摘要
目的:通过观察清金化痰汤对慢性阻塞性肺疾病(COPD)大鼠的自噬调节作用,探讨其对COPD大鼠的炎症反应的影响。方法:采用50只SPF级雄性SD大鼠随机分成5组,分别为正常组、模型组、清金化痰汤高、低剂量组(30,10 g·kg-1),罗红霉素组(0. 017 5 g·kg-1),每组10只,除正常组10只外,余40只用单纯烟熏方法建立COPD大鼠动物模型28 d。苏木素-伊红(HE)染色鉴定模型成立后,5组分别灌胃给药14 d,1次/d,模型组、正常组同体积生理盐水灌胃。末次灌胃1 h后处死大鼠提取气道,采用实时荧光定量聚合酶链式反应(Real-time PCR),蛋白免疫印迹法(Western blot)检测其自噬相关蛋白微管相关蛋白轻链3(LC3),Beclin-1表达;酶联免疫吸附测定(ELISA)检测炎症因子白细胞介素-6(IL-6),IL-8的含量变化。结果:Real-time PCR分析显示,与正常组比较,模型组自噬因子Beclin-1,LC3 mRNA表达均有不同程度升高(P <0. 05);与模型组比较,清金化痰汤处理后能够明显改善COPD大鼠气道上皮细胞自噬反应,自噬表达明显减少(P <0. 05),高剂量组与罗红霉素组比较差异不显著。Western blot结果显示,与正常组比较,模型组自噬蛋白表达明显升高(P <0. 05);与模型组比较,清金化痰汤药物处理组自噬蛋白Beclin-1,LC3表达下降(P <0. 05);清金化痰汤高剂量组与罗红霉素组比较差异不显著。ELISA结果显示,模型组大鼠的炎症水平升高,用药处理后大鼠气道上皮细胞内的炎症因子IL-6,IL-8含量均明显下降(P <0. 05)。结论:清金化痰汤能够减轻COPD大鼠支气管的炎症反应,其机制可能与清金化痰汤抑制气道上皮的自噬水平有关。
Objective:To explore the effect of Qingjin Huatan Tang(QJHTD)on the inflammatory response of chronic obstructive pulmonary disease(COPD)rats by observing the autophagy regulating effect of QJHTD on COPD rats.Method:The 50 SPF grade male rats were randomly divided into 5 groups,with 10 rats in each group.In addition to the normal group,the remaining 40 male rats were randomly divided into 5 groups.After the establishment of the hematoxylin and eosin(HE)staining identification model,the drugs were given to the 5 groups by gavage for 2 weeks,high and low-dose QJHTD groups were give the drug at 30,10 g·kg-1.Roxithromycin positive control group was given the drug at 0.017 5 g·kg-1.The model control group and the normal group were given the same volume of normal saline.At 1 h after the last gavage,the rats were put to death to extract the airway,and the expressions of autophagy microtuble-associated protein light chain 3(LC3),Beclin-1 were detected by Real-time quantitative PCR(Real-time PCR)and Western blot.Changes of inflammatory cytokines interleukin-6(IL-6)and interleukin-8(IL-8)were detected by enzyme linked immunosorbent assay(ELISA).Result:Real-time PCR analysis showed that compared with the normal group,Beclin-1 and LC3 mRNA expressions of autophagy factors in the model group were increased to varying degrees(P<0.05).QJHTD could significantly improve the autophagy response of airway epithelial cells in COPD rats,and the autophagy expression was significantly reduced compared with the model control group(P<0.05),with no significant difference between the high-dose group and the positive control group.Western blot results show that compared with the normal group,the expression of autophagy protein in the model group was significantly increased(P<0.05).compared with model control group,autophagy protein Beclin 1,LC3 expressions of the QJHTD treatment group were decreased(P<0.05).There was no significant difference between the high-dose QJHTD group and the Roxithromycin positive control group.ELISA results showed
作者
吴林娜
赵媚
许光兰
WU Lin-na;ZHAO Mei;XU Guang-lan(The First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,Nanning 530023,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2019年第18期30-35,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金地区基金项目(81760848)
关键词
清金化痰汤
慢性阻塞性肺疾病(COPD)
自噬
Qingjin Huatan Tang(QJHTD)
chronic obstructive pulmonary disease(COPD)
autophagy
