陈英 冯淬灵 李根茂 葛东宇 王骏

摘要：目的 探讨解毒清肺合剂调节慢性阻塞性肺疾病模型大鼠气道黏液高分泌的作用机制。方法 采用气道滴注脂多糖联合烟熏方法建立慢性阻塞性肺疾病模型。40只清洁级Wistar大鼠随机分为空白对照组、模型组、解毒清肺组与克拉霉素组。模型组、解毒清肺组、克拉霉素组分别给予生理盐水、解毒清肺合剂、克拉霉素灌胃，空白对照组正常喂养，连续30 d。实验第31日，处死大鼠提取肺组织，每组随机选取6只，HE染色观察肺组织病理形态及黏液腺体增生，实时荧光定量PCR检测各组大鼠肺组织中性粒细胞弹性蛋白酶（NE）、黏蛋白5AC（MUC5AC）mRNA表达，免疫组化检测肺组织及气道上皮NE、MUC5AC蛋白表达。结果 与空白对照组比较，模型组气道上皮黏液腺体增生、肺组织NE及MUC5AC mRNA表达、气道上皮NE及MUC5AC蛋白表达升高（P<0.05，P<0.01）；与模型组比较，解毒清肺组气道上皮黏液腺体增生显著降低（P<0.01），且作用与克拉霉素组相当；解毒清肺组肺组织NE、MUC5AC mRNA表达显著降低（P<0.01），且作用优于克拉霉素组；解毒清肺组气道上皮MUC5AC蛋白表达降低（P<0.05），且作用与克拉霉素组相当。解毒清肺组和克拉霉素组气道上皮NE蛋白表达与模型组无差异。结论 解毒清肺合剂通过NE下调MUC5AC蛋白表达，调节慢性阻塞性肺疾病气道黏液高分泌。

关键词：解毒清肺合剂；慢性阻塞性肺疾病；气道黏液高分泌；弹性蛋白酶；黏蛋白5AC；大鼠

中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2016）05-0073-05

Effects of Jiedu Qingfei Mixture on Expressions of NE and MUC5AC in Lung Tissue of Rats with Chronic Obstructive Pulmonary Disease CHEN Ying1， FENG Cui-ling1， LI Gen-mao2， GE Dong-yu2， WANG Jun1 （1. Dongzhimen Hosipital， Beijing University of Chinese Medicine， Beijing 100700， China； 2. Beijing University of Chinese Medicine， Beijing 100029， China）

Abstract： Objective To explore the mechanism of Jiedu Qingfei Mixture for airway mucus hypersecretion of rat models with chronic obstructive pulmonary disease （COPD）. Methods Airway instilling lipopolysaccharide combining fuming method was used to establish COPD models. Forty clean level Wistar strain rats were randomly divided into blank control group， model group， Jiedu Qingfei group， and clarithromycin group. Model group， Jiedu Qingfei group， and clarithromycin group were given normal saline， Jiedu Qingfei Mixture， and clarithromycin by gavage respectively， while the blank control group was raised normally for 30 d. All rats were killed on the 31st day for taking lung tissue （6 rats from each group were chosen randomly）. Pathological changes of lung tissue and mucous glands hyperplasia were observed by HE staining method. NE and MUC5AC mRNA expression on lung tissue were detected by RT-PCR method. Protein expressions of NE and MUC5AC on pulmonary tissue and airway epithelium were detected by immunohistochemical method. Results Compared with blank control group， mucous glands hyperplasia on airway epithelium， mRNA expression of NE and MUC5AC in lung tissue， and protein expressions of NE and MUCA5C on airway epithelium in the model group significantly increased （P<0.05， P<0.01）. Compared with model group， mucous glands hyperplasia on airway epithelium in Jiedu Qingfei group significantly decreased （P<0.01）， as same as clarithromycin group； Jiedu Qingfei group showed better effects on down-regulating NE and

MUC5AC mRNA expression in lung tissue compared with clarithromycin group. MUC5AC protein expression on airway epithelium in Jiedu Qingfei group significantly decreased （P<0.05）， as same as clarithromycin group. Jiedu Qingfei group and clarithromycin group had no difference on NE protein expression in airway epithelium compared with model group. Conclusion Jiedu Qingfei group Mixture can reduce airway mucus hypersecretion of COPD by down-regulating MUC5AC expression through neutrophil elastase.

Key words： Jiedu Qingfei Mixture； chronic obstructive pulmonary disease； airway mucus hypersecretion； neutrophil elastase； MUC5AC； 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的慢性呼吸系统疾病。以黏蛋白5AC（Mucin 5AC，MUC5AC）为主的气道黏液高分泌被认为是影响COPD病情和预后的独立危险因素[1]，气道黏液高分泌的机制主要有病原相关分子模式、中性粒细胞弹性蛋白酶（neutrophil elastase，NE）、细胞因子[2]，而NE被认为是目前已知最强促黏液分泌剂[3]。COPD属中医学“肺胀”范畴，痰、热、毒为慢性阻塞性肺疾病急性加重（acute exacerbation of chronic obstructive pulmonary disease，AECOPD）主要病理因素[4]。前期研究表明，解毒清肺合剂可减少AECOPD模型大鼠气道杯状细胞的化生，减少气道黏液高分泌[5]。……