干预支气管哮喘小鼠气道黏液高分泌的研究

Research of gefinitib for the treatment on airway mucus hypersecretion of bronchial asthma in mice

目的探讨表皮生长因子受体(EGFR)阻滞剂吉非替尼抑制支气管哮喘小鼠气道黏液高分泌的作用。方法将BALB/c小鼠随机分为正常对照组、哮喘组和吉非替尼治疗组(干预组)。采用苏木精-伊红(HE)染色法、过碘酸-雪夫(PAS)特殊染色法、免疫组化法、原位末端转移酶标记法分别检测各组小鼠气道炎症及杯状细胞的数量和黏液分泌、p-EGFR蛋白表达、细胞凋亡等状况,western blot法检测各组小鼠肺组织中p-EGFR变化。结果与正常组相比,哮喘组小鼠出现典型的哮喘激发症状,肺组织中出现明显的以淋巴细胞和嗜酸性粒细胞为主的炎症细胞浸润等病理改变;哮喘组小鼠小气道杯状细胞数显著增加,干预组数量明显减少,差异具有统计学意义(P均<0.05);与哮喘组相比,干预组小气道中p-EGFR阳性、Bcl-2阳性杯状细胞数量分别显著减少,细胞凋亡、Bax阳性杯状细胞数量分别显著增加,差异具有统计学意义(P均<0.05)。结论吉非替尼可抑制支气管哮喘小鼠气道黏液高分泌,其机制可能是阻滞EGFR通路,抑制黏液分泌的同时,诱导分泌黏液的杯状细胞凋亡。

Objective To evaluate the treatment effect of gefinitib,a selective for epidermal growth factor receptor （EGFR） tyrosine kinase inhibitor,on the airway mucus hypersecrefion in asthmatic mice.Methods BALB/c mice were randomly divided into a sham-challenged asthmatic group,an asthma group,and a gefinitib treatment group（treatment group）.HE staining detected the chronic inflammation of the airway in mice of all groups.PAS staining detected the cell counting and mucus secretion of airway goblet cells in mice of all groups.Immunohistochemistry method detected p-EGFR,Bax,Bcl-2 protein expression of goblet cells in small bronchus of the last two groups.TUNEL method detected apoptosis of goblet cells in small bronchus of the last two groups.Western blot method detected p-EGFR protein expression of lung tissues in mice of all groups.Results Compared with the sham-challenged asthmatic group,there was more significant inflammatory cell infiltration around the bronchus and perivascular of asthma group.Compared with sham-challenged asthmatic group,there were more goblet cells in the small bronchus of the asthma group,and the number was reduced in the small bronchus of the treatment group（all P 〈 0.05,respectively） Compared with the asthma group,the numbers of goblet cells which positively expressed p-EGFR and Bcl-2 in the small bronchus of the treatment group were reduced significantly,and the numbers of goblet cells which positively expressed Bax and apoptosis were increased significantly （all P 〈 0.05,respectively）.Conclusions Gefinitib can inhibit airway mucus hypersecretion of asthmatic mice,and the mechanism may be that block the EGFR signaling to inhibit the mucus production and to induce apoptosis of goblet cells in the small bronchus.