Passive Cutaneous Anaphylaxis Reaction Research Articles

Platycodin D를 포함하는 도라지 추출물이 IgE/Ag 유도 제 1형 과민반응에 미치는 영향

최근 알러지성 비염, 알러지성 피부염, 알러지성 천식, 알러지성 과민반응 등과 같은 질병이 점차 늘어나고 있으며 그에 따른 효과적인 치료제나 효율적인 치료법이 시급한 실정이다. 본 연구에서 동물모델과 비만세포 탈과립작용에서 Platycodin D가 포함된 도라지 추출물(PG-Platycodin D)의 항알러지 효과를 연구하였다. <TEX>$In$</TEX> <TEX>$vivo$</TEX> 상에서 PG-Platycodin D가 항원-항체 반응에 의한 알러지 반응을 효과적으로 억제하는지 살펴보기 위한 아나필락틱 쇼크 평가에서 Platycodin D의 함량이 1%에서 5%까지 증가한 도라지 추출물일수록 <TEX>$in$</TEX> <TEX>$vivo$</TEX> 수준에서의 알러지반응이 억제되는 것을 확인할 수 있었다. 또한 항원-항체 반응에 의해 매개된 RBL-2H3 비만세포의 탈 과립현상에 대한 PG-Platycodin D의 효과를 알아보기 위한 <TEX>${\beta}$</TEX>-hexosaminidase의 방출량 측정에서 Platycodin D의 함량이 1%에서 5%까지 포함된 도라지 추출물에서 농도가 증가함에 따라 <TEX>${\beta}$</TEX>-hexosaminidase의 방출량이 농도 의존적으로 감소하는 것을 알 수 있었다. 또한, PG-Platycodin D의 처리가 항원-항체 반응에 의해 매개된 RBL-2H3 비만세포 내의 IL-3의 발현이 감소하는 양상을 확인할 수 있었다. 이들의 결과로부터 Platycodin D가 포함된 도라지 추출물이 IL-3의 유전자 발현을 억제함으로써 항원-항체 반응에 의한 탈 과립현상을 억제하여 알러지 작용을 제어하는 가능성을 확인하였다. Allergic response is associated with mast cells, through the release of arachidonic acid, proinflammatory cytokines, and histamine. We investigated the effect of Platycodon grandiflorum including platycodin D (PG-Platycodin D) on allergic responses in an animal model and on mast cell degranulation. PG-Platycodin D suppressed the release of <TEX>${\beta}$</TEX>-hexosaminidase, a type of marker associated with degranulation. PG-Platycodin D efficiently inhibited the passive cutaneous anaphylaxis (PCA) reaction in ICR mice. In addition, molecular analysis demonstrated that PG-Platycodin D inhibited mRNA expression of both IL-3. These results suggest that PG-Platycodin D can inhibit mast cell degranulation through the inhibition of IL-3 mRNA expression, and that PG-Platycodin D may potentially serve as an anti-allergic agent.

PEGylation is effective in reducing immunogenicity, immunotoxicity, and hepatotoxicity of α-momorcharin in vivo

Background and aim: α-momorcharin (α-MMC), a type I ribosome-inactivating protein (RIP) from Momordica charantia, is well known for its antitumor and antivirus activities. However, the immunotoxicity and hepatotoxicity hampers its potential therapeutic usage. In order to reduce its toxicity, we had modified the α-MMC with polyethylene glycol (PEG), and detected the toxicity of the PEGylated α-MMC conjugates (α-MMC-PEG) in vivo.Materials and methods: After α-MMC purified from bitter melon seeds, α-MMC-PEG was constructed with a branched 20 kDa (mPEG) 2-Lys-NHS, the tests of immunogenicity, immunotoxicity, and general toxicity of α-MMC-PEG were conducted in guinea pig and rat.Results: The titer of specific IgG in rats, immunized by α-MMC-PEG, were approximately one-third of those that by α-MMC, all the guinea pigs treated with α-MMC died of anaphylaxis shock within 5 min, while no animals treated with α-MMC-PEG died in the active systemic anaphylaxis (ASA) test. The passive cutaneous anaphylaxis (PCA) reaction of α-MMC-PEG challenge in rats was significantly smaller than that of the α-MMC. The liver damage was greatly released, such as the change of globulin (GLB), aspartate aminotransferase (AST), total bilirubin (TBIL) cholesterol (CHOL), albumin (ALB), and the degree of hepatocyte necrosis in repeated toxicity study.Conclusions: PEGylation is effective in reducing the immunogenicity, immunotoxicity, and hepatotoxicity of α-MMC in vivo.

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

Ethnopharmacological relevanceScutellaria 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 methodsSprague–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 30min 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 conclusionsSB 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.

Mast Cell Restricted Mouse and Human Tryptase·Heparin Complexes Hinder Thrombin-induced Coagulation of Plasma and the Generation of Fibrin by Proteolytically Destroying Fibrinogen

The mouse and human TPSB2 and TPSAB1 genes encode tetramer-forming tryptases stored in the secretory granules of mast cells (MCs) ionically bound to heparin-containing serglycin proteoglycans. In mice these genes encode mouse MC protease-6 (mMCP-6) and mMCP-7. The corresponding human genes encode a family of serine proteases that collectively are called hTryptase-β. We previously showed that the α chain of fibrinogen is a preferred substrate of mMCP-7. We now show that this plasma protein also is highly susceptible to degradation by hTryptase-β· and mMCP-6·heparin complexes and that Lys(575) is a preferred cleavage site in the protein α chain. Because cutaneous mouse MCs store substantial amounts of mMCP-6·heparin complexes in their secretory granules, the passive cutaneous anaphylaxis reaction was induced in the skin of mMCP-6(+)/mMCP-7(-) and mMCP-6(-)/mMCP-7(-) C57BL/6 mice. In support of the in vitro data, fibrin deposits were markedly increased in the skin of the double-deficient mice 6 h after IgE-sensitized animals were given the relevant antigen. Fibrinogen is a major constituent of the edema fluid that accumulates in tissues when MCs degranulate. Our discovery that mouse and human tetramer-forming tryptases destroy fibrinogen before this circulating protein can be converted to fibrin changes the paradigm of how MCs hinder fibrin deposition and blood coagulation internally. Because of the adverse consequences of fibrin deposits in tissues, our data explain why mice and humans lack a circulating protease inhibitor that rapidly inactivates MC tryptases and why mammals have two genes that encode tetramer-forming serine proteases that preferentially degrade fibrinogen.

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

Ethnopharmacological relevanceAcorus calamus Linn. (Araceae) is a traditional herbal plant used for centuries to treat various allergic symptoms including asthma and bronchitis. Aim of the studyThe 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 methodsCell 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. ResultsHot 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. ConclusionHW 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 an ellagic acid glucoside, okicamelliaside, on antigen-mediated degranulation in rat basophilic leukemia RBL-2H3 cells and passive cutaneous anaphylaxis reaction in mice

Degranulation inhibitors in plants are widely used for prevention and treatment of immediate-type allergy. We previously isolated a new ellagic acid glucoside, okicamelliaside (OCS), from Camellia japonica leaves for use as a potent degranulation inhibitor. Crude extracts from leaves also suppressed allergic conjunctivitis in rats. In this study, we evaluated the in vivo effect of OCS using a pure sample and performed in vitro experiments to elucidate the mechanism underlying the extraordinary high potency of OCS and its aglycon. The IC50 values for degranulation of rat basophilic leukemia cells (RBL-2H3) were 14nM for OCS and 3μM for aglycon, indicating that the two compounds were approximately 2 to 3 orders of magnitude more potent than the anti-allergic drugs ketotifen fumarate, DSCG, and tranilast (0.17, 3, and >0.3mM, respectively). Antigen-induced calcium ion (Ca2+) elevation was significantly inhibited by OCS and aglycon at all concentrations tested (p<0.05). Upstream of the Ca2+ elevation in the principle signaling pathway, phosphorylation of Syk (Tyr525/526) and PLCγ-1 (Tyr783 and Ser1248) were inhibited by OCS and aglycon. In DNA microarray-screening test, OCS inhibited expression of proinflammatory cytokines [interleukin (IL)-4 and IL-13], cytokine-producing signaling factors, and prostaglandin-endoperoxidase 2, indicating that OCS broadly inhibits allergic inflammation. During passive cutaneous anaphylaxis in mice, OCS significantly inhibited vascular hyperpermeability by two administration routes: a single intraperitoneal injection at 10mg/kg and per os at 5mg/kg for 7days (p<0.05). These results suggest the potential for OCS to alleviate symptoms of immediate-type allergy.

Inhibitory Effects of Water-Soluble Low-Molecular-Weight β-(1,3-1,6)D-Glucan Isolated fromAureobasidium pullulans1A1 Strain Black Yeast on Mast Cell Degranulation and Passive Cutaneous Anaphylaxis

We investigated the effects of water-soluble low-molecular-weight β-(1,3-1,6) D-glucan isolated from Aureobasidium pullulans 1A1 strain black yeast (LMW-β-glucan) on mast cell-mediated anaphylactic reactions. Although it is known that LMW-β-glucan has anti-tumor, anti-metastatic and anti-stress effects, the roles of LMW-β-glucan in immediate-type allergic reactions have not been fully investigated. We examined whether LMW-β-glucan could inhibit mast cell degranulation and passive cutaneous anaphylaxis (PCA). LMW-β-glucan dose-dependently inhibited the degranulation of both rat basophilic leukemia (RBL-2H3) and cultured mast cells (CMCs) activated by calcium ionophore A23187 or IgE. However, LMW-β-glucan had no cytotoxicity towards RBL-2H3 cells and CMCs. Furthermore, orally administered LMW-β-glucan inhibited the IgE-induced PCA reaction in mice. These results show LMW-β-glucan to be a possible compound for the effective therapeutic treatment of allergic diseases.

Enhancing effects of trichloroethylene and tetrachloroethylene on type I allergic responses in mice

Trichloroethylene (TCE) and tetrachloroethylene (perchloroethylene; PCE) are commonly identified as environmental contaminants of groundwater. Previously, we investigated the enhancing effects of TCE and PCE on antigen-induced histamine release and inflammatory mediator production in rat mast cells. In this study, to examine the potential effect of TCE and PCE on antigen-induced histamine release from mouse mast cells, mouse bone marrow-derived mast cells (BMMC) were sensitized with anti-dinitrophenol (DNP) monoclonal IgE antibody and then stimulated with DNP-BSA containing with TCE or PCE. Both TCE and PCE significantly enhanced antigen-induced histamine release from BMMC. Next we investigated the effects of TCE and PCE on the passive cutaneous anaphylaxis (PCA) reaction in vivo using ICR mice. TCE and PCE significantly enhanced the PCA reaction in a dose-dependent manner. In addition, we examined the enhancing effects of ingesting small amount of TCE and PCE in drinking water on antigen-stimulated allergic responses. After the ICR mice had ingested TCE or PCE in their drinking water for 2 or 4 weeks, we performed the PCA reaction. Both TCE and PCE ingestion enhanced the PCA reaction in a dose-dependent manner for 4 weeks. These results suggest that exposure to TCE and PCE leads to the augmentation of type I allergic responses in many species.

노다케닌의 항 알러지 효과

현재 전 세계적으로 알러지 질환은 급격히 증가하고 있는 추세이다. 기존의 길항제나 항원 특이적 면역 치료법의 부작용으로 인하여 항 알러지 작용에 효과적인 천연 기능성 물질에 대한 연구가 알러지 질환 치료의 유망한 분야로 여겨지고 있다. 비만세포는 히스타민이나 아라키돈 산, 그리고 전 염증성 사이토카인 등을 방출하는 알러지 반응에 주요하게 작용하는 효과세포이다. 우리는 참당귀의 뿌리로부터 추출한 노다케닌의 알러지 반응 억제효과에 대해서 연구하였다. 노다케닌을 이용하여 비만세포에서의 탈 과립현상 억제 효과를 측정한 결과, 비만세포에서 노다케닌이 탈과립의 표지인 <TEX>${\beta}$</TEX>-hexosaminidase의 방출량을 농도 의존적으로 억제하는 것을 관찰하였고, 이러한 탈과립 억제 현상이 다양한 사이토카인 중 IL-4과 TNF-<TEX>${\alpha}$</TEX>의 양을 감소시킴으로써 매개된다는 사실을 RT-PCR을 통해 확인하였다. 또한 쥐에서 수동피부과민반응을 유도하여 노다케닌을 경구투여 했을 때 알러지성 과민반응 증상을 효과적으로 억제하는 것을 관찰하였다. 결과적으로, 이러한 결과들은 노다케닌이 항 알러지 약물의 개발에 잠재적인 후보물질이 될 수 있음을 시사한다. Mast cells are major effector cells associated with allergic responses. They are activated through the release of histamine, arachidonic acid, and proinflammatory cytokines. We investigated the effect of nodakenin, derived from the roots of Angelica gigas Nakai, on mast cell degranulation and on an allergic response in an animal model. We also investigated the effect of nodakenin on expression of multiple cytokines. Nodakenin suppressed the release of <TEX>${\beta}$</TEX>-hexosaminidase, a marker of degranulation, as well as the expression of interleukin IL-4 and TNF-<TEX>${\alpha}$</TEX> mRNA. Nodakenin inhibited the passive cutaneous anaphylaxis (PCA) reaction in ICR mice in a dose-dependent manner. These results suggest that nodakenin can inhibit mast cell degranulation through the inhibition of IL-4 and TNF-<TEX>${\alpha}$</TEX> mRNA expression, and that nodakenin may potentially serve as an anti-allergic agent.

Hot Water Extract of Adzuki (Vigna angularis) Suppresses Antigen‐Stimulated Degranulation in Rat Basophilic Leukemia RBL‐2H3 Cells and Passive Cutaneous Anaphylaxis Reaction in Mice

The hot water extract of adzuki (HWEA), which is produced as a byproduct in the adzuki bean boiling process, has anti-tumor, antioxidative, and anti-diabetic activities. In this study, we fractionated HWEA to 4 fractions using stepwise gradient column chromatography with water and ethanol, and demonstrated the effects of each fraction on antigen (Ag)-stimulated degranulation in rat basophilic leukemia RBL-2H3 cells. The 40% ethanol eluate extract (EtEx.40) showed the strongest inhibition level of these fractions. To reveal the inhibitory mechanisms underlying degranulation by EtEx.40, we investigated intracellular reactive oxygen species (ROS) production, intracellular free Ca²⁺ concentration ([Ca²⁺]i), and early intracellular signaling pathways. Treatment with EtEx.40 markedly inactivated Lyn following Ag stimulation, resulting in the suppressions of intracellular elevation of [Ca²⁺]i and production of ROS. To identify the active compound in EtEx.40, we isolated 7 flavonoids from EtEx.40 and calculated their inhibition levels on Ag-stimulated degranulation. These flavonoids inhibited degranulation by about 25-60%. We further examined the in vivo effects of HWEA or EtEx.40 using a passive cutaneous anaphylaxis (PCA) reaction. Both extracts strongly suppressed the PCA reaction. These findings suggest that HWEA and/or EtEx.40 are beneficial for alleviating type I allergic symptoms.

Inhibitory effect of Lactobacillus plantarum K-1 on passive cutaneous anaphylaxis reaction and scratching behavior in mice

Lactobacillus plantarum K-1 (LP) inhibiting AP-1 (c-Jun) and NF-κB activations was isolated from kimchi, and its inhibitory activity against scratching behavior and passive cutaneous anaphylaxis reaction in mice was investigated. Heat-inactivated LP (heated at 60°C for 30 min) potently inhibited the expression of TNF-α and IL-4 as well as the activation of their transcription factors, NF-κB and c-jun, in phorbol 12'-myristate 13'-acetate-stimulated RBL-2H3 cells. LP (1 × 10(10) CFU per mouse) showed a potent inhibition against passive cutaneous anaphylaxis reaction induced by the IgE-antigen complex in mice, inhibiting it by 87.5%. LP (1 × 10(10) CFU/mouse) inhibited histamine-induced scratching behavior by 58.9% compared to the control group. LP significantly inhibited vascular permeability induced by histamine. The inhibitory activity of LP against vascular permeability was in proportion to its inhibition against scratching behavior. LP potently inhibited histamine-induced cytokine production: it (1 × 10(10) CFU per mouse) inhibited IL-4, IL-1β, and TNF-α expression by 88.9%, 88.6%, and 98.9%, respectively. LP also inhibited IgE level increased by histamine by 85.3%. It inhibited histamine-induced the activations of their transcription factors, NF-κB and c-Jun. Based on these findings, LP may improve allergic diseases, such as anaphylaxis, atopic dermatitis, rhinitis, and pruritus by inhibiting the expression of IgE-switching cytokine IL-4 and proinflammatory cytokines IL-1β and TNF-α via NF-κB and AP-1 signaling pathways.

Pyeongwee-San extract (KMP6): a new anti-allergic effect

The prevalence of allergic diseases is increasing due to rapid industrialization and changes in lifestyle. Pyeongwee-San (KMP6) is a traditional Korean medicine that has been used as a basic prescription for digestive disorders. This study investigated the efficacy of KMP6 and its component hesperidin on experimental allergic models. The anti-allergic effect of KMP6 was studied against a compound 48/80-induced systemic anaphylactic reaction and the ear swelling response. In addition, a human mast cell line (HMC-1) was used to analyze the activity of histidine decarboxylase. Passive cutaneous anaphylaxis (PCA) from immunoglobulin E (IgE) was used. KMP6 and hesperidin inhibited the compound 48/80-induced systemic anaphylactic reaction and the ear swelling response as well as histamine release, intracellular calcium levels and tryptase release from rat peritoneal mast cells. KMP6 inhibited histidine decarboxylase activity in stimulated HMC-1 cells and macrophages. In addition, KMP6 inhibited the PCA reaction induced by IgE as well as the levels of IgE, interleukin (IL)-4, IL-5, IL-6 and IL-13 in serum from mice. These results suggest that KMP6 may exert an anti-allergic effect through not only the inhibition of mast cell degranulation but also the inhibition of histamine synthesis.

주박 에틸아세테이트 추출물의 항알러지 효과

Mast cells, the central effector cells involved in the allergic response, release histamine, arachidonic acid, and proinflammatory cytokines. We investigated the effect of the ethyl acetate fraction (EA), derived from Korean rice wine lees, on RBL-2H3 cell degranulation and passive cutaneous anaphylaxis in an animal model. The EA fraction suppressed the release of beta-hexosaminidase, a marker of degranulation, and the mRNA expression of interleukin-3 (IL-3) and IL-13. EA also successfully suppressed the passive cutaneous anaphylaxis (PCA) reaction in mice in a dose-dependent manner. These results suggest that EA can inhibit mast cell degranulation through the inhibition of IL-3 and IL-13 mRNA expression, and that EA may potentially serve as an anti-allergic agent.

Fritillaria ussuriensisExtract Inhibits the Production of Inflammatory Cytokine and MAPKs in Mast Cells

Fritillaria ussuriensis (FU, derived from the bulbs of various species of the genus Fritillaria, including Fritillaria thunbergii Miq.) is used in herbal medicine to treat conditions such as eczema, skin burns, and frostbite. In this study, we investigated the mechanism of the anti-allergy effect of FU. FU extract (80 mg/kg), orally administered to Sprague-Dawley (SD) rats, significantly inhibited the passive cutaneous anaphylaxis (PCA) reaction. It inhibited the compound 48/80-induced release of histamine from rat peritoneal mast cells in a concentration-dependent manner. Significant inhibitory effects of the FU extract on IL-6, IL-8, and TNF-α (1, 10, and 100 µg/mL) were observed in HMC-1 cells. Treatment with FU attenuated PMA plus A23187-induced phosphorylation of all three MAPKs, especially at concentrations of 10 and 100 µg/mL. Further, it (80 mg/kg) led to significant inhibition of mast-cell accumulation in ear tissue at the chronic phase. These results indicate that it inhibits allergic reactions.

The mechanism of the anti-inflammatory activity of salicylate.

AbstractSalicylate reduced the leakage of circulating dye in the reversed passive cutaneous anaphylaxis reaction in the guinea pig. 2,4-Dinitrophenol, although a more powerful uncoupling reagent than salicylate, had no effect compared to corresponding control animals. In this test in the guinea-pig, salicylate inhibits the increased capillary permeability due to cutaneous anaphylaxis and to histamine, but not to Miles-Wilhelm permeability factor. It is concluded that the effect of salicylate in reducing the increased capillary permeability produced in these reactions does not result from an uncoupling action on oxidative phosphorylation processes. The results suggest that salicylates act by preventing antigen-antibody combinations from exerting their effects on the capillary wall.

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

Cordyceps militaris has been used as a traditional herbal medicine for treating allergy in East Asia. We investigated the anti-allergic efficacy of Cordyceps militaris and its mechanism of action. 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. 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. Our study suggests that GSCME might be used as a therapeutic agent for allergic diseases.

Crucial role for autophagy in degranulation of mast cells

Autophagy plays a crucial role in controlling various biological responses including starvation, homeostatic turnover of long-lived proteins, and invasion of bacteria. However, a role for autophagy in development and/or function of mast cells is unknown. To investigate a role for autophagy in mast cells, we generated bone marrow-derived mast cells (BMMCs) from mice lacking autophagy related gene (Atg) 7, an essential enzyme for autophagy induction. Bone marrow-derived mast cells were generated from bone marrow cells of control and IFN-inducible Atg7-deficient mice, and morphologic and functional analyses were performed. We found that conversion of type I to type II light chain (LC3)-II, a hallmark of autophagy, was constitutively induced in mast cells under full nutrient conditions, and LC3-II localized in secretory granules of mast cells. Although deletion of Atg7 did not impair the development of BMMCs, Atg7(-/-) BMMCs showed severe impairment of degranulation, but not cytokine production on FcεRI cross-linking. Intriguingly, LC3-II but not LC3-I was co-localized with CD63, a secretory lysosomal marker, and was released extracellularly along with degranulation in Atg7(+/+) but not Atg7(-/-) BMMCs. Moreover, passive cutaneous anaphylaxis reactions were severely impaired in mast cell-deficient WBB6F1-W/W(V) mice reconstituted with Atg7(-/-) BMMCs compared with Atg7(+/+) BMMCs. These results suggest that autophagy is not essential for the development but plays a crucial role in degranulation of mast cells. Thus, autophagy might be a potential target to treat allergic diseases in which mast cells are critically involved.

Anti-allergic effects of Perilla frutescens var. acuta Kudo 30% ethanol extract powder

Perilla frutescens var. acuta Kudo (PF) is a traditional Korean medicinal herb for allergic reaction regulation. In the present study, we investigated the effect of 30% ethanol extract powder of PF (EPPF) and rosmarinic acid (RA), the active compound of EPPF on various allergic reactions using in vivo and in vitro models. EPPF and RA significantly inhibited compound 48/48-induced systemic anaphylactic reaction and histamine release (P < 0.05). In addition, EPPF and RA significantly inhibited passive cutaneous anaphylaxis (PCA) in a dose-dependent manner (P < 0.05). These effects were stronger than those of disodium cromoglycate, the reference drug tested. EPPF and RA also significantly inhibited production of inflammatory cytokines, tumor necrosis factor-a interleukin (IL)-6, and vascular endothelial growth factor on the PCA reaction and phorbol 12-myristate 13-acetate and calcium ionophore A23187-stimulated human mast cell line, HMC-1 cells (P < 0.05). Moreover, EPPF and RA showed an inhibitory effect on lipopolysaccharide (LPS)-induced IL-4 production from whole spleen cells. Finally, EPPF and RA significantly decreased IL-4-dependent IgE production by LPS-stimulated whole spleen cells (P < 0.05). In conclusion, these results indicate that EPPF has potent anti-allergic activities.

Tanshinones isolated from the rhizome of Salvia miltiorrhiza inhibit passive cutaneous anaphylaxis reaction in mice

The rhizome of Salvia miltiorrhiza Bunge (SM, family Labiatae), which contains tanshinones as main constituents, has been used as a cardiovascular and anti-inflammatory agent in Chinese medicine. This study aimed to elucidate anti-allergic effects of the root of Salvia miltiorrhiza Bunge (SM, family Labiatae) and its main constituents, tanshinones, against passive cutaneous anaphylaxis (PCA) reaction. PCA reaction was induced by IgE-antigen complex (IAC) in ICR mice. Protein expression of IL-4 and TNF-α in rat basophilic leukemia (RBL)-2H3 cells was performed by enzyme-linked immunosorbent assay and NF-κB and c-jun (AP-1) activation assayed by immunoblot. Tanshinones inhibited the PCA reaction and reduced IL-4 and TNF-α production in mice as well as in IAC-stimulated RBL-2H3 cells. Tanshinones also inhibited NF-κB and AP-1 activation in RBL-2H3 cells stimulated with IAC. Among tested tanshinones, tanshinone I exhibited the most potent inhibition, followed by 15,16-dihydrotanshinone I, tanshinone IIA and cryptotanshinone. SM and tanshinones may ameliorate the PCA reaction by inhibiting the allergic cytokines IL-4 and TNF-α via NF-κB and AP-1 pathways.

Inhibitory Effect of Farnesylthiosalicylic Acid on Mediators Release by Mast Cells: Preferential Inhibition of Prostaglandin D2 and Tumor Necrosis Factor-α Release

There is substantial evidence suggesting that the Ras inhibitor farnesylthiosalicylic acid (FTS) may modulate various aspects of immune function and inflammation in addition to its well known anti-cancer activity. In this regard, we have recently shown that FTS suppresses T lymphocyte-mediated immune responses. Mast cells (MC), the main effector cells in the elicitation of the allergic response, are known to secrete granule-associated mediators and to release prostaglandins and cytokines on FCεRI-cross-linking, thereby contributing to the pathogenesis of allergic diseases. We hypothesized that MC act as an additional target for FTS. In the present work we analyze the effects of FTS on MC degranulation, prostaglandin release, and cytokine release in vitro, and on the elicitation of IgE-mediated MC dependent cutaneous allergic inflammation in vivo. First we have established that FTS inhibited Ras activation in MC. Next, we have shown that FTS preferentially inhibited prostaglandin (PG) D(2) and tumor necrosis factor (TNF)-α release without having any significant effect on MC β-hexosaminidase secretion. In vivo administration of FTS inhibited the late phase of passive cutaneous anaphylaxis reaction. The time course of FTS-induced inhibition in vivo correlated with mediators release and not with degranulation. This data suggests that FTS may have an inhibitory effect on MC mediated allergic inflammation, and thus may be considered as a possible therapeutic modality.