β-石竹烯醇抑制哮喘豚鼠气道炎症及清除氧自由基作用

建立卵白蛋白（OA）致敏的特异性哮喘豚鼠模型，观察β-石竹烯醇对特异性哮喘豚鼠引喘潜伏期、支气管肺泡灌洗液（BALF）白细胞分类计数、支气管形态病理学改变以及血清和肺组织超氧化物歧化酶（SOD）活性、丙二醛（MDA）水平的影响。结果表明，β-石竹烯醇灌胃给药，可显著延长特异性哮喘豚鼠引喘潜伏期，减少BALF中各细胞组分，减轻支气管炎症性病理改变；提高哮喘豚鼠血清和肺组织SOD活性、降低MDA水平。抑制气道炎症和清除氧自由基（OFR）可能是β-石竹烯醇平喘作用的机制之一。     

氮卓斯汀对豚鼠哮喘的拮抗作用及相关机制

目的:研究组胺H1受体拮抗剂——氮卓斯汀(azelastine,AZT)对豚鼠哮喘的拮抗作用,探讨氮卓斯汀拮抗哮喘的可能机制。方法:以卵白蛋白致敏豚鼠,制备哮喘模型,通过豚鼠引喘潜伏期和跌倒率的变化评价氮卓斯汀对哮喘的拮抗作用;分别计数外周血和支气管肺泡灌洗液(BALF)中嗜酸性粒细胞的数量;采用ELISA法检测血清和BALF中IL-4和IL-5的质量浓度。结果:与模型组相比,氮卓斯汀能显著延长致敏豚鼠的引喘潜伏期,降低跌倒率;氮卓斯汀显著减少哮喘豚鼠血清和BALF中嗜酸性粒细胞的数量,降低血清和BALF中IL-4和IL-5的水平。结论:降低IL-4和IL-5的水平,减少血中嗜酸性粒细胞的数量及其在气道中的浸润,可能是组胺H1受体拮抗剂——氮卓斯汀拮抗哮喘的机制之一。     

白细胞介素-1受体拮抗剂拮抗豚鼠哮喘的相关机制

目的：研究白细胞介素-1受体拮抗剂（IL-1ra)拮抗豚鼠哮喘的机制。方法：建立致敏豚鼠模型。吸入不同浓度的的IL-1ra。引喘豚鼠，检测引喘后8d血清，肺组织中，内皮素-1（ET），一氧化氮（NO），以及血液，肺泡灌洗液（BALF）中嗜酸性粒细胞（EOS）的变化情况。结果：IL-1ra可以明显升高血清和肺组织中的NO浓度，降低血清和肺组织中的ET含量，减少血液和肺泡灌洗液中的EOS的数量；高剂量组缓解作用较强。结论：IL-1ra可增加机体内的NO浓度，降低ET的含量，减少EOS浸润，IL-1ra对豚鼠哮喘具有拮抗作用。     

孟鲁司特对哮喘豚鼠气道嗜酸粒细胞炎症的抑制作用

目的　研究白三烯受体拮抗剂孟鲁司特 (mon telukast,MK )对哮喘豚鼠气道嗜酸性粒细胞 (Eosinophil,Eos)炎症的影响 ,探讨孟鲁司特拮抗哮喘气道炎症的可能机制。方法　以卵白蛋白致敏豚鼠制备哮喘模型。用密度梯度分离法分离并计数支气管肺泡灌洗液 (BALF)中不同密度的嗜酸性粒细胞 ;采用TUNEL技术原位检测嗜酸性粒细胞凋亡 ;通过ELISA法检测BALF中IL 5的含量 ;采用荧光酶标记法在PharmaciaUnicap 10 0System中测定BALF中嗜酸性粒细胞阳离子蛋白 (ECP)的水平。结果　孟鲁司特能降低哮喘豚鼠BALF中Eos的数量 ;在孟鲁司特治疗组 ,嗜酸性粒细胞凋亡指数明显升高 ,BALF中IL 5和ECP的含量降低 ,与模型组比较差异均有显著性。结论　降低气道IL 5和ECP的水平 ,促进嗜酸性粒细胞凋亡 ,减少嗜酸性粒细胞浸润 ,可能是白三烯受体拮抗剂孟鲁司特拮抗哮喘气道炎症的一个重要机制。     

小剂量哌替啶对香烟烟雾致豚鼠急性肺气道反应的影响

目的研究小剂量阿片受体激动剂哌替啶(Peth)对香烟烟雾吸入引起的豚鼠急性气道收缩反应和炎性反应的影响。方法观察Peth0.01,0.1和1mg.kg-1对豚鼠自主吸入75%香烟烟雾(含25%O2)60mL后,气道阻力和肺动态顺应性变化的影响及气道组织血管通透性变化的影响;观察Peth0.1mg.kg-1对豚鼠2h内分6次吸入(共360mL)75%浓度的香烟烟雾后,支气管肺泡灌洗液(BALF)中白细胞总数和分类计数改变的影响,测定BALF中一氧化氮(NO)含量。结果Peth能减轻或明显抑制香烟烟雾刺激后气道阻力增高和肺动态顺应性下降的反应,抑制气道组织各段微血管通透性增加的反应,降低BALF中的白细胞总数和中性粒细胞比例,降低BALF中NO的含量。结论小剂量Peth对豚鼠急性神经源性气道收缩反应和炎性反应具有抑制作用。     

孟鲁司特对哮喘豚鼠气道淋巴细胞凋亡及核因子-κB活化的影响

目的:研究白三烯受体阻滞剂孟鲁司特(montelukast,MK)对哮喘豚鼠气道淋巴细胞(lymphocyte,LC)凋亡和核因子-κB(nuclear factor-κB,NF-κB)活化的影响及二者相关性,探讨MK阻滞哮喘气道炎症的可能机制。方法:以卵白蛋白致敏豚鼠制备哮喘模型。用密度梯度离心法分离并计数支气管肺泡灌洗液(BALF)中的淋巴细胞;采用TUNEL技术原位检测淋巴细胞凋亡;通过免疫细胞化学染色技术检测NF-κB胞核阳性的淋巴细胞。结果:与模型组相比,MK能显著降低哮喘豚鼠BALF中LC的数量,升高淋巴细胞凋亡率,降低NF-κB胞核阳性率。淋巴细胞凋亡异常与NF-κB激活增加呈显著负相关。结论:MK抑制气道淋巴细胞NF-κB的激活和促进淋巴细胞凋亡可能是其阻滞哮喘气道炎症的一个重要机制。     

止嗽散及其改进方对哮喘豚鼠作用的比较

目的:比较止嗽散及其改进方对由卵蛋白致敏哮喘豚鼠作用的异同。方法:用Wright染色法计血液和肺泡灌洗液(BALF)中嗜酸性粒细胞(EOS)的数目。采用放射免疫法和硝酸还原酶法分别测定哮喘豚鼠血清和BALF中内皮素-1(ET-1)、一氧化氮(NO)的含量。在光镜和电镜下观察哮喘豚鼠肺组织的病理变化。结果:与止嗽散比较,止嗽散改进方(MZ)能够明显减少血液和BALF中EOS的数目,降低哮喘豚鼠血清和BALF中ET-1、NO的含量,减轻哮喘豚鼠肺组织的病理改变。结论:MZ减少哮喘豚鼠体内EOS数目和降低ET-1、NO含量的作用可能与其加入地龙、杏仁等中药有关。     

孟鲁司特对哮喘豚鼠气道淋巴细胞凋亡及核因子-kB活化的影响

目的：研究白三烯受体阻滞剂孟鲁司特(montelukast，MK)对哮喘豚鼠气道淋巴细胞(lymphocyte．LC)凋亡和核因子-kB(nuclear faclor-kB，NF-kB)活化的影响及二者相关性，探讨MK阻滞哮喘气道炎症的可能机制。方法：以卵白蛋白致敏豚鼠制备哮喘模型。用密度梯度离心法分离并计数支气管肺泡灌洗液(BALF)中的淋巴细胞；采用TUNEL技术原位检测淋巴细胞凋亡；通过免疫细胞化学染色技术检测NF-kB胸核阳性的淋巴细胸。结果：与模型组相比，MK能显著降低哮喘豚鼠BALF中LC的数量．升高淋巴细胸凋亡率．降低NF-kB胸核阳性率。淋巴细胞凋亡异常与NF-kB激活增加呈显著负相关。结论：MK抑制气道淋巴细胸NF-kB的激活和促进淋巴细胞凋亡可能是其阻滞哮喘气道炎症的一个重要机制。     

加味止嗽散有效部位群对过敏性哮喘豚鼠作用的实验研究

目的:观察加味止嗽散有效部位群(MZC)对过敏性哮喘豚鼠血液和支气管肺泡灌洗液(BALF)中嗜酸粒细胞(Eos),内皮素1(ET-1)一氧化氮(NO)含量的影响和对肺组织结构的影响。方法:采用Wright染色法计血液和BALF中Eos的数目。采用放射免疫法和硝酸还原酶法分别测定哮喘豚鼠血清和BALF中ET-1NO的含量。光镜和电镜下观察豚鼠肺组织。结果:哮喘模型组豚鼠体内的Eos,ET1,NO的量明显高于正常对照组(P<0.01),肺组织病理学和超微结构发生明显改变。与哮喘模型组比较,MZC治疗组剂量依赖性地降低豚鼠血液和BALF中Eos,ET-1NO的含量(P<0.05或P<0.01)和减轻肺组织病理改变。结论:MZC平喘作用机制可能与其减少哮喘豚鼠体内Eos数量,降低ET-1NO含量和减轻肺组织结构损伤有关。     

小儿哮喘宁平喘作用的研究

目的初步评价小儿哮喘宁的平喘作用,为临床应用提供科学的实验依据。方法运用乙酰胆碱及组胺致豚鼠哮喘模型、内毒素（lipoplysaccharides,LPS）致小鼠急性肺损伤模型、卵白蛋白（ovalbumin,OVA）致豚鼠过敏性哮喘模型及OVA加LPS致小鼠过敏性哮喘模型,评价小儿哮喘宁的平喘作用。结果小儿哮喘宁20、10、5 g/kg均能延长乙酰胆碱＋组胺所致豚鼠化学刺激性哮喘的引喘潜伏期及OVA致豚鼠过敏性哮喘的引喘潜伏期并减少跌倒动物数;20 g/kg小儿哮喘宁能降低LPS致小鼠急性肺损伤程度;对嗜酸粒细胞（eosinophil,EOS）浸润型过敏性哮喘动物模型的作用优于对中性粒细胞（neutrophil,NEU）浸润型过敏性哮喘动物模型的作用。结论小儿哮喘宁有平喘的作用,该作用可能与其抑制肺水肿、降低气道EOS浸润有关。     

孟鲁司特对哮喘豚鼠气道嗜酸性粒细胞凋亡及Fas mRNA表达的影响

目的研究白三烯受体拮抗剂孟鲁司特(montelukast,MK)对哮喘豚鼠气道嗜酸性粒细胞(eosinophil,Eos)凋亡和Fas mRNA表达的影响。方法以卵白蛋白致敏豚鼠制备哮喘模型。用密度梯度离心法分离并计数支气管肺泡灌洗液(BALF)中的嗜酸性粒细胞；采用TUNEL技术原位检测嗜酸性粒细胞凋亡；通过逆转录-多聚酶链反应(RT-PCR)技术检测嗜酸性粒细胞Fas mRNA的表达。结果孟鲁司特能显著降低哮喘豚鼠BALF中Eos的数量；在孟鲁司特治疗组，嗜酸性粒细胞凋亡指数明显升高，Fas mRNA的表达显著增强，与模型组比较均有显著性差异。结论嗜酸性粒细胞凋亡与Fas mRNA表达增加高度相关；增强气道嗜酸性粒细胞Fas mRNA的表达，促进其凋亡，可能是孟鲁司特拮抗哮喘气道炎症的一个重要机制。     

火把花根片对哮喘豚鼠抗炎机制及其对肺泡灌洗液IL-5受体mRNA表达的影响

目的　探讨火把花根片对哮喘豚鼠抗炎作用机制及其对肺泡灌洗液 (BALF)白介素 5(IL 5)受体mRNA表达的影响。方法　 3 2只健康豚鼠随机分为 4组 :正常对照组 ,哮喘组 ,地塞米松治疗组 ,火把花根片治疗组。后 3组制作成哮喘豚鼠模型 ,再激发用药。测定肺泡灌洗液的细胞总数和细胞分类 ,观察肺组织的病理改变 ,进行BALF的IL 5受体mRNART PCR半定量分析。结果　火把花根片治疗组BALF的嗜酸性粒细胞 (Eos)数较正常组和哮喘组 ,差异均有显著性 ,分别为P <0 0 5和P <0 0 1。肺组织病理切片 ,火把花根片组充血水肿明显 ,但Eos肺组织浸润较哮喘组有明显减少。BALF的IL 5受体mRNART PCR半定量分析显示 ,火把花根片治疗组与地塞米松治疗组差异无显著性 ,P >0 0 5,但和正常组与哮喘组比较差异有显著性 ,P <0 0 1。结论　火把花根片能抑制哮喘豚鼠的气道炎症 ,为临床治疗哮喘提供初步的理论基础     

雷公藤多甙对哮喘豚鼠嗜酸性粒细胞凋亡的影响

目的　探讨雷公藤多甙治疗哮喘的机制。方法　应用原位末端标记技术 (TUNEL)、形态学观察 ,观察雷公藤多甙对哮喘豚鼠体内嗜酸性粒细胞 (Eos)凋亡的影响。结果　 (1)哮喘豚鼠体内Eos凋亡率显著低于正常豚鼠 (P <0 0 5 )。 (2 )哮喘豚鼠应用雷公藤多甙治疗后 ,体内Eos凋亡率显著增加 (P <0 0 1) ,并伴随气道炎症和哮喘反应的减轻。结论　诱导哮喘体内Eos凋亡是雷公藤多甙治疗哮喘的重要机制     

Tryptase-induced airway microvascular leakage in guinea pigs: involvement of tachykinins and leukotrienes.

Tryptase, a serine protease synthesized by and stored in mast cells, is implicated as an important mediator in the pathogenesis of airway inflammation. In this study, tryptase was evaluated for its ability to induce microvascular leakage into the airways of guinea pigs. Dose- and time-dependent increases in airway microvascular leakage were produced by intratracheal tryptase (0.3-3 microg). Intratracheal tryptase (3-30 microg) had no effect on airway tone as measured by pulmonary insufflation pressure. Tryptase-induced airway microvascular leakage was partially blocked by the tachykinin NK1 receptor antagonist CP 99994 [(+)-(2S,3S)-3-(2-methoxybenzylamino)-2-phenylpiperidine] and an inhibitor of leukotriene formation SCH 37224 (1-(1,2-dihydro-4-hydroxy-2-oxo-1-phenyl-1,8-naphthyridin-2-yl)pyrrolidinium, hydroxide inner salt). Neither CP 99994 nor SCH 37224 inhibited tryptase proteolytic activity in-vitro. Pretreatment of guinea pigs with histamine H1 receptor antagonists or a tachykinin NK2 receptor antagonist had no affect on the airway microvascular leakage induced by tryptase. It is speculated that tryptase may be important in the pathogenesis of airway inflammation, particularly in disorders that involve increased airway microvascular leakage such as asthma.     

丁苯酞对卵白蛋白所致支气管哮喘豚鼠的影响

目的　探讨丁苯酞对卵白蛋白所致支气管哮喘豚鼠模型的影响。方法　采用卵白蛋白经致敏与激发两步来复制支气管哮喘豚鼠模型,通过药物干预,观测丁苯酞对哮喘豚鼠的行为学、肺功能、BALF中炎性细胞、肺组织病理学及IgE的影响。结果　30、60、120mg?kg_-1丁苯酞能降低哮喘豚鼠的哮喘行为学评分、肺组织湿干重比值、BALF中炎性细胞及血清T-IgE水平,改善肺功能,缓解肺组织病理学变化(P＜0.05, 0.01)。结论　丁苯酞具有缓解肺部炎症而治疗支气管哮喘的作用。     

Adrenal influences on the inhibitory effects of procaterol, a selective Beta-two-adrenoceptor agonist, on antigen-induced airway microvascular leakage and bronchoconstriction in Guinea pigs

While the guinea pig has been the preferred choice for use as a model of allergic bronchial asthma in the evaluation of anti-asthmatic drugs, it has been shown that antigen-induced bronchoconstriction in guinea pigs is attenuated by epinephrine released from the adrenal gland. In order to investigate the possible influence of the adrenal gland on the effects of antiexudative and bronchodilative drugs on antigen-induced airway responses, we examined the inhibitory effects of procaterol, a selective beta(2)-adrenoceptor agonist, on antigen-induced airway microvascular leakage and bronchoconstriction in adrenalectomized guinea pigs and compared them with the drug's effects in sham-operated animals. Guinea pigs sensitized passively with anti-ovalbumin (OA) guinea-pig serum were adrenalectomized or sham-operated under urethane anesthesia and examined 30 min after surgery in the following experiments. (1) Animals were intravenously administered Evans blue dye to quantify airway plasma exudation, and then OA was inhaled for 10 min while measuring pulmonary inflation pressure, a parameter of bronchoconstriction. Procaterol (1, 3, 10, or 30 microg/kg) or saline (control) was administered into the airways 10 min prior to OA inhalation. The amount of extravasated Evans blue dye in the airways was calculated. (2) Venous blood samples were collected during OA or saline inhalation and plasma catecholamine levels were compared. In control animals, OA-induced increases in both the amount of Evans blue dye and in pulmonary inflation pressure were markedly greater in adrenalectomized animals than in sham-operated animals. Procaterol dose-dependently inhibited OA-induced airway microvascular leakage and bronchoconstriction, and its effects were more potent in adrenalectomized animals (significant at 1 microg/kg and higher) than in sham-operated animals (significant at 10 microg/kg and higher). Although the plasma concentration of epinephrine during OA inhalation was approximately 3 times higher than that during saline inhalation in sham-operated animals, no difference was seen in adrenalectomized animals. In conclusion, while procaterol essentially possesses pronounced inhibitory effects on antigen-induced airway microvascular leakage and bronchoconstriction in guinea pigs, the effects are considerably masked by epinephrine released from the adrenal gland.     

Bronchopulmonary inflammation and airway smooth muscle hyperresponsiveness induced by nitrogen dioxide in guinea pigs.

We investigated whether acute exposure to nitrogen dioxide (NO2) causes major inflammatory responses (inflammatory cell recruitment, oedema and smooth muscle hyperresponsiveness) in guinea pig airways. Anaesthetised guinea pigs were exposed to 18 ppm NO2 or air for 4 h through a tracheal cannula. Bronchoalveolar lavage was performed and airway microvascular permeability and in vitro bronchial smooth muscle responsiveness were measured. Exposure to NO2 induced a significant increase in eosinophils and neutrophils in bronchoalveolar lavage fluid, microvascular leakage in the trachea and main bronchi (but not in peripheral airways), and a significant in vitro hyperresponsiveness to acetylcholine, electrical field stimulation, and neurokinin A, but not to histamine. Thus, this study shows that in vivo exposure to high concentrations of NO2 induces major inflammatory responses in guinea pig airways that mimic acute bronchitis induced by exposure to irritant gases in man.