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吴阳洋,欧阳桂兰,陈新海,游柏稳.固本平喘方对慢性阻塞性肺疾病模型大鼠气道MUC5AC水平的影响[J].湖南中医药大学学报,2024,44(4):565-571[点击复制] |
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| 固本平喘方对慢性阻塞性肺疾病模型大鼠气道MUC5AC水平的影响 |
| 吴阳洋,欧阳桂兰,陈新海,游柏稳 |
| (湖南中医药大学第二附属医院, 湖南 长沙 410000) |
| 摘要: |
| 目的 使用固本平喘方对香烟烟雾暴露联合气管滴注脂多糖（lipopolysaccharide，LPS）、猪胰弹性蛋白酶（porcine pancreatic elastase，PPE）造模大鼠进行干预，以主要分泌性黏蛋白5AC （mucin5AC，MUC5AC）作为评价指标，测定肺组织MUC5AC含量、蛋白表达、mRNA表达，评价各组气道黏液分泌强度。方法 将64只SPF级SD大鼠适应性饲养1周后，随机取52只进行造模，造模成功后取50只随机分为模型组（蒸馏水）、地塞米松组（0.15 mg·kg-1）及固本平喘方高（28 g·kg-1）、中（14 g·kg-1）、低（7 g·kg-1）剂量组，每组10只，另取10只大鼠为对照组（蒸馏水）。15 d后处死大鼠并收集标本。HE染色观察大鼠肺组织病理学变化；ELISA法检测肺组织MUC5AC水平；RT-PCR检测肺组织MUC5AC mRNA表达水平；Western blot检测肺组织MUC5AC蛋白表达；免疫荧光检测肺组织MUC5AC表达分布态势。结果 模型组大鼠肺组织结构显著破坏，肺泡腔内及肺间质见大量炎性浸润；地塞米松组及固本平喘方各剂量组可见肺泡腔受损程度及组织炎性浸润程度减轻。与对照组比较，其余各组MUC5AC含量均升高（P＜0.05）；与模型组比较，地塞米松组及固本平喘方各剂量组MUC5AC含量及MUC5AC mRNA表达均下降（P＜0.05），地塞米松组及固本平喘方高、中剂量组MUC5AC蛋白表达降低（P＜0.05）。肺组织免疫荧光可见，MUC5AC在模型组中呈现强表达，在地塞米松组中呈现较弱表达，在固本平喘方低、中、高剂量组中，表达强度依次减弱。结论 固本平喘方可改善慢性阻塞性肺疾病模型大鼠的肺组织病理损伤，有效减少MUC5AC含量、下调MUC5AC mRNA表达、降低MUC5AC蛋白表达，改善大鼠气道黏液高分泌状态。 |
| 关键词: 慢性阻塞性肺疾病 固本平喘方 黏蛋白5AC 气道黏液状态 |
| DOI：10.3969/j.issn.1674－070X.2024.04.006 |
| 投稿时间：2023-09-11 |
| 基金项目:湖南省中医药管理局重点项目（2021003）。 |
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| Effects of Guben Pingchuan Formula on MUC5AC level in the airway of chronic obstructive pulmonary disease model rats |
| WU Yangyang,OUYANG Guilan,CHEN Xinhai,YOU Bowen |
| (The Second Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410000, China) |
| Abstract: |
| Objective Using Guben Pingchuan Formula (GBPCF) to intervene the rat model established by cigarette smoke exposure combined with tracheal instillation of lipopolysaccharide (LPS) and porcine pancreatic elastase (PPE),taking mucin 5AC (MUC5AC) as the evaluation indicator,so as to measure its content,protein expression,and mRNA expression in the rat lung tissue and evaluate the airway mucus secretion intensity in each group.Methods After one week of adaptive feeding of 64 SPF-level SD rats,52 of them were randomly taken for modeling.After successful modeling,50 model rats were randomized into model group (distilled water),dexamethasone group (0.15 mg·kg-1),and GBPCF high-(28 g·kg-1),medium-(14 g·kg-1),and low-dose groups (7 g·kg-1),with ten rats in each group.Another ten rats were selected as control group (distilled water).After 15 days,the rats were euthanized and specimens were collected.HE staining was used to observe the pathological changes in the lung tissue of rats,ELISA to determine the level of MUC5AC in the lung tissue,RT-PCR to measure MUC5AC mRNA expression level,Western blot to examine MUC5AC protein expression,and immunofluorescence to check the distribution pattern of MUC5AC expression.Results The pulmonary tissue structure of the model group was notably damaged,with a large amount of inflammatory infiltration in the alveolar cavity and pulmonary interstitium.The degree of alveolar damage and tissue inflammatory infiltration was reduced in the dexamethasone group and GBPCF groups of various doses.Compared with the control group,the content of MUC5AC in all other groups increased (P＜0.05).Compared with the model group,the content and mRNA expression of MUC5AC in the dexamethasone group and GBPCF groups of various doses decreased (P＜0.05),and the MUC5AC protein expression in the dexamethasone group,the high-and medium-dose groups of GBPCF decreased (P＜0.05).Immunofluorescence of the lung tissue revealed strong MUC5AC expression in the model group,weaker expression in the dexamethasone group,and successively decreased expressions in the low-,medium-,and high-dose groups of GBPCF.Conclusion GBPCF can alleviate the pathological damage of lung tissue in model rats of chronic obstructive pulmonary disease (COPD),effectively reduce MUC5AC content,downregulate MUC5AC mRNA expression,decrease MUC5AC protein expression,and improve the high secretion state of airway mucus in rats. |
| Key words: chronic obstructive pulmonary disease Guben Pingchuan Formula mucin 5AC airway mucus |
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