射干麻黄药对对支气管哮喘小鼠模型气道炎症及外周血Th1/Th2的影响

目的:观察不同比例射干麻黄药对对支气管哮喘小鼠模型气道中炎症细胞及外周血Th1/Th2平衡的影响.方法:将80只BALB/c小鼠随机分为A正常对照组、B哮喘模型组、C地塞米松组、D麻黄单药组、E射干单药组及不同比例射干麻黄药对组F、G、H),每组10只.采用鸡卵蛋白(OVA)致敏和激发建立小鼠哮喘模型,给予不同药物对小鼠进行干预.干预完成后取支气管肺泡灌洗液(BALF)测定白细胞总数及嗜酸性粒细胞计数,采用ELISA检测小鼠血清中IL-4、IL-5、IL-13及IFNγ的水平,HE染色观察小鼠肺组织病理炎症等改变.结果:射干麻黄药对组可以显著减轻哮喘小鼠的哮喘症状,降低外周血中IL-4、IL-5、IL-13水平、升高IFN-γ水平,并能降低BALF中白细胞总数及嗜酸粒细胞水平.结论:射干麻黄药对可以显著减轻哮喘小鼠气道炎症,抑制炎症介质的释放,纠正Th1/Th2失衡,从而达到治疗哮喘的作用.     

Narirutin inhibits airway inflammation in an allergic mouse model

1. Flavonoids are naturally occurring compounds that possess anti-allergic, anti-inflammatory, antiproliferative and anti-oxidant properties. In the present study, we investigated whether the flavonoid narirutin could reduce airway inflammation in ovalbumin (OVA)-sensitized/challenged NC/Nga mice, a model of allergic eosinophilic airway inflammation. 2. Mice were initially immunized intraperitoneally with OVA on Days 0 and 7 and then challenged with inhaled OVA on Days 14, 15 and 16. In addition, some mice received narirutin orally at doses of 0.1, 1 or 10 mg/kg bodyweight daily on Days 7-16. 3. At 10 mg/kg, but not 0.1 or 1 mg/kg, narirutin significantly diminished OVA-induced airway inflammation caused by infiltration of lung tissue with inflammatory and mucus-producing cells, as well as reduced eosinophil counts in the peripheral blood and bronchoalveolar lavage fluid (BALF), interleukin (IL)-4 levels in BALF and IgE levels in serum. 4. The mechanism of the anti-inflammatory effect of narirutin are likely to be associated with a reduction in the OVA-induced increases of IL-4 and IgE in a murine model of allergic eosinophilic airway inflammation. These findings suggest that narirutin may be an effective new tool in the treatment of bronchial asthma.     

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.     

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.     

Inhibitory effects of Actinidia polygama extract and cyclosporine A on OVA-induced eosinophilia and bronchial hyperresponsiveness in a murine model of asthma

Actinidia polygama is one of the well known herb used in oriental medicine for treatment of anti-inflammatory and many allergic diseases. Anti-asthmatic effects of A. polygama in the development of OVA-induced eosinophilia and hyperresponsiveness in murine model of asthma have not been fully investigated in vivo. Cyclosporine A (CsA) has been shown to inhibit single allergen-induced allergic inflammation such as eosinophilic and lymphocytic infiltration and mRNA expression for interleukin (IL)-4 and IL-5. Asthma is a chronic inflammatory disease of the mucosa and is associated with excess production of Th2 cytokines and eosinophil influx in lung. To clarify the anti-inflammatory and anti-asthmatic effects of A. polygama and CsA, we examined the influence of A. polygama fructus extract (APF) and CsA on the development of pulmonary eosinophilic inflammation in murine model of asthma. Our results have shown that APF and CsA have profound inhibitory effects on the accumulation of eosinophills into airways, with the reduction of eosinophil and total lung leukocyte number by reducing IL-4, IL-5, IL-13 and IgE levels in the BALF. Moreover, APF decreased eosinophil CCR3 expression and CD11b expression in lung cells. These results indicate that APF has a deep inhibitory effect on airway inflammation and hyperresponsiveness in murine model of asthma and play a crucial role as an immunomodulator which possess anti-inflammatory and anti-asthmatic property by modulating the relationship between Th1/Th2 cytokine imbalance.     

Zingiber officinale ameliorates allergic asthma via suppression of Th2-mediated immune response

Abstract

Context: Ginger has been used commonly in the traditional system of medicine for the treatment of respiratory disorders.

Objective: The present study investigates the immunosuppressive activity of ginger by using the mouse model of ovalbumin-induced allergic asthma.

Materials and methods: Treatment with ethanol extract (500?mg/kg) and aqueous extract (720?mg/kg) of rhizomes, and methylprednisolone (5?mg/kg) was initiated 1 week after second sensitization of mice with ovalbumin and continued for 7?d. RT-PCR followed by gel electrophoresis and ELISA were used for the evaluation of mRNA expression levels and protein levels of Th2 type markers, respectively. Lung tissue histopathology was conducted by using H&E and PAS staining.

Results: We observed significant reduction in goblet cell hyperplasia (0.83?±?0.17 and 1.0?±?0.26), infiltration of inflammatory cells in airways (0.67?±?0.33 and 1.0?±?0.37), and edema with vascular congestion (1.0?±?0.26 and 1.2?±?0.17) by both ethanol and aqueous extracts, respectively. A highly significant reduction of total and differential count of eosinophils and neutrophils in BALF, and eosinophil count in blood were also observed. Both extracts significantly inhibited Th2-mediated immune response, which is evident by a decrease in mRNA expression levels of IL-4 and IL-5. Protein levels of IL-4 and IL-5 in BALF, along with total serum IgE levels, were also significantly suppressed by both extracts.

Discussion and conclusion: Our study validated the traditional use of ginger in respiratory disorders and suggests that ginger reduces allergic airway inflammation, possibly by the suppression of Th2-mediated immune response.     

Copaiba oil suppresses inflammation in asthmatic lungs of BALB/c mice induced with ovalbumin

Asthma is a chronic inflammatory disease that represents high hospitalizations and deaths in world. Copaiba oil (CO) is popularly used for relieving asthma symptoms and has already been shown to be effective in many inflammation models. This study aimed to investigate the immunomodulatory relationship of CO in ovalbumin (OVA)-induced allergic asthma. The composition of CO sample analyzed by GC and GC–MS and the toxicity test was performed in mice at doses of 50 or 100 mg/kg (by gavage). After, the experimental model of allergic asthma was induced with OVA and mice were orally treated with CO in two pre-established doses. The inflammatory infiltrate was evaluated in bronchoalveolar lavage fluid (BALF), while cytokines (IL-4, IL-5, IL-17, IFN-γ, TNF-α), IgE antibody and nitric oxide (NO) production was evaluated in BALF and lung homogenate (LH) of mice, together with the histology and histomorphometry of the lung tissue. CO significantly attenuated the number of inflammatory cells in BALF, suppressing NO production and reducing the response mediated by TH2 and TH17 (T helper) cells in both BALF and LH. Histopathological and histomorphometric analysis confirmed that CO significantly reduced the numbers of inflammatory infiltrate in the lung tissue, including in the parenchyma area. Our results indicate that CO has an effective in vivo antiasthmatic effect.     

特异性免疫治疗与糖皮质激素对变异性鼻炎患者Th17和RORγt的影响

目的 分析特异性免疫治疗与糖皮质激素对变异性鼻炎(allergic rhinitis,AR)患者辅助性T细胞17(Th17)及转录因子孤独受体(RORγt)mRNA表达的影响.方法 选取2010年6月—2014年5月收治的50例AR患者作为研究对象,依据临床治疗方案分为A、B两组,每组25例,A组采用糖皮质激素治疗,B组采用特异性免疫治疗方案,分别于治疗前后取患者外周血液,检测Th17细胞阳性率,测定RORγt mRNA表达水平,与正常健康人对照(对照组).测定患者白介素-17(IL-17)、IL-6和IL-23炎性介质水平,同时评估患者临床症状的改善情况.结果 AR患者Th17细胞阳性率、RORγt mRNA表达、IL-6、IL-17及IL-23水平均高于对照组(P<0.05).B组治疗后IL-6、IL-17、IL-23、Th17细胞阳性率和RORγt mRNA表达水平均低于治疗前和A组(P<0.05).两组治疗后鼻部症状评分及鼻炎生活质量表(RQLQ)评分均低于治疗前(P<0.05).治疗后B组鼻部症状评分及RQLQ评分低于A组(P<0.05).结论 AR患者外周血内Th17细胞、RORγt转录因子及相关炎性因子均可能参与AR的发病过程,采用特异性免疫治疗方案,可下调AR患者相关细胞因子及转录因子水平,改善患者鼻炎症状及生活质量.     

Dexamethasone alleviate allergic airway inflammation in mice by inhibiting the activation of NLRP3 inflammasome

Dexamethasone (DEX) is the mainstay treatment for asthma, which is a common chronic airway inflammation disease. However, the mechanism of DEX resolute symptoms of asthma is not completely clear. Here, we aimed to analyze the effect of DEX on airway inflammation in OVA-induced mice and whether this effect is related to the inhibition of the activation of NLRP3 inflammasome. Female (C57BL/6) mice were used to establish the allergic airway inflammation model by inhalation OVA. The number of inflammatory cells in the bronchi alveolar lavage fluid (BALF) was counted by Swiss-Giemsa staining, and the contents of IL-1β, IL-18, IL-5 and IL-17 were detected by ELISA. The degree of inflammatory cells infiltration and mucous cells proliferation in lung tissue were separately observed by H&E and PAS staining. The proteins expression of NLRP3, pro-caspase-1, caspase-1, IL-1β, IL-6 and IL-17 in lung tissue were detected by Western blotting. We found that DEX significantly inhibited OVA-induced inflammatory cells infiltration, airway mucus secretion and goblet cell proliferation in mice. The total and classified numbers of inflammatory cells and the levels of IL-1β, IL-18, IL-5 and IL-17 in the BALF of the experimental group were significantly lower than those of the model group after DEX treatment. DEX also significantly inhibited the activity of NLRP3 inflammasome and reduced the protein contents of Pro-Caspase-1, Caspase-1, Capase-1/Pro-Caspase-1, IL-1β, IL-6 and IL-17 in lung tissues. Our study suggested that DEX alleviates allergic airway inflammation by inhibiting the activity of NLRP3 inflammasome and the levels of IL-1β and IL-18.     

辛伐他汀对慢性阻塞性肺疾病模型大鼠炎性因子表达的影响

目的 研究辛伐他汀对慢性阻塞性肺疾病(COPD)模型大鼠炎性因子表达的影响.方法 将50只大鼠随机分为空白对照组(等体积生理盐水)、模型组(等体积生理盐水)和辛伐他汀高、中、低剂量组(15,10,5 mg/kg).灌胃给药,每日1次,连续灌胃56 d(除烟熏日外).除空白对照组外,其余各组大鼠分别于灌胃第29,43天气...     

Anti-inflammatory effect of amurensin H on asthma-like reaction induced by allergen in sensitized mice

AIM: To explore the anti-inflammatory effects of amurensin H on asthma-like reaction induced by allergen in sensitized mice. METHODS: BALB/c mice were sensitized by ovalbumin (OVA, ip) on d 0 and d 14 and challenged with 1% OVA on d 18 to 22. Mice developed airway eosinophilia, mucus hypersecretion, and elevation in cytokine levels. Mice were administered amurensin H orally at the doses of 49, 70, or 100 mg/kg once every day from d 15 to the last day. Bronchoalveolar lavage fluid (BALF) were collected at 24 h and 48 h after the last OVA challenge. Levels of tumor necrosis factor-alpha (TNF-alpha), interleukin 4 (IL-4), interleukin 5 (IL-5), and interleukin 13 (IL-13) in BALF were measured using ELISA method. Differential cell counts of macrophages, lymphocytes, neutrophils and eosinophils were performed in 200 cells per slide (one slide per animal). Lung tissue sections of 6-mum thickness were stained with Mayer's hematoxylin and eosin for assessment of cell infiltration, mucus production, and tissue damage. RESULTS: Oral administration of amurensin H significantly inhibited OVA-induced increases in total cell counts, eosinophil counts, and TNF- alpha, IL-4, IL-5 and IL-13 levels in BALF. In addition, amuresin H dramatically decreased OVA-induced lung tissue damage and mucus production. CONCLUSION: Amurensin H may have therapeutic potential for the treatment of allergic airway inflammation.     

Genipin inhibits allergic responses in ovalbumin-induced asthmatic mice

Genipin is a natural compound isolated from the fruit of Gardenia jasminoides with various pharmacological effects. In this study, we investigated whether genipin effectively alleviates allergic responses in a murine model of ovalbumin (OVA)-induced asthma. The mice were administered an intraperitoneal injection of OVA on day 0 and 14 to boost the immune response; genipin was then administered from day 18 to 23 by oral gavage. On days 21 to 23, mice were OVA-challenged using am ultrasonic nebulizer, and airway hyperresponsiveness (AHR) was determined on day 24 by plethysmography. Genipin significantly reduced the inflammatory cell count in bronchoalveolar lavage fluids (BALF) and AHR, which were accompanied by lower interleukin-5 (IL-5), IL-13 and OVA-specific immunoglobulin (Ig) E levels in the BALF or serum from OVA-induced asthmatic mice. In histology, genipin significantly decreased airway inflammation and mucus hypersecretion in OVA-induced asthmatic mice. Additionally, genipin inhibited OVA-induced increases in the expression of inducible nitric oxide synthase and cyclooxygenase-2 proteins. Further, genipin reduced the activity and protein levels of matrix metalloproteinase-9 in lung tissue from OVA induced asthmatic mice. Overall, genipin effectively alleviated the asthmatic inflammatory response in an OVA-induced asthmatic model. Therefore, our results suggest that genipin has therapeutic potential for treating asthma.     

Protective effect of platycodin D on liver injury in alloxan-induced diabetic mice via regulation of Treg/Th17 balance

Platycodin D is a major pharmacological constituent of Platycodi Radix with immunomodulatory activity. The present study was designed to investigate how platycodin D (PLD) reveals liver injury in diabetic mice and its mechanism. Fifty mice were divided into five groups randomly: control group, model group, rosiglitazone (ROG, 10 mg/kg) group, PLD (50 mg/kg) group, and PLD (100 mg/kg) group. Diabetes was induced with the injection of alloxan monohydrate (150 mg/kg) subcutaneously, and animals with blood glucose level of ≥ 250 mg/dl were considered as diabetic mice. After the first day of diabetes induction, the treatments were performed for 8 weeks. Then the animals were anaesthetized, and blood and liver samples were also collected for further assay. PLD significantly decreased the serum levels of glucose, insulin, interleukin-6 (IL-6), interleukin-1β, tumor necrosis factor-α (TNF-α), and interleukin (IL)-17A and increased IL-10 level in serum. PLD effectively downregulated aspartate transaminase (AST), alanine aminotransferase (ALT), total cholesterol (TC), and triglycerides (TG) in liver. PLD also attenuated liver histological change. In addition, PLD significantly attenuated IL-17A and IL-10 levels in vitro, flow cytometry (FCM) studies also showed that PLD remarkably inhibited Th17 cells and significantly increased Treg cells in liver tissues and spleen cells. Western blot demonstrated PLD inhibited the phosphorylation of JAK and STAT-3 and the expression of RORγt and increased the expression of Foxp3. The findings showed that PLD exerts beneficial effects on alloxan-induced liver injury in mice.     

An anti-inflammatory role for a phosphoinositide 3-kinase inhibitor LY294002 in a mouse asthma model

Phosphoinositide 3-kinase (PI3K) exhibits broad functional effects in immune cells. We investigated the role of PI3K in allergic airway inflammation using LY294002, a specific PI3K inhibitor, in a mouse asthma model. BALB/c mice were sensitized and challenged with ovalbumin (OVA), and developed airway eosinophilia, mucus hypersecretion, elevation in cytokine levels, and airway hyperresponsiveness. Intratracheal administration of LY294002 significantly inhibited OVA-induced increases in total cell counts, eosinophil counts, and IL-5, IL-13, and eotaxin levels in bronchoalveolar lavage fluid. Histological studies show that LY294002 dramatically inhibited OVA-induced lung tissue eosinophilia and airway mucus production. In addition, LY294002 significantly suppressed OVA-induced airway hyperresponsiveness to inhaled methacholine. Western blot analysis of whole lung lysates shows that LY294002 markedly attenuated OVA-induced serine phosphorylation of Akt, a direct downstream substrate of PI3K. Taken together, our findings suggest that inhibition of PI3K signaling pathway can suppress T-helper type 2 (Th2) cytokine production, eosinophil infiltration, mucus production, and airway hyperresponsiveness in a mouse asthma model and may have therapeutic potential for the treatment of allergic airway inflammation.