Insamhodo-tang, a traditional Korean medicine, regulates mast cell-mediated allergic inflammation in vivo and in vitro

Aim of the study

Insamhodo-tang (IHT) has traditionally been used in Korea to treat a variety of diseases, including chronic cough, tuberculosis, and chronic bronchitis. However, the anti-allergic and anti-inflammatory effects of IHT and its molecular mechanisms have yet to be clearly elucidated. In this study, we attempted to evaluate the effects of IHT on mast cell-mediated allergy inflammation in vitro and in vivo.

Materials and methods

We investigated to ascertain the pharmacological effects of IHT on both compound 48/80-induced and 2,4-dinitrofluorobenzene (DNFB)-induced allergic reactions under in vivo conditions. Additionally, to find a possible explanation for the anti-inflammatory mechanisms of IHT, we evaluated the regulatory effects of IHT on the level of inflammatory mediators in phorbol 12-myristate 13-acetate plus calcium ionophore A23187 (PMACI)-stimulated human mast cells (HMC-1).

Results

The finding of this study demonstrated that IHT reduced compound 48/80-induced systemic anaphylactic shock, DNFB-induced dermatitis, and ear swelling responses in mice. Additionally, IHT inhibited the production of interleukin (IL)-6, IL-8, and TNF-α, as well as the activation of nuclear factor-κB and caspase-1 in PMACI-stimulated HMC-1.

Conclusion

Collectively, the findings of this study provide us with a novel insight into the pharmacological actions of IHT as a potential molecule for use in the treatment of allergic inflammation diseases.     

Inula japonica extract inhibits mast cell-mediated allergic reaction and mast cell activation

Ethnopharmacological relevance

The flowers of Inula japonica (Inulae Flos) have long been used in traditional medicine for the treatment of bronchitis, digestive disorders, and inflammation. However, the mechanisms underlying its anti-inflammatory effects remain yet to be elucidated. The objectives of this study were 1) to assess the anti-allergic activity of the ethanol extract of flowers of Inula japonica extract (IFE) in vivo, 2) to investigate the mechanism of its action on mast cells in vitro, and 3) to identify its major phytochemical compositions.

Materials and methods

The anti-allergic activity of IFE was evaluated using mouse bone marrow-derived mast cells (BMMCs) in vitro and a passive cutaneous anaphylaxis (PCA) animal model in vivo. The effects of IFE on mast cell activation were evaluated in terms of degranulation, eicosanoid generation, Ca²⁺ influx, and immunoblotting of various signaling molecules.

Results

IFE inhibited degranulation and the generation of eicosanoids (PGD₂ and LTC₄) in stem cell factor (SCF)-stimulated BMMCs. Biochemical analysis of the SCF-mediated signaling pathways demonstrated that IFE inhibited the activation of multiple downstream signaling processes including mobilization of intracellular Ca²⁺ and phosphorylation of the mitogen-activated protein kinases (MAPKs), PLCγ1, and cPLA₂ pathways. When administered orally, IFE attenuated the mast cell-mediated PCA reaction in IgE-sensitized mice. Its major phytochemical composition included three sesquiterpenes, 1-O-acetylbritannilactone, britanin and tomentosin.

Conclusions

This study suggests that IFE modulates eicosanoids generation and degranulation through the suppression of SCF-mediated signaling pathways that would be beneficial for the prevention of allergic inflammatory diseases. Anti-allergic activity of IFE may be in part attributed particularly to the presence of britanin and tomentosin as major components evidenced by a HPLC analysis.     

Effect of Sophora flavescens Aiton extract on degranulation of mast cells and contact dermatitis induced by dinitrofluorobenzene in mice

Ethnopharmacological relevance

The dried root of Sophora flavescens Aiton (Sophorae radix, SR) has long been used in traditional medicine for the treatment of fever and swelling in eastern countries.

Materials and methods

The present study investigated the anti-allergic and anti-inflammatory effects of SR using 1-fluoro-2,4-dinitrofluorobenzene (DNFB)-induced contact dermatitis mouse model and in vitro using RBL-2H3 cells.

Results

In mice, the topical application of 10 mg/mL of SR effectively inhibited enlargement of ear thickness and weight induced by repeated painting with DNFB. Topical application of SR also inhibited hyperplasia, edema, spongiosis and infiltration of mononuclear cells in ear tissue. In addition, production levels of interferon-gamma and tumor necrosis factor-alpha were decreased by SR in vivo. Finally, the release of histamine and β-hexosaminidase, and migration were inhibited by treatment with SR.

Conclusions

These data indicate the potential of SR in treating patients with allergic skin diseases and also suggest that related mechanisms are involved in anti-inflammatory action on the Th 1 skewing reaction and inhibition against recruitment and degranulation of mast cells.     

Anti-allergic activity of German chamomile (Matricaria recutita L.) in mast cell mediated allergy model

Ethnopharmacological relevance

Chamomile is most popular used medicinal plant and extensively consumed as a tea or tisanes. Traditionally this plant was used for treatment of many ailments such as allergy disorders and inflammatory mediated diseases.

Aim of the study

We investigated the effects of anti-allergic activity of Matricaria recutita L. on mast cell mediated allergic models.

Materials and methods

The protective effect of methanol extract of Matricaria recutita against compound 48/80 induced anaphylaxis and pruritis models for acute phase of hypersensitivity reactions were carried out. The late phase hypersensitivity reactions by compound 48/80 induced mast cell degranulation and histamine release from blood along with serum nitric oxide (NO), rat peritoneal fluid nitric oxide (NO) and bronchoalveolar fluid nitric oxide (NO) levels were measured.

Results

The methanol extract of Matricaria recutita L. showed inhibitory effects on anaphylaxis induced by compound 48/80 and significant dose dependent anti-pruritis property was observed by inhibiting the mast cell degranulation. Mast cell membrane stabilization activity was also observed in compound 48/80 induced mast cell activation. Dose dependent reduction in the histamine release, along with decreased release of serum, rat peritoneal and BAL fluid nitric oxide (NO) levels were observed.

Conclusion

These results suggest that the methanol extract of Matricaria recutita showed potent anti-allergic activity by inhibition of histamine release from mast cells.     

Topical application of an ethanol extract prepared from Illicium verum suppresses atopic dermatitis in NC/Nga mice

Ethnopharmacological relevance

Illicium verum is a traditional herbal medicine with anti-inflammatory properties used in Asia. However, its usefulness in the treatment of allergic diseases remains unclear. This study evaluated the anti-inflammatory and antiallergic effects of I. verum extract (IVE) in a mouse model of atopic dermatitis.

Materials and methods

We investigated the effects of IVE on compound 48/80-induced histamine release, and phorbol 12-myristate13-acetate and calcium ionophore A23187-stimulated cytokines secretion in MC/9 mast cells. Atopic dermatitis was induced in NC/Nga mice by exposure to extract of house dust mite (Dermatophagoides farinae). After a topical application of IVE on ear and skin lesions, we evaluated the severity of skin symptoms, ear thickness, inflammatory cell infiltration, and serum levels of immunoglobulin E (IgE), histamine, interleukin (IL)-6, and intercellular adhesion molecule (ICAM)-1. In addition, we determined the expression of IL-4, IL-6, tumor necrosis factor (TNF)-α, interferon (IFN)-γ thymus- and activation-regulated chemokine (TARC), regulated on activation, normal T cell expressed and secreted (RANTES), ICAM-1, and vascular cell adhesion molecule (VCAM)-1 in ear tissues.

Results

IVE inhibited secretion of histamine, IL-4, IL-6, and TNF-α from mast cells in a dose-dependent manner. Topical application of IVE significantly reduced dermatitis scores, ear thickness, and serum levels of IgE, histamine, IL-6, and ICAM-1. Histopathological analysis demonstrated decreased epidermal thickening and dermal infiltration by inflammatory cells. In the ear lesions, IVE treatment reduced expression of IL-4, IL-6, TNF-α, TARC, RANTES, ICAM-1, and VCAM-1, but not IFN-γ.

Conclusions

These results indicate that IVE inhibits atopic dermatitis-like skin lesions by suppressing the expression of cytokines, chemokines, and adhesion molecules. These results suggest that IVE may be a potential therapeutic candidate for atopic dermatitis.     

Curine inhibits mast cell-dependent responses in mice

Ethnopharmacological relevance

Curine is a bisbenzylisoquinoline alkaloid and the major constituent isolated from Chondrodendron platyphyllum, a plant that is used to treat inflammatory diseases in Brazilian folk medicine. This study investigates the effectiveness of curine on mast cell-dependent responses in mice.

Materials and methods

To induce mast cell-dependent responses, Swiss mice were subcutaneously sensitized with ovalbumin (OVA—12 μg/mouse) and Al(OH)₃ in a 0.9% NaCl solution. Fifteen days later, the animals were challenged with OVA through different pathways. Alternatively, the animals were injected with compound 48/80 or histamine, and several parameters, including anaphylaxis, itching, edema and inflammatory mediator production, were analyzed. Promethazine, cromoglycate, and verapamil were used as control drugs, and all of the treatments were performed 1 h before the challenges.

Results

Curine pre-treatment significantly inhibited the scratching behavior and the paw edema induced by either compound 48/80 or OVA, and this protective effect was comparable in magnitude with those associated with treatment with either cromoglycate or verapamil. In contrast, curine was a weak inhibitor of histamine-induced paw edema, which was completely inhibited by promethazine. Curine and verapamil significantly inhibited pleural protein extravasations and prostaglandin D₂ (PGD₂) and cysteinyl leukotrienes (CysLTs) production following allergen-induced pleurisy. Furthermore, like verapamil, curine inhibited the anaphylactic shock caused by either compound 48/80 or an allergen. In in vitro settings, these treatments also inhibited degranulation as well as PGD₂ and CysLT production through IgE-dependent activation of the mast cell lineage RBL-2H3.

Conclusion

Curine significantly inhibited immediate allergic reactions through mechanisms more related to mast cell stabilization and activation inhibition than interference with the pro-inflammatory effects of mast cell products. These findings are in line with the hypothesis that the alkaloid curine may be beneficial for the treatment of allergic disorders.     

Mast cell stabilization and antihistaminic potentials of Curculigo orchioides rhizomes

Aim of the study

To investigate the mast cell stabilization and antihistaminic activities of the rhizomes of Curculigo orchioides (COR). Extract of Curculigo orchioides Gaertn. (Fam. Amaryllidaceae) has been reported to possess immunostimulant, and anti-inflammatory potentials. In Indian traditional system of medicine it is also used as anti-asthmatic and anti-inflammatory.

Materials and methods

Estimation of histamine release is key parameter for evaluating any target for its anti-allergic potential. The stabilization potential of the alcoholic extract of COR (100–400 mg/kg) against mast cell degranulation was studied on isolated mice peritoneal mast cells. The antihistaminic activity was performed by determining the mortality rate of mice upon exposure to compound 48/80 and effect on inhibition of histamine release upon degranulation.

Results

The raised number of intact mast cells intimates that the COR stabilized the mast cell degranulation (60.96 ± 1.96%) and percentage antihistaminic potential of the extract (63.58 ± 1.8 inhibition at dose of 400 mg/kg) and it virtues further work towards the isolation of phytoconstituents from this plant.

Conclusion

This finding provides evidence that COR inhibits mast cell-derived immediate-type allergic reactions and mast cell degranulation.     

Treatment of perennial allergic rhinitis using Shi-Bi-Lin,a Chinese herbal formula

Aim of the study

Shi-Bi-Lin (SBL) is modified from the classic formula Cang-Er-Zi-San which has been used to treat chronic rhinitis, paranasal sinusitis and allergic rhinitis by herbal practitioners. The present study aimed at patients with moderate to severe perennial allergic rhinitis. SBL which has been shown effective in treating a guinea-pig model of allergic rhinitis, was evaluated for its efficacy and safety.

Materials and methods

126 allergic rhinitis patients were recruited in a double-blind randomized control trial. Half of the patients received SBL capsules and the others half received placebo for 4 weeks. Symptoms scores, physician's evaluation, nose examination, quality of life, adverse effects, serum cytokines were evaluated before and after treatment.

Results

SBL was found to be safe and effective in relieving some symptoms of perennial allergic rhinitis, improving the nose condition, and enhancing some domains of quality of life when compare to placebo, (p < 0.05). In the 2 weeks follow up after treatment completion, the SBL enjoyed a prolongation of symptom control (p = 0.05).

Conclusion

SBL relieved symptoms of nose blockage among patients with perennial allergic rhinitis, and some aspects of the quality of life were also improved. The improvement was sustained for at least 2 weeks after treatment. No serious adverse events were encountered.     

The hydroalcoholic extract of Cassia alata (Linn.) leaves and its major compound rhein exhibits antiallergic activity via mast cell stabilization and lipoxygenase inhibition

Ethnopharmacological relevance

Leaves of Cassia alata (family: Caesalpiniaceae) are ethnomedically claimed as anti-asthmatic. In the current study we aimed to investigate the anti-allergic activities of hydro-methanolic extract of Cassia alata (Linn.) and its constituents rhein and kaempferol on triple antigen/sheep serum-induced mast-cell degranulation in rats.

Materials and methods

Antiallergic activity of hydroalcoholic extract of Cassia alata along with its two components rhein and kaempferol was evaluated using in vivo mast cell stabilization assay. Inhibitory effect on lipoxygenase (LOX) enzyme was also evaluated in vitro. Further chemical standardization of Cassia alata extract was done using rhein and kaempferol by HPTLC-densitometric method.

Results

The hydroalcoholic extract of Cassia alata significantly inhibited mast cell degranulation at 200 mg/kg dose. Both chemical constituents rhein and kaempferol also showed potent (>76%) inhibition of mast-cell degranulation at 5 mg/kg. Extract and rhein inhibited LOX enzyme with IC₅₀ values of 90.2 and 3.9 μg/mL, respectively, whereas kaempferol was inactive.

Conclusion

Our results suggest that Cassia alata exhibit anti-allergic activity through mast cell stabilization and LOX inhibition. Thus, Cassia alata or its active constituents could be potential alternative treatment for allergic diseases.     

Effects of taraxasterol on ovalbumin-induced allergic asthma in mice

Ethnopharmacological relevance

Taraxasterol was isolated from the Chinese medicinal herb Taraxacum officinale which has been frequently used as a remedy for inflammatory diseases. In the present study, we determined the in vivo protective effect of taraxasterol on allergic asthma induced by ovalbumin (OVA) in mice.

Materials and methods

Mice were sensitized and challenged with OVA, and were orally treated daily with taraxasterol at 2.5, 5 and 10 mg/kg from day 23 to 27 after sensitization. The number of inflammatory cells in bronchoalveolar lavage fluid (BALF) was determined. Th2 cytokine interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-13 (IL-13) production in BALF and OVA-specific immunoglobulin E (IgE) production in sera were measured using ELISA. Histological changes in lung tissues were examined using hematoxylin and eosin (H&E) and periodic acid-Schiff staining (PAS). Airway hyperresponsiveness (AHR) to inhaled methacholine was assessed.

Results

Taraxasterol dramatically decreased the total inflammatory cell and main inflammatory cell counts, reduced the production of Th2 cytokine IL-4, IL-5, IL-13 in BALF and OVA-specific IgE in sera, and suppressed AHR in a dose-dependent manner. Histological studies demonstrated that taraxasterol substantially suppressed OVA-induced inflammatory cells infiltration into lung tissues and goblet cell hyperplasia in airways.

Conclusions

This finding suggests that taraxasterol protects against OVA-induced allergic asthma in mice.     

Inhibitory effects of Acorus calamus extracts on mast cell-dependent anaphylactic reactions using mast cell and mouse model

Ethnopharmacological relevance

Acorus calamus Linn. (Araceae) is a traditional herbal plant used for centuries to treat various allergic symptoms including asthma and bronchitis.

Aim of the study

The present study was focused to provide a pharmacological basis for the traditional use of Acorus calamus in allergic symptoms using the mast cell-dependent anaphylactic reactions in in vitro and in vivo models.

Materials and methods

Cell viabilities were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Dinitrophenyl-human serum albumin (DNP-HSA) induced β-hexosaminidase and interleukin (IL)-4 productions in IgE-sensitized rat basophilic leukaemia (RBL-2H3) cells were measured by enzymatic assay and enzyme-linked immunosorbent assay (ELISA). Passive cutaneous anaphylaxis (PCA) reaction mouse model was implemented for in vivo studies.

Results

Hot water (HW), butylene glycol (BG), hexane (HE) and steam distilled (SD) extracts of Acorus calamus showed different cytoxicity levels evaluated in RBL-2H3 cells. Sub-toxic doses of HW extract suppressed the β-hexosaminidase secretion and IL-4 production significantly and dose dependently in DNP-HSA induced IgE-sensitized RBL-2H3 cells compared to other extracts of Acorus calamus. Further, in vivo studies also revealed that the HW extract significantly inhibited the PCA reaction in mouse compared to the normal control group.

Conclusion

HW extract of Acorus calamus most effectively inhibited degranulation and IL-4 secretion in DNP-HSA-stimulated RBL-2H3 cells and also reduced the mast cell-mediated PCA reaction in mouse, providing a therapeutic evidence for its traditional use in ameliorating allergic reactions.     

Inhibitory effects of adlay bran (Coix lachryma-jobi L. var. ma-yuen Stapf) on chemical mediator release and cytokine production in rat basophilic leukemia cells

Ethnopharmacological relevance

Adlay (Job's tears, Coix lachryma-jobi L. var. ma-yuen Stapf) has long been used in China to treat rheumatism.

Aim of the study

We investigated the anti-allergic effects of adlay bran on rat basophilic leukemia (RBL)-2H3 cells.

Materials and methods

To evaluate the anti-allergic effects of adlay bran, the release of histamines and cytokines were measured using ELISA. To explore the mechanism of these effects, the protein expression levels were determined using western blotting.

Results

A 40.8 μg/mL concentration of the ethyl acetate fraction of the ethanolic extracts of adlay bran (ABE-EtOAc) effectively inhibited mast cell degranulation. The 40–100% EtOAc/Hex subfractions of ABE-EtOAc inhibited histamine release with an IC₅₀ of 71–87 μg/mL. Moreover, the ABE-EtOAc subfractions suppressed the secretion of interleukin (IL)-4, IL-6 and tumor necrosis factor-α in the RBL-2H3 cells, indicating that adlay bran can inhibit cytokine secretion in the late phase of the allergic reaction. In addition, adlay bran reduced the intracellular production of reactive oxygen species, inhibited the phosphorylation of Akt and decreased the expression of protein kinase C. Furthermore, six phenolic acids and one flavone were isolated. Of these compounds, luteolin showed the most potent inhibitory activity (IC₅₀ = 1.5 μg/mL).

Conclusion

Adlay bran extract reduced the release of histamines and cytokines and suppressed the production of Akt. These combined effects influenced the signal transduction in RBL-2H3 cells, thereby revealing the mechanisms of the anti-allergic effects of adlay.     

Effect of an extract of Andrographis paniculata leaves on inflammatory and allergic mediators in vitro

Aim of study

Andrographis paniculata has been known to possess widespread traditional application in the treatment of allergy and inflammatory diseases. In the current study, we sought to examine the effects of an extract of Andrographis paniculata leaves on inhibition of lipopolysaccharide (LPS) induced [nitric oxide (NO), prostaglandin E₂ (PGE₂), interleukin-1beta (IL-1 beta), and interleukin-6 (IL-6)] and calcimycin (A23187) induced [leukotriene B₄ (LTB₄), thromboxane B₂ (TXB₂) and histamine] mediators in diverse cell based models.

Materials and methods

Effect of an extract of Andrographis paniculata leaves (AP) was studied on inhibition of LPS induced NO, PGE₂, IL-1 beta and IL-6 in J774A.1 murine macrophages; A23187 induced LTB₄ and TXB₂ in HL-60 promyelocytic leukemic cells and histamine in RBL-2H3 rat basophilic leukemia cells.

Results and conclusion

AP illustrated significant alleviation of pro-inflammatory, inflammatory, and allergic mediators. However, no inhibition was observed against histamine release. This outcome has been summed up to deduce that AP is fairly potent in attenuating the inflammation by inhibiting pro-inflammatory (NO, IL-1 beta and IL-6), inflammatory (PGE₂ and TXB₂) and allergic (LTB₄) mediators.     

Cordia verbenacea and secretion of mast cells in different animal species

Ethnopharmacology relevance

Different plant species from Cordia genera are used in folk medicine as anti-inflammatory medication throughout the tropical and subtropical regions of the world. In Brazil, Cordia verbenacea is a medicinal plant known as “erva-baleeira”. The alcoholic extracts, decoctions and infusions with leaves of C. verbenacea are used in Brazilian traditional medicine for treatment of cough, pneumonia, parasitic diseases and, especially, the inflammatory processes. Anti-inflammatory activity was already demonstrated; however, molecular mechanisms of action are not completely understood. Considering the importance of histamine in early events of inflammation and in allergic diseases, we evaluated the effect of ethanol extract of leaves of C. verbenacea on histamine release (in vitro and in vivo studies) from different types of mast cells induced by chemical agents using several species of rodents.

Materials and Methods

The extraction and quantification of histamine were performed by using an automatic fluorometric continuous flow system.

Results

The extract of C. verbenacea (30 μg/ml) reduced the in vitro secretion of histamine from rat mast cells induced by ionophore A23187, concanavalin A and compound 48/80, respectively, to 22.1 ± 2.2%, 24.3 ± 2.5% and 21.4 ± 2.1%. At the same concentration, the extract also inhibited the secretion of histamine from mast cells of guinea pig induced by ionophore A23187 to 33.3 ± 2.2%, and mast cells of hamster induced by ionophore A23187 and concanavalin A to 15.8 ± 2.5% and 10.8 ± 2.6%, respectively. The oral treatment with the extract (300 mg/kg) also inhibited the secretion of histamine induced by A23187 about to 36.3 ± 3.2% in rats.

Conclusions

C. verbenacea inhibits the in vitro secretion of histamine from mast cells of different animal species, as well as the secretion of mast cells from animals treated with the extract, which gives not only the proven anti-inflammatory effect of the plant, but also anti-allergic effect, opening new possibilities for future anti-allergic herbal medicine.     

The inhibitory activity of atractylenolide Ш, a sesquiterpenoid,on IgE-mediated mast cell activation and passive cutaneous anaphylaxis (PCA)

Ethnopharmacological relevance

AT Ш, a sesquiterpenoid, is the major component of Atractylodes japonica Koidz that has been used as a traditional oriental medicine.

Aim of the study

We investigated the anti-allergic activity of AT Ш and its mechanism of action.

Materials and methods

The released amount of β-hexosaminidase in mast cells, a key parameter of degranulation, was measured. Anti-allergic potential of AT Ш was evaluated using passive cutaneous anaphylaxis in vivo. The anti-allergic mechanism of AT Ш was investigated by immunoblotting analysis, RT-PCR and measurement of [Ca²⁺]i in mast cells.

Results

AT Ш significantly inhibited IgE/Ag-mediated degranulation with an IC₅₀ value (36±4 μM) in RBL-2H3 cells without affecting cell viability. It also suppressed IgE/Ag-mediated passive cutaneous anaphylaxis (PCA) response with an ED₅₀ value (65±41 mg/kg) in vivo. AT Ш suppressed the production of interleukin (IL-4) and tumor necrosis factor (TNF)-alpha mRNAs more potent than the Src-family kinase inhibitor PP2 in RBL-2H3 cells at all concentrations. In order to elucidate the anti-allergic mechanisms of AT Ш in mast cells, we examined the activated levels of signaling molecules. AT Ш inhibited the phosphorylation of Lyn, Fyn, Syk, LAT, PLCγ, Gab2, Akt, p38, and JNK kinases expression. IgE/Ag-mediated [Ca²⁺]i elevation was significantly inhibited by AT Ш.

Conclusions

Our study suggests that AT Ш might be used as a therapeutic agent for allergic diseases.     

The ethyl acetate extract of Cordyceps militaris inhibits IgE-mediated allergic responses in mast cells and passive cutaneous anaphylaxis reaction in mice

Ethnopharmacological relevance

Cordyceps militaris has been used as a traditional herbal medicine for treating allergy in East Asia.

Aim of the study

We investigated the anti-allergic efficacy of Cordyceps militaris and its mechanism of action.

Materials and methods

β-Hexosaminidase release of mast cells, a key parameter of degranulation, was evaluated. Anti-allergic potential of Cordyceps militaris was studied using passive cutaneous anaphylaxis (PCA) in vivo. The anti-allergic mechanism of Cordyceps militaris was investigated by immunoblotting analysis, RT-PCR and other biological approaches in mast cells.

Results

GSCM EtOAc extract (GSCME) inhibited antigen-induced degranulation with a IC50 value (28.5 μg/ml) in RBL-2H3 cells and antigen-induced passive cutaneous anaphylaxis (PCA) response with a ED50 value (665 mg/kg) in vivo. The release of interleukin (IL-4) and tumor necrosis factor (TNF-α were decreased by GSCME in RBL-2H3 cells. In order to elucidate the anti-allergic mechanisms of GSCME in mast cells, we examined the activated levels of signaling molecules. GSCME inhibited the phosphorylation Syk, ERK, p38 and JNK expression. Identified genistein, daidzein, genistein 7-O-β-d-glucoside 4″-O-methylate, genistein 4′-O-β-d-glucoside 4″-O-methylate, glycitein 7-O-β-d-glucoside 4″-O-methylate, daidzein 7-O-β-d-glucoside 4″-O-methylate and adenosine in GSCME, inhibited antigen-induced degranulation in RBL-2H3 cells.

Conclusions

Our study suggests that GSCME might be used as a therapeutic agent for allergic diseases.     

Anti-allergic and anti-inflammatory effects of bakkenolide B isolated from Petasites japonicus leaves

Aims of the study

To elucidate the anti-allergic and anti-inflammatory effects of Petasites genus, we studied the effects of several compounds isolated from Petasites japonicas leaves.

Materials and methods

Bakkenolide B was isolated from Petasites japonicus leaves. Antigen-induced degranulation was measured in RBL-2H3 mast cells by measuring β-hexosamidase activity. Induction of inducible nitric oxide synthase and cyclooxygenase 2 was measured by Western blotting in peritoneal macrophages. Ovalbumin-induced asthma model was used for in vivo efficacy test of bakkanolide B.

Results

We found that bakkenolide B, a major component of the leaves, concentration-dependently inhibited RBL-2H3 mast cell degranulation. Bakkenolide B also inhibited the gene inductions of inducible nitric oxide synthase and cyclooxygenase 2 in mouse peritoneal macrophages. Furthermore, in an ovalbumin-induced asthma model, bakkenolide B strongly inhibited the accumulation of eosinophils, macrophages, and lymphocytes to bronchoalveolar lavage fluid.

Conclusion

Bakkenolide B has suppressive properties for allergic and inflammatory responses and may be utilized as a potent agent for the treatment of asthma.     

Anti-inflammatory and anti-allergic effects of Agrimonia pilosa Ledeb extract on murine cell lines and OVA-induced airway inflammation

Ethnopharmacological evidence

Agrimonia pilosa Ledeb (Rosaceae, AP) has long been used as a traditional medicine in Korea and other Asian countries to treat various diseases.

Aim of the study

In the present study, the anti-inflammatory and anti-allergic effects of AP extract in in vitro cell lines and in vivo mouse model of inflammation and the molecular mechanisms involved were reported.

Materials and methods

Using Raw 264.7 murine macrophages the effects of methanol extract of AP in lipopolysaccharide (LPS)-induced production of inflammatory mediators were measured. Further IgE-DNP-induced interleukin (IL)-4 production and degranulation in RBL-2H3 rat basophilic cell lines was also estimated. To investigate the anti-asthmatic effect of AP in vivo, airway inflammation in ovalbumin (OVA)-induced mouse model was used.

Results

AP attenuated the production of inflammatory mediators such as NO, PGE₂ and pro-inflammatory cytokines in LPS-induced Raw 264.7 cells. Further, AP inhibited IL-4 production and degranulation in IgE-DNP-induced RBL-2H3 cells. Furthermore, AP attenuated the infiltration of immune cells into lung, cytokines production in broncho-alveolar lavage fluid (BALF) and airway-hyperresponsiveness (AHR) on OVA-induced mouse model of inflammation.

Conclusion

Our results showed that AP attenuated the activation of macrophages, basophils, and inhibited the OVA-induced airway inflammation. The molecular mechanisms leading to AP's potent anti-inflammatory and anti-allergic effects might be through regulation of TRIF-dependent and Syk-PLCγ/AKT signaling pathways, suggesting that AP may provide a valuable therapeutic strategy in treating various inflammatory diseases including asthma.