解毒清肺合剂对慢性阻塞性肺疾病气道黏液高分泌模型大鼠EGFR/MAPK信号转导通路的影响

Effects of Jiedu Qingfei Formula on EGFR/MAPK signaling pathway in rat model of chronic obstructive pulmonary disease with airway mucus hypersecretion

目的:探讨解毒清肺合剂调节慢性阻塞性肺疾病(COPD)大鼠气道黏液高分泌的作用机制。方法:40只清洁级Wistar大鼠随机分为空白对照组(A)、模型组(B)、解毒清肺组(C)、克拉霉素组(D)。B、C、D组大鼠给与气管内滴注LPS联合烟熏的方法建立COPD气道黏液高分泌模型。B、C、D组连续30d分别给予0.9%氯化钠溶液、解毒清肺合剂(8m L/kg)、克拉霉素(83.3mg/kg)灌胃,空白对照组正常喂养。实验第31天,处死大鼠提取肺组织,每组随机选取6只,采用实时荧光定量PCR检测肺组织表皮生长因子受体(EGFR)、黏蛋白5AC(MUC5AC)基因表达,阿尔新兰-过碘酸雪夫法观察气道上皮杯状细胞数目,肺组织及气道上皮EGFR、ERK、JNK、p38、MUC5AC蛋白的表达。结果:模型组大鼠肺组织杯状细胞数目、EGFR蛋白、ERK、JNK、p38、MUC5AC m RNA及MUC5AC蛋白表达较空白对照组显著升高(P<0.01);解毒清肺组和克拉霉素组杯状细胞数目、ERK、JNK、p38、MUC5AC m RNA及MUC5AC蛋白较模型组显著降低(P<0.05,P<0.01),克拉霉素组EGFR m RNA表达较模型组显著升高(P<0.01);解毒清肺组EGFR m RNA及MUC5AC m RNA表达较克拉霉素组显著降低(P<0.01,P<0.05),EGFR蛋白表达较克拉霉素组显著升高(P<0.01)。结论:解毒清肺合剂可能通过抑制ERK、JNK、p38MAPK信号通路,减轻COPD气道黏液高分泌。

Objective： To explore effect of Jiedu Qingfei Formula on EGFR/MAPK signaling pathway in rat model of chronic obstructive pulmonary disease with airway mucus hypersecretion. Methods： Forty Wistar rats were randomly divided into control group（A）, model group（B）, Jiedu Qingfei Formula group（C） and Clarithromycin group（D）. The model of COPD with airway mucus hypersecretion was established by dripping LPS in bronchus combined with smudging. Rats in all groups except for the control group separately received the normal saline, Jiedu Qingfei Formula and Clarithromycin by gavage, continuously for 30 days. All rats were sacrificed to collect the lung tissue on the 31 th day of experiment. The m RNA expression of EGFR and MUC5 AC were detected by real-time PCR. Alcian blue periodic acid Schiff staining method was used to observe hyperplasia of goblet cells on airway epithelium. Immunohistochemical method was used to detect the proteins expression of EGFR, ERK, JNK, p38 and MUC5 AC on pulmonary tissue and airway epithelium. Results： Compared with the control group, goblet cells on airway epithelium in the model group presented the hyperplasia. Compared with the control group, the proteins expression of EGFR and MUC5 AC in the model group were significantly increased, as well as the m RNA expression of ERK, JNK, p38 and MUC5 AC. Compared with the model group, the hyperplasia of goblet cells in the Jiedu Qingfei Formula group and Clarithromycin group was less observed. Compared with the model group, the protein expression of MUC5 AC in the two treatment groups was significantly decreased, as well as the m RNA expression of ERK, JNK, p38 and MUC5 AC. The m RNA expression of EGFR in the Clarithromycin group was significantly higher than that in the model group. Compared with the Clarithromycin group, the m RNA expression of EGFR and MUC5 AC in the Jiedu Qingfei Formula decreased, while the protein of EGFR increased. Conclusion： Jiedu Qingfei Formula could relieve the airway mucus hypersecretion of chronic obstructive pulmonary disease by the inhibition of ERK, JNK, p38 and MAPK signaling pathway.