烟雾暴露大鼠气道胰岛素样生长因子1及骨桥蛋白的表达变化

目的观察烟雾暴露与戒烟大鼠气道胰岛素样生长因子1(IGF-1)与骨桥蛋白(OPN)的表达变化,探讨烟雾暴露大鼠气道损伤的机制。方法将雄性SD大鼠40只随机分为5组:对照组(C组)、烟雾暴露1个月组(S1组)、烟雾暴露1个月及戒烟1个月组(Q1组)、烟雾暴露3个月组(S2组),烟雾暴露3个月及戒烟1个月组(Q2组),HE染色观察肺组织病理学改变;免疫组化染色检测IGF-1及OPN蛋白在气道的表达;RT-PCR检测肺组织IGF-1及OPN mRNA的表达。结果与C组比较,S1、S2组支气管上皮细胞IGF-1、OPN蛋白及mRNA表达均增强(P<0.05),Q1、Q2组支气管上皮细胞的IGF-1、OPN蛋白及mRNA表达低于各吸烟组(P<0.05),支气管IGF-1蛋白与OPN蛋白呈正相关(r=0.558,P<0.05)。结论 IGF-1、OPN参与烟雾暴露大鼠的气道重塑,早期戒烟有助于逆转气道重塑。     

低剂量茶碱对气道黏液高分泌影响的研究

目的探讨低剂量茶碱对大鼠气道黏液高分泌的影响及其作用机制.方法32只SD大鼠随机分为空白对照组、模型组、茶碱自身对照组、茶碱干预组,造模后第21天处死大鼠,采集肺组织标本和血标本,用苏木精-伊红(HE)染色、阿辛兰-过碘酸雪夫染色(AB-PAS),观察气道上皮细胞黏蛋白的分泌;免疫组化检测肺内气道杯状细胞的Muc5ac蛋白表达和肺内气道的核转录因子-κB(NF-κB);实时荧光定量PCR(Real-time PCR)法检测肺组织Muc5acmRNA;放射免疫法检测肺组织匀浆cAMP含量;采用高效液相色谱法检测血浆茶碱浓度.结果丙烯醛雾化可导致大鼠出现明显的气道黏液高分泌表现,用低剂量茶碱干预可显著降低气道AB/PAS染色阳性、Muc5ac蛋白及mRNA的表达和NF-κB的入核率,但未能使肺组织cAMP含量恢复.结论低剂量茶碱可抑制丙烯醛所致的气道黏液高分泌,其机制可能与抑制NF-κB的活化有关;未显示通过抑制cAMP起降解作用.     

利多卡因对大鼠气道黏膜表面结构的影响

目的:探讨利多卡因对大鼠气道黏膜表面结构的影响.方法:将25只Wistar大鼠随机分为正常对照组5只、利多卡因组20只,经大鼠气管穿刺注入利多卡因,并在2、24、48、72 h 4个时间点处死大鼠5只.取气管后1/3段的腹侧部,做扫描电镜观察大鼠气道黏膜表面结构.结果:利多卡因组均出现不同程度的黏液分泌增加,黏稠度增加,纤毛倒伏、粘着在一起、排列紊乱、部分断裂缺失.结论:利多卡因可对大鼠气道黏膜表面结构造成损害,但其可自我修复.     

易喘平胶囊对哮喘模型小鼠气道黏液高分泌的抑制作用

目的:观察易喘平胶囊(补肾活血法)对哮喘气道黏液高分泌的抑制作用及机制。方法:将32只雄性BALB/c小鼠随机分为正常对照组、哮喘组、地塞米松组和易喘平胶囊组(n=8),制备卵蛋白致敏的哮喘模型,高碘酸希夫(PAS)法检测各组小鼠气道黏膜杯状细胞数量及黏液分泌情况,逆转录聚合酶链反应(RT PCR)检测肺组织黏液蛋白MUC5AC mRNA的水平,检测支气管肺泡灌洗液(BALF)中嗜酸粒细胞数及细胞总数,酶联免疫吸附法(ELISA)检测肺组织匀浆液中白细胞介素4(IL4)和γ干扰素(IFNγ)的含量。结果:与哮喘组相比,易喘平胶囊组和地塞米松组小鼠小支气管上皮杯状细胞数量明显减少,管腔内黏液量也减少;同时后两组的MUC5AC mRNA水平也显著降低(P均<0.01)。易喘平胶囊组和地塞米松组BALF中嗜酸粒细胞数和细胞总数,以及肺组织中IL4含量均低于哮喘组(P均<0.01),而IFNγ含量有所提高(P均<0.01)。结论:易喘平胶囊对哮喘气道黏液高分泌具有一定抑制作用,调节辅助T淋巴细胞Th1/Th2的平衡、降低气道炎症程度是其发挥作用的机制之一。     

N-乙酰半胱氨酸联合小剂量阿奇霉素对慢性阻塞性肺疾病稳定期气道黏液高分泌的治疗效果

目的探讨N-乙酰半胱氨酸、小剂量阿奇霉素联合治疗慢性阻塞性肺疾病(COPD)稳定期气道黏液高分泌的效果。方法回顾性分析2017年11月至2018年11月78例COPD稳定期气道黏液高分泌患者的临床资料,依据用药方案分为观察组与对照组,每组39例。观察组给予N-乙酰半胱氨酸、小剂量阿奇霉素联合治疗,对照组给予常规治疗。结果观察组的炎症指标和肺功能指标较对照组组佳,差异有统计学意义(P 0. 05)。结论 N-乙酰半胱氨酸、小剂量阿奇霉素联合治疗气道黏液高分泌的效果显著,能改善COPD稳定期患者的预后质量。     

慢性阻塞性肺疾病急性加重期患者的非人工气道管理

慢性阻塞性肺疾病(COPD)患者的气道病理改变主要为炎症细胞浸润表层上皮,黏液分泌腺增大和杯状细胞增多使黏液分泌增加,在此基础上出现相应的黏液高分泌、纤毛功能失调、气流受限、肺过度充气、气体交换异常等病理生理学改变,黏液高分泌和纤毛功能失调导致慢性咳嗽及多痰[1].因而气道管理在COPD患者中显得尤为重要.细菌感染是导致COPD患者急性加重的主要诱因,在急性加重期痰液量增加明显,粘稠不易咳出,进一步加重将导致人工气道的建立,而急性加重期加强气道管理可以有效地降低人工气道的建立,在改善患者预后的同时减轻人工气道建立所带来的痛苦及经济负担.     

气道廓清技术在气道黏液高分泌相关疾病中的应用现状

介绍临床常用气道廓清技术的原理及使用方法,剖析其在呼吸科气道黏液高分泌相关疾病中的应用现状,为临床应用及气道廓清技术的进一步研究提供参考依据。     

平喘方对支气管哮喘模型大鼠气道顺应性影响研究

目的研究平喘方防治支气管哮喘的作用机理。方法通过对SD大鼠用卵蛋白等腹腔注射致敏、雾化吸入激发，建立支气管哮喘大鼠模型。于哮喘激发后28d观察空白对照组、哮喘模型组，平喘方组、地塞米松组各组大鼠气道顺应性及肺组织病理变化。结果模型组较空白对照组Co显著减小（P〈0．01），Cp显著增高（P〈0．01），气遗顺应性下降；光镜下，模型大鼠肺内细支气管上皮部分损伤脱落，细支气管壁上平滑肌细胞增生，胶原纤维增多。平喘方组较模型组Co显著增加（P〈0．01），Cp显著减少（P〈0．01）．气道顺应性改善；光镜下，平喘方组大鼠支气管壁上胶原纤维减少。结论平喘方能明显改善气道顺应性，改善腔原沉积，改善气道重建。     

复方甲氧那明胶囊联合克拉霉素治疗21例感染后咳嗽伴有气道黏液高分泌患者的临床疗效观察

选取2012年12月²013年12月在我院接受治疗的42例感染后咳嗽合并气道黏液高分泌的患者,按照抽签法将其分为治疗组和对照组各21例,治疗组采用复方甲氧那明胶囊与克拉霉素联合治疗,对照组单纯采用复方甲氧那明胶囊治疗。治疗7d后,观察两组患者的临床效果及咳嗽的严重程度。结果治疗组患者临床总有效率为85.71%(18/21),对照组临床总有效率为66.67%(14/21),两组比较差异有统计学意义(P<0.05)。两组患者均没有不良反应发生。克拉霉素与复方甲氧那明联合治疗伴有气道黏液高分泌的感染后咳嗽,能够有效缓解患者的咳嗽症状。     

低渗液体外培养对气道上皮黏液分泌的影响及与水通道蛋白-5表达的相关性研究

目的 探讨体外气液界面培养气道上皮在低渗液条件下黏液分泌与水通道蛋白-5（AQP5）表达间的关系，以进一步阐明渗透浓度、AQP5在慢性气道黏液高分泌过程中的地位和作用。方法利用体外气液界面细胞培养模型，进行兔气道上皮细胞原代培养。培养1周后，培养液渗透浓度换为235mmol／L（实验A组）、255mmol／L（实验B组）、270mmol／L（实验C组）和282mmol／L（对照组）。培养12h后，采用逆转录-聚合酶链反应（RT—PCR）检测各组AQP5mRNA表达，采用蛋白质免疫印迹法检测上清液中黏蛋白5AC（MUC5AC）分泌量。结果 倒置显微镜下各培养组细胞成多层生长。各实验组中MUC5AC分泌量均高于对照组（P〈0．001），其中以实验A组为最高；而AQP5 mRNA均明显低于对照组（P〈0．001），以实验A组为最低；实验组AQP5mRNA表达与MUC5AC呈显著负相关（r=-0.77，P〈0.001）。结论 在气液界面细胞培养模型中，低渗液条件下MUC5AC分泌量显著增加，较好地模拟了慢性气道炎症黏液高分泌的体内环境；AQP5mRNA表达与MUC5AC呈显著负相关。低渗环境以及由此引起的AQP5mRNA表达可能参与慢性阻塞性肺疾病黏液高分泌的形成过程。     

血必净注射液对卵蛋白致敏小鼠气道MUC5AC及Th1／Th2细胞因子表达的影响

目的研究血必净注射液对卵蛋白（OVA）致敏小鼠气道MUC5AC及Th1/Th2细胞因子表达的影响,探讨干预气道黏液高分泌的机制。方法卵蛋白腹腔注射致敏小鼠,32只小鼠随机分组为：对照组、哮喘组、地塞米松治疗组和血必净治疗组,每组8只,酶联免疫吸附试验（ELISA）检测小鼠肺泡灌洗液（BALF）中IL-4、IFN-γ的水平变化,实时RT-PCR检测小鼠肺组织MUC5AC mRNA表达变化。结果 （1）与对照组小鼠气道MUC5A mRNA表达（0.377±0.021）相比,哮喘组（1.103±0.087）明显增多（P〈0.01）;地塞米松治疗组（0.403±0.038）及血必净治疗组（0.437±0.031）小鼠气道MUC5A mRNA表达较哮喘组明显降低（P〈0.01）;（2）与对照组小鼠BALF表达IL-4[（22.812±1.978）ng/L]及IFN-γ[（101.232±9.664）ng/L]比较,哮喘组BALF表达IL-4[（87.234±6.901）ng/L]水平升高（P〈0.01）,而IFN-γ表达[（47.231±3.887）ng/L]明显降低（P〈0.01）;与哮喘组比较,地塞米松治疗组（[36.289±3.012）ng/L]及血必净治疗组IL-4水平[（38.112±2.761）ng/L]均显著降低（P〈0.01）;结论血必净注射液降低IL-4和MUC5AC的表达,提示在抑制气道黏液高分泌及控制哮喘方面具有意义。     

T细胞免疫球蛋白-黏蛋白-1对卵蛋白致敏小鼠气道MUC5AC及Th1/Th2细胞因子表达的影响

目的 研究T细胞免疫球蛋白-黏蛋白-1（TIM-1）对卵蛋白（OVA）致敏小鼠气道MUC5AC及Th1/Th2细胞因子表达的影响,探讨气道黏液高分泌的机制.方法 OVA腹腔注射致敏小鼠,随机分组为：正常对照组（A组）、哮喘组（B组）和哮喘＋ TIM-1抗体处理组（C组）,每组8只,酶联免疫吸附试验（ELISA）检测小鼠肺泡灌洗液（BALD中IL-4、IFN-γ的水平变化,实时RT-PCR检测外周血单个核细胞（PBMCs） TIM-1 mRNA及气道MUC5AC mRNA表达.结果 （1）与A组小鼠外周血PBMCs表达TIM-1 mRNA（0.618±0.043）相比,B组（1.129±1.101）及C组（0.898±0.071）TIM-1 mRNA表达明显增多（P＜0.01）,且C组明显低于B组（P＜0.01）;（2）B组和C组小鼠BALF中IL-4表达分别为（67.123±3.341） ng/L和（44.217±2.201）ng/L,较A组[（23.211±2.142） ng/L]明显增多（P＜0.01）,且C组明显低于B组（P＜0.01）;B组及C组小鼠BALF表达IFN1分别为（53.475±4.776） ng/L和（87.116±5.611）ng/L,均低于A组[（124.624±9.292） ng/L] （P ＜0.01）,而C组与B组相比,有所增加（P＜0.01）;（3）B组（1.004±0.081）及C组（0.673±0.029）小鼠气道MUC5A mRNA表达与A组（0.314±0.027）相比,明显增加（P＜0.01）,且C组低于B组（P＜0.01）;（4）小鼠外周血PBMCs TIM-1 mRNA表达与IL-4、MUC5AC表达呈正相关,而与IFN-γ表达呈负相关.结论 抑制TIM-1表达可减少IL-4和MUC5AC的分泌,提高IFN-γ表达,提示在调节黏液高分泌及哮喘治疗中具有意义.     

Epithelial tethering of MUC5AC-rich mucus impairs mucociliary transport in asthma

The development of pathologic mucus, which is not readily cleared from the airways, is an important contributor to the morbidity and mortality associated with asthma. It is not clear how the major airway mucins MUC5AC and MUC5B are organized within the mucus gel or how this gel contributes to airway obstruction in asthma. Here, we demonstrated that mucus plugs from individuals with fatal asthma are heterogeneous gels with distinct MUC5AC- and MUC5B-containing domains. Stimulation of cultured human bronchial epithelial cells with IL-13, a key mediator in asthma, induced the formation of heterogeneous mucus gels and dramatically impaired mucociliary transport. Impaired transport was not associated with defects in ciliary function but instead was related to tethering of MUC5AC-containing mucus gel domains to mucus-producing cells in the epithelium. Replacement of tethered mucus with untethered mucus restored mucociliary transport. Together, our results indicate that tethering of MUC5AC-containing domains to the epithelium causes mucostasis and likely represents a major cause of mucus plugging in asthma.     

Physiology of airway mucus clearance

Respiratory tract secretions consist of mucus, surfactant, and periciliary fluid. The airway surface fluid is present as a bilayer, with a superficial gel or mucous layer and a layer of periciliary fluid interposed between the mucous layer and the epithelium. A thin layer of surfactant separates the mucous and periciliary fluid layers. The mucous layer extends from the intermediate airway to the upper airway and is approximately 2-10 microm thick in the trachea. Airway mucus is the secretory product of the goblet cells and the submucosal glands. It is a nonhomogeneous, adhesive, viscoelastic gel composed of water, carbohydrates, proteins, and lipids. In health, the mucous gel is primarily composed of a 3-dimensional tangled polymer network of mucous glycoproteins or mucin. Mucin macromolecules are 70-80% carbohydrate, 20% protein, and 1-2% sulfate bound to oligosaccharide side chains. The protein backbones of mucins are encoded by mucin genes (MUC genes), at least 8 of which are expressed in the respiratory tract, although MUC5AC and MUC5B are the 2 principal gel-forming mucins secreted in the airway. Mucus is transported from the lower respiratory tract into the pharynx by air flow and mucociliary clearance. Expectorated sputum is composed of lower respiratory tract secretions along with nasopharyngeal and oropharyngeal secretions, cellular debris, and microorganisms. Disruption of normal secretion or mucociliary clearance impairs pulmonary function and lung defense and increases risk of infection. When there is extensive ciliary damage and mucus hypersecretion, airflow-dependent mucus clearance such as cough becomes critically important for airway hygiene.     

Azithromycin inhibits MUC5AC production induced by the Pseudomonas aeruginosa autoinducer N-(3-Oxododecanoyl) homoserine lactone in NCI-H292 Cells

The features of chronic airway diseases, including chronic bronchitis, cystic fibrosis, bronchiectasis, and diffuse panbronchiolitis, include chronic bacterial infection and airway obstruction by mucus. Pseudomonas aeruginosa is one of the most common pathogens in chronic lung infection, and quorum-sensing systems contribute to the pathogenesis of this disease. The quorum-sensing signal molecule [N-(3-oxododecanoyl) homoserine lactone (3O-C(12)-HSL)] not only regulates bacterial virulence but also is associated with the immune response. In this study, we investigated whether 3O-C(12)-HSL could stimulate the production of a major mucin core protein, MUC5AC. The effect of a macrolide on MUC5AC production was also studied. 3O-C(12)-HSL induced NCI-H292 cells to express MUC5AC at both the mRNA and the protein levels in time- and dose-dependent manners. A 15-membered macrolide, azithromycin, inhibited MUC5AC production that was activated by 3O-C(12)-HSL. 3O-C(12)-HSL induced extracellular signal-regulated kinase (ERK) 1/2 and I-kappa B phosphorylation in cells, and this induction was suppressed by azithromycin. 3O-C(12)-HSL-induced MUC5AC production was blocked by the ERK pathway inhibitor PD98059. Our findings suggest that the P. aeruginosa autoinducer 3O-C(12)-HSL contributes to excessive mucin production in chronic bacterial infection. Azithromycin seems to reduce this mucin production by interfering with intracellular signal transduction.     

Upper airway mucus deposition in lung tissue of burn trauma victims

Previous study in an ovine model of smoke inhalation and burn (S + B) injury has shown distal migration of upper airway mucus. This study examines the localization of an upper airway gland specific mucus, mucin 5B (MUC5B) in lung autopsy tissues of burn-only injury and in victims of S + B injury. We hypothesize that victims with S + B injury would exhibit increased distal migration of MUC5B than that seen in victims of burn-only injury. Autopsy lung tissue from victims of burn injury alone (n = 38) and combined S + B injury (n = 22) were immunostained for MUC5B. No normal lung tissues were included in the study. Semiquantitative analysis of the extent of MUC5B in bronchioles and parenchyma was performed on masked slides. Irrespective of injury conditions, all victims showed MUC5B in bronchioles. Mucin 5B was seen in the parenchyma except in two burn victims. No statistically significant difference was seen in the mean bronchiolar and parenchyma MUC5B scores between S + B and burn-only victims (P > 0.05). No strong statistical correlation of MUC5B scores with days postinjury or to the number of ventilatory days was evident. The percentage of pneumonia, identified histologically, was also similar between study groups. This study did not confirm our results in an ovine model of S + B injury. In contrast, virtually all pediatric burn victims, regardless of concomitant inhalation injury, showed MUC5B in their bronchioles and parenchyma. Increased mucus synthesis and/or impaired mucociliary function may contribute to the pulmonary pathophysiology associated with burn injury.