Sensitized Guinea Pigs Research Articles

The preventive effects of natural adjuvants, G2 and G2F on tracheal responsiveness and serum IL-4 and IFN-Γ (th1/th2 balance) in sensitized guinea pigs

OBJECTIVE:The effects of natural adjuvants on lung inflammation and tracheal responsiveness were examined in sensitized guinea pigs.METHODS:The responses of guinea pig tracheal chains and the serum levels of interleukin-4 and interferon-gamma were examined in control pigs and three other groups of guinea pigs: the sensitized group and two other sensitized groups treated with either adjuvant G2 or adjuvant G2F (n = 7 for each group). Sensitization of the animals was achieved by injection and inhalation of ovalbumin.RESULTS:The results showed that sensitized animals had increased tracheal responsiveness and increased serum levels of interleukin-4 and interferon-gamma compared to controls (p<0.05 to p<0.001). Treatments with either G2 or G2F prevented the increase in tracheal responsiveness and serum interleukin-4 (p<0.01 to p<0.001). However, the serum levels of interferon-gamma and the interleukin-4-to-interferon-gamma ratio was increased in the treated groups (p<0.001 for all cases).CONCLUSIONS:These results indicate important preventive effects of two natural adjuvants, particularly G2, on the changes in tracheal responsiveness, serum cytokines and the interleukin-4-to-interferon-gamma ratio (T helper 1/T helper 2 balance) in sensitized guinea pigs.

Identifying the allergenicity of maize pollen in Iran.

Maize is a member of the Poaceae family, capable of producing large amounts of pollen grains which may constitute important allergens in spring and summer. The aim of this study was to determine the protein content of maize pollen and its allergenicity in guinea pigs. The morphology of maize pollens was determined using light microscopy and scanning electron microscopy. The size of separated proteins was obtained by SDS-PAGE. A group of animals were immunized with maize pollen extract and the others were kept as control. After 40 days, the skin prick test was done in animals after blood sampling for counting the eosinophils. The allergenisity of proteins was identified by immunoblotting of transferred bonds using sera from sensitized guinea pigs. Pollen grains showed a spherical, monoporate structure with the scabrate exine surface. The SDS-PAGE indicated a major band of about 50 kD.We also showed increase in flare and wheal diameter following skin prick test in sensitized guinea pigs along with an elevated number of eosinophils. The presence of group 13 allergen (Zea m13) with molecular weight of ~ 50 kD was found in immunoblotting results. This study showed one protein in maize pollen extract that could be considered as an allergen belonging to group 13 of allergen categories. However, further investigations should be scheduled for precise analysis of the proteins. This allergen can be used for diagnostic or therapeutic purposes (vaccination approaches) in allergic asthma patients.

Androgens are bronchoactive drugs that act by relaxing airway smooth muscle and preventing bronchospasm.

Changes in the androgen levels in asthmatic men may be associated with the severity of asthma. Androgens induce a nongenomic relaxation in airway smooth muscle, but the underlying mechanisms remain unclear. The aim of this study was to investigate the potential bronchorelaxing action of testosterone (TES) and its metabolites (5α- and 5β-dihydrotestosterone (DHT). A preventive effect on ovalbumin (OVA)-induced bronchospasm was observed in sensitized guinea pigs for each androgen. Androgens were studied in response to bronchoconstrictors: carbachol (CCh) and KCl in isolated trachea rings with and without epithelium from non-sensitized and sensitized animals as well as on OVA-induced contraction. Androgens concentration-dependently abolished the contraction in response to CCh, KCl, and OVA. There were significant differences in the sensitivity to the relaxation induced by each androgen. 5β-DHT was more potent for relaxing KCl-induced contraction, while TES and 5α-DHT were more potent for CCh- and OVA-induced contraction. No differences were found in preparations with and without epithelium or in the presence of a nitric oxide (NO) synthase inhibitor or an inhibitor of K(+) channels. These data indicate the absence of involvement of the epithelium-, NO- and K(+) channels-dependent pathway in androgen-induced relaxation. However, in dissociated tracheal myocytes loaded with the calcium-binding fluorescent dye Fura -2, physiological concentrations of androgens decreased the KCl-induced [Ca(2+)]i increment. 5β-DHT was the most potent at decreasing KCl-induced [Ca(2+)]i increment and preventing bronchospasm. We suggest that androgen-induced brochorelaxation was mediated via decreased Ca(2+) influx through L-type Ca(2+)channels but additional Ca(2+) entry blockade may be involved. Molecular changes in androgen structure may determine its preferential site of action.

Unsweetened Natural Cocoa Has Anti-Asthmatic Potential

Unsweetened natural cocoa powder is enriched with nutraceutical abundance of anti-asthmatic compounds theobromine and theophylline. Cocoa powder, which is prepared after removal of the cocoa butter, contains about 1.9&#x0025; theobromine and 0.21&#x0025; caffeine. Anecdotal reports indicate that regular consumption of unsweetened natural cocoa powder (UNCP), a common practice in Ghana, West Africa, has the potential to reduce the tendency of asthmatic episodes. In the present paper we studied the effect of regular ingestion of aqueous extract of UNCP on hematological and histopathological changes that occur in ovalbumin (OVA)-sensitized guinea pigs. OVA-sensitized guinea pigs were challenged with aerosolized OVA 1 hour after ingestion of 300 mg/kg (low dose) or 600 mg/kg (high dose) of UNCP for 35 consecutive days. Histopathological and haematological changes in the OVA-sensitized guinea pigs were evaluated. Both negative and positive controls with distilled water and prednisolone, respectively, were used. OVA-sensitized guinea pigs demonstrated concentration-independent reduction in immune response to aerosolized OVA. There were no histo-architectural changes in the bronchiolar smooth muscles of the treated groups. Unsweetened natural cocoa powder has potential anti-asthmatic properties when administered orally at the doses tested.

Investigation into the mechanism of action of essential oil of Pistacia integerrima for its antiasthmatic activity

Ethnopharmacological relevancePistacia integerrima J.L. Stewart ex Brandis locally known as Karkatashringi is an important medicinal plant whose galls are valued in traditional medicine used in India for the treatment of asthma, chronic bronchitis, phthisis, diarrhea, fever, other ailments for the respiratory tract, and as antispasmodic, carminative, antiamoebic and anthelmintic. However, in vitro and in vivo investigations providing new insights into its pharmacological properties have not been thoroughly investigated yet. The present investigation aimed to elucidate the probable mechanism of antiasthmatic action of essential oil of Pistacia integerrima J.L. Stewart ex Brandis galls (EOPI). MethodsEOPI was tested using in vitro studies such as antioxidant activity, mast cell degranulation, angiogenesis, isolated guinea pig ileum preparation and soyabean lipoxidase enzyme activity. In vivo studies included lipopolysaccharide-induced bronchial inflammation in rats and airway hyperresponsiveness in ovalbumin in sensitized guinea pigs using spirometry. ResultsEOPI (5–30µg/ml) inhibits 5-lipoxidase enzyme activity with IC50 of 19.71µg/ml and DPPH scavenging activity up to 100µg/ml with maximum inhibition of 44.93±2.53% at 100µg/ml. Pre-treatment with EOPI inhibited erythropoietin-induced angiogenesis. It showed dose dependent (10, 30 and 100µg/ml) anti-allergic activity by inhibiting compound 48/80 induced mast cell degranulation to an extent 19.08±0.47%. The finding that essential oil induced inhibition of transient contraction of acetylcholine in calcium free medium, and relaxation of S-(-)-Bay 8644-precontracted isolated guinea pig ileum jointly suggests that the L-subtype Cav channel is involved in spasmolytic action of EOPI.Treatment with EOPI dose dependently (7.5, 15 and 30mg/kg i.p.) inhibited lipopolysaccharide-induced increase in total cell count, neutrophil count, nitrate–nitrite, total protein, albumin levels in bronchoalveolar fluid and myeloperoxidase levels in lung homogenates. Roflumilast was used as a standard. EOPI reduced the respiratory flow due to gasping in ovalbumin sensitized guinea pigs. ConclusionThe study demonstrates the effectiveness of essential oil of Pistacia integerrima J.L. Stewart ex Brandis galls in bronchial asthma possibly related to its ability to inhibit L-subtype Cav channel, mast cell stabilization, antioxidant, angiostatic and through inhibition of 5-lipoxygenase enzyme.

Phytochemical profiles and biological activity evaluation of Zanthoxylum bungeanum Maxim seed against asthma in murine models

Ethnopharmacological relevanceZanthoxylum bungeanum Maxim seed (ZBMS) has been used in Traditional Chinese Medicine (TCM) as an ingredient of polyherbal formulations for the treatment of inflammation and asthma. The aim of this study was to analyze the major composition and to evaluate the anti-asthma activity of ZBMS. Materials and methodsSome murine models including acetylcholine/histamine-induced asthma, ovalbumin-induced airway inflammation, ear edema and toe swelling measurement, citric acid-induced cough, and anti-stress abilities were investigated to fully study the anti-asthma activity of ZBMS.GC chromatography was also performed to analyze the major fatty acid composition of ZBMS. ResultsThe results demonstrated that the major fatty acid composition of ZBMS includes oleic acid (20.15%), linoleic acid (26.54%), and α-linolenic acid (30.57%), which was the leading component of ZBMS, and that the total fatty acid content of ZBMS was 77.27%. The murine models demonstrated that ZBMS displays a protective effect on guinea pig sensitization, a dose-dependent inhibition of the increases in RL and decreases in Cdyn, which resulted in the relief of auricle edema and toe swelling in mice and anti-stress activity. ConclusionOur results validate the traditional use of ZBMS for the treatment of asthma and other inflammatory joint disorders, and suggest that ZBMS has potential as a new therapeutic agent for asthma management.

The effect of safranal, a constituent of Crocus sativus (saffron), on tracheal responsiveness, serum levels of cytokines, total NO and nitrite in sensitized guinea pigs

BackgroundThe effect of safranal (one of the constituents of Crocus sativus) on ovalbumin (OVA) sensitized guinea pigs was examined. MethodsOne group of sensitized guinea pigs were given drinking water alone (group S), three groups drinking water containing three concentrations of safranal and one group contain dexamethasone (S+D). Tracheal responses (TR) of the animals to methacholine as effective concentration causing 50% of maximum response (EC50 M), TR to 0.1% OVA, relative to contraction induced by 100μM methacholine, IL-4, IFN-γ, total NO and nitrite levels in serum were measured. ResultsThe TR to both methacholine and OVA, the level of total NO, nitrite and IL-4 significantly increased but IFN-γ and IFN-γ/IL-4 ratio was decreased in group S compared controls (p<0.05 to p<0.001). The TR to both methacholine and OVA in treated animals with dexamethasone and all concentrations of safranal were significantly decreased compared to S group (p<0.01 to p<0.001). The level of serum IL-4 in treated guinea pigs was significantly decreased but IFN-γ and IFN-γ/IL-4 ratio was increased compared to S group (p<0.01 to p<0.001). The levels of total NO and nitrite were significantly decreased in treated groups compared to sensitized group (p<0.05 to p<0.001). ConclusionThese results showed a preventive effect for safranal on tracheal responses and serum cytokine, total NO and nitrite levels as well as increased Th1/Th2 balance in sensitized guinea pigs.

The Effect of Natural Adjuvants on Pathological Changes in Sensitized Guinea Pig Lungs

Background:Anti-inflammatory effect of natural adjuvants has been reported. Lung inflammation is the most characterized pathological feature in asthma.Objectives:The effects of three natural adjuvants (PC, G2, and G2F registered as a patent in the Iranian Patent Office) on sensitized guinea pigs lungs were examined in the present study.Materials and Methods:Lung pathological changes were examined in control and five groups of randomly divided guinea pigs including: sensitized animals (S, receiving normal saline, 0.5 ml i.p.); sensitized animals treated with adjuvant PC; G2F (0.1 ml i.p. for both cases); G2 (0.4 ml i.p.); and PC + G2 (receiving both PC and G) adjuvants (twice a week for 4 weeks for all groups). Sensitization of animals was done by injection and inhalation of ovalbumin (OA).Results:All pathological changes in S group including the eosinophil infiltration (scoring 3.28 ± 0.28), lymphocyte infiltration (2.82 ± 0.26), local epithelial necrosis (2.71 ± 0.47) and mucosal plug (2.75 ± 0.37) were significantly higher than control group (0.64 ± 0.18, 1.36 ± 0.24, 0.36 ± 0.18 and 0.28 ± 0.18 for eosinophil infiltration, lymphocyte infiltration, epithelial necrosis and mucosal plug respectively, P < 0.001 for all cases). Treatment with all adjuvants improved all pathological changes significantly (P < 0.05 to P < 0.001).Conclusions:These results indicate preventive effects of all natural adjuvants (especially G2) on pathological changes of the lung in sensitized guinea pigs.

Effect of Fluticasone and Salmeterol on Tracheal Responsiveness to Ovalbumin and Lung Inflammation, Administrated during and after Sensitization

The effect of duration of administration of fluticasone propionate and salmeterol on tracheal responsiveness to ovalbumin and total and differential white blood cell in sensitized guinea pig was examined. Six groups of guinea pigs (n = 7) were sensitized to ovalbumin. Three groups of them were subjected to inhaled fluticasone propionate and salmeterol, one group during sensitization (A), one group after that (for 18 days, B), and the other one during sensitization but with 18 days delay before measurements (C). Three other groups were treated with placebo in the same manner. The tracheal responsiveness to ovalbumin and total and differential white blood cells of three placebo groups were significantly higher than those of control group (P < 0.001 for all cases). Tracheal responsiveness to ovalbumin and total and differential white blood cell in treated groups with fluticasone propionate and salmeterol were significantly decreased compared to those of placebo groups (nonsignificant to P < 0.001). The improvement in all variables in treatment groups A and C were more pronounced than group B. The results showed that fluticasone propionate and salmeterol had a prevention effect on tracheal hyperresponsiveness to ovalbumin and lung inflammation which was more pronounced when administered during than after sensitization.

Nutraceutical with Anti-Inflammatory Activity for the Management of Airway Remodeling in Bronchial Asthma: &amp;lt;i&amp;gt;Kalanchoe integra&amp;lt;/i&amp;gt; Var. Crenata (Andr.) Cuf Leaf Extract

Background: Kalanchoe integra is widely used in folklore medicine as an antiasthmaticagent. Previous studies have shown the ameliorating effect of Kalanchoe integra leaf extract [KILE] on bronchial hyperesponsiveness and inflammation.Further, the stabilizing effect of Kalanchoe sp on mast cell degranulation,suggests that Kalanchoe species are suitable candidates for allergicasthma therapy. This study is designed to investigate the anti-asthmaticpotential of KILE and monitor the accompanying histopathological andimmunobiochemical changes that occur in an animal model of bronchial asthmausing ovalbumin sensitized guinea pigs. Method: Thirty male guinea pigs weredivided into five groups of six animals each. Bronchial asthma was simulated inguinea pigs using ovalbumin. Both low dose (300 mg/kg) and high dose extract(900 mg/kg) were administered daily for 42 days. Prednisolone (2.5 mg/kg) wasthe standard drug used. Results: Guinea pigs in all KILE treated groups maintained the integrity of their airwaystructures: bronchial folds and walls, alveoli, alveolar ductsand sacs. KILE and prednisolone caused a reduction in immune parameters (p 0.001), extent of bronchoconstriction, bronchialwall thickness and goblet cell accumulation in the sensitized guineapigs. Conclusion: This study demonstrates the anti-asthmatic potential of KILE duringprolonged administration by the oral route.

Critical Role of Protein Kinase C (PKC) in the Onset of Airway Hypersensitivity in Ova-Sensitized Guinea Pig Model of Asthma

Background and Objectives: Protein kinase C (PKC) activation plays an important role in activation of T-lymphocytes in asthma. Airway hypersensitivity is one of the main characteristic features of asthma, the mechanism of onset of which is not clearly understood. Therefore, the objective was to elucidate the role of PKC in etiopathogenesis of airway hypersensitivity in asthma. Methods: Male guinea pigs (n = 30) were sensitized with ovalbumin and day of initial allergen-specific immune response determined by intradermal test, airway hypersensitivity, BALF cytology and lung histopathology. Total PKC activity, PKC isoenzymes and phosphoinositides were assessed in airway smooth muscles (ASM) and peripheral blood lymphocytes. Results: Intradermal test revealed that day 9 was the earliest time of allergen-specific response and onset of airway hypersensitivity to ovalbumin. It was associated with significant increase in total and differential (lymphocytes and eosinophils) BALF counts and grade I peribronchiolar chronic lymphocytic inflammation in lung. On day 14, grade II infiltration of lymphocytes and eosinophils with onset ofstructural remodelingofproximal and distal airways was seen. Total PKC activity, expression of PKCα, PKCe and phosphoinositides increased significantly in ASM and lymphocytes on day 9 and were maximum on day 14. There was no change in PKC-τ expression. Conclusions: Activation of PKC, particularly PKCα and PKCe, mediated signal transduction pathway plays a critical role in lymphocyte infiltration and onset of airway hypersensitivity, airway remodeling and asthma pathophysiology. The present study is the first one on the mechanism of the etiopathogenesis of the disease, which shows a direct evidence of the role of PKC mediated pathway in the initiation and onset of airway hypersensitivity in ovalbumin sensitized guinea pig model.

Lead exposure affects inflammatory mediators, total and differential white blood cells in sensitized guinea pigs during and after sensitization

Background: Serum inflammatory mediators and white blood cells (WBC) counts in the blood of sensitized and lead exposed guinea pigs were evaluated. Methods: Guinea pigs were randomly allocated into control (C), sensitized (S) and sensitized groups exposed to three lead concentrations (0.1, 0.2 and 0.4 M) during (DS) and after sensitization (PS), (n = 6 for each group). Animals were sensitized by intra-peritoneal injection and aerosol inhalation of ovalbumin (OA). Serum total protein, PLA2, IgE, histamine, total and differential WBC counts of blood were evaluated. Results: Serum PLA2, total protein, IgE, histamine, total and differential WBC counts in all three sensitized groups were significantly increased, but the percentage of lymphocyte was decreased compared to control group (p < 0.05 to p < 0.001). Serum total protein and total WBC number in all animals exposed to lead as well as the eosinophil and histamine in animals exposed to highest lead concentration and IgE in sensitized groups exposed to lead after sensitization were significantly higher, but the lymphocyte in animals exposed to two higher lead concentrations was lower than non exposed sensitized group (p < 0.05 to p < 0.001). The changes in all parameters in lead exposed animals after sensitization were higher than those during sensitization which was statistically significant for total WBC count in the animals exposed to low lead concentration (p < 0.05). Conclusion: Inhaled lead can increase serum total protein, PLA2, IgE and histamine levels, total and most differential WBC counts in sensitized animal which was more pronounced in animals exposed to lead after compared to those during sensitization.

Development of Automated Conjunctival Hyperemia Analysis Software

Conjunctival hyperemia is observed in a variety of ocular inflammatory conditions. The evaluation of hyperemia is indispensable for the treatment of patients with ocular inflammation. However, the major methods currently available for evaluation are based on nonquantitative and subjective methods. Therefore, we developed novel software to evaluate bulbar hyperemia quantitatively and objectively. First, we investigated whether the histamine-induced hyperemia of guinea pigs could be quantified by image analysis. Bulbar conjunctival images were taken by means of a digital camera, followed by the binarization of the images and the selection of regions of interest (ROIs) for evaluation. The ROIs were evaluated by counting the number of absolute pixel values. Pixel values peaked significantly 1 minute after histamine challenge was performed and were still increased after 5 minutes. Second, we applied the same method to antigen (ovalbumin)-induced hyperemia of sensitized guinea pigs, acquiring similar results except for the substantial upregulation in the first 5 minutes after challenge. Finally, we analyzed human bulbar hyperemia using the new software we developed especially for human usage. The new software allows the automatic calculation of pixel values once the ROIs have been selected. In our clinical trials, the percentage of blood vessel coverage of ROIs was significantly higher in the images of hyperemia caused by allergic conjunctival diseases and hyperemia induced by Bimatoprost, compared with those of healthy volunteers. We propose that this newly developed automated hyperemia analysis software will be an objective clinical tool for the evaluation of ocular hyperemia.

Prostaglandin D2 receptord-type prostanoid receptor 2 mediates eosinophil trafficking into the esophagus

Eosinophilic esophagitis is characterized by eosinophil-predominant inflammation in the esophagus. How eosinophils migrate and infiltrate into the esophagus, however, is less clear. Our previous study demonstrated that mast cell activation led to eosinophil infiltration in the esophagus. Prostaglandin D2 (PGD2) is an important mediator released from activated mast cells. The present study aims to determine whether PGD2 induces eosinophil infiltration into the esophagus via a d-type prostanoid receptor 2 (DP2) receptor-dependent mechanism. Using an in vivo guinea pig model, PGD2, d-type prostanoid receptor 1 (DP1) agonist, or DP2 agonist were injected into the esophagus. Esophageal tissues were removed 2 hours after injections and proceeded to either hematoxylin-eosin (HE) staining or immunofluorescent staining of eosinophil major basic protein (MBP) to compare each treatment-induced eosinophil infiltration in the esophagus. In a separate study, ovalbumin (OVA)-sensitized guinea pigs were pretreated with either DP2 or DP1 antagonists, followed by inhalation of OVA to induce mast cell activation. Esophageal tissues were then processed for immunofluorescent staining of MBP. PGD2 injection in the esophagus led to an increase of eosinophil infiltration in esophageal epithelium at the injection site as revealed by HE staining. Increased infiltration of eosinophils was further confirmed by the increased presence of MBP-labeled immunopositive (MBP-LI) cells in esophageal epithelium. Injection with DP2 agonist 15(R)-PGD2, but not DP1 agonist BW 245C, mimicked the PGD2-induced response. In OVA-sensitized animals, antigen inhalation increased MBP-LI cells in esophageal epithelium. Pretreatment with DP2 antagonist BAY-u3405, but not DP1 antagonist BW 868C, inhibited the antigen inhalation-induced increase of MBP-LI cells in esophageal epithelium. These data support the hypothesis that PGD2 induces eosinophil trafficking into the esophageal epithelium via a DP2-mediated pathway, suggesting a role of DP2 antagonist in the prevention of eosinophilic esophagitis.

Antagonist profile of ibodutant at the tachykinin NK2 receptor in guinea pig isolated bronchi

In this study we have characterized the pharmacological profile of the non-peptide tachykinin NK2 receptor antagonist ibodutant (MEN15596) in guinea pig isolated main bronchi contractility. The antagonist potency of ibodutant was evaluated using the selective NK2 receptor agonist [βAla8]NKA(4-10)-mediated contractions of guinea pig isolated main bronchi. In this assay ibodutant (30, 100 and 300nM) induced a concentration-dependent rightward shift of the [βAla8]NKA(4-10) concentration-response curves without affecting the maximal contractile effect. The analysis of the results yielded a Schild-plot linear regression with a slope not different from unity (0.95, 95% c.l. 0.65–1.25), thus, indicating a surmountable behavior. The calculated apparent antagonist potency as pKB value was 8.31±0.05. Ibodutant (0.3–100nM) produced a concentration-dependent inhibition of the nonadrenergic–noncholinergic (NANC) contractile response induced by electrical field stimulation (EFS) of intrinsic airway nerves in guinea pig isolated main bronchi. At the highest concentration tested (100nM) ibodutant almost abolished the EFS-induced bronchoconstriction (95±4% inhibition), the calculated IC50 value was 2.98nM (95% c.l. 1.73–5.16nM). In bronchi from ovalbumin (OVA) sensitized guinea pigs ibodutant (100nM) did not affect the maximal contractile response to OVA, but completely prevented the slowing in the fading of the motor response induced by phosphoramidon pretreatment linked to the endogenous neurokinin A release. Altogether, the present study demonstrates that ibodutant is a potent NK2 receptor antagonist in guinea pig airways.

Selenium-containing lycium barbarum polysaccharides antioxidant research

Anti-inflammatory, anti oxidant and effect of Crocus sativus (C. sativus) on Th1/Th2 balance were described previously.The preventive effects of the extract of Crocus sativus on tracheal responsiveness and plasma levels of IL-4, IFN-γ, total NO and nitrite were examined on sensitized guinea pigs.Five groups of sensitized guinea pigs to ovalbumin (OVA), were given drinking water containing three concentrations of the extract of Crocus sativus, dexamethasone (S+D) or alone (group S). Tracheal responses (TR) of control animals (group C) and sensitized guinea pigs (n=6, for each group) to methacholine, OVA and the levels of IL-4, IFN-γ, total NO and nitrite in serum were examined.The TR to both methacholine and OVA, the levels of serum IL-4, total NO and nitrite in S guinea pigs were significantly increased but that of IFN-γ and IFN-γ/IL-4 ratio (Th1/Th2 balance) were decreased compared to the controls (p<0.05 to p<0.001). In the treated animals with dexamethasone and all concentrations of the extract, TR to both methacholine and OVA, IL-4, total NO and nitrite were significantly decreased but IFN-γ and IFN-γ/IL-4 ratio increased compared to S group (p<0.05 to p<0.001). The effects of the highest concentration of the extract was greater than those of other concentrations and the effect of dexamethasone (p<0.05 to p<0.01).These results not only showed a preventive effect of C. sativus extract on tracheal responses and serum levels of inflammatory mediators in sensitized guinea pigs but also showed increased Th1/Th2 balance.

The impact of Zataria multiflora Boiss extract on in vitro and in vivo Th1/Th2 cytokine (IFN-γ/IL4) balance

Ethnopharmacological relevanceAnti-inflammatory, anti-oxidant and effect of Zataria multiflora on Th1/Th2 balance were previously described. Different therapeutic effects of this plant have been described in Iranian traditional medicine. To evaluate the immune modulatory effects of Zataria multiflora on Th1/Th2 balance, which may be implicated in inflammatory disorders, in vitro and in vivo studies were carried out. Materials and methodsThe effects of three concentrations of the extract, dexamethasone, and saline on interleukin 4 (IL-4) and interferon γ (IFN-γ) gene expression were evaluated in phytohemagglutinin (PHA) stimulated and non-stimulated human peripheral blood mononuclear cells (hPBMCs). RNA was extracted from the hPBMCs to make cDNA for real time PCR relative quantification. Furthermore, the effect of the extract on serum level of IL-4 and IFN-γ was assessed in ovalbumin (OA) sensitized guinea pigs (n=6 for each group). ResultsDexamethasone showed significant inhibitory effect on both IFN-γ and IL-4 gene expression and serum level of the cytokines and significantly enhanced IFN-γ/IL-4 ratio (p<0.05–p<0.001). The extract inhibited IL-4 and enhance IFN-γ gene expression and IFN-γ/IL-4 ratio too (p<0.05–p<0.001). In sensitized animals also serum level of IL-4 were significantly decreased after treatment with both dexamethasone and extract, but serum level of IFN-γ and IFN-γ/IL-4 ratio were significantly increased due to extract treatment (p<0.01 for medium and p<0.001 for high concentration). ConclusionsThese results indicated consistent in vitro and in vivo data for selective immune modulatory effect of the extract of Zataria multiflora which increased IFN-γ, decreased IL-4, and enhanced the ratio of IFN-γ to IL-4 (Th1/Th2 balance). Therefore, the extract of Zataria multiflora may have therapeutic value in inflammatory responses such as allergy, autoimmunity and infectious diseases associated with Th1/Th2 imbalance.

The Extract of Crocus sativus and Its Constituent Safranal, Affect Serum Levels of Endothelin and Total Protein in Sensitized Guinea Pigs.

The effect of the extract of Crocus sativus and its constituent, safranal on inflammatory markers in sensitized guinea pigs was examined. Ovalbumin (OA) sensitized guinea pigs were given drinking water alone (group S), or drinking water containing three concentrations of safranal, three concentrations of extract and one concentration of dexamethasone, (n=6, for all groups) and serum levels of endotheline-1 (ET-1) and total protein (TP) were assessed. Serum levels of ET-1 and TP in group S were significantly higher than control group (P<0.01 for ET-1 and P<0.001 TP). Treatment of S animals with dexamethasone, most concentrations of the extract and safranal significantly reduced serum levels of ET-1 and TP compared to group S (P <0.01 to P <0.001). The effects of one concentration of the extract and safranal were significantly higher than dexamethasone (P <0.05 to P <0.01). A preventive effect of the extract of C. sativus and its constituent safranal on serum inflammatory markers in sensitized guinea pigs was shown.

Evaluation of anti-allergic and anti-anaphylactic activity of ethanolic extract of Zizyphus jujuba fruits in rodents

The present study reports the anti-allergic activity of ethanolic extract of Zizyphus jujuba Mill., Rhamnaceae, and its possible mode of action. The effect of extract of Z. jujuba at different doses (250, 500 and 1000mg/kg, orally) was simulated on studied animal models of asthma and allergy: a) milk induced eosinophilia and leukocytosis; b) compound 48/80 induced mast cell degranulation; and, c) active and passive cutaneous anaphylaxis. In addition, extract of Z. jujuba's effect on sensitized guinea pig ileum (ex vivo) and tracheal chain preparations (invitro) were investigated. Treatment with extract of Z. jujuba at all doses significantly: prevented the milk-induced eosinophilia and compound 48/80 induced degranulation of mesenteric mast cells; decreased passive cutaneous and active anaphylactic reactions. In addition, extract of Z. jujuba inhibited acetylcholine as well as histamine induced tracheal chain contraction, and also antigen induced contraction of sensitized guinea pig ileum (Shultz- Dale inhibition test). Furthermore, it exhibited also free radicals scavenging activity (in vitro). The observed anti-allergic and anti-anaphylactic activity of extract of Z. jujuba may be largely through the stabilization of mast cells by the membrane presence of phytoconstituents (steroidal saponins and flavonoids).

A selective antagonist of histamine H4 receptors prevents antigen‐induced airway inflammation and bronchoconstriction in guinea pigs: involvement of lipocortin‐1

Among the pathogenic mechanisms of asthma, a role for oxidative/nitrosative stress has been well documented. Recent evidence suggests that histamine H₄ receptors play a modulatory role in allergic inflammation. Here we report the effects of compound JNJ 7777120 (JNJ), a selective H4 receptor antagonist, on antigen-induced airway inflammation, paying special attention to its effects on lipocortin-1 (LC-1/annexin-A1), a 37 kDA anti-inflammatory protein that plays a key role in the production of inflammatory mediators. Ovalbumin (OA)-sensitized guinea pigs placed in a respiratory chamber were challenged with antigen. JNJ (5, 7.5 and 10 mg.kg⁻¹) was given i.p. for 4 days before antigen challenge. Respiratory parameters were recorded. Bronchoalveolar lavage (BAL) fluid was collected and lung specimens taken for further analyses 1 h after antigen challenge. In BAL fluid, levels of LC-1, PGD2 , LTB4 and TNF-α were measured. In lung tissue samples, myeloperoxidase, caspase-3 and Mn-superoxide dismutase activities and 8-hydroxy-2-deoxyguanosine levels were measured. OA challenge decreased LC-1 levels in BAL fluid, induced cough, dyspnoea and bronchoconstriction and increased PGD2 , LTB4 and TNF-α levels in lung tissue. Treatment with JNJ dose-dependently increased levels of LC-1, reduced respiratory abnormalities and lowered levels of PGD2 , LTB4 and TNF-α in BAL fluid. Antigen-induced asthma-like reactions in guinea pigs decreased levels of LC-1 and increased TNF-α and eicosanoid production. JNJ pretreatment reduced allergic asthmatic responses and airway inflammation, an effect associated with LC-1 up-regulation.