陈英　冯淬灵　李根茂

摘要：目的 观察清金化痰汤对慢性阻塞性肺疾病（COPD）模型大鼠表皮生长因子受体（EGFR）/MAPK信号通路的影响，探讨其干预COPD气道黏液高分泌的作用机制。方法 气管内滴注LPS联合烟熏方法建立COPD气道黏液高分泌模型。实验大鼠随机分为空白组、模型组、清金化痰汤组、克拉霉素组，每组10只。空白组正常饲养，其余3组分别给予生理盐水、清金化痰汤、克拉霉素片灌胃，每日1次，连续30 d。各组大鼠于实验第31日分别处死，HE染色观察肺组织病理形态及黏液腺体增生情况，实时荧光定量PCR检测肺组织EGFR、MUC5AC基因表达，免疫组化检测肺组织及气道上皮P-EGFR、P-ERK、P-JNK、P-p38、MUC5AC蛋白表达。结果 与空白组比较，模型组大鼠气道上皮黏液腺体增生及P-EGFR、P-ERK、P-JNK、P-p38、MUC5AC蛋白表达显著升高（P<0.01），MUC5AC mRNA表达升高（P<0.05）；与模型组比较，清金化痰汤组大鼠气道上皮黏液腺体增生及P-p38、P-ERK、MUC5AC蛋白表达显著降低（P<0.05，P<0.01），P-JNK蛋白表达显著升高（P<0.01）；清金化痰汤组肺组织EGFR、MUC5AC mRNA表达显著降低（P<0.01）。结论 清金化痰汤可能通过抑制EGFR下游ERK、p38信号通路，干预COPD气道黏液高分泌。

关键词：清金化痰汤；慢性阻塞性肺疾病；气道黏液高分泌；表皮生长因子受体/MAPK信号通路；大鼠

DOI：10.3969/j.issn.1005-5304.2016.10.014

中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2016）10-0056-07

Abstract： Objective To observe the effects of Qingjin Huatan Decoction on EGFR/MAPK signaling pathway of airway mucus hypersecretion rats with chronic obstructive pulmonary disease （COPD）. Methods Intratracheal instillation of LPS combined with smudging method was used to establish COPD airway mucus hypersecretion rat models. Experimental rats were randomly divided into blank group， model group， Qingjin Huatan Decoction group and clarithromycin group. The blank group was normally fed， while the other three groups were given NS， Qingjin Huatan Decoction， and clarithromycin respectively for gavage， once a day for 30 days. All rats were killed on the 31st day， and pathological changes of lung tissue and mucous glands hyperplasia were observed by HE staining method. The gene expressions of EGFR and MUC5AC in lung tissue were detected by RT-PCR method. The protein expressions of P-EGFR， P-ERK， P-JNK， P-p38 and MUC5AC in pulmonary tissue and airway epithelium were detected by immunohistochemical method. Results Compared with the blank group， mucous glands hyperplasia on airway epithelium， protein expressions of P-EGFR， P-ERK， P-JNK， P-p38 and MUC5AC on airway epithelium significantly increased in the model group （P<0.01）； gene expression of MUC5AC of lung tissue increased （P<0.05）. Compared with the model group， mucous glands hyperplasia on airway epithelium， P-p38， P-ERK and MUC5AC protein expression on airway epithelium in Qingjin Huatan Decoction group significantly decreased （P<0.05， P<0.01）； the protein expression of P-JNK increased significantly （P<0.01）. EGFR and MUC5AC mRNA in lung tissue in Qingjin Huatan Decoction group decreased significantly （P<0.01）. Conclusion Qingjin Huatan Decoction can reduce airway mucus hepersecrection of COPD by inhibiting ERK and p38 signal pathway on EGFR downstream.

Key words： Qingjin Huatan Decoction； chronic obstructive pulmonary disease； airway mucus hypersecretion； EGFR/MAPK signaling pathway； rats

慢性阻塞性肺疾病（chronic obstructive pulmonary disease，COPD）是一种严重危害人类健康，加重家庭及社会负担的慢性呼吸系统疾病。气道黏液高分泌是COPD的重要病理特征，在COPD发病率和死亡率方面起着重要作用，干预气道黏液高分泌已成为目前治疗COPD的研究热点[1-2]。研究表明，细胞表皮生长因子受体（EGFR）及其下游MAPK信号通路与气道黏液高分泌密切相关[3-4]。前期研究表明，清金化痰汤可以显著抑制COPD模型大鼠EGFR基因表达，抑制气道黏液高分泌[5]。本实验观察清金化痰汤对COPD模型大鼠EGFR/MUC5AC信号通路的影响，探讨其干预COPD气道黏液高分泌的作用机制。

1 材料与方法