Antisense oligonucleotides-induced local blockade of T-bet expression leads to airway inflammation in rats

AIM: To explore whether local blockade of T-box expressed in T cells (T-bet) expression in the lungs could lead to airway inflammation. METHODS: Twenty-four rats were randomly divided into 4 groups: saline group, ovalbumin (OVA)-sensitized group, nonsense group, and the antisense group. The OVA-sensitized rats were sensitized and challenged with OVA, and the rats in the nonsense and antisense groups were subjected to an aerosol delivery of the nonsense and antisense oligonucleotides (AS-ODN) of T-bet (0.1%, w/v). The levels of interferon-gamma (IFN-gamma), interleukin-4 (IL-4), and IL-5 in the bronchoalveolar lavage fluid (BALF) were detected by ELISA, and the mRNA and the protein expression of T-bet and GATA-3 genes were examined by in situ hybridization and Western blot analysis, respectively. RESULTS: The expression of T-bet mRNA and protein in the lungs of the rats in the antisense group were inhibited effectively. The lungs of the rats in the antisense and OVA-sensitized groups showed eosinophil and lymphocyte inflammatory infiltration, and eosinophilia located predominantly around the airways. The number of GATA-3 mRNA-positive cells and the level of GATA-3 protein in the lungs of the rats in the antisense and the OVA-sensitized groups significantly increased. The level of IL-4 and IL-5 in the BALF in the antisense and OVA-sensitized groups were elevated, but the level of IFN-gamma decreased markedly. CONCLUSION: Antisense ODN-induced local blockade of T-bet expression leads to airway inflammation with a selective alteration in patterns of cytokine expression and recruitment of eosinophil cells similar to that in the OVA-sensitized animals.     

当归对阴虚哮喘BALB/c小鼠Th2优势免疫应答的影响

目的 探讨当归对阴虚哮喘BALB/c小鼠Th2细胞优势免疫应答的影响。方法 采用注射卵清白蛋白(ovalbumin,OVA)致敏、吸入OVA激发的方法来复制BALB/c小鼠哮喘模型,实验后期给予甲状腺素以复制阴虚哮喘模型,通过观测哮喘行为学、血清及支气管肺泡灌洗液(broncho-alveolar lavage fluid,BALF)中IL-4、IL-13的水平、BALF及肺组织中炎性细胞的含量、肺组织中GATA-3蛋白表达水平,探讨当归的平喘作用及对Th2细胞优势免疫应答的影响。结果 当归可明显减少阴虚哮喘小鼠哮喘发作的次数及其发作时的症状,改善肺组织病理学而减少肺组织及BALF中炎性细胞的数量,降低血清及BALF中IL-4、IL-13的水平,抑制肺组织中GATA-3蛋白的表达(P<0.05或0.01);同时,当归与地塞米松配伍后对嗜酸粒细胞、IL-4及GATA-3的抑制作用具有一定的协同作用(P<0.05或0.01)。结论 当归具有平喘作用,抑制细胞因子IL-4、IL-13及转录因子GATA-3的表达而缓解Th2优势免疫应答可能是其作用机制之一。     

CpG ODN对小鼠哮喘模型气道炎症及信号转导子和转录激活子6（STAT6）表达的影响

目的：研究寡核甘酸免疫刺激序列（CpG oligonucleotides,CpG ODN）对急性支气管哮喘小鼠气道炎症及信号转导子和转录激活子6（STAT6）表达的调控作用。方法：将32只雄性BALB／c小鼠随机分为对照组、哮喘组、地塞米松干预组和CpG ODN干预组,建立急性哮喘气道炎症模型。对支气管肺泡灌洗液（BALF）进行细胞总数、嗜酸粒细胞（EOS）分类计数;取左叶肺组织切片做HE染色观察气道和肺组织炎症改变;用免疫组织化学法检测气道上皮组织中STAT6表达。结果：哮喘组小鼠BALF中细胞总数、EOS绝对值显著高于对照组（P〈0．05）,地塞米松组和CpG ODN组上述炎症细胞计数比哮喘组明显减少（P〈0．05）。HE染色示地塞米松组和CpG ODN组小鼠肺组织炎症程度较哮喘组减轻。STAT6在气道上皮细胞表达,哮喘组气道上皮细胞中STAT6表达较对照组增强,地塞米松组和CpG ODN组STAT6表达与哮喘组比较有明显减少（P〈0．05）,较对照组增加。支气管上皮细胞STAT6蛋白表达与BALF中的EOS绝对值呈显著正相关（r=0．748）,而地塞米松组和CpG ODN组之间的作用无统计学差异（P〉0．05）。结论：应用地塞米松和CpG ODN均可在一定程度上减轻哮喘小鼠的气道炎症,CpG ODN可能通过下调肺组织中STAT6蛋白的过度表达,从而抑制依赖于STAT6/IL-4通路的急性气道炎症,减轻哮喘的症状。     

钩藤碱固体脂质纳米粒对哮喘小鼠miR-155/p38 MAPK轴的影响

目的　观察钩藤碱固体脂质纳米粒（Rhy-SLN）对支气管哮喘模型小鼠微小RNA-155（miR-155）/p38丝裂原活化蛋白激酶（p38 MAPK）轴的影响。方法　15只BALB/c小鼠随机均分为正常对照组、模型组和Rhy-SLN组。Rhy-SLN组小鼠行鼻内氢氧化铝致敏操作前以Rhy-SLN（50 mg/kg）灌胃;正常对照组和模型组给予等量生理盐水。干预结束后,肺泡灌洗液涂片观察嗜酸粒细胞的数量;HE染色观察小鼠肺组织病理改变情况;酶联免疫吸附试验（ELISA）法检测小鼠免疫球蛋白E（IgE）、白细胞介素（IL）-13水平;羟脯氨酸测试盒检测小鼠肺组织中羟脯氨酸水平;Werstern blot检测小鼠肺组织（α-SMA）、胶原蛋白Ⅰ（collagen Ⅰ）、p38 MAPK、p-p38 MAPK蛋白表达水平;荧光定量聚合酶链反应（qPCR）检测小鼠肺组织miR-155 mRNA表达水平。结果　与模型组比较,Rhy-SLN组可以降低哮喘小鼠肺泡灌洗液中嗜酸粒细胞的数目、血清中IgE和肺泡灌洗液中IL-13的水平（P＜0.05）;减轻肺组织炎性细胞浸润;降低肺组织中α-SMA、collagenⅠ、羟脯氨酸和p-p38 MAPK的表达（P＜0.05）;上调小鼠肺组织中miR-155的表达水平（P＜0.05）。结论　Rhy-SLN可缓解小鼠哮喘,其机制可能与调节miR-155/p38 MAPK轴,降低气道的炎症反应有关。     

Effect of topical immunomodulators on acute allergic inflammation and bronchial hyperresponsiveness in sensitised rats

We examined the effects of different immunomodulators administered topically on asthmatic responses in a rat model of asthma. Sensitised Brown-Norway rats were administered rapamycin, SAR943 (32-deoxorapamycin), IMM125 (a hydroxyethyl derivative of D-serine(8)-cyclosporine), and budesonide by intratracheal instillation 1 h prior to allergen challenge. Allergen exposure induced bronchial hyperresponsiveness, accumulation of inflammatory cells in bronchoalveolar lavage fluid, and also an increase in eosinophils and CD2+, CD4+ and CD8+ T cells in the airways. Interleukin-2, interleukin-4, interleukin-5, interleukin-10, and interferon-gamma mRNA expression was upregulated by allergen exposure. Budesonide abolished airway inflammation, suppressed the mRNA expression for interleukin-2, interleukin-4, and interleukin-5 (P<0.03), and bronchial hyperresponsiveness (P<0.05). IMM125 suppressed airway infiltration of eosinophils, and CD8+ T cells (P<0.02), and prevented the upregulated mRNA expression for interleukin-4, interleukin-5, and interferon-gamma (P<0.02). Rapamycin suppressed CD8+ T cell infiltration in airway submucosa (P<0.03), and mRNA expression for interleukin-2 (p<0.002), while SAR943 suppressed interleukin-2, interleukin-4, and interferon-gamma mRNA (P<0.05). IMM125, rapamycin and SAR943 did not alter airway submucosal CD2+ and CD4+ T cell infiltration, and bronchial hyperresponsiveness. CD8+ T cells, in contrast to CD4+ T cells, are more susceptible to the inhibition by IMM125 and rapamycin, which also caused greater suppression of Th1 compared to Th2 cytokine mRNA expression. In this acute model of allergic inflammation, differential modulation of Th1 and Th2 cytokines may determine the effects of various immunomodulators on airway inflammation and bronchial hyperresponsiveness.     

p38蛋白激酶抑制剂SB203580对支气管哮喘小鼠气道炎症和Th2类细胞因子的影响

目的观察特异性p38蛋白激酶(p38 MAPK)抑制剂SB203580对哮喘小鼠气道炎症和Th2类细胞因子的影响。方法BALB/c小鼠30只随机分成3组,即正常对照组、哮喘模型组和SB203580干预组。通过原位分子杂交和酶联免疫吸附法(ELISA)检测肺组织IL-4、IL-5 mRNA和支气管肺泡灌洗液(BALF)中白细胞介素(IL-4、IL-5)含量的变化,并观察BALF中炎症细胞和肺组织病理学改变。结果哮喘模型组小鼠BALF中炎症细胞计数和IL-4、IL-5含量以及肺组织IL-4、IL-5mRNA的表达较正常对照组明显升高,差异具有显著性(P<0.01);SB203580干预组小鼠上述指标较哮喘模型组小鼠明显降低,差异亦具有显著性(P<0.01),肺组织病理学改变明显减轻。结论SB203580能降低气道炎症细胞的聚集和炎症介质的表达。抑制p38 MAPK的活性可能成为哮喘治疗的新途径。     

Diosgenin, a steroidal sapogenin, enhances antigen-specific IgG2a and interferon-gamma expression in ovalbumin-sensitized BALB/c mice

The effect of diosgenin, the most abundant sapogenin in Chinese yam, on humoral immunity was investigated. Ovalbumin (OVA)-sensitized and challenged BALB/c mice were administered daily with diosgenin for 34 days. The production of OVA-specific serum IgG2a was significantly enhanced by diosgenin treatment, whereas total IgE and OVA-specific IgG1, IgG2a and IgM were unaffected. In parallel with the enhancement of IgG2a, OVA-induced IFN-gamma secretion and mRNA expression were markedly elevated in splenocytes of diosgenin-treated mice, whereas IL-4 expression was unaltered. Furthermore, the expression of T-bet, but not of GATA-3, in splenocytes was up-regulated by diosgenin administration. However, diosgenin treatment did not modulate IL-4 mRNA expression and inflammatory cell infiltration in the lung of OVA-sensitized and challenged mice. Collectively, these data suggest that diosgenin regulates the systemic immune response towards the Th1 direction in response to OVA sensitization. The present study provides evidence to show that intake of diosgenin modulates certain aspects of acquired immunity, including the enhancement of antigen-specific IgG2a and IFN-gamma expression, which may be mediated through the up-regulation of Th1 differentiation.     

柚皮素通过抑制NF-κB信号通路减轻哮喘大鼠气道炎症反应#br#

摘要：目的　探讨柚皮素对哮喘大鼠气道炎症反应的影响及其作用机制。方法　利用卵清蛋白（OVA）诱导建立哮喘大鼠模型,实验分为正常对照组、哮喘模型组、柚皮素100 mg/kg及柚皮素200 mg/kg组,实验结束后麻醉脱颈处死大鼠;Giemsa染色检测支气管肺泡灌洗液（BALF）中炎性细胞的数量及分类;HE染色观察肺组织病理学变化;酶联免疫吸附测定试验（ELISA）检测血清中核因子κB（NF-κB）的含量和BALF中辅助性T2（Th2）细胞因子的含量;免疫组化和实时定量PCR（qPCR）检测哮喘大鼠肺部NF-κB蛋白及mRNA的表达水平。脂多糖（LPS）诱导建立A549炎症细胞模型,柚皮素处理细胞后免疫荧光和Western blot检测A549细胞中NF-κB蛋白的表达水平。结果　柚皮素100 mg/kg和柚皮素200 mg/kg组均可减少单核细胞、中性粒细胞、嗜酸性粒细胞、淋巴细胞对哮喘大鼠肺部和支气管的浸润,并可改善哮喘大鼠肺泡的生理结构,减少支气管黏膜上皮脱落,促进支气管假复层及纤毛结构修复。柚皮素可降低肺部NF-κB p65、NF-κB p50蛋白和mRNA的表达（P＜0.05）,降低血清中NF-κB p65、NF-κB p50和BALF中Th2细胞因子白细胞介素（IL）-4、IL-5、IL-13的含量（P＜0.05）。在体外LPS诱导的炎症A549细胞中,柚皮素可降低NF-κB p65、NF-κB p50蛋白的表达,上调IκBα的表达（P＜0.05）。结论　柚皮素可有效减轻哮喘大鼠肺部和支气管的炎症反应,其潜在的作用机制可能与抑制NF-κB信号通路有关。     

CCR3 monoclonal antibody inhibits airway eosinophilic inflammation and mucus overproduction in a mouse model of asthma

AIM: To explore the effect of a rat anti-mouse CC-chemokine receptor-3 (CCR3) monoclonal antibody (CCR3 mAb) on airway eosinophilia and mucus overproduction in asthmatic mice. METHODS: An asthma model was sensitized and challenged by ovalbumin (OVA) in male C57BL/6 mice. Asthmatic mice were given dual administration (intraperitoneal injection and aerosol inhalation) of CCR3 mAb or nonspecific rat IgG (ns-IgG). The number of total and differential inflammatory cells in the bronchial alveolar lavage fluid (BALF) was counted. Eosinophils number, the goblet cell percentage (GCP) and airway mucus index (AMI) were measured in the lung tissues. Interleukin (IL)-5 levels in the BALF were examined. The expression of MUC5AC and the epidermal growth factor receptor (EGFR) mRNA in the lung tissues was detected by semi-quantitative RT-PCR. The results were compared among the groups. RESULTS: CCR3 mAb significantly suppressed the increased eosinophils in the BALF and lung tissues in OVA-challenged mice compared with ns-IgG-treated mice. IL-5 levels in the BALF in CCR3 mAb and ns-IgG administration mice exhibited no obvious changes relative to OVA-challenged asthmatic mice. CCR3 mAb reduced the increased GCP and AMI after OVA challenge and decreased the enhanced expression of MUC5AC and EGFR mRNA in lung tissues in asthmatic animals. CONCLUSION: CCR3 mAb can significantly inhibit airway eosinophilia and mucus overproduction in asthmatic mice. Blockage of CCR3 may represent a new strategy to asthma therapy.     

Anti-asthmatic effect of schizandrin on OVA-induced airway inflammation in a murine asthma model

Asthma comprises a triad of reversible airway obstruction, bronchial smooth muscle cell hyperreactivity to bronchoconstrictors, and chronic bronchial inflammation. Clinical and experimental findings have established eosinophilia as a sign of allergic disorders. In the present investigation, we evaluated the anti-asthmatic effects of schizandrin and its underlying mechanisms in an in vivo murine asthmatic model. To accomplish this, female BALB/c mice were sensitized and challenged with ovalbumin (OVA), and examined for the following typical asthmatic reactions: increased numbers of eosinophils and other inflammatory cells in bronchoalveolar lavage fluid (BALF); production of Th1 cytokines (such as tumor necrosis factor (TNF)-α in BALF); production of Th2 cytokines (such as interleukin IL-4 and IL-5) in BALF; presence of total and OVA-specific immunoglobulins (Ig)E in serum; presence of oxidative stress; hyperplasia of goblet cells in the lung; and marked influx of inflammatory cells into the lung. Our results collectively show that schizandrin exerts profound inhibitory effects on accumulation of eosinophils into the airways and reduces the levels of IL-4, IL-5, IFN-γ, and TNF-α in BALF. Additionally, schizandrin suppresses the production of reactive oxygen species (ROS) in a dose-dependent manner, and inhibits goblet cell hyperplasia and inflammatory cell infiltration in lung tissue. Thus, schizandrin has anti-asthmatic effects, which seem to be partially mediated by reduction of oxidative stress and airway inflammation, in a murine allergic asthma model. These results indicate that schizandrin may be an effective novel therapeutic agent for the treatment of allergic asthma.     

Edaravone, a novel antidote against lung injury and pulmonary fibrosis induced by paraquat?

Edaravone (MCI-186) is a potent free radical scavenger used clinically to treat acute brain infarction. Its antioxidant ability maybe also do favor to protect against lung injury. In this study, we evaluated whether edaravone could protect against lung injury and pulmonary fibrosis in paraquat-treated rats. Rats were divided into four groups (Control group, Edaravone group, Paraquat intoxication group and Paraquat+Edaravone group) and sacrificed on day 1, 3, 5, 7, 14 and 28. Lung tissue and bronchoalveolar lavage fluid (BALF) were collected and analyzed biochemically and histologically. Paraquat intoxication significantly increased malondialdehyde (MDA), hydroproline, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) levels in lung tissue and BALF, and also increased mRNA expression of transforming growth factor-β1 (TGF-β1), matrix metalloproteinase-2 (MMP-2), and tissue inhibitor of metalloproteinase-1 (TIMP-1) in lung tissue, but decreased SOD and GSH-PX activity in lung tissue and BALF. Histological examination of paraquat-treated rats showed acute injury with interstitial edema and widespread inflammatory cell infiltration in the alveolar space and septum, as well as fibrosis. After edaravone treatment, levels of MDA, IL-6, TNF-α and hydroproline decreased, but SOD and GSH-PX activity in lung tissue and BALF increased. In addition, the mRNA expression of TGF-β1, MMP-2, and TIMP-1 down-regulated. Histological examination showed that edaravone decreased interstitial edema, inflammatory cell infiltration and prevented the process of pulmonary fibrosis.     

川芎嗪对致敏大鼠气道炎症气道重塑的影响和作用机制

目的探讨川芎嗪通过调节GATA-3和白细胞介素(IL)-5,抑制致敏大鼠气道炎症和气道重塑的分子机制。方法32只Wistar大鼠随机分成健康组、哮喘组、川芎嗪组和地塞米松(DXM)组,每组8只。苏木素-伊红(HE)染色行肺组织病理学检查、嗜酸性粒细胞计数及气道平滑肌(ASM)厚度测定;免疫组织化学两步法测定肺组织GATA-3和IL-5的表达;行肺组织GATA-3、IL-5表达与气道炎症反应间的相关性分析。结果川芎嗪和地塞米松可有效减少哮喘肺组织嗜酸性粒细胞数量和减轻平滑肌厚度,与哮喘组比较,差异有统计学意义(P<0.05);两个干预组肺组织中GATA-3和IL-5的表达较哮喘组明显减少,差异有统计学意义(P<0.05);肺组织GATA-3和IL-5表达与肺组织中嗜酸性粒细胞计数及平滑肌厚度呈正相关。结论川芎嗪可降低哮喘大鼠肺组织GATA-3和IL-5的表达,有效抑制哮喘的气道炎症,改善气道壁重塑。     

Administration of mycobacterial Ag85A and IL-17A fusion protein attenuates airway inflammation in a murine model of asthma

Interleukin (IL)-17A contributes to the development of asthma, especially in severe asthma which has characteristic neutrophil infiltration in airways. However, IL-17A-blocking antibody could escalate T helper (Th) 2 cytokines, such as IL-13, IL-4 in murine models. We aimed at determining the effect of mycobacterial Ag85A and IL-17A fusion protein—Ag85A-IL-17A on airway inflammation in a murine model of asthma. IL-17A recombinant protein fused mycobacterial immunodominant antigen Ag85A was constructed, expressed and purified. The fusion protein was then administrated into BALB/c mice and its anti-inflammatory effects in the infiltration of inflammatory cells, Th2/Th17 cytokines in BALF, histopathological changes of lung tissues as well as chemokines in lung tissues were evaluated in the murine model of asthma. We found that administration of mycobacterial Ag85A and IL-17A fusion protein induced IL-17A specific immunoglobulin (Ig)G in sera and significantly decreased IL-17A and IL-6 levels in bronchoalveolar lavage fluid (BALF). Ag85A-IL-17A vaccinated mice also showed marked reduction in the infiltration of inflammatory cells in peribronchiolar region and significant decrease in total cells, eosinophil cells and neutrophil cells in BALF. The increased levels of IL-13 and IL-4 in BALF of ovalbumin-sensitized mice were significantly reduced by the administration of Ag85A-IL-17A. Furthermore, CD3⁺CD4⁺IL-13⁺ splenocytes stimulated with OVA and CXCL1 mRNA, CCL2 mRNA and GATA-3 mRNA expressed in lung tissues were decreased markedly in Ag85A-IL-17A vaccinated group. Our results demonstrate remarkable antiallergic effects of Ag85A-IL-17A in a murine model of asthma and it may have protective effects on allergic asthma.     

来氟米特对胶原性关节炎大鼠T细胞漂移的影响

目的 探讨来氟米特对胶原性关节炎大鼠中Th1和Th2细胞漂移的影响.方法 建立胶原性关节炎模型;来氟米特2mg/kg灌胃给药;HE染色行膝关节滑膜病理学检查并评分;称重大鼠体重和脾脏重量,计算脾脏指数;RT-PCR法检测脾淋巴细胞中T-bet mRNA和GATA-3 mRNA的表达.ELISA法检测Con A刺激的脾淋巴细胞分泌Th1和Th2型细胞因子IFN-γ和IL-4的量.结果 与模型组比较,来氟米特(2mg/kg,灌胃给药15天)明显降低CIA大鼠病理组织学评分,降低脾脏指数.与关节炎模型组比较,药物干预组GATA-3 mRNA的表达增高、IL-4的分泌增多,T-bet mRNA的表达降低、IFN-γ的分泌减少.结论 来氟米特对胶原性关节炎具有治疗作用,该作用可能和维持T细胞亚群的平衡有关.     

致敏大鼠脑皮层和肺气道中干扰素—γ和白介素—4相关性的变化

目的:探讨致敏大鼠抗原攻击后脑皮层和肺气道中干扰素-γ(IFN-γ)和白介素-4(IL-4)出现的相关性变化.方法:观察致敏大鼠吸入抗原诱导的支气管肺灌洗液(BALF)和肺组织切片炎症变化,用ELISA法测定BALF和脑皮层IFN-γ和IL-4水平变化.结果:抗原攻击组BALF中的炎症细胞数目明显高于对照未攻击组(P<0.05).地塞米松(DXM,0.5mg/kg,ip)明显减少BALF中的白细胞总数,几乎完全抑制嗜酸性粒细胞(EOS)和淋巴细胞的聚集,但增加中性粒细胞数目.抗原攻击组的组织学检查积分(EOS浸润、粘膜水肿和上皮损伤)也明显高于对照未攻击组(P<0.05).DXM(0.5 mg/kg,ip)减少支气管和细支气管的EOS数目,改善粘膜水肿和上皮损伤.致敏大鼠抗原攻击后,BALF中的IFN-γ水平降低伴随IL-4升高导致了IFN-γ/IL-4比例下降.与此同时,脑皮层匀浆中也出现相似的改变.DXM(0.5mg/kg,ip)能反转BALF和脑皮层匀浆中的IFN-γ/IL-4比例下降.结论:致敏大鼠抗原攻击后脑皮层和肺气道中的IFN-γ和IL-4出现相关性变化.     

脐带间充质干细胞减轻哮喘小鼠气道炎症的研究

目的研究脐带间充质干细胞对鸡卵清蛋白诱导的哮喘小鼠模型气道炎症的抑制作用。方法将48只6⁸周龄雌性BALB/c小鼠随机分为四组,单纯哮喘组(OVA组),干细胞治疗组(MSC组),地塞米松治疗组(DM组)及生理盐水对照组(NS组)。23 d后,支气管肺泡灌洗液(BALF)细胞计数总细胞数、嗜酸粒细胞数;取肺组织行病理切片HE染色观察气道周围炎细胞浸润情况。同时免疫组织化学法观察IL-17蛋白在各组小鼠肺组织中的表达。结果同其他各组相比,单纯哮喘组肺泡灌洗液和肺组织中有大量炎性细胞浸润,而地塞米松治疗组、干细胞治疗组气道炎细胞浸润明显减轻(P<0.05),两个治疗组之间无明显差异(P>0.10)。与此结果相一致,MSC组、DM组中,炎性因子IL-17表达较OVA组明显减少(P<0.05),而DM组IL-17表达与MSC组相比无明显差异(P>0.10)。结论脐带间充质干细胞能够明显抑制鸡卵清蛋白诱导的哮喘小鼠气道炎症。     

氨溴索对哮喘大鼠炎性因子的影响及其作用机制

目的:研究氨溴索对哮喘大鼠气道炎症的影响及其可能的作用机制。方法:将30只雄性SD大鼠随机分为3组:对照组、模型组、治疗组。模型组、治疗组卵清蛋白(OVA)致敏。治疗组致敏同时给予腹腔注射氨溴索50 mg.kg-1.d-1。各组动物于最后一次雾化吸入后24 h收集支气管肺泡灌洗液(BALF)。行BALF中炎症细胞总数、PMN计数及细胞分类计数。取BALF上清液,应用酶联免疫吸附试验法测定IL-8水平。HE染色观察肺组织病理改变,免疫组织化学技术观察NF-κBp65、MMP-9、MPO在肺组织中的表达。结果:(1)模型组显示支气管、血管周围大量炎性细胞浸润,治疗组显示支气管痉挛及炎性细胞浸润明显减轻。(2)模型组BALF中炎性细胞及PMN计数、IL-8显著高于对照组(P<0.01)。治疗组BALF中炎性细胞及PMN计数、IL-8显著低于模型组(P<0.01)。(3)模型组肺组织中NF-κBp65、MMP-9、MPO的表达水平显著高于对照组(P<0.01)。治疗组大鼠肺组织中NF-κBp65、MMP-9、MPO的表达水平显著低于模型组(P<0.01)。结论:氨溴索可能是通过抑制转录因子NF-κB的活化,继而下调IL-8、MMP-9、MPO等的表达,减少PMN为主的炎症细胞在气道内的浸润,对哮喘患者发挥治疗作用。     

黄芪注射液对哮喘大鼠p38蛋白激酶和白细胞介素-5表达的影响

目的：探讨黄芪注射液对哮喘大鼠p38蛋白激酶（p38 MAPK）和白细胞介素-5（IL-5）表达的影响。方法：应用鸡卵清蛋白（OVA）腹腔注射致敏和反复超声雾化吸入复制大鼠哮喘模型。40只大鼠随机分成5组：正常对照组,哮喘模型组和黄芪注射液低、中、高剂量组（2.5、5.0、10.0mL/kg）。分别采用酶联免疫吸附法（ELISA）、原位分子杂交方法和蛋白质印迹检测支气管肺泡灌洗液（BALF）IL-5含量、肺组织IL-5 mRNA和磷酸化p38 MAPK表达的变化,并观察BALF中炎症细胞计数、分类以及肺组织病理学变化。结果：哮喘模型组大鼠BALF中炎症细胞计数、IL-5含量和肺组织中IL-5 mRNA及磷酸化p38 MAPK表达均较正常对照组显著增加（P〈0.01）;黄芪干预组的上述改变较哮喘模型组显著降低（P〈0.01）,肺组织病理学损伤程度明显减轻,黄芪注射液低、中、高剂量组之间差异无统计学意义。肺组织磷酸化p38 MAPK表达水平与BALF中嗜酸性粒细胞（EOS）计数和IL-5、IL-5 mRNA含量之间分别呈显著正相关（r=0.62、0.69、0.74,P〈0.01）。结论：p38 MAPK可能参与了支气管哮喘的发病过程。黄芪对哮喘的治疗作用可能部分与抑制p38 MAPK的磷酸化活化、降低炎症介质释放、减轻炎症细胞浸润有关。     

环孢素A气雾吸入对哮喘大鼠气道炎症的影响

目的研究环孢素A(CsA)气雾吸入对抗原诱导的大鼠过敏性气道炎症的作用。方法用卵白蛋白(OA)致敏大鼠,2周后气雾吸入CsA(5，10，20 g·L^-1),每天1次，连续7 d。大鼠致敏后d 20和d 21用OA(10 g·L^-1，每天1次)攻击，观察第2次OA攻击24 h后支气管肺泡灌洗液及外周血中嗜酸性粒细胞的数量和支气管肺组织病理学改变情况，测定支气管肺泡灌洗液中TNF-α含量。结果CsA气雾吸入能明显降低支气管肺泡灌洗液及外周血中嗜酸性粒细胞的数量，减轻肺组织中炎症细胞特别是嗜酸性粒细胞的浸润,减轻组织水肿及上皮损伤等气道炎症状况,降低支气管肺泡灌洗液中TNF-α含量。结论CsA气雾吸入对大鼠过敏性气道炎症具有抑制作用，其作用机制与细胞因子TNF-α释放减少有关。     

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。