Cellular changes in bronchoalveolar lavage fluid in hyperoxia-induced lung injury

It is well known that high concentration oxygen exposure is a model of acute lung injury (ALI). However, controversy exists over the mechanism. This study was designed to clarify the cellular characteristics in bronchoalveolar lavage fluid (BALF) and body weight loss of rats exposed to oxygen(> 90%). Young male Wistar rats, aged 6 weeks, were divided into three groups: (1) room air group (exposed to room air, n = 22); (2) hyperoxia < 48 h group (exposed to over 90% oxygen for less than 48 h, n = 18); (3) hyperoxia 66–72 h group (exposed to over 90% oxygen for 66–72 h group, n = 7). Compared to the room air group, the total cell counts in the hyperoxia 66–72 h group decreased, whereas the neutrophils increased significantly. The body weights of the rats exposed to room air continued to increase. However, the body weights of oxygen-exposed rats increased slightly on the first day and weight loss was seen from the second day. All rats were noted to have bilateral pleural effusion in the hyperoxia 66–72 h group. The data suggests that (1) an increase in neutrophil count is an evident feature of hyperoxia-induced lung injury; (2) high concentration oxygen exposure can give rise to anorexia and malnutrition, which may play a role in hyperoxia-induced lung injury. Blocking neutrophil influx into lung tissue in the early phase and improving malnutrition are two effective methods to reduce hyperoxic lung injury.     

氰戊菊酯染毒大鼠肺灌洗液中细胞与生化效应

给大鼠一次气管注入氰戊菊酯悬液(FS)以及中毒室内吸入氰戊菊酯(FA),通过分析BALF中细胞与生化组分评价肺脏的毒性反应。结果表明PAM_s数下降,PMN数、Alb和NANA含量增加,LDH、ALP和ACP活性升高,提示氰戊菊酯引起肺细胞生物膜损伤、弥漫性肺泡炎和肺水肿。氰戊菊酯的阈剂量和阈浓度分别为0.93 mg/kg和200 mg/m~3,阈下剂量和阈下浓度为0.19 mg/kg和40 mg/m~2。研究结果为探讨氰戊菊酯的肺脏毒性及其机理,为制订其卫生标准提供依据。     

支气管肺泡灌洗后兔肺组织的病理观察

目的观察支气管肺泡灌洗后兔肺组织病理变化。方法将实验用新西兰兔31只予以气管插管并分为生理盐水10ml(A组,n=11)、亚胺培南(50mg/10ml)(B组,n=8)和氟康唑(800μg/10ml)(C组,n=12)3组。灌洗2h后处死动物,采用常规石蜡组织切片和HE染色法对兔肺组织进行病理观察。结果肺细支气管腔和部分肺泡腔内可见淡红色液体,并由此导致局限性肺气肿和/或肺不张。肺细支气管管腔内和管壁积有散在的中性粒细胞;支气管黏膜上皮坏死、剥脱。3组上述病变组间无明显差异。结论与生理盐水灌洗比较,亚胺培南或氟康唑局部肺泡灌洗并未增加局部病理损伤。     

Histopathological observation of rabbits lung tissue after bronchoalveolar lavage

目的 观察支气管肺泡灌洗后兔肺组织病理变化.方法 将实验用新西兰兔31只予以气管插管并分为生理盐水10 ml(A组,n=11)、亚胺培南(50 mg/10 ml) (B组,n=8)和氟康唑(800 μg/10 ml) (C组,n=12)3组.灌洗2h后处死动物,采用常规石蜡组织切片和HE染色法对兔肺组织进行病理观察.结果 肺细支气管腔和部分肺泡腔内可见淡红色液体,并由此导致局限性肺气肿和/或肺不张.肺细支气管管腔内和管壁积有散在的中性粒细胞;支气管黏膜上皮坏死、剥脱.3组上述病变组间无明显差异.结论 与生理盐水灌洗比较,亚胺培南或氟康唑局部肺泡灌洗并未增加局部病理损伤.     

Varespladib inhibits secretory phospholipase A2 in bronchoalveolar lavage of different types of neonatal lung injury

Secretory phospholipase A2 (sPLA2), which links surfactant catabolism and lung inflammation, is associated with lung stiffness, surfactant dysfunction, and degree of respiratory support in acute respiratory distress syndrome and in some forms of neonatal lung injury. Varespladib potently inhibits sPLA2 in animal models. The authors investigate varespladib ex vivo efficacy in different forms of neonatal lung injury. Bronchoalveolar lavage fluid was obtained from 40 neonates affected by hyaline membrane disease, infections, or meconium aspiration and divided in 4 aliquots added with increasing varespladib or saline. sPLA2 activity, proteins, and albumin were measured. Dilution was corrected with the urea ratio. Varespladib was also tested in vitro against pancreatic sPLA2 mixed with different albumin concentration. Varespladib was able to inhibit sPLA2 in the types of neonatal lung injury investigated. sPLA2 activity was reduced in hyaline membrane disease (P < .0001), infections (P = .003), and meconium aspiration (P = .04) using 40 μM varespladib; 10 μM was able to lower enzyme activity (P = .001), with an IC(50) of 87 μM. An inverse relationship existed between protein level and activity reduction (r = 0.5; P = .029). The activity reduction/protein ratio tended to be higher in hyaline membrane disease. Varespladib efficacy was higher in vitro than in lavage fluids obtained from neonates (P < .001).     

Alteration of bronchoalveolar lavage cell populations following bleomycin treatment in mice

Clinical use of the antitumor antibiotic bleomycin (BLM) is associated with the development of interstitial pulmonary fibrosis. Shortly after acute lung damage caused by intratracheal (it) administration of BLM to experimental animals there is an influx of inflammatory cells which are believed to modulate the process of fibrosis. This study was undertaken to determine what subpopulations of lymphocytes were in the bronchoalveolar lavage (BAL) cell population of C57BL/6J mice at various times after a single it dose of BLM and to determine whether BAL T-lymphocytes were activated after BLM treatment. The BAL lymphocyte population was analyzed by differential cell analysis and flow cytometry utilizing monoclonal antibodies specific for lymphocyte subpopulations. The majority of lymphocytes in the BAL of control and BLM-treated mice were T-lymphocytes, with less than 10% being B-cells. During the first 7 days after BLM the number of Lyt-2+ T-cells exceeded L3T4+ T-cells while in control mice the reverse was observed. The percentage of BAL lymphocytes expressing the IL-2 receptor did not change significantly at 3 and 7 days after BLM treatment, but was significantly decreased at Day 14. In contrast, the total number of lymphocytes expressing the IL-2 receptor was increased at all time points investigated. These results demonstrated that the majority of lymphocytes in the BAL were T-cells and that while the percentage of activated lymphocytes did not increase following BLM treatment, the absolute number did and this increased number of activated lymphocytes may be important in the disease process.     

急性双光气染毒兔肺泡灌洗液表面张力的变化

目的　通过对日本大耳兔双光气肺损伤后肺泡灌洗液蛋白和表面张力变化的测定 ,探讨肺泡表面活性物质 (PS)在双光气肺损伤机制中的作用。方法　纯种日本大耳白兔 40只 ,性别不拘 ,体重为 (3 0 4± 0 78)kg,随机分为对照组 (1 0只 )、即刻组 (5只 )、2h组 (5只 )、4h组 (5只 )、8h组 (5只 )、1 2h组 (5只 ) ;处理前 1 2h停喂饲料和水 ;染毒的浓度为 (30 0± 2 0 ) μg L ,染毒时间为 2 0min。染毒后 ,对于按不同时间分组的各组实验动物分别于 0、2、4、8和 1 2h处死。处死后立即取出肺脏 ,按 30ml kg体重的剂量 ,用肺灌洗液分次灌洗支气管肺泡。灌洗液离心 ,80 0r min× 1 0min ,取上清备用。结果　发现肺泡灌洗液蛋白含量随着染毒后时间的延长而逐渐增加 (P <0 0 1 ) ,并存在一定的线性相关 ,直线方程为 :^y =0 92 1 2 +0 842 8×T ,r=0 952 0 (^y :蛋白总量 ,T :时间 )。肺泡灌洗液最大表面张力变化不明显。肺泡灌洗液收缩时最小表面张力升高 ,迟滞环面积减小 ,且在染毒后立即就迅速下降 (P <0 0 1 )。同时 ,随着染毒后动物中毒时间的延长 ,最大表面张力和最小表面张力的差距也逐渐缩小。由此可见 ,肺泡表面张力改变 ,除了与磷脂发生变化有关外 ,还与蛋白质大量渗入、以及相关活性蛋白成分进入肺     

The fate of antioxidant enzymes in bronchoalveolar lavage fluid over 7 days in mice with acute lung injury

Characterization of lung injury is important if timely therapeutic intervention is to be used properly and successfully. In this study, lung injury was defined as the progressive formation of pulmonary edema. Our model gas was phosgene, a pulmonary edemagenic compound. Phosgene, widely used in industry, can produce life-threatening pulmonary edema within hours of exposure. Four groups of 40 CD-1 male mice were exposed whole-body to either air or a concentration x time (c x t) amount of 32-42 mg/m(3) (8-11 ppm) phosgene for 20 min (640-840 mg x min/m(3)). Groups of air- or phosgene-exposed mice were euthanized 1, 4, 8, 12, 24, 48, or 72 h or 7 days postexposure. The trachea was excised, and 800 micro l saline was instilled into the lungs and washed back and forth 5 times to collect bronchoalveolar lavage fluid (BALF). The antioxidant enzymes glutathione peroxidase (GPx), glutathione reductase (GR), superoxide dismutase (SOD), total glutathione (GSH), and protein were determined at each time point. Phosgene exposure significantly enhanced both GPx and GR in phosgene-exposed mice compared with air-exposed mice from 4 to 72 h, p < or = 0.01 and p < or = 0.005, respectively. BALF GSH was also significantly increased, p < or = 0.01, from 4 to 24 h after exposure, in comparison with air-exposed. BALF protein, an indicator of air/blood barrier integrity, was significantly higher than in air-exposed mice 4 h to 7 days after exposure. In contrast, BALF SOD was reduced by phosgene exposure from 4 to 24 h, p < or = 0.01, versus air-exposed mice. Except for protein, all parameters returned to control levels by 7 days postexposure. These data indicate that the lung has the capacity to repair itself within 24-48 h after exposure by reestablishing a functional GSH redox system despite increased protein leakage. SOD reduction during increased leakage may indicate that barrier integrity is affected by superoxide anion production.     

儿童肺泡灌洗液RT-PCR检测对支原体感染的辅助诊断价值探讨

目的 探讨支原体感染儿童肺泡灌洗液RT-PCR检测的早期诊断和病情评估。方法 选取2016年1月至12月安徽省儿童医院收治的235例感染肺炎支原体（MP）的患儿,检测重症肺炎和轻症肺炎肺泡灌洗液MP-DNA和血清MP-IgM,根据2项指标的差异和MP-DNA拷贝量的高低,分析MP-DNA与病情的关系。结果 男患儿MP-IgM阳性率低于女患儿,>3~14岁患儿MP-DNA阳性率高于1~3岁患儿,入院前病程≤ 7 d患儿MP-IgM阳性率低于入院前病程>7 d患儿,差异有统计学意义（P<0.05）。重症肺炎患儿中,MP-DNA高拷贝量占85.71%,高于轻症肺炎患儿,差异有统计学意义（P<0.05）;重症肺炎的MP-DNA拷贝量高于轻症肺炎患儿,差异有统计学意义（P<0.05）。结论 RT-PCR法检测支原体肺炎患儿肺泡灌洗液MP-DNA拷贝量可为快速确诊和判断患儿病情提供参考。     

Fibreoptic bronchoscopy and bronchoalveolar lavage in the investigation of the immunocompromised lung.

The records of 18 immunocompromised patients with recent onset of pulmonary disease who had fibreoptic bronchoscopy and bronchoalveolar lavage over a two year period (1989-90) were reviewed. The underlying diseases were human immunodeficiency virus (HIV) infection (n = 7), organ transplantation (n = 9), and chemotherapy for malignancy (n = 2). Four patients were receiving prophylactic therapy and 12 had been started on empirical therapy for infection. Patients proceeded to bronchoscopy either because of atypical disease presentation or failure to respond to empirical therapy. Bronchoscopy with bronchoalveolar lavage was diagnostic in 13/18 (72%) patients and provided clinically useful information in 16/18 (89%). There was one diagnostic failure (6%); Pneumocystis carinii pneumonia in an HIV positive patient receiving nebulised pentamidine prophylaxis was missed. Transbronchial biopsies were not routinely performed and provided additional diagnostic information in only 1/6 (17%) patients. Overall, the commonest diagnoses were Pneumocystis carinii pneumonia (61%) and cytomegalovirus pneumonitis (28%). There were no complications of the procedures. In this highly selected setting of diagnostic or therapeutic uncertainty, fibreoptic bronchoscopy with bronchoalveolar lavage remains an effective and safe technique for evaluating pulmonary disease in immunocompromised patients.     

Inhalation toxicity of methyl isocyanate: biochemical and cytological profile of bronchoalveolar lavage fluid in rats

Acute pulmonary injury in rats following a single inhalation exposure to methyl isocyanate (3.2 mg l-1) was reflected by alterations in the biochemical and cytological constituents of bronchoalveolar lavage fluid (BALF) over a period of 30 days. Total protein, sialic acid and lactic acid contents of BALF were increased followed by a gradual decline to normalcy between day 3 and day 30 post-exposure. The activity of lactic dehydrogenase also increased progressively with time. The cellularity of BALF was increased significantly and primarily comprised of polymorphonuclear neutrophils at 8 days. The adherence of macrophages was unchanged but their viability was lowered at 30 days post-exposure. The results indicate the significance of bronchoalveolar lavage analysis in monitoring pulmonary toxicity by methyl isocyanate, which is characterized by the hypoxic condition and reduced cellular defence. Some toxic manifestations are potentially reversible with time after cessation of exposure.     

Some further properties of human pulmonary mast cells recovered by bronchoalveolar lavage and enzymic dispersion of lung tissue

The properties of human pulmonary mast cells obtained by bronchoalveolar lavage (BAL) and enzymic dispersion of lung tissue have been compared with those of basophil leucocytes. On challenge with anti-human IgE, the pulmonary cells released both histamine and the newly generated mediators prostaglandin D2 (PGD2) and leukotriene C4 (LTC4). In contrast, the blood leucocytes released histamine but very little leukotriene and no prostanoid. Interestingly, both basophil leucocytes and BAL cells released histamine spontaneously in a hyperosmolar environment whereas dispersed lung (DL) cells showed limited reactivity under these conditions. The possible clinical significance of these findings in human bronchial asthma is discussed.     

实时荧光定量PCR检测肺泡灌洗液中白细胞吞噬细菌种类对ICU内HAP或VAP的诊断价值

目的 探讨实时荧光定量PCR(RT-PCR)检测肺泡灌洗液中白细胞吞噬细菌种类对ICU内医院获得性肺炎(HAP)或呼吸机相关性肺炎(VAP)患者的诊断价值。方法 根据临床肺部感染评分(CPIS),将ICU内92例存在人工气道的患者分为CPIS>6分组(65例)和CPIS≤6分组(27例)。采用CD45抗原及磁微粒分离法获取肺泡灌洗液中白细胞,提取细胞内DNA,进行RT-PCR扩增,检测肺泡灌洗液中白细胞内鲍曼不动杆菌、铜绿假单胞菌、肺炎克雷伯菌、金黄色葡萄球菌和大肠埃希菌。结果 CPIS>6分组RT-PCR与痰培养细菌检测结果无统计学差异(P>0.05),两种方法吻合度较高,一致性有统计学差异(Kappa=0.846,P<0.05);RT-PCR检测CPIS>6分组细菌阳性的灵敏度、特异度和正确指数分别为93.55%、91.18%和0.85。而CPIS≤6分组RT-PCR与痰培养细菌检测结果有统计学差异(P<0.05),两种方法吻合度较差,一致性无统计学差异(Kappa=0.065,P>0.05)。结论 采用RT-PCR检测肺泡灌洗液中白细胞吞...     

Some properties of mast cells obtained by human bronchoalveolar lavage

The properties of human pulmonary mast cells obtained by enzymic dispersion of whole lung and by bronchoalveolar lavage (BAL) have been compared with those of the basophil leucocyte. The latter cell types responded with release of histamine to challenge with anti-human IgE but the dispersed cells reacted only after passive sensitisation with serum from an atopic donor. Disodium cromoglycate inhibited the release of histamine from both types of pulmonary mast cell although the characteristics of the inhibition were different in the two cases. The drug was ineffective against the basophil. Increased numbers of mast cells were recovered by lavage of asthmatic subjects and these cells responded to immunological challenge with an enhanced release of histamine. The possible clinical significance of these findings in human bronchial asthma is discussed.     

Comparative sensitivity of measurements of lung damage made by bronchoalveolar lavage after short-term exposure of rats to ozone

Consequences of exposure of rats for 2 days or less to O3 at various concentrations between 0.12 and 0.96 ppm were measured using several assays performed on bronchoalveolar lavage fluid. Changes in apparent lung permeability were assessed by measurement of recovery of labelled bovine serum albumin in lung lavage fluid after intravenous injection ("permeability index"). The relative sensitivity of this assay was compared with the sensitivity of measurements of changes in protein and of enzyme content in lavage fluid. Permeability index increased in an exposure concentration-dependent manner after 6 or 24 h of exposure to O3 at or above levels of 0.4 ppm. Permeability index was also increased after 2 days of exposure to 0.2 ppm of O3. The activities of lactate dehydrogenase, acid phosphatase, and N-acetyl-beta-D-glucosaminidase in lung lavage fluid were less sensitive indicators of O3 damage than was altered permeability index. Increased lactate dehydrogenase activity could only be detected after continuous exposure of rats for at least 1 day to 0.64 (or higher) ppm of O3, while acid phosphatase and N-acetyl-beta-D-glucosaminidase activities were increased after exposure of rats to O3 at 0.4 ppm or above for 1 day. Activities of these enzymes were not increased after 6 h of exposure to 0.64 ppm of O3 or after 2 days of exposure to 0.2 ppm. Increased lavage protein content was the most sensitive measurement of the consequences of O3 exposure to rats in these protocols. The lavagable protein content increased after exposure of rats to O3 for 6 h at 0.4 ppm and for 1 or 2 days of exposure to 0.12 ppm, the current peak hourly National Ambient Air Quality standard for O3. While the biological significance of these observations remains to be determined, measurement of lavage protein content is a simple, sensitive indicator of acute changes in the lung caused by exposure to environmentally relevant concentrations of O3.     

The comparison of a fibrogenic and two nonfibrogenic dusts by bronchoalveolar lavage

Analysis of bronchoalveolar lavage fluid (BALF) appears to be a sensitive approach to characterizing an acute inflammatory response within the lung. More work, however, is needed to determine if analyses of BALF endpoints can predict chronic responses (i.e., fibrosis). The objective of the present study was to compare the dose and temporal pulmonary response of a known fibrogenic agent, silica, and two known nonfibrogenic agents, aluminum oxide and titanium dioxide. Animals were instilled with silica (0, 0.2, 1.0, or 5.0 mg/100 g body wt), titanium dioxide (1.0 or 5 mg/100 g body wt), aluminium oxide (1.0 or 5.0 mg/100 g body wt) or saline. Animals (n = 5/group) were terminated 1, 7, 14, 28, and 63 days following instillation, and the BALF was characterized by biochemical and cellular assays. Histopathological changes were determined at 60 days after exposure. The biochemical results demonstrated BALF levels of lactate dehydrogenase (LDH), beta-glucuronidase (BG), N-acetylglucosaminidase (NAG), and total protein (TP) increased in a dose-related fashion at the earlier time points for all test materials, with the magnitude of change being greatest for silica. The temporal response for these parameters was significantly different for the two classes of materials. With time, the response for the fibrogenic dust steadily increased, while the levels for the nonfibrogenic dusts decreased toward normal values during the 2-month study period. Of the cellular changes, total cell numbers, neutrophils, and lymphocyte numbers were the most sensitive markers of the pulmonary response. As shown with the biochemical parameters, the cellular response to silica increased with time while that of the nuisance dusts did not. It was also found that, similar to inhalation studies, high doses of a nuisance dust may result in toxicity/inflammation. This toxicity at high dose levels emphasizes the importance of choosing relevant doses when comparing potentially fibrogenic and nonfibrogenic dusts. In conclusion, the persistent and progressive changes seen in the biochemical (LDH, TP, BG, NAG) and cellular parameters (total cells, neutrophils and lymphocytes) following silica administration correlated with the fibrotic response which occurred after exposure to this material. The less dramatic and transient changes seen with aluminum oxide and titanium dioxide correlated with the inert nature of these nuisance dusts. The results of this study indicate evaluation of BALF may provide a means to predict the chronic pulmonary response to a material.     

Inhalation toxicity of diborane in rats assessed by bronchoalveolar lavage examination

 Changes in bronchoalveolar lavage fluid (BALF) and blood were examined to assess the toxic effects of diborane (B₂H₆, CAS: 19287-45-7) on the lung. Male Wistar rats were exposed to diborane at 20 ppm (intended concentration) for 4 h (phase I study) to evaluate time-course changes up to 14 days, and at 10 or 1 ppm (intended concentrations) to assess the dose-effect relationship after 3 days (phase II study). BALF parameters [leukocyte counts, α₁-antitrypsin (α₁-AT), superoxide dismutase (SOD), total protein, phospholipids etc.] were examined and biochemical and histopathological studies were also carried out. In the phase I study, neutrophils (%) in BALF increased on the day of exposure and then decreased gradually for 3 days. Rapid and marked increases in α₁-AT and SOD activity in BALF were detected on the day of exposure, and phospholipids had sharply increased on day 3. After 14 days, these parameters in the exposed rats had returned to their background level and α₁-AT decreased significantly. In the phase II study, total protein, α₁-AT activity and phospholipids in BALF showed dose-dependent increases, and serum α₁-AT activity increased significantly. Alveolar capillary and alveolar cell damage were confirmed in rats exposed to 20 ppm, 10 ppm or 1 ppm diborane for 4 h by evaluating the parameters examined. The protection system appeared to start operating immediately after exposure, and the recovery mechanism seemed to start operating 1 day after exposure and cease by day 14. The no-observed-effect level could not be observed. Received: 14 February 1995 / Accepted: 30 May 1995     

Ranking toxicity of industrial dusts by bronchoalveolar lavage fluid analysis.

Female wistar rats were inoculated intratracheally with 10 mg/ml suspensions of various dusts, viz: quartz, fly ash, mica and corundum in physiological saline. Biochemical markers of bronchoalveolar lavage fluid (BALF) were analysed 8 days after the instillation of the dusts. Elevated levels of proteins, sialic acid and phospholipid contents and the activity of lactate dehydrogenase correlated well with the degree of the known fibrogenic potential of different dusts in the lungs in the following order, quartz greater than fly ash greater than mica greater than corundum. beta-glucuronidase activity, was however, only elevated in the quartz inoculated group of rats. It is suggested that biochemical constituents of BALF analysed shortly after the exposure to different dusts can be useful to mirror alterations in the tissue response to mineral dusts.     

支气管肺泡灌洗在弥漫性肺实质疾病预后评估中的价值

目的探讨支气管肺泡灌洗(BALF)在弥漫性肺实质疾病(DPLD)预后评估中的价值。方法回顾分析27例弥漫性肺实质疾病患者临床资料及BALF结果,对该组患者进行随访,观察BALF中细胞成分与预后的关系。结果 IPF5例,COP4例,NSIP7例,HP5例,CVD-ILD6例,病情缓解13例,加重3例,死亡11例;淋巴细胞比例与中性粒细胞比例与生存时间的COX回归分析显示淋巴细胞百分比为保护因素(OR=0.950,95%可信区间0.913～0.990,P=0.018)。中性粒细胞百分比为危险因素(OR=1.023,95%可信区间0.998～1.042,P=0.064)。结论 BALF在DPLD诊断及预后评估中具有重要价值,淋巴细胞增高为保护因素,中性粒细胞增高为危险因素。