Phlomis umbrosa root inhibits mast cell-dependent allergic reactions and inflammatory cytokine secretion

The effect of an aqueous extract of Phlomis umbrosa Turcz. (Labiatae) root (PUAE) on mast cell-dependent allergic reactions and inflammatory cytokine secretion were investigated. PUAE (0.01-1 g/kg) inhibited compound 48/80-induced systemic allergic reaction. When PUAE was employed in a systemic allergic reaction test, the plasma histamine levels were reduced in a dose-dependent manner. PUAE (0.1 and 1 g/kg) also significantly inhibited the local allergic reaction activated by anti-dinitrophenyl (DNP) IgE. PUAE (0.001-1 mg/mL) dose-dependently inhibited the histamine release from rat peritoneal mast cells activated by compound 48/80 or anti-DNP IgE. PUAE (0.01-1 mg/mL) inhibited the secretion of interleukin (IL)-1beta in phorbol 12-myristate 13-acetate plus calcium ionophore A23187-stimulated human mast cell line (HMC-1) cells. PUAE (1 mg/mL) inhibited the gene expression and production of the main inflammatory cytokine, TNF-alpha, in HMC-1 cells. These results provide evidence that PUAE may be beneficial in the treatment of allergic diseases.     

Shini-san inhibits mast cell-dependent immediate-type allergic reactions

Shini-San has been used for treatment of allergic disease in Korea. However, its effect in experimental models remains unknown. The mast cell plays a pivotal role in initiating allergic response by secreting intracytoplasmic granular mediators such as histamine. The present report describes an inhibitory effect of Shini-San on mast cell-mediated immediate-type allergic reactions. Topical application of compound 48/80 can induce an ear swelling response in normal (WBB6F1(-)+/+) mice but not in congenic mast cell-deficient WBB6F1-W/WV mice. Shini-San inhibited concentration-dependent mast cell-dependent ear swelling response induced by compound 48/80 in normal mice. Shini-San inhibited concentration-dependent passive cutaneous anaphylaxis induced by anti-dinitrophenyl (DNP) immunoglobulin E (IgE) in rats by topical application. Shini-San also inhibited in concentration-dependent fashion the histamine release from the rat peritoneal mast cells by compound 48/80 or anti-DNP IgE. Moreover, Shini-San had a significant inhibitory effect on compound 48/80-induced systemic anaphylactic reaction. These results indicate that Shini-San inhibits immediate type allergic reactions by inhibition of mast cell degranulation in vivo and in vitro.     

The anti‐anaphylactoid effects of Piperine through regulating MAS‐related G protein‐coupled receptor X2 activation

Mast cells play an important role in inflammatory and allergic diseases. MAS‐related G protein‐coupled receptor X2 (MRGPRX2) is a novel G protein‐coupled receptor in mast cells that mediates drug‐induced anaphylactoid reactions. Piperine has been reported to have anti‐inflammatory and anti‐allergic pharmacological activities. However, whether the pharmacological effects are regulated by MRGPRX2 has not yet been reported. The purpose of this study was to assess the anti‐anaphylactoid effect of Piperine and to explore its potential mechanism. The anti‐anaphylactoid effect of Piperine was assessed by an in vivo mouse hindpaw extravasation model. Mast cell intracellular calcium mobilization was measured by a calcium imaging assay. An enzyme immunoassay was used to evaluate the release of pro‐inflammatory factors from stimulated mast cells. Activated mast cell related signals were assessed by western blot. A cell membrane chromatography assay was used to determine the binding characteristics of Piperine and MRGPRX2. The results showed that Piperine suppressed mast cell intracellular Ca²⁺ mobilization, inhibited cytokines and chemokines release, and down‐regulated the phosphorylation level of phospholipase Cγ1, protein kinase C, inositol 1,4,5‐triphate receptor, P38, protein kinase B, and ERK. Meanwhile, Piperine can bind to MRGPRX2 as a specific antagonist. Hence, Piperine can be served as a novel therapeutic drug candidate for MRGPRX2‐mediated anaphylactoid reactions.     

Antiallergic effect of KOB03, a polyherbal medicine, on mast cell-mediated allergic responses in ovalbumin-induced allergic rhinitis mouse and human mast cells

Ethnopharmacological relevance

KOB03 is a polyherbal medicine consisting of five different herbs and has commonly been used for the treatment of various allergic diseases. However, its precise anti-allergic effect and mechanism remain unknown.

Aim of the study

The aim of this study was to investigate the effect of KOB03 on allergic responses through the regulation of mast-cell mediated allergic inflammation.

Materials and methods

To determine the effect of KOB03 on mast cell-mediated allergic reactions, we investigated the parameter changes of in vivo models such as compound 48/80-induced systemic anaphylaxis and ovalbumin (OVA)-induced allergic rhinitis, and the release of allergic inflammatory mediators such as histamine, immunoglobulin (Ig) E, and inflammatory cytokines via the MAPKs and NF-kappaB pathways.

Results

The oral administration of KOB03 at doses of 100 and 200 mg/kg inhibited histamine release and mortality in compound 48/80-induced anaphylactic rats. KOB03 also improved rhinitis symptoms, inhibited the histopathological changes of nasal mucosa, and decreased the serum levels of histamine, OVA-specific IgE and TNF-α in OVA-induced allergic rhinitis in mice. In vitro, KOB03 suppressed compound 48/80-induced histamine release by blocking mast cell degranulation. In addition, KOB03 inhibited the production of inflammatory cytokines such as TNF-α, IL-1β, IL-6 and IL-8 in PMA/A23187-stimulated HMC-1 mast cells by suppressing their gene expression and blocking the ERK1/2 and p38 MAPK and NF-κB pathways.

Conclusions

These results suggest that KOB03 has an anti-allergic effect by modulating mast cell-mediated allergic responses in allergic rhinitis.     

Cheongyeolsaseuptang inhibits production of TNF-alpha, IL-6 and IL-8 as well as NF-kappa B activation in human mast cells

Traditional Korean medicine, Cheongyeolsaseuptang (CYSST) has been widely applied as a treatment of rheumatoid arthritis (RA) in Korea. However, its effect in experimental models remains unknown. Recent reports suggest that in patients with RA, synovial mast cells increase in number and show signs of activation and production of cytokines. In this study, we investigated the effect of CYSST on production of cytokines by activated human mast cell line, HMC-1. When CYSST (1mg/ml) was added, the production of tumor necrosis factor-alpha, interleukin (IL)-6, and IL-8 was significantly inhibited about 37, 33.6, and 48%, respectively on phorbol 12-myristate 13-acetate (PMA) plus calcium ionophore A23187-stimulated HMC-1 cells. In addition, CYSST inhibited PMA plus A23187-induced activation of nuclear factor-kappaB. These findings may help understanding the mechanism of action of this medicine leading to control activated mast cells on inflammatory condition like RA.     

Blockade of IL-6 secretion pathway by the sesquiterpenoid atractylenolide III

Atractylenolide III (1) is the major bioactive component of Atractylodes lancea. The aim of this study was to analyze the effect on the regulation of interleukin (IL)-6 secretion pathway caused by 1. This sesquiterpenoid inhibited the secretion and expression of IL-6 in phorbol 12-myristate 13-acetate- and calcium ionophore A23187-stimulated human mast cells (HMC)-1. In addition, 1 inhibited histamine release in stimulated HMC-1 cells. In stimulated HMC-1 cells, 1 suppressed activation of p38 mitogen-activated protein kinase, C-Jun-N-terminal protein kinase, and nuclear factor-κB. In addition, 1 suppressed the activation of caspase-1 and the expression of receptor interacting protein-2. These results provide new insights that atractylenolide III (1) may control immunological reactions by regulating the cellular functions of IL-6 in mast cells.     

广西甜茶的抗过敏作用研究

目的：研究广西甜茶（Rubus suavissimus S.Lee）提取物的抗过敏作用并观察其对大鼠腹腔肥大细胞释放组胺的影响,探讨可能的作用机制。方法：采用2,4-二硝基氟苯（DNFB）及绵羊红细胞（SRBC）诱发的小鼠迟发型过敏反应,小鼠异种被动皮肤过敏反应（PCA）,观察连续口服给予广西甜茶提取物的抗过敏作用。利用高效液相-库仑阵列电化学检测系统（HPLC-ECD）测定Compound 48/80刺激大鼠腹腔肥大细胞释放组胺,评价广西甜茶提取物对肥大细胞释放组胺的影响。结果：与模型组相比,连续口服广西甜茶提取物125、250及500 mg/kg均能显著抑制由DNFB诱发小鼠耳肿胀及血管通透性的增高。此外,广西甜茶提取物可以减轻小鼠异种被动皮肤过敏反应及对SRBC诱发小鼠迟发型过敏反应足跖肿胀也显示有一定程度的抑制作用。体外实验证明广西甜茶提取物对Compound 48/80刺激大鼠腹腔肥大细胞释放组胺呈剂量依赖性抑制作用。结论：广西甜茶具有显著的抗过敏作用,其作用机制可能与抑制肥大细胞释放组胺有关。     

Antiallergic effects of Scutellaria baicalensis on inflammation in vivo and in vitro

Ethnopharmacological relevance

Scutellaria baicalensis (SB) is one of the most widely used medicinal herbs for the treatment of inflammation. In this study, we investigated the antiallergic effect of SB in vivo and in vitro.

Materials and methods

Sprague–Dawley (SD) rats received intradermal injections of anti-DNP IgE at each of three dorsal skin sites. Forty-eight hours later, each rat received an injection of DNP-HSA in saline containing 4% Evans blue through the dorsal vein of the penis. One hour before injection, SB extract was administered orally. The dorsal skin of the rats was removed and the pigment area measured. In addition, rat peritoneal mast cells (RPMCs) were cultured and purified to investigate histamine release. In vitro, human mast cells (HMC-1) were pretreated with SB extract for 30 min before stimulation with phorbol 12-myristate 13-acetate (PMA) plus A23187. The effects on pro-inflammatory cytokine expression and mitogen activated protein (MAP) kinase expression were investigated using TNF-α and IL-8 assays, and Western blotting analysis of HMC-1 cells.

Results and conclusions

SB treatment inhibited the passive cutaneous anaphylaxis reaction compared to the control group, and histamine release decreased significantly following treatment of RPMCs with SB. In HMC-1 cells, SB restored IL-8 and TNF-α expression and inhibited MAP kinase expression in compound 48/80-induced HMC-1 cells. These data suggest that SB may prove to be a useful anti-inflammatory agent through its downregulation of the expression of various inflammatory mediators.     

Effect of Vitex rotundifolia on immediate-type allergic reaction

We investigated the effect of aqueous extract of Vitex rotundifolia (L.) (Verbenaceae) fruits (VRFE) on the immediate-type allergic reactions in vivo and in vitro. VRFE (10(-4)-1.0 g/kg) dose-dependently inhibited systemic allergic reaction induced by compound 48/80. When VRFE was employed in a systemic allergic reaction test, the plasma histamine levels were reduced in a dose-dependent manner. VRFE (5x10(-1) and 1.0 g/kg) inhibited passive cutaneous anaphylaxis activated by anti-dinitrophenyl (DNP) IgE. VRFE (10(-3)-1.0 mg/ml) also dose-dependently inhibited the histamine release from the rat peritoneal mast cells (RPMC) by compound 48/80 or anti-DNP IgE. Moreover, VRFE (10(-3) mg/ml) had a significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor-alpha production from RPMC. These results suggest that VRFE may be beneficial in the regulation of immediate-type allergic reaction.     

The inhibitory effect of Houttuynia cordata extract on stem cell factor-induced HMC-1 cell migration

Hottuynia cordata Thunb (Saururaceae; HC) is known as a therapeutic drug that has been used in traditional oriental medicine for the treatment of allergy. Mast cells play an important role in a variety of inflammatory diseases, and specifically asthma and atopy. In the present study, we investigated the effect of HC extracts on the migration of the human mast cell line, HMC-1, in response to stem cell factor (SCF). Treatment with HC extracts at a concentration of 10mug/ml for 24h showed no significant decrease in the survival rate of the HMC-1 cells. SCF showed the typical bell-shape curve for the HMC-1 cell chemoattraction with the peak of the curve at the SCF concentration of 100ng/ml. HC-1, which was the whole plant (Houttuynia cordata) extracted with 80% EtOH, and HC-3, which was the residue successively partitioned with EtOAc, both had inhibitory effects on HMC-1 cell movement. After the treatment with 10mug/ml HC-1 extract for 6 and 24h, the chemotactic index (CI) of HMC-1 cells decreased up to 74 and 63%, respectively. HC-3 extract treatment for 6 and 24h lowered the CI to 72 and 44%, respectively. The HC-1 and HC-3 extracts had no inhibitory effect on the mRNA and surface protein expressions of c-kit, SCF receptor. SCF mediated the chemotaxis signaling via NF-kappaB activation, and both extracts inhibited the activation. Therefore, our results indicate that HC-1 and HC-3 extracts decrease the chemotactic ability of HMC-1 cells in response to SCF by inhibiting the NF-kappaB activation, and these substances may be useful for treating mast cell-induced inflammatory diseases.     

Inhibition of mast cell-dependent immediate-type hypersensitivity reactions by purple bamboo salt

Purple bamboo salt is a specially processed salt according to the traditional recipe using normal salt and bamboo etc. It has been used as a folk medicine for the purpose of prevention and treatment of various diseases in Korea. This report describes an inhibitory effect of purple bamboo salt on mast cell-mediated immediate-type allergic reactions. Purple bamboo salt (0.01-1 microg per site) significantly inhibited the ear swelling response induced by intradermal injection of compound 48/80 in mice. Purple bamboo salt (0.01-1 mg/ml) dose-dependently inhibited the histamine release from the rat peritoneal mast cells (RPMCs) by compound 48/80. Purple bamboo salt (0.01-1 g/kg) also dose-dependently inhibited the passive cutaneous anaphylaxis (PCA) by oral administration. Our results provide evidence that purple bamboo salt will be beneficial in the regulation of immediate-type of allergic reactions.     

仙蓟化斑胶囊对肥大细胞脱颗粒及组胺释放影响的研究

[目的]研究仙蓟化斑胶囊抗变态反应的作用机制,为后期实验及其临床治疗过敏性紫癜提供依据。[方法]制备致敏的大鼠及抗卵蛋白血清,给予不同浓度药物,采用大鼠被动皮肤过敏反应(PCA)实验,测定大鼠皮肤蓝斑直径,同时采用大鼠颅骨骨膜肥大细胞脱颗粒实验、大鼠肥大细胞释放组胺实验,测定脱颗粒百分率和组胺释放水平。[结果]仙蓟化斑胶囊可使蓝斑直径变小、可降低肥大细胞脱颗粒百分率、降低其组胺释放,并呈明显的剂量依赖关系。[结论]仙蓟化斑胶囊有抗变态反应的作用,明显抑制PCA、肥大细胞脱颗粒及组胺的释放。     

Suppressive effects of Houttuynia cordata Thunb (Saururaceae) extract on Th2 immune response

ETHNOPHARMACOLOGICAL RELEVANCE: Houttuynia cordata Thunb (Saururaceae), known as 'E-Sung-Cho' in Korea, has been traditionally used for the treatment of herpes simplex, chronic sinusitis, and allergy. AIM OF THE STUDY: To investigate the inhibitory activity of Houttuynia cordata Thunb fractions (HcFs) on the T helper 2 (Th2) immune response, we evaluated the alternation of the release of Th2-type cytokines and chemokines such as interleukin (IL)-4 and IL-5, and thymus and activation-regulated chemokine (TARC/CCL17). MATERIALS AND METHODS: Ethanol fraction was obtained from dried and powdered whole plants of Houttuynia cordata Thunb using ethanol. The residue was diluted with water and was then successively partitioned with n-hexane, EtOAc and BuOH. HcFs include ethanol, n-hexane, EtOAc, BuOH and water fractions. RT-PCR and ELISA were performed to measure mRNA and protein expression of cytokines. RESULTS: HcFs inhibited the expression of IL-4 and IL-5 in response to phorbol 12-myristate 13-acetate (PMA) and calcium ionophore (CaI) in Jurkat T cells and the human mast cell line, HMC-1. IL-4- and tumor necrosis factor-alpha (TNF-alpha)-induced TARC production was blocked by HcFs in skin fibroblast CCD-986sk cells, particularly by the ethanol extract of Hc. Stimulants included in PMA, phytohemagglutinin (PHA) and CaI, increased the mRNA level of CC chemokine receptor 4 (CCR4), a receptor of TARC, in Jurkat T cells, and the ethanol extract of HcF weakly blocked the increased mRNA level. However, the stimulants and ethanol extract had no effect on the CCR4 protein level. The ethanol extract inhibited TARC-induced migration, as well as basal migration of Jurkat T cells. CONCLUSIONS: This study may show the usefulness of HcFs in the ethnopharmacological treatment of Th2-mediated or allergic inflammation, through the down-regulation of the production of Th2 cytokines and TARC, as well as cell migration.     

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.     

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.     

Effects of magnolialide isolated from the leaves of Laurus nobilis L. (Lauraceae) on immunoglobulin E-mediated type I hypersensitivity in vitro

Ethnopharmacological relevance

Laurus nobilis L. (Lauraceae) has been used for folk medicines in the Mediterranean area and Europe to treat various disorders including skin inflammation (dermatitis) and asthma.

Aim of the study

Our aim was to investigate the scientific evaluation of the compounds from Laurus nobilis L. on immuniglobulin E (IgE)-mediated type I hypersensitivity responses in vitro such as atopic dermatitis and asthma.

Methods and materials

Seven compounds were isolated and examined for the mast cell stabilizing effect on IgE-sensitized RBL-2H3 mast cells by measuring the β-hexosaminidase activity. In addition, the effects on interleukin (IL)-4 production and IL-5-dependent Y16 early B cell proliferation were investigated as well as their cytotoxic effects on RBL-2H3 cells.

Results

Among the seven isolated compounds, magnolialide attenuated the release of β-hexosaminidase from RBL-2H3 cells with an IC₅₀ value of 20.2 μM, while the other compounds revealed no significant effects at concentrations tested. Furthermore, magnolialide significantly inhibited the IL-4 release with an IC₅₀ value of 18.1 μM and IL-4 mRNA expression with an IC₅₀ value of 15.7 μM in IgE-sensitized RBL-2H3 cells. In addition, the inhibition of IL-5-dependent proliferation of early B cells (Y16 cells) by magnolialide was demonstrated with an IC₅₀ value of 18.4 μM.

Conclusion

These results suggest that the magnolialide might be a candidate for the treatment of IgE-mediated hypersensitivity responses such as atopic dermatitis and asthma by inhibiting mast cell degranulation, the IL-4 production, and IL-5-dependent early B cell proliferation, key factors in the development and amplification of type I hypersensitivity reactions.     

Proanthocyanidin‐rich Pinus radiata bark extract inhibits mast cell‐mediated anaphylaxis‐like reactions

Mast cells play a critical role in the effector phase of immediate hypersensitivity and allergic reactions. Pinus radiata bark extract exerts multiple biological effects and exhibits immunomodulatory and antioxidant properties. However, its role in mast cell‐mediated anaphylactic reactions has not been thoroughly investigated. In this study, we examined the effects of proanthocyanidin‐rich water extract (PAWE) isolated from P. radiata bark on compound 48/80‐induced or antidinitrophenyl (DNP) immunoglobulin E (IgE)‐mediated anaphylaxis‐like reactions in vivo. In addition, we evaluated the mechanism underlying the inhibitory effect of PAWE on mast cell activation, with a specific focus on histamine release, using rat peritoneal mast cells. PAWE attenuated compound 48/80‐induced or anti‐DNP IgE‐mediated passive cutaneous anaphylaxis‐like reactions in mice, and it inhibited histamine release triggered by compound 48/80, ionophore A23187, or anti‐DNP IgE in rat peritoneal mast cells in vitro. Moreover, PAWE suppressed compound 48/80‐elicited calcium uptake in a concentration‐dependent manner and promoted a transient increase in intracellular cyclic adenosine‐3′,5′‐monophosphate levels. Together, these results suggest that proanthocyanidin‐rich P. radiata bark extract effectively inhibits anaphylaxis‐like reactions.