MUC1抑制COPD中烟草烟雾诱导的黏液高分泌

目的：探究黏蛋白1(MUC1)对烟草烟雾（CS）暴露诱导的慢性阻塞性肺疾病（COPD）中MUC5AC和MUC5B的表达影响。方法：（1）动物实验：健康SPF级雄性C57BL/6小鼠20只,随机分为空白对照组（n=10）和CS暴露组（n=10）,采用CS暴露24周的方法建立COPD小鼠模型。建模结束后,检测小鼠肺功能;肺组织切片进行HE和PAS染色评估肺组织病理改变及气道杯状细胞增生情况;Western blot法检测肺组织MUC1蛋白水平;ELISA法检测支气管肺泡灌洗液中白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)、MUC5AC和MUC5B的表达水平,以评估小鼠肺部炎症及黏液分泌情况。（2）细胞实验：用烟草烟雾提取物（CSE）处理人支气管上皮细胞系16HBE,通过Western blot、RT-qPCR和免疫荧光染色检测MUC1、MUC5AC和MUC5B的表达。用MUC1小干扰RNA(siRNA)和MUC1过表达质粒转染16HBE细胞以沉默和过表达MUC1,确定MUC1对MUC5AC和MUC5B表达的影响。结果：（1）CS组小鼠表现为类似COPD的病变,包括肺气肿、肺功能下...     

通光散对慢性阻塞性肺疾病模型大鼠气道阻力和气道炎症的影响

目的 建立慢性阻塞性肺疾病(COPD)大鼠模型,观察通光散对COPD模型大鼠气道阻力和气道炎症的影响.方法 60只雄性SD大鼠按随机数字表法分为正常对照组、模型组、阳性对照组和通光散组,每组15只.模型组、阳性对照组和通光散组采用熏烟和气管给予内毒素脂多糖的复合方法建立COPD大鼠模型.第15天开始各组大鼠给予每周5次药物干预,通光散组给予通光散汤3 ml灌胃;阳性对照组给予羧甲司坦150 mg/kg灌胃;正常对照组和模型组给予生理盐水3ml灌胃.第90天实验结束,进行大鼠有创气道阻力测定和肺组织病理检测.结果 正常对照组、模型组、阳性对照组和通光散组大鼠有创气道阻力分别为(0.227±0.027) cm H2O· ml-1·s-1、(0.425±0.117)cm H2O·ml-1·s-1、(0.263±0.043) cm H2O· m l-1·s-1、(0.269±0.050)cm H2O·ml-1· s-1(1 cm H2O=0.098 kPa).模型组大鼠气道阻力高于其他3组(均P＜0.05),而其余3组之间差异均无统计学意义(均P＞0.05).病理检测显示,模型组大鼠肺组织有严重的支气管组织和肺泡组织病理改变,而通光散组和阳性对照组的病变程度均较轻.结论 通光散可以降低COPD模型大鼠的气道阻力,抑制气道炎性反应.     

气道黏液高分泌机制的研究进展

气道黏液高分泌主要表现为气道黏液理化性质的改变,包括以MUC5AC为主的黏液组分改变和由黏液腺细胞为主过渡到以杯状细胞为主的分泌黏液细胞的改变。香烟烟雾为主的吸入性有害物和细菌感染是诱导气道黏液高分泌的主要刺激物,而热休克蛋白、信号传导通路和黏蛋白MUC5AC表达的增加是气道黏液高分泌的主要机制。     

N-乙酰半胱氨酸联合阿奇霉素对慢性阻塞性肺疾病模型大鼠氧化应激的影响

 目的:探讨N-乙酰半胱氨酸(NAC)联合阿奇霉素(AZI)对慢性阻塞性肺疾病(COPD) 模型大鼠氧化应激的影响。方法:将60只雄性Wistar大鼠随机分为5组,即正常对照(control)组、模型(model)组、AZI治疗组、NAC治疗组和联合治疗组(AZI+NAC组)。采用烟熏联合气管内滴入脂多糖的方法诱导大鼠COPD模型。NAC组和AZI组大鼠每日烟熏前30 min分别给予NAC和AZI灌胃,AZI+NAC组则给予NAC和AZI联合灌胃。第31天行肺功能检测后处死大鼠,提取支气管肺泡灌洗液(BALF)进行细胞计数,并采用ELISA法测定BALF中白细胞介素-8(IL-8)、白细胞介素-17(IL-17)和肿瘤坏死因子-α(TNF-α)的含量。制作肺组织切片及肺匀浆,测定肺匀浆超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和丙二醛(MDA)水平。结果:与control组对比,其余4组均出现肺功能的下降,组织病理提示炎症细胞浸润、肺泡破坏等表现。与control组比较,其余4组BALF中白细胞总数、单核巨噬细胞、中性粒细胞和淋巴细胞均显著增高(P<0.05);与model组、AZI组、NAC组比较,AZI+NAC组BALF中白细胞总数、中性粒细胞和淋巴细胞均显著下降(P<0.05)。与model组对比,AZI组、NAC组和AZI+NAC组IL-8、IL-17、TNF-α和MDA含量均显著降低(P<0.05),SOD和GSH-Px活性均显著增高(P<0.05);与AZI组和NAC组比较,AZI+NAC组IL-8、IL-17、TNF-α和MDA含量均下降,SOD和GSH-Px活性均增高,差异有统计学显著性(P<0.01)。结论:NAC和AZI均能减轻COPD模型大鼠肺部炎症和氧化损伤,两者联合能增强抗氧化作用,可能更适合COPD的临床治疗。     

大鼠慢性阻塞性肺疾病模型的建立及其气道和肺组织M受体的水平

慢性阻塞性肺疾病(COPD)是一种重要的慢性呼吸系统疾病，其基础和临床研究越来越广泛，目前国内虽然有慢性支气管炎(慢支)动物模型的文献报道，但均未能证实其有气流阻塞，因此，不能称之为COPD。我们采用组织学检查和肺功能测定的方法对吸入高浓度SO2气体诱导的大鼠COPD模型进行了鉴定，并对其气道和肺组织M受体进行了检测。1 材料和方法1.1 动物：选体重80～120g雄性Wistar大鼠17只，随机分为2组，COPD组8只，正常对照组9只。将COPD组大鼠暴露于250ppm的SO2气体，5h/d,5d/w，共7周。1.2 肺功能测定：乌拉坦腹腔注射麻醉大鼠，…     

槲皮素基于EGFR信号通路减轻大鼠气道黏液高分泌北大核心CSCD

目的:探讨槲皮素对LPS诱导的大鼠气道黏液高分泌的作用及其分子机制。方法:将36只SD大鼠随机分为:对照组、LPS组、槲皮素低、中、高剂量组、AG1478组;气道内滴注LPS建立大鼠气道黏液高分泌模型。HE染色观察肺组织的病理变化;AB-PAS染色观察杯状细胞增生和黏液分泌情况;ELISA检测肺组织中MUC5AC浓度;采用免疫组织化学法(IHC)检测MUC5AC、EGFR、PKC、NF-κB表达;RT-qPCR检测肺组织MUC5AC及EGFR mRNA表达;Western blot检测p-EGFR、EGFR、p-PI3K、PI3K、p-PKC、PKC、p-AKT、AKT、NF-κB表达。结果:与LPS组相比,槲皮素低、中、高剂量组病理变化逐渐减轻,肺组织炎症积分和AB-PAS阳性着色面积逐渐降低(P<0.05);槲皮素能减少肺组织MUC5AC、EGFR、PKC、NF-κB表达,降低MUC5AC含量,并下调MUC5AC及EGFR的mRNA水平(P<0.05);槲皮素能减少p-EGFR/EGFR、p-PI3K/PI3K、p-PKC/PKC、p-AKT/AKT、NF-κB蛋白表达(P<0.05)。结论:气管内滴注LPS能成功诱导大鼠气道黏液高分泌模型,槲皮素对LPS诱导大鼠气道黏液高分泌具有拮抗作用,其作用机制可能与抑制EGFR信号通路,调控p-EGFR/EGFR、p-PI3K/PI3K、p-PKC/PKC、p-AKT/AKT、NF-κB表达,下调MUC5AC表达有关。     

慢性阻塞性肺疾病小鼠模型制备方法的比较研究

目的:采用香烟烟雾(cigarette smoke,CS)暴露、肺炎克雷伯杆菌(Klebsiella pneumoniae,KP)感染、聚肌胞苷酸(polyinosinic-polycytidylic acid,Poly I:C)滴鼻、CS暴露联合KP感染和CS暴露联合Poly I:C滴鼻5种方法,建立并比较慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)小鼠模型。方法:将288只雄性BALB/c小鼠随机分为正常组(normal组)、CS组、KP组、CS+KP组、Poly I:C组及CS+Poly I:C组,每组48只。第1～8周造模,分别于第4、8、16和24周末取材。观察小鼠肺组织平均肺泡数(mean alveolar number,MAN)、肺泡平均截距(mean linear intercept,MLI)、肺功能呼气峰流速(peak expiratory flow,PEF)和50%潮气量呼气流量(50%tidal volume expiratory flow,EF50)的变化,检测肺组织中肿瘤坏死因子α(tumor nec...     

大鼠慢性阻塞性肺疾病模型的建立

慢性阻塞性肺疾病（ ｃｈｒｏｎｉｃ ｏｂｓｔｒｕｃｔｉｖｅ ｐｕｌ－ｍｏｎａｒｙ ｄｉｓｅａｓｅ, ＣＯＰＤ）是具有气流阻塞特征的慢性支气管炎和／或肺气肿［１］,由于其发病率高,严重影响患者的劳动能力和生活质量,是倍受重视与广泛研究的疾病。为便于开展ＣＯＰＤ的研究,本实验在Ｒｅｉｄ［２］建立慢性支气管炎大鼠模型方法的基础上,采用ＳＯ２熏吸法,通过检测模型大鼠肺功能,确认有气流阻塞发生而建立的ＣＯＰＤ实验模型。 材料和方法 一、实验动物 ４周龄的健康雄性Ｗｉｓｔａｒ大鼠４５只,体重１００～２００ｇ,随机分…     

气管滴注大气细颗粒物构建大鼠慢性阻塞性肺疾病模型

目的:采用气管滴注大气细颗粒物(PM2.5)建立慢性阻塞性肺疾病(COPD)肺功能损伤大鼠模型,并对该模型进行评价.方法:SD大鼠随机分为正常对照组、2.5 mg/kg PM2.5组、5 mg/kg PM2.5组和10 mg/kg PM2.5组.按1 mL/kg气管滴注PM2.5溶液,连续5次.采用小动物呼吸肺功能测量...     

Th17细胞在慢性阻塞性肺疾病肺部炎症中的作用

由于长期吸烟的刺激,活化的辅助性T细胞17（Th17）能浸润到肺组织并参与肺泡壁的破坏及肺气肿的形成.Th17细胞在IL-23的作用下,分泌IL-17、IL-21、IL-22等多种细胞因子,其中IL-17可以促进气道中性粒细胞的募集和激活,IL-21在维持、增强Th17细胞的数量及功能的同时,能增加CD8＋细胞的数量及其细胞毒活性,在吸烟诱导的慢性阻塞性疾病（COPD）肺部连续炎症中发挥持续放大效应.在不同的炎症环境中,Th17细胞及其分泌的细胞因子与Th1细胞、调节性T细胞（Treg）以及多种参与固有免疫应答的细胞间相互作用和调节,共同参与COPD的发病,并构成了固有免疫与适应性免疫之间的桥梁.     

Exacerbations of chronic obstructive pulmonary disease and chronic mucus hypersecretion

Chronic obstructive pulmonary disease (COPD) exacerbations are an important cause of the considerable morbidity and mortality found in COPD. COPD exacerbations increase with increasing severity of COPD, and some patients are prone to frequent exacerbations leading to hospital admission and readmission. These frequent exacerbations may have considerable impact on quality of life and activities of daily living. Factors that increase the risk for COPD exacerbations are associated with increased airway inflammation caused by common pollutants and bacterial and/or viral infections. These inflammatory responses cause mucus hypersecretion and, thereby, airway obstruction and associated exacerbations. While chronic mucus hypersecretion is a significant risk factor for frequent and severe exacerbations, patients with chronic mucus hypersecretion have a lower rate of relapse after initial treatment for acute exacerbation. The benefit of antibiotics for treatment of COPD exacerbations is small but significant. While the mechanisms of actions are not clear, mucolytic agents reduce the number of days of disability in subjects with exacerbations. Reducing mucous cell numbers in small airways could be a useful way to reduce chronic mucus hypersecretion. Our studies suggest that programmed cell death is crucial in the resolution of metaplastic mucous cells, and understanding these mechanisms may provide novel therapies to reduce the risk of COPD exacerbations.     

FIZZ1在吸烟大鼠COPD模型肺组织的表达及与气道炎症的相关性研究

目的:探讨FIZZ1在吸烟大鼠慢性阻塞性肺疾病(COPD)模型肺组织内的表达及其与COPD慢性气道炎症的相关性。方法:选用70只雄性Wistar大鼠,采用单纯吸入香烟烟雾的方法制造大鼠COPD模型,HE染色观察大鼠肺组织病理变化鉴定造模成功。采用免疫组化定性分析及Western blot定量检测COPD大鼠肺组织中FIZZ1在不同时点的蛋白表达。对支气管肺泡灌洗液(BALF)行炎症细胞计数。酶联免疫吸附法(ELISA)检测不同时点BALF及血清中白细胞介素4(IL-4)和肿瘤坏死因子α(TNF-α)的含量。结果:HE染色显示模型组大鼠在第20周之后炎症反应呈慢性过程,并逐渐呈现COPD的病理特征。免疫组化结果显示在模型组中,FIZZ1蛋白表达随着炎症反应的加剧而显著增强,且在COPD病变组织处表达相对较强;Western blot进一步证实了免疫组化的检测结果(P0.05)。与对照组比较,模型组大鼠BALF中炎症细胞总数、中性粒细胞绝对数和淋巴细胞绝对数在第4周开始均显著升高(P0.05)。一定范围内,与对照组比较,模型组大鼠BALF及血清中炎症因子IL-4和TNF-α升高(P0.05)。结论:FIZZ1作为一种肺组织特异性炎症介质,可能参与了COPD炎症反应的发生与发展,其机制可能与诱导炎症细胞浸润和炎症因子分泌有关。     

Immune response in chronic obstructive pulmonary disease

Chronic obstructive pulmonary disease (COPD) is a major public health problem because of its high prevalence, rising incidence and associated socio-economic cost. The inhalation of toxic particles and gases, mostly tobacco smoke, is the main risk factor for COPD. Yet, not all smokers are equally susceptible to these toxic effects and only a percentage of them develop the disease (so-called ‘susceptible smokers’). This, in combination with the observation that COPD shows familial aggregation, suggests that the genetic background of the smoker is a key element in the pathogenesis of the disease. On the other hand, it is well established that ‘susceptible’ smokers exhibit an enhanced inflammatory response of the lung parenchyma as compared with ‘resistant’ smokers (i.e., those who manage to maintain lung function within the normal age range despite their habit). Importantly, in COPD patients this inflammatory response does not resolve after quitting smoking, again at variance with resistant smokers. All in all, these observations suggest that the pathogenesis of COPD may involve, in some patients, an autoimmune component which contributes to the enhanced and persistent inflammatory response that characterizes the disease. Here we: i) review briefly the pathobiology of COPD; ii) present the available scientific evidence supporting a potential role for autoimmunity in COPD; iii) propose a three-step pathogenic hypothesis in the transition from smoking to COPD; and iv) discuss potential implications for the diagnosis and treatment of this frequent, growing, devastating and costly disease.     

胸腺五肽辅助治疗慢性阻塞性肺病的疗效观察

目的观察胸腺五肽辅助治疗慢性阻塞性肺病(COPD)的疗效。方法将78例COPD患者随机分为2组,对照组给予常规治疗,治疗组加用胸腺五肽,观察比较2组患者症状体征缓解时间、平均住院时间以及3个月内再入院率。结果治疗组症状体征缓解时间、平均住院时间均较对照组显著缩短(P<0.05),治疗组3个月内再入院率较对照组显著降低(P<0.05)。结论胸腺五肽可增强COPD患者的免疫功能,缩短平均住院时间,降低再发风险,提高患者生活质量。     

慢性阻塞性肺疾病的全身效应

 慢性阻塞性肺疾病（Chronic Obstructive Pulmonary Disease,COPD）是一种常见病、多发病,其典型表现为气流受限和气道重塑,导致肺的结构与功能的改变。由于COPD的患者多种炎症介质的激活使患者处于系统性炎症状态,使得COPD不仅仅是呼吸系统的疾病,同时还包含许多肺外表现,即全身效应。常见的全身性表现包括：骨骼肌萎缩、恶液质、骨质疏松;而慢性炎症使COPD的患者缺血性心脏病、肺动脉高压、糖尿病等疾病的患病率大大增加。     

Prediction of exacerbation onset in chronic obstructive pulmonary disease patients

The objective of this study was to develop an algorithm for prediction of exacerbation onset in Chronic Obstructive Pulmonary Disease (COPD) patients based on continuous self-monitoring of physiological parameters from telehome-care monitoring. 151 physiological parameters of COPD patients were monitored on a daily/weekly basis for up to 2 years. Data were segmented in 30-day periods leading up to an exacerbation (exacerbation episode) and starting from a 14-day recovery period post-exacerbation (control episode) and tested in 6 intervals to predict exacerbation onset using k-nearest neighbour (k?=?1, 3, 5). A classifier with sensitivity of 73%, specificity of 74%, positive predictive value of 69%, negative predictive value of 78% and an accuracy of 74% was achieved using data intervals consisting of 5 days. Intelligent processing of physiological recordings have potential for predicting exacerbation onset.     

Persistence and genetic adaptation of Pseudomonas aeruginosa in patients with chronic obstructive pulmonary disease

ObjectivesIt is unclear whether recurrent sputum culture with Pseudomonas aeruginosa from patients with chronic obstructive pulmonary disease (COPD) is caused by intermittent airway carriage by different P. aeruginosa lineages or persistent carriage by the same lineage, and whether lineages genetically adapt during carriage.MethodsWhole-genome sequencing was performed for P. aeruginosa isolates sampled longitudinally from sputum cultures in patients with COPD who were enrolled in an ongoing randomized controlled trial (clinicaltrials.gov: NCT03262142).ResultsA total of 153 P. aeruginosa isolates were sequenced for 23 patients during 365 days of follow-up. Recurrent presence of P. aeruginosa was seen in 19 patients (83%) and was caused by persistence of the same clonal lineage in all but one patient. We identified 38 genes mutated in parallel in two or more lineages, suggesting positive selection for adaptive mutations. Mutational enrichment analysis revealed genes important in antibiotic resistance and chronic infections to be more frequently mutated.DiscussionRecurrent P. aeruginosa was common and carried for a prolonged time after initial detection in the airways of patients with COPD. Recurrence was caused by persistence of the same clonal lineage and was associated with genetic adaptation. Trial data on possible clinical benefits of attempting antibiotic eradication of P. aeruginosa in COPD are warranted.