吉非替尼抑制博莱霉素诱导的小鼠肺纤维化

Gefitinib prevents bleomycin-induced lung fibrosis in mice

目的:观察吉非替尼对博莱霉素诱导的小鼠肺纤维化的抑制作用。方法:将40只SPF级雌性BALB/c小鼠分为4组:对照组(气管滴入生理盐水)、单纯口服吉非替尼组(吉非替尼灌胃200 mg/kg)、纤维化组(气管滴入博莱霉素3 mg/kg)、纤维化吉非替尼干预组(气管滴入博莱霉素+吉非替尼灌胃20 mg/kg)。实验第14 d杀鼠取肺,左肺石蜡切片行HE染色与Masson染色,免疫组化检测总表皮生长因子受体(EGFR)及磷酸化EG-FR;取右肺检测羟脯氨酸含量。结果:纤维化吉非替尼干预组肺病理损伤较纤维化组减轻,气道上皮下胶原沉积及肺羟脯氨酸含量减少(P<0.05),气道上皮及肺间质细胞磷酸化EGFR表达评分下降(P<0.05)。单纯口服吉非替尼组小鼠气道上皮下未见明显胶原沉积,肺羟脯氨酸含量及磷酸化EGFR表达评分与对照组相比无显著差异(P>0.05)。结论:吉非替尼灌胃能显著抑制博莱霉素诱导的小鼠肺纤维化,大剂量(200 mg/kg)吉非替尼灌胃未引起明显肺纤维化。

AIM： To evaluate the effect of epidermal growth factor receptor-tyrosine kinase inhibitor（EGFR-TKI） on lung fibrosis induced by bleomycin in mice.METHODS： Forty BALB/c female mice were randomly divided into 4 groups： the mice in control group were given vehicle orally with administering of saline intratracheally;the mice in Ge200 group were given gefitinib（a tyrosine kinase inhibitor） at dose of 200 mg/kg orally with administering of saline intratracheally;the mice in BLM group were given vehicle orally with administering of bleomycin intratracheally;the mice in BLM＋Ge20 group was given gefitinib at dose of 20 mg/kg orally with administering of bleomycin intratracheally.All animals were sacrificed by abdominal aortic bleeding 14 days after treatments.The left lung was stained with hematoxylin,eosin and Masson＇s trichrome respectively for the pathological examination.Total EGFR and phosphorylated EGFR were detected by immunohistochemistry.The tissues of right lung were sampled and the contents of hydroxyproline（HYP） were measured to quantitate the lung fibrosis.RESULTS： Gefitinib prevented lung fibrosis induced by bleomycin with significantly reducing lung collagen accumulation and the level of HYP in BLM＋Ge20 group（P0.05）.The phosphorylation of EGFR in lung mesenchymal cells and epithelial cells was inhibited by treating gefitinib after intratracheal administration of bleomycin（P0.05）.Furthermore,in those mice that did not receive bleomycin treatment（Ge200 group）,gefitinib neither induced the lung fibrosis nor increased the expression of p-EGFR.CONCLUSION： These findings suggest that,in the preclinical setting,gefitinib has a protective effect on lung fibrosis induced by bleomycin.Gefitinib at high dose（200 mg/kg） dose not induces lung fibrosis.