The Fermented Soy Product ImmuBalanceTM Suppresses Airway Inflammation in a Murine Model of Asthma

The fermented soy product ImmuBalance contains many active ingredients and its beneficial effects on some allergic diseases have been reported. We hypothesized that ImmuBalance could have potential effects on airway inflammation in a murine model of asthma. Mice sensitized and challenged with ovalbumin developed airway inflammation. Bronchoalveolar lavage fluid was assessed for inflammatory cell counts and levels of cytokines. Lung tissues were examined for cell infiltration and mucus hypersecretion. Oral administration of ImmuBalance significantly inhibited ovalbumin-induced eosinophilic inflammation and decreased Th2 cytokine levels in bronchoalveolar lavage fluid (p < 0.05). In addition, lung histological analysis showed that ImmuBalance inhibited inflammatory cell infiltration and airway mucus production. Our findings suggest that supplementation with ImmuBalance may provide a novel strategy for the prevention or treatment of allergic airway inflammation.     

CTRP9抑制幼年哮喘小鼠气道平滑肌细胞增殖和炎性反应的研究

目的 研究C1q/肿瘤坏死因子相关蛋白9(CTRP9)对哮喘幼鼠气道平滑肌细胞(airway smooth muscle cells,ASMCs)增殖和气道炎症的作用。方法 ELISA检测正常、哮喘儿童和小鼠血清中CTRP9含量;原代培养哮喘小鼠ASMCs,经pcDNA3.1-CTRP9转染后,MTT检测ASMCs增殖,ELISA测定TNF-α和IL-6含量,Western 印记检测TLR4和NF-κB p65蛋白表达以及NF-κB p65磷酸化水平;pcDNA3.1-CTRP9转染哮喘小鼠,HE染色观察肺组织炎性细胞的浸润程度;收集小鼠支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF),光镜下检测嗜酸性粒细胞、巨噬细胞和中性粒细胞数目;检测小鼠肺部TNF-α、IL-6、TLR4和NF-κB的表达。结果 哮喘儿童和小鼠血清中CTRP9含量低于正常组;CTRP9能抑制ASMCs增殖、炎症因子分泌和TLR4/NF-κB通路活化;CTRP9也能抑制哮喘小鼠肺部炎性细胞浸润、各炎性反应细胞数目、炎症因子分泌和TLR4/NF-κB通路活化。结论 CTRP9能抑制幼年哮喘小鼠ASMCs增殖和炎性反应。     

Lactobacillus gasseri suppresses Th17 pro-inflammatory response and attenuates allergen-induced airway inflammation in a mouse model of allergic asthma

Probiotics are normal inhabitants of the gastrointestinal tract of man and are widely considered to exert a number of beneficial effects in many diseases. But the mechanism by which they modulate the immune system is poorly understood. The present study was planned to explore the anti-allergic effect of Lactobacillus gasseri on a mouse model of allergic asthma. Dermatophoides pteronyssinus (Der p) sensitised and challenged BALB/c mice were orally administered via oral administration with three different doses of L. gasseri (low, 1 × 10(6) colony-forming units (CFU); medium, 2 × 10(6) CFU; high, 4 × 10(6) CFU), in 700 μl of PBS daily, starting from 2 weeks before Der p sensitisation for 4 weeks. After the allergen challenge, airway responsiveness to methacholine, influx of inflammatory cells to the lung, and cytokine levels in bronchoalveolar lavage (BAL) fluids and splenocytes culture were assessed. Our results showed that oral administration of a high dose of L. gasseri (4 × 10(6) CFU) decreased airway responsiveness to methacholine, attenuated the influx of inflammatory cells to the airways and reduced the levels of TNF-α, thymus and activation-regulated chemokine (TARC) and IL-17A in BAL fluids of Der p-sensitised and -challenged mice. Moreover, L. gasseri decreased IL-17A production in transforming growth factor-α and IL-6 stimulated splenocytes and cell numbers of IL-17 producing alveolar macrophages in L. gasseri-treated mice as compared to non-treated, Der p-sensitised and -challenged mice. In conclusion, oral administration with L. gasseri can attenuate major characteristics of allergen-induced airway inflammation and IL-17 pro-inflammatory immune response in a mouse model of allergic asthma, which may have clinical implication in the preventive or therapeutic potential in allergic asthma.     

封闭胸腺基质淋巴生成素受体对超敏原引起的气道炎症性应答的作用

目的为阐明胸腺基质淋巴生成素受体（Thymic stromal lymphopoietin receptor,TSLPR）在超敏原引起的气道炎症应答中的作用,并在小鼠模型中探讨局部封闭TSLPR用以缓解哮喘的可能性。方法对TSLPR抗体或同型抗体预处理,且以鸡卵白蛋白（OVA）诱导的各组小鼠,分别行气道浸润细胞的分类和记数、H＆E和PAS的肺组织染色分析;并以酶联免疫吸附测定法（ELISA）检测支气管肺泡灌洗液中炎性细胞因子;进一步分析OVA激发的小鼠树突状细胞（DCs）的迁移能力和成熟状态。结果在小鼠OVA致敏前施以抗TSLPR抗体可显著减少气道粘液的分泌、嗜酸性粒细胞和淋巴细胞浸润;同时引发IL-4、IL-5水平的明显下降。而作为其机制之一,TSLPR的中和作用可阻止超敏原诱导的DCs的成熟和迁移。结论封闭TSLPR介导的信号通路可缓解哮喘小鼠的气道炎症反应,有望成为一项新的防治气道变应性疾病策略。     

Inhibition of allergen-induced airway inflammation and hyperreactivity by recombinant lactic-acid bacteria

Recombinant lactic-acid bacteria (LAB) are able to inhibit allergen-specific T-cell responses. In this study, we examined whether oral feeding of recombinant LAB was able to suppress allergen-induced airway inflammation and hyperreactivity (AHR) in a murine model. Animals were intraperitoneally sensitized with Dermatophagoides pteronyssinus group-5 allergen (Der p 5) and orally treated with recombinant LAB containing a plasmid-encoded Der p 5 gene or placebo on day 7 and day 14 for three days consecutively. Twenty-one days after sensitization, mice underwent inhalational challenging. Der p 5-specific immunological responses including changes to specific immunoglobulin G and E (IgE) levels, the presence of cells in the bronchoalveolar lavage fluid (BALF), and AHR were assessed following this inhalational challenge. We demonstrated that oral feeding of recombinant LAB could significantly decrease the synthesis of Der p 5-specific IgE, and AHR. Furthermore, following such treatment, we also noted that both neutrophils and eosinophils had infiltrated the BALF to a significantly lower extent, when compared to the vehicle-treated group. Neither recombinant allergen nor LAB alone was able to suppress allergen-induced immune responses. Our findings suggest that treatment with recombinant LAB at a low dose can suppress allergen-induced airway allergic inflammation, this providing a basis for developing a novel therapeutic method for allergic airway diseases.     

Cis-9,trans-11-conjugated linoleic acid inhibits allergic sensitization and airway inflammation via a PPARgamma-related mechanism in mice

Milk consumption from early childhood on has been found to be inversely correlated with allergic sensitization and the onset of bronchial asthma. We tested whether cis-9,trans-11-conjugated linoleic acid (c9,t11-CLA), naturally occurring in milk fat, may prevent allergic sensitization and inhibit airway inflammation in a murine asthma model. BALB/c mice were fed a diet enriched in 1 wt% of c9,t11-CLA or a control diet 7 d prior to and for 32 d during sensitization [d 1 and 14, 100 mg/L ovalbumin (OVA) in adjuvant vs. PBS] and airway challenges (d 28-30, 1% OVA in PBS vs. PBS). Subgroups of mice were coadministered 20 micromol/L of the selective PPARgamma antagonist GW9662 during each OVA challenge. C9,t11-CLA feeding resulted in significantly reduced IgE production and allergen-induced in vivo airway hyperresponsiveness. Further, less mucous plugging of segmental bronchi and significantly reduced interleukin-5 and eosinophils were determined in bronchoalveolar lavage fluids of c9,t11-CLA-fed mice. C9,t11-CLA feeding prevented the downregulation of PPARgamma mRNA in the lung tissues observed after allergen sensitization and airway challenges in control mice. The inhibitory effects of c9,t11-CLA on airway inflammation were partially prevented by coadministration of GW9962. Further, c9,t11-CLA feeding resulted in a significantly lower concentration of the eicosanoid precursor, arachidonic acid, in tissue lipids. These findings demonstrate that dietary c9,t11-CLA can reduce allergic airway inflammation, most likely via a PPARgamma-related mechanism and by reducing eicosanoid precursors. They give new insights into the fatty acid-mediated mechanism of immunomodulation and may represent a step toward an attractive novel strategy in the dietary prevention and treatment of allergic asthma.     

Oral administration of allergen extracts from mugwort pollen desensitizes specific allergen-induced allergy in mice

Clinically, sublingual immunotherapy (SLIT) using allergen extracts effectively alleviates the symptoms of allergic rhinitis and asthma. We hypothesized that oral administration of a high-dose of allergen extracts imitates SLIT, which may prevent IgE-related responses in allergic diseases. In the present study, we investigated the effects of oral administration of allergen extracts from mugwort pollen (MP) on allergen-induced inflammation and airway hyperresponsiveness (AHR) in an allergic mouse model. After administration of MPdrop containing Art v 1 and Art v 4 extracts derived from MP specifically in MP-sensitized mice, the effects of MPdrop on AHR, inflammatory cell accumulation, cytokine production in the bronchoalveolar lavage fluid and lung tissue, and serum IgE and IgG levels were investigated. The results indicated that MPdrop not only prevented the AHR in response to methacholine in a dose-dependent manner but also significantly reduced the total serum and allergen-specific IgE levels. All of the maximal effects were achieved at a dose of 100μg/(kgd) and were comparable to the effects of dexamethasone at a dose of 0.5mg/(kgd). Furthermore, oral administration of MPdrop dose-dependently elevated allergen-specific serum IgG2a levels, reduced total and allergen-specific IgE levels and normalized the imbalance between the Th1 cytokine IL-12 and Th2 cytokines IL-4 and IL-5. Finally, oral administration of MPdrop significantly reduced goblet cell hyperplasia and eosinophilia in the MP-sensitized allergic mouse model. These data suggest that MPdrop effectively improves specific allergen-induced inflammation and AHR in MP-sensitized and -challenged mice and provides the rationale for clinical use of MPdrop in the specific allergen-induced asthma.     

Influence of age on the development of immunological lung response in intrauterine undernourishment

OBJECTIVE: We investigated the effect of intrauterine undernourishment on some features of asthma using a model of allergic lung inflammation in rats. The effects of age at which the rats were challenged (5 and 9 wk) were also evaluated. METHODS: Intrauterine undernourished offspring were obtained from dams that were fed 50% of the nourished diet of counterparts and were immunized at 5 and 9 wk of age. They were tested for immunoglobulin E anti-ova titers (by passive cutaneous anaphylaxis), cell count in the bronchoalveolar fluid, leukotriene concentration, airway reactivity, mucus production, and blood corticosterone and leptin concentrations 21 d after immunologic challenge. RESULTS: Intrauterine undernourishment significantly reduced the antigen-specific immunoglobulin E production, inflammatory cell infiltration into airways, mucus secretion, and production of leukotrienes B(4)/C(4) in the lungs in both age groups compared with respective nourished rats. The increased reactivity to methacholine that follows antigen challenge was not affected by intrauterine undernourishment. Corticosterone levels increased with age in the undernourished rats' offspring, but not in the nourished rats' offspring. Undernourished offspring already presented high levels of corticosterone before inflammatory stimulus and were not modified by antigen challenge. Leptin levels increased with challenge in the nourished rats but not in the undernourished rats and could not be related to corticosterone levels in the undernourished rats. CONCLUSION: Intrauterine undernourishment has a striking and age-dependent effect on the offspring, reducing lung allergic inflammation.     

变应性鼻炎大鼠TNFAIP3基因在鼻腔上皮细胞的表达

目的 研究致敏-治疗过程中肿瘤坏死因子α诱导蛋白3(tumor necrosis factor alpha-induced protein 3,TNFAIP3)基因在大鼠鼻腔黏膜上皮的表达,探讨TNFAIP3基因在变应性鼻炎免疫进程中的作用.方法 本实验于在2019年1月—2020年1月开展,将24只Wistar大鼠,...     

互隔交链孢霉诱导小鼠气道变态反应性炎症动物模型的构建

目的研究使用互隔交链孢霉孢子构建C57BL/6小鼠肺部变态反应性炎症模型的方法。方法模型组、阳性对照组和阴性对照组各12只小鼠。模型组以互隔交链孢霉孢子悬液致敏和激发小鼠,分别于第0、5、10天腹腔注射霉菌孢子悬液,第14天开始连续4天鼻滴霉菌孢子悬液。阳性对照组以鸡卵蛋白(OVA)代替霉菌孢子,阴性对照组以磷酸缓冲液(PBS)代替霉菌孢子。进行肺组织病理检查,支气管肺泡灌洗液(BALF)中细胞计数分类和总蛋白浓度测定,ELISA测定BALF中的白细胞介素-4(IL-4)和干扰素-γ(IFN-γ)及血浆中的总IgE和特异性IgE,测定小鼠气道阻力。结果模型组和阳性对照组可见明显炎症细胞浸润,特别是嗜酸性粒细胞,而阴性对照组则未见。BALF中细胞总数和嗜酸性粒细胞比例较阴性对照组明显升高(P<0.05);BALF上清中IL-4、总蛋白水平较阴性对照组明显升高(P<0.05),IFN-γ水平较阴性对照组降低(P<0.05)。血浆中总IgE和特异性IgE较阴性对照组明显升高(P<0.05)。小鼠气道阻力较阴性对照相比有明显升高(P<0.05)。结论使用互隔交链孢霉孢子致敏和激发C57BL/6小鼠成功的建立了小鼠气道变态反应性炎症模型。     

Nasal and bronchial responses to flour-inhalation in subjects with occupationally induced allergy affecting the airway

Objectives: The aim of the study was to follow the similarities and differences, of cellular and mediator changes and mucosal/vascular permeability in the upper and lower airway after specific and nonspecific bronchial provocation, in bakers with diagnosed occupationally induced allergy affecting the airway. In addition, the authors try to find whether there is a relationship between cellular changes in nasal and bronchoalveolar lavage, and bronchial hyperreactivity. Methods: The study participants were 10 bakers with occupational bronchial asthma and allergic rhinitis. All patients were sensitized to investigated allergen-flour. Nasal- and bronchoalveolar lavage techniques were used to evaluate the changes of the cellular and mediator response (tryptase, eosinophil cationic protein, ECP) and albumin level after specific (flour) and placebo provocation. In addition, bronchial hyperreactivity for histamine, and forced expiratory volume in 1 s (FEV₁) were measured after the challenge. Results: There was a significant increase in the percentage of eosinophils, basophils and albumin in nasal and bronchoalveolar lavage of occupationally sensitized bakers. A statistically significant increase in the percentage of neutrophils in bronchoalveolar lavage was observed only 24 h after the allergen challenge. The level of tryptase in nasal lavage was significantly higher during the early allergic response. The levels of ECP in both nasal and bronchoalveolar lavage were significantly increased during the late allergic response. There were also severe bronchial reactions and increase of bronchial hyperreactivity for histamine in occupationally sensitized bakers in the late phase of allergic reaction. Conclusion: Eosinophils and basophils proved to be the predominant cells in nasal and bronchoalveolar lavage of patients with occupationally induced bronchial asthma and rhinitis. The prolonged increase of albumin level seems also to be a good predictor of protracted nasal and bronchial inflammation. The results obtained confirmed that tryptase and ECP are good markers for monitoring mast cell and eosinophil degranulation during the allergic reaction. Increase of airway responsiveness reflects an eosinophil and basophil contribution to airway allergic response. Received: 24 June 1999 / Accepted: 25 April 2000     

磷酸酰肌醇-3激酶抑制剂对小鼠气道炎症影响的实验研究

目的观察磷酸酰肌醇-3激酶(PI3K)抑制剂LY294002对卵白蛋白(OVA)诱导哮喘小鼠气道炎症的影响。方法Balb/c小鼠30只,随机分为三组,每组10只:PI3K抑制剂治疗组(LY组),OVA组和正常对照组(NC组)。通过OVA多次腹腔注射致敏和反复雾化激发,建立哮喘小鼠模型。末次抗原激发后48h收集支气管肺泡灌洗液(BALF)和骨髓标本,计数细胞总数和嗜酸性粒细胞(Eos),并行肺组织学检查。结果与OVA组比较,LY组BALF中细胞总数及Eos绝对数明显减少(p<0.05);肺组织中细支气管、血管周围Eos浸润得到控制,气道粘液分泌减少。结论PI3K抑制剂(LY294002)对卵白蛋白(OVA)致敏的哮喘小鼠气道炎症具有抑制作用。     

A whey-based glutathione-enhancing diet decreases allergen-induced airway contraction in a guinea-pig model of asthma

Since an allergen-induced early asthmatic reaction is likely to be accompanied by oxidative stress and since levels of the endogenous antioxidant glutathione can be enhanced by a whey-based diet (undenatured whey protein concentrate, UWPC), it was investigated whether UWPC could alleviate allergen-induced lung contractions. Guinea pigs were fed water or UWPC twice a day starting at day - 3 up to day 20. The animals were sensitised to ovalbumin or received saline on day 0. Serum samples were taken at several days after sensitisation to measure allergen-specific IgG. On day 20, lungs were isolated and perfused with buffer containing the allergen ovalbumin. Airway contractions were assessed, and mediators and indicators for oxidative stress were measured in the lung effluent. Moreover, glutathione levels were determined in the liver. The indicator of oxidative stress and airway contractile mediator, 8-iso-PGF(2α), was increased upon ovalbumin challenge in ovalbumin-sensitised groups. Furthermore, thiobarbituric acid-reactive substances (TBARS) were increased as well. Sensitisation with ovalbumin increased IgG levels from day 12 up to day 20, which were not influenced by the UWPC diet. In contrast, the UWPC diet significantly enhanced glutathione levels in the liver. Moreover, the UWPC diet significantly reduced the ovalbumin-induced anaphylactic response by 45 % and decreased PGE2 levels by 55 % in the effluent fluid. We show for the first time that during anaphylaxis, there is acute oxidative stress in the respiratory tract. The UWPC diet did not influence the sensitisation response to the allergen but did increase endogenous glutathione levels. The UWPC diet profoundly reduces allergen-induced airway constrictions, which opens new avenues for dietary management of allergic diseases.     

Liposomal retinoic acids modulate asthma manifestations in mice

Signaling of all-trans retinoic acid (ATRA) through nuclear retinoid acid (RA) receptors regulates several biological functions in airway epithelial cells, eosinophils, and immune cells, yet its impact on different in vivo aspects of pulmonary allergic reaction remains elusive. We compared the effect of a treatment with liposomally encapsulated ATRA (Lipo-ATRA) in a mouse model of ovalbumin (OVA)-induced T helper (Th) 2-type responses and airway remodeling. Daily intraperitoneal injections of 10 mg/kg Lipo-ATRA, at the time of each of the 2 systemic sensitizing injections, increased OVA-induced Immunoglobulin E synthesis, bronchoalveolar lavage (BAL) eosinophilia, and accumulation of IL-5, transforming-growth factor beta1, fibronectin, eotaxin/chemokine (C-C motif) ligand 11 (eotaxin/CCL11) and regulated upon activation, normal T expressed and secreted chemokine (C-C motif) ligand 5. In contrast, Lipo-ATRA, administered during each of the 4 intranasal OVA challenges, did not affect these variables. Regardless of the treatment regimen, Lipo-ATRA augmented mucin levels in BAL fluid and reduced lung total collagen content. In vitro incubation of mouse splenocytes or purified spleen cluster of differentiation (CD) 4-positive T lymphocytes, with ATRA, increased, respectively, OVA- and anti-CD 3 antibody-induced IL-4 and IL-5 production and inhibited IFNgamma release. These findings demonstrate that, when given during systemic sensitization, Lipo-ATRA exacerbates allergic immune and inflammatory responses, most likely by promoting Th2 development.     

小檗碱对流感病毒感染小鼠免疫功能的影响

目的探究小檗碱对流感病毒感染小鼠免疫功能及炎症反应的调节作用。方法选取60只小鼠分为正常对照组、流感病毒组和小檗碱组,20只/组;除正常对照组外各组使用流感病毒滴鼻制成流感病毒小鼠模型;使用1 mg/ml小檗碱给小鼠灌胃0.4 ml/d。检测肺泡灌洗液中白细胞总数及各类白细胞数;苏木精-伊红(HE)染色观察小鼠肺组织病理改变情况;采用流式法检测辅助性T细胞(Th)1/Th2及Th17/调节性T细胞(Treg)比例;检测血清中白细胞介素(IL)-6、IL-4和肿瘤坏死因子(TNF)-α水平;使用蛋白质免疫印迹法检测视黄酸诱导基因蛋白-1(RIG-1)、线粒体抗病毒信号蛋白(MAVS)及核因子(NF)-κB蛋白表达情况。结果HE染色显示,流感病毒组肺组织炎性细胞增多、肺泡结构被破坏,小檗碱组小鼠炎性细胞数目减少。相比正常对照组,流感病毒组小鼠肺泡灌洗液中白细胞总数及各类白细胞数、Th1/Th2、Th17/Treg、IL-6、TNF-α、RIG-1蛋白、MAVS蛋白及NF-κB蛋白水平均升高,IL-4水平降低(P<0.05);相比流感病毒组,小檗碱组小鼠肺泡灌洗液中白细胞总数及各类白细胞数、Th1/Th2、Th17/Treg、IL-6、TNF-α、RIG-1蛋白、MAVS蛋白及NF-κB蛋白水平均降低,IL-4水平升高(P<0.05)。结论小檗碱可以通过影响RLH信号传导通路的活化,调节T淋巴细胞亚群比例,同时抑制炎症反应来实现抗流感病毒的作用。     

不同剂量流感病毒感染小鼠模型免疫应答的比较性研究

目的 比较不同剂量流感病毒感染小鼠后肺组织急性损伤情况,初步探讨感染剂量与机体抗病毒的固有、适应性免疫应答的关系。方法 用流感病毒A/PR/8/34（PR8;H1N1）构建小鼠感染102 PFU和103 PFU剂量模型,HE法检测肺组织损伤情况,HA法和免疫荧光染色法检测肺组织病毒滴度和感染程度,RT - PCR法检测肺组织炎性因子干扰素α（interferon - α, IFN - α）、干扰素β（interferon - α, IFN - β）、白细胞介素1β（interleukin - 1β, IL - 1β）、白细胞介素6（interleukin - 6, IL - 6）、肿瘤坏死因子α（tumor necrosis factor - α, TNF - α）及Toll样受体7（Toll - like receptor - 7, TLR7）、髓样分化因子88（myeloid differentiation primary response gene 88, MyD88）、肿瘤坏死因子受体相关蛋白6（TNF receptor associated factor 6, TRAF6）和核因子κB（nuclear factor κB, NF - κB）mRNA水平,免疫组化法对肺组织TLR7、MyD88、TRAF6和NF - κB蛋白定位。流式细胞术检测肺组织、支气管肺泡灌洗液（bronchoalveolar lavage fluid, BALF）和淋巴结中CD4+、CD8+T细胞比例。结果 与102 PFU组比较,103 PFU组小鼠肺组织病毒滴度显著升高（t = 3.071,P = 0.008）,炎症因子IFN - α、IFN - β、IL - 1β、IL - 6、TNF - α mRNA水平上升（t = 4.816,P = 0.003;t = 4.104,P = 0.006;t = 3.992,P = 0.007;t = 4.049,P = 0.007;t = 3.737,P = 0.009）,TLR7、MyD88、TRAF6和NF - κB mRNA水平有不同程度的升高（t = 2.905,P = 0.027;t = 3.854,P = 0.008;t = 3.760,P = 0.009;t = 4.312,P = 0.005）及蛋白表达显著增加（t = 4.712,P = 0.003;t = 4.008,P = 0.007;t = 4.398,P = 0.005;t = 4.564,P = 0.004）,BALF、肺组织和淋巴结中CD4+T细胞比例呈上升趋势（t = 3.771,P = 0.009;t = 2.463,P = 0.049;t = 3.386,P = 0.015）,BALF和肺组织CD8+T细胞比例上调（t = 5.297,P = 0.002;t = 3.502,P = 0.013）。结论 病毒载量影响机体固有和适应性免疫应答状态,通过上调 TLR7 - MyD88 - TRAF6 - NF - κB信号通路促进下游炎症反应,并启动CD4+T和CD8+T细胞发挥抗病毒效应。     

Fisetin Suppresses the Inflammatory Response and Oxidative Stress in Bronchial Epithelial Cells

Fisetin is isolated from many fruits and vegetables and has been confirmed to improve airway hyperresponsiveness in asthmatic mice. However, whether fisetin reduces inflammatory response and oxidative stress in bronchial epithelial cells is unclear. Here, BEAS-2B human bronchial epithelial cells were treated with various concentrations of fisetin and then stimulated with tumor necrosis factor-α (TNF-α) or TNF-α/interleukin-4. In addition, ovalbumin-sensitized mice were treated with fisetin to detect inflammatory mediators and oxidative stress expression. Fisetin significantly reduced the levels of inflammatory cytokines and chemokines in TNF-α-stimulated BEAS-2B cells. Fisetin also attenuated intercellular adhesion molecule-1 expression in TNF-α-stimulated BEAS-2B cells, suppressing THP-1 monocyte adhesion. Furthermore, fisetin significantly suppressed airway hyperresponsiveness in the lungs and decreased eosinophil numbers in the bronchoalveolar lavage fluid of asthmatic mice. Fisetin decreased cyclooxygenase-2 expression, promoted glutathione levels, and decreased malondialdehyde levels in the lungs of asthmatic mice. Our findings indicate that fisetin is a potential immunomodulator that can improve the pathological features of asthma by decreasing oxidative stress and inflammation.     

Dose-dependent thiol and immune responses to ovalbumin challenge in Brown Norway rats

Dose-dependent specific antibody production, antigen-dependent pulmonary inflammation, and thiol changes in the lung and associated lymph nodes were examined in a Brown Norway rat model of pulmonary sensitization. Cysteine (CYSH), glutathione (GSH), and markers of inflammation in bronchoalveolar lavage fluid (BALF) were measured following ovalbumin (OVA) inhalation challenge. Alveolar macrophages (AM) and pulmonary-associated lymph node cells (LNC) were isolated and intracellular CYSH and GSH assessed. OVA-specific IgE and IgG antibodies were quantified from sera. A dose-dependent biphasic response was noted with respect to OVA-specific IgE. OVA-specific IgG concentrations were maximal at 68 mg (OVA)/m3. OVA challenge to sensitized rats induced increases in BALF albumin, total protein, lactate dehydrogenase, CYSH and GSH that were independent of serum antibody concentrations. AM thiols were modestly elevated at low OVA challenge doses, but sharply reduced at the higher OVA challenge doses. In contrast, both thiols were dose dependently elevated in BALE CYSH, but not GSH, was elevated in LNC of OVA challenged rats. In summary, antigen exposure caused a dose-dependent alteration of inflammatory, thiol and immune parameters in OVA sensitized and challenged rats. Changes in thiol levels did not correlate with antibody responses. While the results of the present study do not support a functional role for thiols in the immune response, it is important to note the dose-dependent dramatic alteration seen in thiols following sensitization and challenge.