Skullcapflavone II attenuates ovalbumin-induced allergic rhinitis through the blocking of Th2 cytokine production and mast cell histamine release

Allergic rhinitis is a common heterogeneous chronic upper airway disorder and is an IgE-mediated inflammation characterized by one or more nasal symptoms such as sneezing, itching, nasal discharge, rhinorrhea, post nasal drainage and nasal blockage. In the present study, the effects of skullcapflavone II (SCFII) on upper airway inflammation, Th2 cytokines, and NF-κB signaling in an ovalbumin (OVA)-induced allergic rhinitis (AR) murine model in vivo were investigated. OVA-induced AR mice increased nasal symptoms, eosinophils and mast cells infiltration into nasal cavity, OVA-specific IgE/IgG1 and histamine in serum, Th2 cytokines including IL-13 and GATA3, and NF-κB signaling in NALF and lung homogenate. Interestingly, treatment of SCFII reduced the levels of OVA-specific IgE/IgG1 and histamine in serum, of Th2 cytokines and of NF-κB signaling in the NALF and the lung homogenate, and histopathological changes in the nasal tissue and the lung. Also, dexamethasone suppressed such increases. The results of this study suggested that SCFII may ameliorate allergic inflammation of upper airway in AR mice model by blocking the Th2 cytokine production, the NF-κB signal pathway and the mast cell histamine release. Taken together, we suggest that SCFII may be used as a therapeutic agent for patients with Th2-mediated or mast cell-mediated allergic diseases.     

Human cathelicidin CAP18/LL-37 changes mast cell function toward innate immunity

The antimicrobial peptide LL-37 is generated from skin keratinocytes during infection of Gram-negative bacteria and exerts a microbicidal effect. LL-37 also causes functional changes in mast cells. Mast cells in the skin are involved in the innate immune system response against microbial infections via Toll-like receptors (TLRs), such as TLR4, which that is known to recognize lipopolysaccharide (LPS), a bacterial component. Thus, in the present study, we examined the effects of LL-37 on the expression of TLRs and the generation of cytokines on mast cells, and considered functional changes in the host defense system against bacteria. We observed that LL-37 increased the level of TLR4 mRNA and TLR4 protein, and that LL-37 induced the release of IL-4, IL-5 and IL-1beta from mast cells. Cross-interaction between LL-37-triggered TLR4 augmentation and LL-37-inducible cytokine generation was also examined. Although the up-regulation of LL-37-inducible Th2 cytokines was cancelled by LPS, the augmentation of pro-inflammatory cytokine production was still observed. These findings indicate that LL-37 co-existing with the bacterial component switches mast cell function and directs human mast cells toward innate immunity. In conclusion, LL-37 may be a candidate modifier of the host defense against bacterial entry by serving as an alarm for sentinels such as mast cells.     

Functional molecules in allergic bronchial asthma

Bronchial asthma is considered to be a chronic airway inflammatory disease, characterized by airway obstruction, airway eosinophilic inflammation, and airway hyperresponsiveness (AHR) to a variety of stimuli. AHR is thought to be an important symptom, because the severity of the disease is generally correlated with the degree of AHR. Recent clinical studies have demonstrated the involvement of airway inflammation in the development of allergen-induced AHR, although, the mechanism of allergen-induced AHR has not been fully elucidated and remains controversial. In vivo animal models might provide important information on this point. We have established a mouse model of allergic asthma, which is characterized by airway eosinophilia, IgE production, T helper type 2 (Th2) cytokine production in the airway, and AHR, and investigated the role of inflammatory cells and functional molecules. Results from gene-knockout and mutant mice demonstrated the involvement of T cells, mast cells, prostanoids, and Th2 cytokines including interleukin (IL)-4 and IL-5 in the development of allergen-induced airway inflammation and AHR. In contrast, treatment with anti-IL-4 monoclonal antibody (mAb) or anti-IL-5 mAb during allergen inhalation did not inhibit allergen-induced AHR, although the combination of these mAbs clearly inhibited the enhanced responsiveness. These data indicate that it is a better strategy for control of the disease to inhibit or suppress multifunctional molecules like corticosteroids rather than to inhibit a single factor, because bronchial asthma is a multifactorial disease.     

Stimulation of cysteinyl leukotriene production in mast cells by heat shock and acetylsalicylic acid

Immunoglobulin (Ig) E-dependent activation of mast cells is central to the allergic response. The engagement of IgE-occupied receptors initiates a series of molecular events that causes the release of preformed, and de novo synthesis of, allergic mediators. Cysteinyl leukotrienes are able to contract airway smooth muscle and increase mucus secretion and vascular permeability and recruit eosinophils. Mast cells have also recently been recognized as active participants in innate immune responses. Heat stress can modulate innate immunity by inducing stress proteins such as heat-shock proteins (HSPs). We previously demonstrated that treatment of mast cells with heat shock or acetylsalicylic acid results in an increase of TNF-alpha and IL-6 release. This effect was paralleled by expression of HSP70. In the current study, we further investigated the effects of heat shock and acetylsalicylic acid on the activation of mast cells and the release of cysteinyl leukotrienes. In mouse mast cells, derived from a culture of bone marrow cells, responsiveness to heat shock, acetylsalicylic acid and exogenous or endogenous HSP70 was monitored by measuring leukotriene C4 release. We show that after heat shock treatment and exposure to acetylsalicylic acid leukotriene production was increased. Moreover, exogenous rHSP70 also induced leukotriene production. Because it has been reported that leukotriene production in mast cells may be mediated by Toll like receptor (TLR) activation, and HSP70 also activates TLRs signaling, we further explored these issues by using mast cells that are not able to produce HSP70, i.e. heat shock factor-1 (HSF-1) knockout cells. We found that in HSF-1 knockout bone marrow derived mast cells, heat shock and acetylsalicylic acid failed to induce release of leukotrienes. Moreover, in wild type cells the surface expression of TLR4 was attenuated, whereas the intracellular expression was up-regulated. We conclude that heat shock and acetylsalicylic acid induce the production and release of heat shock proteins from mast cells, which in turn stimulate leukotriene synthesis through activation of TLR4.     

Suppression of allergic reactions by royal jelly in association with the restoration of macrophage function and the improvement of Th1/Th2 cell responses

We studied the immunomodulatory effects of royal jelly (RJ), the principal food source of the queen honeybee. In this study, suppression of allergic reactions by RJ was investigated in DNP-KLH immunized mice (DNP-KLH mice). Oral administration of RJ (1 g/kg) to DNP-KLH mice significantly decreased the serum levels of antigen-specific Ig E and significantly inhibited DNP-KLH mediated-histamine release from mast cells, resulting in the suppression of immediate hypersensitivity reactions of ear skin. In DNP-KLH mice, IFN-gamma (Th1 cytokine) production from CD4+ T cells was suppressed and IL-4 (Th2 cytokine) production from CD4+ T cells was increased as compared to normal mice. On the other hand, RJ improved the balance of Th1/Th2 cell responses from Th2-dominant to Th1-dominant. RJ significantly increased GSH levels in macrophages from DNP-KLH mice. In addition, the administration of RJ to DNP-KLH mice increased IL-12 p40 mRNA expression and NO production, and decreased PG E2 production from macrophages as compared to untreated DNP-KLH mice. These results suggested that RJ suppressed antigen-specific Ig E production and histamine release from mast cells in association with the restoration of macrophage function and improvement of Th1/Th2 cell responses in DNP-KLH mice.     

The active contribution of Toll-like receptors to allergic airway inflammation

Epithelia lining the respiratory tract represent a major portal of entry for microorganisms and allergens and are equipped with innate and adaptive immune signaling receptors for host protection. These include Toll-like receptors (TLRs) that recognize microbial components and evoke diverse responses in cells of the respiratory system. TLR stimulation by microorganism-derived molecules activates antigen presenting cells, control T helper (Th) 1, Th2, and Th17 immune cell differentiation, cytokine production by mast cells, and activation of eosinophils. It is clear that TLR are involved in the pathophysiology of allergic airway diseases such as asthma. Dendritic cells (DCs), a kind of antigen presenting cells, which play a key role in the induction of allergic airway inflammation, are privileged targets for pathogen associated molecular patterns (PAMPs). During the allergic responses, engagement of TLRs on DCs determines the Th2 polarization of the T cells. TLR signaling in mast cells increases the release of IL-5, and TLR activation of airway epithelial cells forces the generation of proallergic Th2 type of cytokines. Although these responses aim to protect the host, they may also result in inflammatory tissue damage in the airway. Under certain conditions, stimulation of TLRs, in particular, TLR9, may reduce Th2-dependent allergic inflammation by induction of Th1 responses. Therefore, understanding the complex regulatory roles of TLRs in the pathogenesis of allergic airway inflammation should facilitate the development of preventive and therapeutic measures for asthmatic patients.     

Marine brevetoxin induces IgE-independent mast cell activation

Brevetoxins (PbTx) are sodium channel neurotoxins produced by the marine dinoflagellate Karenia brevis during red tide blooms. Inhalation of PbTx in normal individuals and individuals with pre-existing airways disease results in adverse airway symptoms including bronchoconstriction. In animal models of allergic inflammation, inhalation of PbTx results in a histamine H₁-mediated bronchoconstriction suggestive of mast cell activation. How mast cells would respond directly to PbTx is unknown. We thus explored the activation of mouse bone marrow–derived mast cells (BMMCs) following exposure to purified PbTx-2. Following in vitro exposure to PbTx-2, we examined cellular viability, mast cell degranulation (β-hexosaminidase release), intracellular Ca²⁺ and Na⁺ flux, and the production of inflammatory mediators (IL-6). PbTx-2 induced significant cellular toxicity within 24 h as measured by LDH release and Annexin-V staining. However, within 1 h of exposure, PbTx-2 induced BMMC degranulation and an increase in IL-6 mRNA expression independent of the high-affinity IgE receptor (FcεRI) stimulation. Activation of BMMCs by PbTx-2 was associated with altered intracellular Ca²⁺ and Na⁺ levels. Brevenal, a naturally produced compound that antagonizes the activity of PbTx, prevented changes in intracellular Na⁺ levels but did not alter activation of BMMCs by PbTx-2. These findings demonstrate that PbTx-2 activates mast cells independent of FcεRI providing insight into critical events in the pathogenesis and a potential therapeutic target in brevetoxin-induced airway symptoms.     

Anti-inflammatory effect of thymoquinone in a mouse model of allergic lung inflammation

Thymoquinone (TQ), the main active constituent of the volatile oil extracted from Nigella sativa's seeds, has been reported to have an anti-inflammatory and immune stimulatory effect on bronchial asthma and inflammation. However, little is known about the factors and mechanisms underlying these effects. In the present study, we examined the effect of TQ on airway inflammation in a mouse model of allergic asthma. Intraperitoneal injection of TQ before airway challenge of ovalbumin (OVA)-sensitized mice resulted in a marked decrease in lung eosinophilia and the elevated Th2 cytokines observed after airway challenge with OVA antigen; both in vivo, in the bronchoalveolar lavage (BAL) fluid and in vitro, following stimulation of lung cells with OVA. TQ also decreased the elevated serum levels of OVA-specific IgE and IgG1. Histological examination of lung tissue demonstrated that TQ significantly inhibited allergen-induced lung eosinophilic inflammation and mucus-producing goblet cells. While TQ showed a significant effect in inhibiting IL-4, IL-5 and IL-13 and some effect in inducing IFN-gamma production in the BAL fluid, it did show a slight effect on in vitro production of IL-4 by cultured lung cells stimulated with OVA antigen. These data suggest that TQ attenuates allergic airway inflammation by inhibiting Th2 cytokines and eosinophil infiltration into the airways; thus demonstrating its potential anti-inflammatory role during the allergic response in the lung.     

The effects of D3R on TLR4 signaling involved in the regulation of METH-mediated mast cells activation

Accumulating studies have revealed that the dopamine D3 receptor (D3R) plays an important role in methamphetamine (METH) addiction. However, the action of D3R on METH-mediated immune response and the underlying mechanism remain unclear. Mast cells (MCs) are currently identified as effector cells in many processes of immune responses, and MC activation is induced by various stimuli such as lipopolysaccharide (LPS). Moreover, CD117 and FcεRI are known as MC markers due to their specific expression in MCs. To investigate the effects of D3R on METH-mediated alteration of LPS-induced MCs activation and the underlying mechanism, in this study, we examined the expression of CD117 and FcεRI in the intestines of wild-type (D3R^+/+) and D3R-deficient (D3R^−/−) mice. We also measured the production of MC-derived cytokines, including TNF-α, IL-6, IL-4, IL-13 and CCL-5, in the bone marrow-derived mast cells (BMMCs) of WT and D3R^−/− mice. Furthermore, we explored the effects of D3R on METH-mediated TLR4 and downstream MAPK and NF-κB signaling induced by LPS in mouse BMMCs. We found that METH suppressed MC activation induced by LPS in the intestines of D3R^+/mice. In contrast, LPS-induced MC activation was less affected by METH in D3R^−/− mice. Furthermore, METH altered LPS-induced cytokine production in BMMCs of D3R^+/+ mice but not D3R^−/− mice. D3R was also involved in METH-mediated modulation of LPS-induced expression of TLR4 and downstream MAPK and NF-κB signaling molecules in mouse BMMCs. Taken together, our findings demonstrate that the effect of D3R on TLR4 signaling may be implicated in the regulation of METH-mediated MCs activation induced by LPS.     

Lipopolysaccharide from Porphyromonas gingivalis stimulates rat mast cells to cysteinyl leukotriene generation and upregulates Toll-like receptor -2 and -4 expression

Mast cells are found in all tissues of the oral cavity and it is suggested that they take part in the development of oral inflammation. As Porhyromonas gingivalis is widely recognized as a major pathogen in the development and progression of gingivitis and periodontitis, the aim of our study is to determine the effect of P. gingivalis lipopolysaccharide (LPS) on mast cell degranulation, cysteinyl leukotriene (cysLT) generation, and migration, as well as Toll-like receptor (TLR)-2 and -4 expression. Experiments were carried out in vitro on rat peritoneal mast cells. LPS-induced mast cell histamine release was estimated by a spectrofluorometric method and cysLT generation by ELISA test. Mast cell migration in response to this antigen was examined according to Boyden's modified method and TLR expression was determined by flow cytometry. We found that P. gingivalis LPS did not induce mast cell degranulation and histamine release. However, activation of mast cells with this bacterial antigen resulted in generation and release of significant amounts of cysLTs. We also documented that LPS from P. gingivalis did not stimulate mast cell migration, even in the presence of laminin, whereas it strongly upregulated TLR2 and TLR4 expression on mast cells. Observations that P. gingivalis LPS activates mast cells to generate and release proinflammatory mediators such as cysLTs and modulates TLR2 and TLR4 expression indicates that these cells might be involved in the emergency of inflammatory processes evolved in response to P gingivalis infection.     

Histamine production via mast cell-independent induction of histidine decarboxylase in response to lipopolysaccharide and interleukin-1

Histamine modulates immune responses. There are at least two ways histamine might be supplied: one is its release from cells that pool pre-formed histamine and the other is its de novo formation via induction of histidine decarboxylase (HDC). Lipopolysaccharide (LPS) and the proinflammatory cytokine interleukin (IL)-1 induce a marked elevation of HDC activity in various tissues or organs. To examine the contribution of mast cells to HDC induction in mice given LPS or IL-1, we examined the effects of LPS and IL-1 on HDC activity and/or histamine content in various organs (liver, lung, spleen or bone marrow) in mast cell-deficient mice (W/Wv), their normal littermates (+/+) and BALB/c mice deficient in IL-1alpha, IL-1beta and tumor necrosis factor (TNF)-alpha (IL-1alpha beta/TNFalphaKO mice). In non-stimulated mice, the histamine in the lung and spleen was contained largely within mast cells. The LPS-stimulated increase in HDC activity in a given organ was similar between +/+ and W/W(v) mice, and between IL-1alpha beta/TNFalphaKO BALB/c and control BALB/c mice, and led to increases in histamine. In W/Wv and +/+ mice, IL-1alpha also elevated HDC activity. These results suggest that (i) in liver, lung and spleen, either the major cells supplying histamine via HDC induction in response to LPS and IL-1 are not mast cells, or mast cells are not a prerequisite for the induction of HDC; (ii) the cells in which HDC is induced by LPS and IL-1 are similar or identical in a given organ; and (iii) neither IL-1 nor TNF-alpha is a prerequisite for the induction of HDC by LPS.     

Fritillaria cirrhosa, Anemarrhena asphodeloides, Lee-Mo-Tang and cyclosporine a inhibit ovalbumin-induced eosinophil accumulation and Th2-mediated bronchial hyperresponsiveness in a murine model of asthma

Asthma is a chronic inflammatory disorder of the airways characterized by excess production of Th2 cytokines and eosinophil accumulation in the lungs. Fritillaria cirrhosa, Anemarrhena asphodeloides and Lee-Mo-Tang are well-known herbs used in oriental medicine for the treatment of asthma and bronchial inflammation. To clarify the anti-asthmatic effects of Fritillaria cirrhosa bulbus, Anemarrhena rhizoma and Lee-Mo-Tang, we examined the development of pulmonary eosinophilic accumulation, control of Th2 cytokine, immunoglobulin E (IgE) and histamine productions in a murine model of asthma. Eosinophil cell proliferation was performed by [(3)H]thymidine uptake, eosinophilic accumulation. Cell counts in bronchoalveolar lavage fluid were investigated by means of fluorescence activated cell sorter analysis and control of Th2 cytokine, IgE and histamine productions were investigated by RT-PCR and ELISA. Moreover, lung tissue was histologically analysed. The suppressive effects of Fritillaria cirrhosa bulbus, Anemarrhena rhizoma and Lee-Mo-Tang on eosinophil recruitment and airway inflammation were demonstrated throughout the reduction of eosinophil numbers. This result correlated with a marked reduction IL-5, IL-13 and IL-4 levels in the bronchoalveolar lavage fluid. Ovalbumin-specific IgE levels were also decreased in serum. Fritillaria cirrhosa bulbus, Anemarrhena rhizoma and Lee-Mo-Tang have deep inhibitory effects on airway inflammation by suppression of Th2 cytokines (IL-4, IL-5 and IL-13), IgE, histamine production, reduction eosinophilic accumulation and increase of interferon-gamma production.     

Mouse strain differences in eosinophilic airway inflammation caused by intratracheal instillation of mite allergen and diesel exhaust particles

Response differences by different strains of mice towards house dust mites (Dermatophagoides farinae) or diesel exhaust particles (DEP) were investigated. Mouse strains BALB/c, ICR and C3H/He received 1 micro g of D. farinae or 1 microg of D. farinae + 50 microg of DEP intratracheally four times at 2-week intervals. Dermatophagoides farinae treatment caused the recruitment of eosinophils and lymphocytes. The order of magnitude of the eosinophilic airway inflammation was BALB/c < ICR < C3H/He mice. The protein levels of eotaxin and IL-5 in lung tissues correlated with the manifestations of eosinophilic airway inflammation by D. farinae administration. Diesel exhaust particles aggravated the manifestation of the eosinophilic inflammation through goblet cell proliferation in the airway and enhanced the local expression of eotaxin and IL-5 in all three strains of mice. The levels of eotaxin and IL-5 in lung tissues corresponded to the pathological changes caused by D. farinae + DEP. The increasing order of production levels of antigen-specific IgG1 by D. farinae or D. farinae + DEP was BALB/c < ICR < C3H/He mice. The significant adjuvant effect of DEP on IgG1 production was observed in the C3H/He mice (P < 0.05). These results suggest that the murine strain differences in the production of eosinophilic airway inflammation by D. farinae + DEP are related to differences in local expression of IL-5 and eotaxin. The enhancing effects of DEP may be mediated by a cytokine increase in the local expression. Antigen-specific IgG1 may be an important immunoglobulin in the pathogenesis of allergic asthma enhanced by DEP.     

Enhancement of antigen-induced eosinophilic inflammation in the airways of mast-cell deficient mice by diesel exhaust particles

The present study was conducted to clarify the involvement of mast cells in the exacerbating effect of diesel exhaust particles (DEP) toward allergic airway inflammation and airway hyperresponsiveness (AHR). Airway inflammation by the infiltration of cosinophils with goblet cell proliferation and AHR, as well as by the production of antigen-specific IgG1 and IgE, in plasma were examined using mast cell-deficient mice (W/W^v) and normal mice (W/W⁺). Both groups of mice received ovalbumin (OVA) or OVA+DEP intratracheally. The eosinophilic airway inflammation and goblet cell proliferation promoted by OVA were significantly greater in W/W⁺ than in W/W^v. A similar result was observed in AHR, but was not significant among both groups of mice. DEP enhanced OVA induced-allergic airway inflammation, goblet cell proliferation, and development of AHR in W/W^v, but not in W/W⁺. DEP decreased production of antigen-specific IgG1 and IgE in both groups of mice. Mast cells were observed in the submucosal layer of the main bronchus in W/W^v. The number of mast cells was significantly decreased by OVA treatment. The results indicate that mast cells are not necessary to enhance airway damage and development of AHR in W/W^v by DEP. However, mast cells may be required for the OVA-induced cosinophilic inflammation, airway damage with goblet cell proliferation, and AHR in W/W⁺.     

Effect of interleukin 3 on the differentiation and histamine content of cultured bone marrow mast cells.

Mouse bone marrow hematopoietic stem cells were isolated from mouse femur bone and cultured in RPMI 1640 supplemented medium with 20 units/ml of the purified T-cell lymphokine, interleukin 3 (IL-3), IL-3 was uniquely able to induce the proliferation and differentiation of mature mast cells in vitro. The sparse granulation of the bone marrow-derived mast cells (BMMC) can be seen by day 5, progressing to definable mast cells by day 7, the mast cells appear morphologically mature and comprise a 96% pure population after 14 days of the culture. The monocytes macrophages, eosinophils and neutrophils disappeared by day 9. After 4 weeks of tissue culture, mast cells are fully mature and completely granulated at 98% cell purity. The BMMC are mononuclear, oval or round in shape and appear smaller than rat peritoneal mast cells. BMMC are stable over 3-5 months in conditioned medium. The homogeneous mast cell population possesses membrane receptors and mediators, such as histamine in their metachromatic granules. The histamine content of BMMC in culture between 2 to 4 weeks rose from 1.43 to 1.82 pg/cell. Moreover, the percentage of histamine release caused by 0.1 microM and 1.0 microM ionophore A23187 was 15% and 35%, respectively. By contrast, the histamine releasing activity of 0.01% and 0.001% compound 48/80 were 12 +/- 2% and 59 +/- 7% respectively. The granular density, histamine content and histamine release activity of BMMC are different from that of peritoneal mast cells.     

Natural helper cells are associated with the exacerbated airway inflammation seen during RSV reinfection of neonatally primed mice

Infection with respiratory syncytial virus (RSV) in neonatal mice causes more aggressive airway disease when the mice are reinfected with the same virus as adults. However, the underlying mechanisms responsible for this phenomenon are not entirely defined. Natural helper (NH) cells are considered a key factor for virus-induced or exacerbated airway inflammation and airway hyper-responsiveness by producing type 2 cytokines. To confirm whether NH cells are involved in the aggravated lung pathology seen during reinfection, BALB/c mice were initially infected as neonates and reinfected in adulthood. We observed that neonatal RSV infection resulted in an enhanced infiltration of eosinophils and neutrophils in the lungs, in parallel with a significant increase in the levels of IL-5 and IL-13 in bronchoalveolar lavage fluids on day 2 after reinfection. It seems likely that pulmonary NH cells may play a role in the occurrence, since mice first infected at 1 wk of age developed an additional increase in the number of NH cells as well as IL-5- and IL-13-producing NH cells in the lungs than those first infected as young adults. In fact, an elevated expression of mRNAs for IL-5 and IL-13 in pulmonary NH cells was detected in mice first infected as neonates. Furthermore, adoptive transfer of NH cells into neonatal mice was able to boost eosinophilic infiltration as well as the production of type 2 cytokines in the lungs after reinfection at adulthood. In contrast, the expression of mRNA for the type 1 cytokine IFN-γ was down-regulated markedly by adoptive transfer of NH cells. Thus, these results suggest that Th2-type NH cells may play a role in the exacerbated airway inflammation seen during RSV reinfection of neonatally primed mice.     

An inhibitor peptide of toll-like receptor 2 shows therapeutic potential for allergic conjunctivitis

Allergic conjunctivitis (AC) is an inflammatory disease of the conjunctiva, which is characterized by antigen challenge and toll-like receptor 2 (TLR2) activation. Here, a designed small peptide ZY12 was found to contain therapeutic potential in staphylococcal enterotoxin B (SEB)-induced AC model. ZY12 treatment showed the remission of clinical signs, plasma total IgE levels, number of mast cells and the proportion of degranulated mast cell in AC mice. Levels of Th2 cytokines (IL-4, IL-5, IL-13) in the lymph nodes or spleen were significantly decreased while those of Th1 cytokine (IFN-γ) were increased in ZY12 treated group, suggesting a protective role of ZY12 in AC by mediating the balance of Th1/Th2 cytokines. Importantly, ZY12 significantly inhibited TLR2 expression in conjunctival tissue. Combined its therapeutic effects with TLR2 inhibitory function, ZY12 might be an ideal candidate for the development of new therapeutic agent for allergic disease.     

Modulatory effect of anisodamine on airway hyper-reactivity and eosinophilic inflammation in a murine model of allergic asthma

Anisodamine, a peripheral muscarinic receptor antagonist, is a naturally occurring atropine derivative that has been isolated, synthesized and characterized by scientists in China. In the present investigation, we evaluated the modulatory effects of anisodamine on airway hyper-reactivity and inflammation in a murine model of allergic asthma. Asthma model was induced successfully by ovalbumin. The activation of cells, airway eosinopilia, cytokine production, and airway function were examined. Our results collectively show that anisomanine could significantly suppress the accumulation of eosinophils into the airways and dramatically inhibited the histological changes in OVA-induced mice. Additionally, anisodamine could restore the Th1/Th2 balance in BALF by downregulating the level of Th2 cell-associated cytokine IL-4 (p<0.01) and upregulating the level of Th1 cell-associated cytokine IFN-γ (p<0.01). In addition, pretreatment with anisodamine also showed strong suppression of allergen-induced bronchial hyper-reactivity with maximum contraction decreasing from 0.45 ± 0.02 g to 0.28 ± 0.03 g (p<0.01). These results suggested the modulatory effects of anisodamine on Th1/Th2 balance by enhancing Th1-related and suppressing Th2-related parameters, as well as its potential application in airway hyper-reactivity and eosinophilic inflammation.