炎症的 Th2 细胞因子抑制剂。”J Allergy Clin Immunol。

Takeyama K, Nohara M, Tamaoki J, Kondo M, Isono K, Nagai A.: "Restoration from goblet, cell hyperplasia to normal epithelial phenotype in ovalbumin-sensitized rats."Am J Respir Crit Care Med. 167(Suppl). A468 (2003)

Takeyama K、Nohara M、Tamaoki J、Kondo M、Isono K、Nagai A.：“卵清蛋白致敏大鼠从杯状、细胞增生恢复到正常上皮表型。”Am J Respir Crit Care Med。