金泰冲剂对哮喘豚鼠血浆及支气管肺泡灌洗液中一氧化氮与内皮素的影响

目的:观察金泰冲剂对哮喘豚鼠血浆及支气管肺泡灌洗液(BALF)中一氧化氮(NO)与内皮素(ET)的影响.方法:用卵蛋白成功诱发豚鼠哮喘,观察金泰冲剂对哮喘豚鼠血浆及支气管肺泡灌洗液中ET及NO的水平.结果:哮喘豚鼠支气管肺泡灌洗液中NO、ET水平较正常组无明显异常,而血清NO、BALF中ET水平较正常组升高,金泰冲剂可有效降低以上两者水平.结论:金泰冲剂可降低BALF中ET水平,推测其可通过调节ET水平而改善钙离子浓度、磷脂代谢而达到功效.     

纤维支气管镜检查结合支气管肺泡灌洗对慢性咳嗽的诊断意义

目的:探讨支气管肺泡灌洗对慢性咳嗽的诊断意义.方法:对经常规方法检查不能明确病因的56例慢性咳嗽患者行经纤维支气管镜肺泡灌洗术(broncho-alveolar lavage,BAL)、刷检、活体组织检查和涂片检查,收集支气管肺泡灌洗液(broncho alveolar lavage fluid,BALF)行细菌培养加药物敏感试验,同时将BALF离心沉淀物送检找抗酸杆菌、癌细胞(并行经口痰找抗酸杆菌、癌细胞).结果:56例病因不明的患者,经上述检查后最终确诊38例,诊断率68%.其中,口痰细菌检出阳性率5%,镜检及BALF检出阳性率68%,两组比较差异有统计学意义(P＜0.01).口痰找抗酸杆菌、癌细胞均阴性,BALF离心沉淀物找到抗酸杆菌9例,找到癌细胞8例.结论:纤维支气管镜检结合肺泡灌洗,可提高用常规方法检查病因不明的慢性咳嗽的确诊率.     

雾化吸入BCG对哮喘小鼠气道炎性作用的影响

目的探讨卡介苗（BCG）雾化吸入对变应性哮喘的治疗作用及其与支气管肺泡灌洗液（BALF）中γ-干扰素（IFN-γ）的关系。方法昆明小鼠32只，随机分成三组，正常对照组，哮喘模型组及治疗组。治疗组又分为BCG治疗组和地塞米松治疗组。治疗组与哮喘模型组一起制作哮喘模型。观察小鼠气道壁炎症变化，并收集支气管肺泡灌洗液（BALF），测定BALF中细胞总数，嗜酸性粒细胞和淋巴细胞数，用酶联免疫法测定BALF中IFN-γ的含量。结果哮喘模型组BALF中细胞总数和嗜酸细胞数显著高于正常对照组（P〈0．01），BALF中IFN-γ水平显著低于正常组（P〈0．01）；BCG治疗组、地塞米松治疗组BALF中细胞数、EOS细胞数显著低于哮喘组，而IFN-γ水平显著高于哮喘组（P〈0．01）。BCG治疗组各项指标与地塞米松治疗组相比差异无统计学意义（P〉0．05）。结论雾化吸入BCG能明显抑制哮喘小鼠气道的炎性反应，吸入BCG对哮喘小鼠有治疗作用。     

瘦素在肥胖哮喘小鼠气道重构中的作用

目的探讨脂肪因子瘦素在支气管哮喘气道重构中的作用。方法高脂饮食制造肥胖模型,经腹腔注射与雾化吸入卵蛋白(OVA)制作慢性哮喘模型。45只雌性C57/6J小鼠随机分为对照组,单纯哮喘组,肥胖哮喘组,每组15只,于末次激发48 h内处死小鼠;计数支气管肺泡灌洗液(BALF)中细胞总数及分类,肺组织HE观察各组小鼠肺组织病理变化及ELISA法测定血清中瘦素水平。结果肥胖哮喘组小鼠BALF中白细胞总数及嗜酸性粒细胞比例以及气道管壁厚度和气道平滑肌厚度均较单纯哮喘组和对照组显著增加(P0.05)。对照组血清瘦素为(1 438.54±208.14)pg/mL,明显低于单纯哮喘和肥胖哮喘组[(1 677.75±214.36)pg/mL,(2 114.20±189.15)pg/mL,P0.05)],血清瘦素水平与气管壁厚度和气道平滑肌厚度呈正相关(P0.05)。结论肥胖通过瘦素参与了哮喘气道重构过程,纠正瘦素失衡有望成为肥胖哮喘患者治疗的新方向。     

支气管肺泡灌洗液抗酸菌涂片诊断可疑肺结核52例分析

目的 :探讨支气管肺泡灌洗液涂片查抗酸菌对可疑肺结核的诊断价值。方法 :采用OLYMPUSB30型纤维支气管镜在相应病变肺段所属支气管分二次注入无菌生理盐水 5 0ml,负压吸引回收液 ,经过滤离心 ,取沉淀物涂片行抗酸染色。结果 :支气管肺泡灌洗液涂片抗酸染色阳性率 36 .5 % ,加术     

胃饥饿素治疗小鼠支气管哮喘的作用机制及对细胞焦亡的影响

目的 探讨胃饥饿素对小鼠支气管哮喘的治疗作用及对细胞焦亡的影响.方法 将30只小鼠随机分为对照组、模型组[卵清蛋白(OVA)造模]和治疗组(OVA联合胃饥饿素),每组10只.构建小鼠哮喘模型,应用病理学和乙酰甲胆碱的反应性评估气道损伤情况.采用酶联免疫吸附法(ELISA)测定肺泡灌洗液(BALF)中肿瘤坏死因子-α(T...     

经支气管镜支气管肺泡灌洗在外科术后肺部感染病原学诊断中的应用

目的 探讨经支气管镜支气管肺泡灌洗(bronchoalveolar lavage,BAL)在外科术后合并肺部感染患者病原学诊断中的应用价值.方法 96例外科术后合并肺部感染患者接受床头支气管镜下吸痰,并行支气管肺泡灌洗,对灌洗液进行病原学检查.结果 96例患者共行灌洗液培养109例次,获得104例次有效病原学检查结果...     

n-3多不饱和脂肪酸对哮喘小鼠白三烯B4及5脂氧合酶基因表达的影响

目的：探讨n-3多不饱和脂肪酸（n-3PUFA）对哮喘小鼠支气管肺泡灌洗液（bronchoalveolar lavage fluid，BALF）中白三烯B4（LTB4）水平、肺组织5脂氧合酶（5-LO）mRNA表达的影响及其可能的机制.方法：建立小鼠卵蛋白哮喘模型．分别灌胃不同剂量的n-3PUFA干预．用ELISA的方法检测BALF中LTB4含量；用RT—PCR的方法检测肺组织中5-LO mRNA表达的变化．结果：哮喘小鼠BALF中LTB4水平、肺组织5-LO基因表达水平较正常对照组显著升高（P〈0．01）；灌胃n-3PUFA干预的3个剂量组与模型组相比，LTB4含量、5-LO基因表达水平下降（P〈0．05）.结论：LTB4在气道里的高浓度及5LO基因表达的上调在哮喘小鼠发病中起重要作用，n—3PUFA可以降低哮喘小鼠BALF中LTB4的浓度，抑制肺组织5-LO mRNA表达，从而起到对抗哮喘气道炎症的作用.     

糖皮质激素对哮喘小鼠肺部炎症和支气管肺泡灌洗液中白介素-25、干扰素-γ的影响

目的:通过观察各实验组小鼠肺部炎症情况和检测支气管肺泡灌洗液(BALF)中白介素-25(IL-25或IL-17E)和干扰素-γ(IFN-γ)的水平,探讨糖皮质激素对小鼠支气管哮喘的治疗作用机制.方法:清洁级BALB/c小鼠随机分为正常组、哮喘组、地塞米松组和布地奈德组.以卵清蛋白致敏激发法建立哮喘小鼠模型.显微镜下观察肺组织病理切片;收集BALF,计算嗜酸性粒细胞(EOS)绝对计数百分比;用ELISA法测定BALF中IL-25和IFN-γ的水平,并做相关性分析.结果:哮喘组小鼠BALF中白细胞总数、EOS数目和百分比分别与正常组、地塞米松组和布地奈德组相比均明显增加(P<0.01).在正常组、地塞米松组和布地奈德组之间对这3个指标进行两两比较差异均无显著性(P>0.05).哮喘组小鼠BALF中IL-25和IFN-γ水平分别与正常组、地塞米松组和布地奈德组相比均减低(P<0.05),在正常组、地塞米松组和布地奈德组之间对这两个指标进行两两比较差异均无显著性(P>0.05).各组小鼠BALF中IL-25和IFN-γ水平都呈正相关关系.结论:糖皮质激素可减轻肺部炎症,促进IL-25和IFN-γ的表达,抑制哮喘症状.     

瘦素在肥胖哮喘小鼠气道重构中的作用

目的探讨脂肪因子瘦素在支气管哮喘气道重构中的作用。方法高脂饮食制造肥胖模型,经腹腔注射与雾化吸入卵蛋白（OVA）制作慢性哮喘模型。45只雌性C57／6J小鼠随机分为对照组,单纯哮喘组,肥胖哮喘组,每组15只,于末次激发48h内处死小鼠;计数支气管肺泡灌洗液（BALF）中细胞总数及分类。肺组织HE观察各组小鼠肺组织病理变化及ELISA法测定血清中瘦素水平。结果肥胖哮喘组小鼠BALF中自细胞总数及嗜酸性粒细胞比例以及气道管壁厚度和气道平滑肌厚度均较单纯哮喘组和对照组显著增加（P〈0．05）。对照组血清瘦素为（1438．54&#177;208．14）pg／mL,明显低于单纯哮喘和肥胖哮喘组[（1677,75&#177;214．36）pg／mL,（2114．20&#177;189．15）pg／mL,P〈0,05）],血清瘦素水平与气管壁厚度和气道平滑肌厚度呈正相关（P〈0．05）。结论肥胖通过瘦素参与了哮喘气道重构过程,纠正瘦素失衡有望成为肥胖哮喘患者治疗的新方向。     

橙皮苷对过敏性哮喘小鼠肺组织炎症及肺泡灌洗液中细胞因子水平影响

目的研究橙皮苷对过敏性哮喘小鼠肺组织炎症及肺泡灌洗液中细胞因子水平影响。方法实验分为:对照组、模型组、实验1组、实验2组和实验3组。模型组、实验1组、实验2组和实验3组构建哮喘模型,实验1组、实验2组和实验3组在实验第25天、26天、27天灌胃橙皮苷(浓度分别为:12 mg/kg、24 mg/kg、36 mg/kg),对照组给予等量的磷酸盐缓冲液(PBS)代替。对小鼠进行哮喘发作情况行为学评分和肺组织炎症评分,酶联免疫吸附(ELISA)检测肺泡灌洗液中白细胞介素-4(IL-4)、白细胞介素-5(IL-5)、白细胞介素-13(IL-13)、干扰素γ(IFN-γ)、白细胞介素-10(IL-10)水平,同时对肺泡灌洗液中炎性细胞数目进行分类计数,Western blot检测肺组织中NF-κBp65蛋白水平。结果与对照组比较,模型组小鼠哮喘发作情况行为学评分和肺组织炎症评分升高,肺泡灌洗液中IL-4、IL-5、IL-13水平也升高,肺泡灌洗液中炎性细胞总数、中性粒细胞数目、嗜酸性粒细胞数目、淋巴细胞数目升高,肺组织中NF-κBp65蛋白水平升高。橙皮苷灌胃处理后过敏性哮喘小鼠哮喘发作情况行为学评分、肺组织炎症评分降低,肺泡灌洗液中炎性细胞总数、中性粒细胞数目、嗜酸性粒细胞数目、淋巴细胞数目减少,IL-4、IL-5、IL-13水平降低,IFN-γ水平升高,IL-10水平也升高,与未经橙皮苷处理的哮喘小鼠比较,差异有统计学意义(P<0.05),呈剂量依赖性。结论橙皮苷能够呈剂量依赖性减轻过敏性哮喘小鼠炎症,调节肺泡灌洗液中细胞因子表达。     

Keratinocyte growth factor pretreatment is associated with decreased macrophage inflammatory protein-2alpha concentrations and reduced neutrophil recruitment in acid aspiration lung injury

A two-hit model of acid aspiration was used to examine the effect of keratinocyte growth factor (KGF) on chemokine levels and neutrophil recruitment into the lung. Mice were subjected to cecal ligation and puncture and then either KGF or saline, intratracheally (i.t.). Forty-eight hours later, the mice were given i.t. acid. After 8 h, neutrophil counts in bronchoalveolar lavage (BAL) fluid were significantly decreased in animals pretreated with KGF (23 +/- 4 x 10(3)/mouse) compared with saline (74 +/- 2 x 10(3)/mouse). In addition, the BAL fluid IL-6 levels were decreased in the KGF-treated group (88+/- 44 pg/mL) compared with the saline group (166 +/- 34 pg/mL). To examine the mechanism behind the KGF-induced reduction in neutrophil influx, the murine chemokines KC and macrophage inflammatory protein (MIP)-2alpha were measured. KC levels in plasma and BAL fluid were not significantly different between the treatment groups. Likewise, levels of MIP-2alpha in plasma were not affected by KGF treatment. However, 8 h after acid aspiration, MIP-2alpha concentrations were significantly lower in the KGF-treated group. The ratio of MIP-2alpha in BAL fluid versus plasma was lower in the KGF group (0.72 +/- 0.28) than in the saline group at 3 h (2.23 +/- 0.93) and also significantly lower in the KGF group (3.02 +/- 0.78) compared with the saline group (6.23 +/- 1.19) at 8 h. In this study, KGF pretreatment after acid aspiration was associated with reduced neutrophil recruitment into the lung and a decrease in MIP-2alpha gradients between BAL fluid and plasma.     

白细胞介素-25与支气管哮喘小鼠发病的关系

目的探讨IL-25与支气管哮喘发病的关系。方法将30只健康雌性Balb/c小鼠随机分为3组：分别为卵白蛋白（OVA）致敏并激发的哮喘模型组（A组）,OVA）致敏、激发并予糖皮质激素治疗组（B组）及正常对照组（C组）,每组10只。于末次激发后24h后各组小鼠摘眼球取血,ELISA法检测其血清IL-25、IFN-γ及IL-4水平。收集支气管肺泡灌洗液（BALF）,行嗜酸性粒细胞（EOS）计数,HE染色观察各组肺组织病理改变。RT-PCR法检测各组肺组织IL-25mRNA和IL-4mRNA表达。结果 1、B组多数小鼠未见明显行为学异常反应,肺组织病理损害较A组明显减轻,BALF中EOS计数（126.2±10.8）×103/L亦显著低于A组（324.7±13.6）×103/L（P〈0.01）。2、A组小鼠血清IL-25、IL-4水平[（80.66±3.06）pg/ml,（124.7±4.13）pg/m]均高于C组[（46.25±1.90）pg/ml,（32.73±2.20）pg/ml],IFN-γ水平[（45.25±5.34）pg/ml]低于C组[（80.56±2.49）pg/ml]其差异均有统计学意义（P〈0.01）。3、A组小鼠肺组织IL-25mRNA和IL-4mRNA表达均高于C组（P〈0.01）,B组小鼠肺组织IL-25mRNA和IL-4mRNA表达均低于A组（P〈0.01）。A组小鼠肺组织IL-25mRNA表达量与BALF中EOS计数呈正相关（r=0.901,P〈0.01）。结论 IL-25参与了哮喘小鼠的发病,在哮喘的发病中起促炎作用,其促进哮喘发病的作用可以被激素抑制。     

半乳凝素-3对哮喘豚鼠肺组织嗜酸性粒细胞凋亡及Bcl-2蛋白表达的影响

目的：探讨半乳凝素-3对哮喘豚鼠肺组织嗜酸性粒细胞凋亡及Bcl-2蛋白表达的影响。方法：36只豚鼠随机分为哮喘组、治疗组及对照组,每组12只。哮喘组及治疗组用卵白蛋白致敏和激发制作哮喘模型,对照组同期予生理盐水。于致敏第15～21日激发前1h治疗组豚鼠予半乳凝素-3腹腔注射,剂量0.5mg/kg,每日1次,连续7d。测定3组豚鼠支气管肺泡灌洗液（BALF）中IL-5含量、嗜酸性粒细胞计数,苏木素-伊红染色观察肺组织病理变化,TUNEL法检测肺组织嗜酸性粒细胞的凋亡,免疫组织化学检测肺组织Bcl-2蛋白的表达。结果：治疗组豚鼠BALF中IL-5含量和嗜酸性粒细胞计数均低于哮喘组（P〈0.05）;其肺组织炎症反应明显轻于哮喘组,肺组织嗜酸性粒细胞计数低于哮喘组,嗜酸性粒细胞凋亡指数高于哮喘组;肺组织Bcl-2蛋白表达低于哮喘组（P〈0.05）。与对照组比较,治疗组BALF中白细胞计数、嗜酸性粒细胞计数、嗜酸性粒细胞占白细胞的比例、IL-5水平均升高（P〈0.05或0.01）;肺组织嗜酸性粒细胞计数、嗜酸性粒细胞凋亡指数均显著升高（P均〈0.01）;肺组织Bcl-2蛋白表达比较差异无统计学意义（P〉0.05）。结论：半乳凝素-3可促进哮喘豚鼠BALF中嗜酸性粒细胞凋亡,减轻肺组织炎症反应,这可能与其降低哮喘豚鼠BALF中IL-5含量、下调肺组织Bcl-2蛋白表达有关。     

Recombinant human soluble tumor necrosis factor-alpha receptor fusion protein partly attenuates ventilator-induced lung injury

Ventilator-induced lung injury is mediated, at least in part, by TNF-alpha. We determined the effect of a recombinant human soluble TNF receptor fusion protein (etanercept) on mechanical ventilation (MV)-induced changes in a murine ventilator-induced lung injury model. After pretreatment with etanercept or placebo, C57Bl/6 mice were anesthetized and randomized to MV with either low tidal volumes (VT, approximately 7.5 mL/kg) or high VT ( approximately 15 mL/kg) for 5 h. Instrumented but spontaneously breathing mice served as controls. End points were lung wet-to-dry ratios, lung histopathology scores, protein levels, neutrophil cell counts and thrombin-antithrombin complex levels in bronchoalveolar lavage fluid (BALF), and cytokine levels in lung homogenates. The number of caspase 3-positive cells was used as a measure for apoptosis. Etanercept treatment attenuated MV-induced changes, in particular, in MV with high VT. Compared with placebo, etanercept reduced the number of neutrophils in BALF and thrombin-antithrombin complex levels in BALF and cytokine levels in lung homogenates. Lung wet-to-dry ratios, histopathology scores, and local protein levels in BALF, however, were not influenced by etanercept treatment. The number of caspase 3-positive cells was significantly higher in etanercept-treated animals. Inhibition of TNF by etanercept attenuates, in part, MV-induced changes.     

血管内皮细胞生长因子和诱导型一氧化氮合酶在支气管哮喘气道重塑中的表达及意义

目的:探讨哮喘气道重塑中血管内皮细胞生长因子(VEGF)和诱导型一氧化氮合酶(iNOS)的表达及意义。方法:32只大鼠中随机取8只作对照组,其余为哮喘组。哮喘组又分为哮喘3d(8只)、7d(8只)和14d(8只)组。对照组大鼠1d和8d各腹腔注射生理盐水2mL,15d开始用37℃生理盐水雾化吸入,1次/d,10min/次,共3d。各哮喘组用卵蛋白粉(OVA)10mg、氢氧化铝20mg制成2mL悬液代替生理盐水作腹腔注射,1%OVA代替生理盐水分别雾化吸入3、7和14d。末次激发24h后行肺泡灌洗,回收肺泡灌洗液,测VEGF和iNOS的浓度。取右下肺组织切片作常规HE染色,测定气道内周径(Pi)、外周径(Pe),并计算管壁面积(WA)和气道平滑肌面积(SMC-A)。结果:各哮喘组气道中VEGF和iNOS浓度、WA/Pi和SMC-A/Pi均高于对照组,而且VEGF和iNOS浓度分别与WA/Pi和SMC-A/Pi呈正相关。结论:VEGF和iNOS可能参与哮喘气道重塑。     

哮喘患者血清和支气管肺泡灌洗液中白细胞介素12水平检测及意义

目的观察急性期哮喘患者治疗前后血清和支气管肺泡灌洗液（BALF）中白细胞介素12（IL-12）水平的变化,研究其与疾病的关系。方法使用酶联免疫吸附测定（ELISA）法检测58例急性期哮喘患者（试验组）治疗前、后IL-12水平;健康非吸烟的自愿献血者30例,为血清对照组,检测其血清IL-12的水平;以胸痛为主诉进行BALF检查证实为健康者12例,作为BALF对照组,检测其IL-12水平的水平。结果急性期哮喘患者血清和BALF中IL-12水平明显低于缓解期（治疗后）和健康对照组（P〈0.05）,缓解期血清和BALF中IL-12水平较健康对照组比较,差异无统计学意义（P〉0.05）。结论提示急性期哮喘患者IL-12分泌减少,不能有效抑制炎症或促炎症细胞因子的合成及释放,亦即不能有效抑制炎症反应,可能是导致或加重气道炎症的原因之一。动态测定IL-12水平是判断支气管哮喘病情变化的有益指标。     

ELISA of ceruloplasmin and alpha-2-macroglobulin in paired bronchoalveolar lavage fluid and serum samples

Bronchoalveolar lavage fluid (BALF) was analysed to obtain information on leakage of proteins from the blood into the respiratory lumen and on local synthesis. Albumin, ceruloplasmin and alpha-2-macroglobulin were measured in paired BALF and serum samples from patients with acute pneumonitis or asthma. Ceruloplasmin (CP) and alpha-2-macroglobulin (A2M) were measured by ELISAs thereby avoiding concentration of BALF. The quotients 10(3) ([protein]BALF)/(protein]serum), Qprotein, were calculated as well as the relative coefficients of excretion, RCE: Qprotein/Qalbumin. The QCP and QA2M increased parallel to Qalbumin in patients with pneumonitis and QCP increased parallel with Qalbumin in the asthma patients. This indicates that abnormal leakage of proteins from the blood rather than local synthesis cause the increased concentrations of these proteins in the BALF. Increased values for the RCE of CP and A2M were observed at normal Qalbumin. We therefore conclude that the determination of CP and A2M yields more detailed information on leakage of proteins from the blood into the airway compartment than that of albumin.