KR101914656B1 - Composition for preventing or treating allergic diseases comprising scirpus planiculmis extract - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating allergic disease containing a Scirpus planiculmis extract, showing effectiveness in preventing or treating allergic disease associated with type 1 and mast cells including at least one disease selected from the group consisting of bronchial asthma, allergic rhinitis, urticaria, anaphylaxis, food allergies, atopic dermatitis, allergic non-hemolytic transfusion reactions, and allergies to drugs such as penicillin, ampicillin and insulin. The pharmaceutical composition of the present invention exhibits anti-allergic effects by reversibly inhibiting degranulation of mRNA of interleukin-4 (IL-4) and tumor necrosis factor (TNF)-alpha in antigen-stimulated mast cells.

Description

Technical Field [0001] The present invention relates to a composition for preventing or treating allergic diseases,

The present invention relates to novel scarps planiculmis, SP) relates to for the prevention and treatment of allergic disease, a composition comprising the extract, and more particularly to compositions for the prevention and treatment of type 1 and mast cell-related allergic diseases including sentinel extract the new seomma magnetic .

Immune responses in IgE-mediated hypersensitivity disorders, including asthma, atopic dermatitis and food allergies, have been extended to innocuous environmental substances. As a result, the number of patients with allergies has increased exponentially worldwide over the past several decades.

It is the mast cell that plays an essential role in various allergic diseases, especially IgE-mediated hypersensitivity reactions. Mast cells are predominantly located in tissues close to the external environment, such as skin, airways and small intestine. When the antigen (Ag) binds to the IgE-sensitized mast cell, the mast cell quickly expresses allergen mediators such as histamine and protease, as well as some of the tumor necrosis factor (TNF) -a and interleukin It releases an inflammatory cytokine and induces an allergic reaction.

Stimulation of FcεRIs by Ag on mast cells initially activates several Src family kinases (SFKs), including Lyn, Fyn and Fgr. The cytoplasmic Syk then binds to the immunoreceptor tyrosine-based activated motif (ITAM) of the FcεRIγ chain to activate and stimulate other important downstream signaling proteins, including LAT and phospholipase C (for degranulation of mast cells) PLC) < / RTI > Thus, activation of Syk is essential for secretion of histamine, allergenic cytokines and other allergen mediators in Ag-stimulated mast cells, and it can be shown that Syk can be an important therapeutic target protein for the treatment of allergic diseases .

Currently there are some treatments for allergic diseases, but this is not ideal. Drug treatments for the treatment of allergic diseases, such as steroids and antihistamines, can significantly alleviate the symptoms of allergic diseases, but have caused unwanted side effects. For example, antihistamines can induce sedation, dizziness and loss of concentration, and steroids can cause diabetes, bone weakness, poor wound healing, and poor resistance to infection. Thus, alternative and supplemental herbal medicines have recently been proposed as additional treatment options for allergic diseases, and currently eupatilin (Stillen < RTI ID = 0.0 >  Dong-A Pharmaceutical) and Joins (SK Chemical) are on the market for treatment of gastric ulcers and arthritis, respectively.

SP belongs to the Poales order and the Cyperaceae family, and is mainly grown in coastal wetlands. SP is widely found in Northeast Asia including Korea, Japan and China. The rhizome of the SP is called a "threonine" and has traditionally been used to treat bronchiectasis, dysmenorrhea, chest pain, and dyspepsia. However, their pharmacological effects and their mechanism of action, especially for allergic reactions, have not yet been studied.

Korean Patent Application No. 2015-0121694 (filed on March 23, 2017) Korean Registered Patent No. 10-1103958 (Registered on January 13, 2012) Korean Patent Application No. 2006-0114866 (filed on May 26, 2008)

D. D. < / RTI > Metcalfe, et al., Mekori, Physiological Reviews, vol. 77, no. 4, pp. 1033.1079,1997. M. Akdis, et al., Journal of Allergy and Clinical Immunology, vol. 119, no. 4, pp. 780-791, 2007. A. Yukselen, and S. G. Kendirli, World Journal of Clinical Cases, vol. 2, no. 12, pp. 859-865, 2014.

In order to avoid the side effects of drug treatment for allergic diseases, the present inventors have searched for herbicide candidates from 100 different natural plant extracts, of which the spermatozoa have an inhibitory effect on IgE-mediated allergic reactions in vitro and in vivo . Accordingly, it is an object of the present invention to provide a pharmaceutical composition for the prevention or treatment of an allergic disease including a Sadomasu sp. Extract.

The present invention provides a pharmaceutical composition for the prevention or treatment of an allergic disease comprising a Sosaum lance extract.

In one embodiment, the Soxhlet extract is selected from the group consisting of alcohols consisting of methanol, ethanol, isopropanol and butanol, ether, methyl acetate, ethyl acetate, acetone; It may be extracted with at least one solvent selected from the group consisting of hexane, glycerin, propylene glycol, 1,3-butylene glycol, diethyl ether, dichloromethane, sorbitol, methylene chloride and hot water.

In one embodiment, the dose of the Soxhlet extract in the pharmaceutical composition may be from 100 to 2000 mg / kg.

In one embodiment, the allergic disease may be a type 1 and an allergic disease associated with a mast cell.

In one embodiment, the allergic disease is selected from the group consisting of bronchial asthma, allergic rhinitis, urticaria, anaphylaxis, atopic dermatitis, food allergy, allergy-free blood transfusions and drug allergy ≪ RTI ID = 0.0 > and / or < / RTI >

In one embodiment, the prevention or treatment of the allergic disease is through the inhibition of degranulation inhibition, tumor necrosis factor alpha (TNF-a) production inhibition, and interleukin-4 (IL-4) Lt; / RTI >

In one embodiment, the pharmaceutical composition may further comprise at least one additive selected from the group consisting of a pharmaceutically acceptable carrier, filler, extender, binder, wetting agent, disintegrant, surfactant, diluent and excipient.

In one embodiment, the pharmaceutical composition is in the form of an oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol; External application; Suppositories, and sterile injection solutions.

The present invention also provides a food composition for the amelioration of an allergic disease, which comprises a Sodium Laurate Extract.

In one embodiment, the food composition is selected from the group consisting of excipients, diluents, preservatives, bactericides, antioxidants, coloring agents, coloring agents, bleaches, seasonings, sweeteners, flavoring agents, swelling agents, enhancers, improvers, emulsifiers, thickeners, stabilizers, An antifoaming agent, a solvent, a release agent, an insect repellent, and a quality improving agent.

The pharmaceutical composition comprising the extract of the present invention can inhibit IgE-mediated degranulation and cytokine production through inhibition of Syk and Syk-mediated signals in mast cells, thereby achieving prevention or treatment of allergic diseases have.

1, Figure 1A is a graph depicting degranulation inhibition of Ag-stimulated mast cells in SP (Figure IA) and its reversible effect (Figure IB). 2.0 × 10 ⁵ cells / tube of RBL-2H3 cells and bone marrow-derived mast cells BMMC were sensitized with 20 ng / ml DNP-specific IgE in 24-well plates for more than 12 hours (FIG. 1A). The cells were preincubated for 30 min with or without SP (10, 30 or 100 μg / ml) or PP2 (10 μM) and then stimulated with 25 ng / ml DNP-BSA for 10 min. RBL-2H3 cells were also preincubated for 30 minutes with or without SP (100 μg / ml) or PP2, and washed three times to recover the degranulation inhibition, indicating that the inhibitory effect was reversible (Fig. 1B). Cells were stimulated with 25 ng / ml DNP-BSA for 10 min and release of beta-hexosaminidase was measured at 450 nm. Values are expressed as mean ± SEM of three separate experiments. Significant differences compared to the Ag-alone group were * p <0.05 and ** p <0.01.
Figure 2 shows the inhibitory effect of SP on gene expression of TNF-a and IL-4. RBL-2H3 cells (1.0 × 10 ⁶ cells / well) were sensitized for 12 hours or more by 20ng / ㎖ DNP- specific IgE in six-well plates. The cells were preincubated for 30 min with or without SP (10, 30 or 100 μg / ml) or PP2 (10 μM) and then stimulated with 25 ng / ml DNP-BSA for 15 min. FIG. 2A shows RT-PCR results for observing gene expression of TNF-α and IL-4, and FIG. 2B shows a graph quantifying the degree of inhibition of gene expression. Densitometric values (mean ± SEM) for three separate experiments are shown. A significant difference compared to the Ag-alone group was indicated by * p < 0.05.
Fig. 3 shows the results of inhibition of phosphorylation of Syk and sub-molecules of SP in mast cells. 2.0 × 10 ⁶ cells / tube of RBL-2H3 cells and BMMC were sensitized by 6 ng / well plate with 20 ng / ml DNP-specific IgE for at least 12 hours. The cells were preincubated for 30 minutes with or without SP (10, 30 or 100 占 퐂 / ml) or PP2 (10 占)) and then stimulated with 25ng / ml DNP-BSA for 10 minutes. FIG. 3A shows Western blotting results for cell lysates, and FIG. 3B shows a graph quantifying the results.
Figure 4 shows the in vivo PCA response of SP and the inhibitory effect of degranulation on mast cells. Twelve hours after intravenous injection of 0.5 [mu] g DNP-specific IgE into each ear, 250 mg of DNP-BSA containing 5 mg / ml Evans blue was injected into the tail of the mice. SP (100, 300 or 1000 mg / kg) or cetirizine (CZ, 20 mg / kg) was orally administered one hour before DNP-BSA injection. One hour after injection of DNP-BSA, IgE-primed ears were cut out and the Evans blue dye content was measured at 620 nm. 4A is a photograph showing the ear, FIG. 4B is a quantification graph of the extracted Evans blue dye, FIG. 4C is a result of tissue staining of the ear, FIG. 4D is a graph showing the total number of mast cells and degranulation- This is a result. Values are expressed as mean ± SEM of three separate experiments. In the degranulation, the significant differences as compared to the Ag-alone group were shown as * p < 0.05 and ** p < 0.01.
Figure 5 is a schematic hypothesis for the mechanism of inhibition of mast cell activation of SP. According to this, phosphorylation of Syk by antigen stimulation activates the signals of the sub-molecules, for example, LAT, PLC-γ, which results in the movement of Ca ^{2 +} and MAP kinases. SP has mast cell inhibitory effect by inhibiting Syk activation.

The present invention provides a pharmaceutical composition for the prevention or treatment of allergic diseases including Scirpus planiculmis ( SP) extract.

In one embodiment, the extraction of the Soxhm material is carried out using an alcohol, ether, methyl acetate, ethyl acetate, acetone, methanol, ethanol, isopropanol and butanol; The solvent may be extracted with one or more solvents selected from the group consisting of hexane, glycerin, propylene glycol, 1,3-butylene glycol, diethyl ether, dichloromethane, sorbitol, methylene chloride and hot water, It can also be extracted by other solvents used for crude drug extraction. Preferably by methanol.

In one embodiment, the dose of the Soxhlet extract in the composition may be in the range of 100 to 2000 mg / kg, preferably 200 to 1000 mg / kg, but is not limited to, the symptoms of the allergic disease, And the like.

In one embodiment, the allergic disease refers to a disease caused by a reaction that occurs when an allergen is injected into a person having an IgE antibody. A wheal occurs at the injection site and erythema appears around it. The reaction reaches its peak at 15-30 minutes and disappears after about an hour. The allergies are also referred to as immediate allergies or anaphylaxis-type allergies since the time it takes for the symptoms to appear after contact with the antigen is short.

Before the antigen enters the body, IgE binds to the FcεRI receptor, an IgE-highly compatible receptor present on the surface of basophils and mast cells. However, when allergen binds to IgE antibodies, these cells degranulate and release chemical mediators, including histamine, leukotriene (LT). Their common physiological effects include airway constriction, capillary dilatation, capillary permeability hyperactivity, and airway mucus secretion hyperactivity. The biological response caused by this chemical is called type 1 allergy. An allergic antigen is not an infectious agent, but an immune response against substances that do not cause an immune response (eg, pollen or house dust mite). People with type 1 allergic disease show "hypersensitivity" to this.

In one embodiment, the allergic disease may be at least one disease selected from the group consisting of bronchial asthma, atopic dermatitis, allergic rhinitis, urticaria, anaphylaxis, food allergies, allergic non-hemolytic transfusions and drug allergies, It does not.

In one embodiment, the prevention or treatment of the allergic disease may be by inhibition of degranulation, tumor necrosis factor alpha (TNF-a) production inhibition and interleukin-4 (IL-4) production inhibition of mast cells.

In one embodiment, the pharmaceutical composition may further comprise at least one additive selected from the group consisting of a pharmaceutically acceptable carrier, filler, extender, binder, wetting agent, disintegrant, surfactant, diluent and excipient, But is not limited thereto. In addition, the pharmaceutically acceptable carrier may include saline, sterilized water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and the like.

In one embodiment, the pharmaceutical composition is in the form of an oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol; External application; But are not limited to, one or more formulations selected from the group comprising suppositories and sterile injectable solutions.

The present invention also provides a food composition for the amelioration of an allergic disease, which comprises a Sodium Laurate Extract.

The food composition of the present invention can be used as a health functional food. The term " functional food " as used herein refers to a food prepared and processed by using raw materials and ingredients having useful functions in the human body. The term " functional " For the purpose of obtaining a beneficial effect for health use.

In one embodiment, the food composition is selected from the group consisting of excipients, diluents, preservatives, bactericides, antioxidants, coloring agents, coloring agents, bleaches, seasonings, sweeteners, flavoring agents, swelling agents, enhancers, improvers, emulsifiers, thickeners, stabilizers, But are not limited to, at least one food additive selected from the group consisting of antifoaming agents, solvents, release agents, insect repellents, and quality improvers.

In one embodiment, the food composition may be prepared and processed into formulations such as, but not limited to, tablets, capsules, powders, granules, liquids,

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrating the present invention, and the scope of the present invention is not limited to these examples.

[Example]

&Lt; Materials and methods >

1. Reagents

A single clone dinitrophenol (DNP) -specific IgE antibody, DNP-bovine serum albumin (BSA), calcium ionomycin, Evans blue, cetirizine and toluidine blue, - purchased from Sigma-Aldrich. Antibodies for phosphorylation and protein expression detection of Syk, LAT, PLCγ1, ERK1 / 2, JNK, p38 and Akt were purchased from Cell Signaling Technology Inc. Cell culture reagents containing minimum essential medium (MEM) containing Earle's salts, Roswell Park Memorial Institute (RPMI) 1640 medium, and fetal bovine serum (FBS) were purchased from Gibco / Life Technologies Technologies Inc.).

2. Plant Extract

SP extract (sample number: 037-059) was collected from Wando-gun, Jeollanam-do on November 3, 2009 and was obtained from the Plant Extract Bank (extract.kribb.re.kr) of KRIBB. The sample was methanol-extracted from whole plants and then dissolved in dimethylsulfoxide (DMSO).

3. Animals

Five week old male BALB / c mice were obtained from the Orient Bio Experimental Animal Center. Isolation and in vivo of BMMC A mouse was used for the manual skin anaphylaxis (PCA) study. All animal studies were approved by the Institutional Animal Care and Use Committee (IACUC) and conducted in accordance with established guidelines.

4. Preparation of mast cell and cell culture

Rat basophilic leukemia (RBL) -2H3 cells were obtained from the American Type Culture Collection (ATCC) and grown in MEM supplemented with glutamine, antibiotics and 15% FBS. Bone marrow-derived mast cells (BMMC) from male BALB / c mice were injected intramuscularly with 4 mM L-glutamine, 100 units / ml penicillin, 100 μg / ml streptomycin, 0.1 mM non-essential amino acids, 1 mM sodium pyruvate, 25 mM HEPES, 10 Ml FBS, 0.05 mM beta-mercaptoethanol, and 10 ng / ml IL-3. The BMMC was used for in vitro testing 4-6 weeks later.

5. Measurement of? -Hexa minidase release in RBL-2H3 cells and BMMC

The secretion of β-hexa minidase was measured to examine the inhibitory effect on degranulation, which is an index of type 1 allergic reaction. Was treated over 12 hours all of RBL-2H3 cells, and are such that the BMMC 20ng / ㎖ DNP--specific IgE in a 24-well plate (2.0 × 10 ⁵ cells / well). The cells were washed twice and resuspended to a final volume of 0.2 ml buffer per well. The buffer used in this experiment was 1,4-piperazine diethanesulfonic acid (PIPES) -buffered medium (25 mM PIPES, pH 7.2, 119 mM NaCl, 5 mM KCl, 0.4 mM MgCl2 for RBL- _{2, 1 mM CaCl 2, 5.6} mM glucose and 0.1% fatty acid-included FRACTION V bovine serum albumin (BSA)); And 20 mM Tyrode buffer (20 mM HEPES, pH 7.4, 135 mM NaCl, 5 mM KCl, 1.8 mM CaCl ₂ , 1 mM MgCl ₂ , 5.6 mM glucose and 0.1% BSA) for BMMC. Cells were pretreated with SP (10, 30 or 100 μg / ml) or PP2 (10 μM) for 30 min and stimulated with 25 ng / ml DNP-BSA for 10 min. Degranulation of mast cells was determined by measuring the ratio of β-hexa minidase released in the supernatant to total β-hexa minidase in buffer and cell eluate.

6. Western blot analysis

Cells were stimulated with 25 ng / ml Ag at the indicated time and cooled on ice to stop stimulation. Total protein extracts were obtained from Lee et al . Blood , vol. 108, no. 3, pp. 956-964, 2006. Briefly, cells were resuspended in 100 용 lysis buffer (20 mM HEPES, pH 7.5, 150 mM NaCl, 1% Nonidet P-40, 10% glycerol, 60 mM octyl β- 10 mM NaF, 1 mM Na ₃ VO ₄ , 1 mM phenylmethylsulfonyl fluoride, 2.5 mM p-nitrophenyl phosphate, 0.7 mg / ml pepstatin, and protease-inhibitor cocktail tablets). Protein content in cell extracts was quantified by the Bradford assay (absorbance at 595 nm). The eluate was denatured at 100 &lt; 0 &gt; C for 5 minutes in 4x Laemmli buffer. For Western blotting, proteins were separated using sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene fluoride membrane (Bio-Rad Laboratories) . The membrane was blocked in 5% BSA-containing TBS-T buffer for 1 hour and then incubated with the primary antibody. The membrane was washed and incubated with a horseradish peroxidase-labeled secondary antibody. These blots were processed using an ImageQuant (TM) LAS 4000 system (GE Healthcare Life Sciences).

Reverse transcriptase-polymerase chain reaction (RT-PCR)

Total RNA was extracted from RBL-2H3 cells using an Easy-spin ^TM total RNA extraction kit (iNtRON Biotechnology branch), and the RNA was extracted from the RBL-2H3 cells according to the manufacturer's protocol using a SuperScript first-strand synthesis system (Invitrogen) Respectively. RT-PCR was carried out at 94 ° C for 45 seconds, 55 ° C for 45 seconds, and 72 ° C for 60 seconds for 30 cycles. The primers used were as follows:

Rat TNF-α Forward: 5-CACCACGCTCTTCTGTCTACTGAAC-3;

Rat TNF-alpha reverse direction: 5-CCGGACTCCGTGATGTCTAAGTACT-3;

Rat IL-4 Forward: 5-ACCTTGCTGTCACCCTGTTC-3;

Rat IL-4 reverse direction: 5-TTGTGAGCGTGGACTCATTC-3;

Rat GAPDH (glyceraldehyde-3-phosphate dehydrogenase) Forward direction: 5-GTGGAGTCTACTGGCGTCTTC-3;

Rat GAPDH Reverse direction: 5-CCAAGGCTGTGGGCAAGGTCA-3.

8. Manual Skin Anaphylaxis (PCA)

The PCA test procedure is described in JH Lee, et al., Clinical and Experimental Allergy , vol. 38, no. 5, pp. 794-804, 2008. &lt; / RTI &gt; Briefly, 0.5 [mu] g of DNP-specific IgE was injected intradermally into the ear of the mouse. After 24 hours, either SP (100, 300 or 1000 mg / kg) or cetirizine (20 mg / kg) was orally administered to mice. One hour later, mice were treated intravenously with 250 mg antigen (DNP-BSA) in 250 ml PBS containing 4% Evans blue dye (Sigma-Aldrich). One hour later, the mice were anesthetized and the ears were cut out to determine the amount of dye present. The ear dyes were extracted by treating each mouse ear at 63 째 C for more than 12 hours in 700 쨉 l of formamide. Absorbance was measured at 620 nm by a microplate reader (Tencana).

9. Statistical Analysis

Data were expressed as mean ± SEM from three or more experiments. Statistical analysis was performed using the one-place ANOVA and the unaffected Student t-test. All statistical calculations were performed using Sigma Stat software (Systat Software, Inc.) and statistically significant at p <0.05.

<Result>

1. Inhibitory effect of SP on degranulation in Ag-stimulated RBL-2H3 cells and BMMC

(1) The stimulation of high affinity IgE receptors (FcεRIs) by IgE and Ag induces an allergic response by releasing allergenic mediators from the cytoplasmic granules of mast cells. The inventors have screened 100 different plant extracts for use in the degranulation test of mast cells to find herbal ant-allergenic drug candidates. In the screening process, the present inventors found that the outpost extract of SP showed a maximal inhibitory effect (about 94%) on mast cell degranulation (see Tables 1 to 4).

(2) To further test the inhibitory effect of SP on granule release of activated mast cells, we used two mast cells, RBL-2H3 and BMMC. It was observed by SP that degranulation from RBL-2H3 cells and BMMC was inhibited in a concentration-dependent manner and had IC ₅₀ values of 17.5 and 23.5 μg / ml, respectively (FIG. 1A). In addition, the inhibitory effect of SP on the degranulation of mast cells disappeared by washing cells after treatment, indicating that the inhibitory effect of SP is reversible (Fig. 1B).

2. Effect of SP on cytokine production in mast cells

(1) Activation of mast cells increases the secretion of cytokines IL-4 and TNF-α, which is very important for late allergic reactions. Thus, we investigated the effect of SP on the inhibition of Fc [epsilon] RI-mediated inflammatory cytokine production in Ag-stimulated mast cells.

(2) Treatment with SP significantly inhibited the production of Ag-induced TNF-a and IL-4 in mast cells in a concentration-dependent manner (Figure 2A). It was confirmed that the effect of SP at 100 占 퐂 / ml was equivalent to the effect of PP2, which is a common Src family kinase inhibitor, by approximately 13-fold reduction in expression of TNF-α and 5-fold in IL-4 expression ).

3. Effect of SP on activation of Syk, LAT and lower signal proteins in Ag-stimulated mast cells

(1) The present inventors investigated how SP inhibits the activation of mast cells. The aggregation of receptors by Ag activates Syk kinase, a pivotal signaling protein for mast cell activation, which in turn activates a number of lower signaling proteins. From the results of the present invention, it was found that SP significantly inhibited activation of Syk, LAT, PLCγ1 and Akt in Ag stimulated RBL-2H3 mast cells and BMMC. MAP kinase (e.g., ERK1 / 2, JNK and p38) plays an important role in the production of inflammatory cytokines including TNF-a and IL-4 (see FIG. 5) And p38 in a concentration-dependent manner (Fig. 3).

(2) These results indicate that SP inhibits activation of mast cells through activation of the Syk and Syk-dependent sub-signaling proteins LAT, PLCγ1, Akt and MAP kinase.

4. Effect of SP on Ag-induced manual skin anaphylaxis (PCA) in mice

(1) PCA is one of the most frequently used animal models to assess anti-allergic activity in vivo. In this method, since the antibody binds to the mast cell of the biotissue and the antigen binds to the antibody to expand the vascular permeability when injected into the animal and then the corresponding antigen and the dye are intravenously injected into the animal, And the reaction of the pigment to gather. The degree of allergic inhibition can be evaluated by the extent to which the blue pigment of Evans Blue is lightened.

(2) In the present invention, the PCA reaction is inhibited by SP in a dose-dependent manner (FIGS. 4A and 4B). At a dose of 1,000 mg / kg, the inhibitory effect of SP is similar to that of cetirizine (20 mg / kg), which is an anti-allergic drug, and SP can also be expected to be used as an anti-allergic herbal medicine .

(3) Tissue staining also showed that SP inhibited the degranulation of mast cells in the ear tissue of Ag-induced PCA mice (Fig. 4C and 4D).

<Discussion>

IgE-dependent activation of mast cells plays a key role in the allergic response. When stimulated by Ag, mast cells rapidly release a chemical mediator including histamine, tryptase, chemokine and proinflammatory cytokine stored in cytoplasmic granules to induce an allergic reaction. The results of the present invention show that SP inhibits Ag-stimulated degranulation of mast cells in vitro and in vivo, and inhibits allergic responses in PCA mice. In addition, it was observed that inhibition of mast cell degranulation by SP was reversible. These results suggest that SP may be a good candidate for anti-allergy medicines.

When mast cells are stimulated by Ag, Syk is first activated after activation of Src family kinase (SFK), followed by activation of LAT, PI ₃ K / Akt and MAP kinase. The cells in Ca ^{+ 2} is moving, it stimulates the degranulation and the release of other allergic mediators, including the cytokines and eicosanoids (Fig. 5). Thus, Syk inhibitors may be a major strategy for allergic diseases. In the present invention, SP can inhibit Syk activation in both RBL-2H3 cells and BMMC (FIGS. 4A and 4B), suggesting that the anti-allergic effect of SP inhibits Ag-induced activation of Syk in mast cells It can be deduced that it is mediated.

MAP kinases such as ERK, JNK and p38 play an important role in the production and synthesis of inflammatory cytokines, and these inflammatory cytokines, such as TNF-a and IL-4, are important for the development of allergic inflammation. SP inhibits the expression levels of TNF-? And IL-4 (FIG. 2), and the activation of three MAP kinases, ERK1 / 2, JNK and p38 in a dose-dependent manner (FIG. 3). These results also indicate that SP inhibits the activation of MAP kinase, thereby inhibiting the cytokine-mediated allergic response to the development of allergic diseases.

In the human body, when an allergic antigen enters the body, B cells produce IgE in the presence of atopic disease. IgE then binds to FcεRI receptors on mast cells. The same allergen will then bind to mast cells, leading to the release of granules and allergy mediators. PCA has been used as a conventional model for IgE-mediated type 1 hypersensitivity reactions in mice. As a result of the present invention, it can be inferred that SP effectively inhibits the PCA response in a dose-dependent manner (Fig. 4), from which SP inhibits mast cell degranulation in vivo and thus has an inhibitory effect on allergic reactions there was. In summary, the inventors first observed that SP inhibits mast cell activation and the mechanism of action of SP by Ag in vitro and in vivo. SP inhibited the production of mast cell degranulation and pro-inflammatory cytokines (i.e., TNF-a and IL-4) by inhibiting the activation of Syk and Syk-mediated signal transduction (Fig. 5). Thus, SP can be usefully used in IgE-mediated allergic diseases. However, further studies on toxicity and active ingredients of SP will be needed.

<Conclusion>

The present invention first demonstrated the anti-allergic activity of SP in vitro and in vivo. SP is believed to inhibit IgE-mediated degranulation and cytokine production through inhibition of Syk and Syk-mediated signal transduction in mast cells, and it is anticipated that an anti-allergic herbal drug using SP extract will be developed in the future .

Claims (10)

Saeseom maejagi (Scirpus The present invention relates to a pharmaceutical composition for preventing or treating an allergic disease, The method according to claim 1,
Wherein the Sadomasu extract comprises an alcohol comprising methanol, ethanol, isopropanol and butanol; At least one solvent selected from the group consisting of ethers including diethyl ether, methyl acetate, ethyl acetate, acetone, hexane, glycerin, propylene glycol, 1,3-butylene glycol, dichloromethane, sorbitol, methylene chloride, . &Lt; / RTI &gt; The method according to claim 1,
Wherein the dose of the extract is from 100 to 2000 mg / kg. The method according to claim 1,
Wherein said allergic disease is type 1 or an allergic disease related to mast cell. 5. The method of claim 4,
Wherein said allergic disease is selected from the group consisting of bronchial asthma, allergic rhinitis, urticaria, anaphylaxis, food allergy, atopic dermatitis, allergy-free blood transfusions and drug allergies. Lt; RTI ID = 0.0 &gt; 1, &lt; / RTI &gt; The method according to claim 1,
Wherein the prevention or treatment of the allergic disease is caused by inhibition of degranulation of the mast cell, inhibition of tumor necrosis factor alpha (TNF-a) production and inhibition of interleukin-4 (IL-4) production. The method according to claim 1,
Wherein the pharmaceutical composition further comprises at least one additive selected from the group consisting of pharmaceutically acceptable carriers, fillers, extenders, binders, wetting agents, disintegrants, surfactants, diluents and excipients . The method according to claim 1,
Oral formulations comprising powders, granules, tablets, capsules, suspensions, emulsions, syrups or aerosols; External application; &Lt; / RTI &gt; wherein the composition is at least one selected from the group consisting of suppositories and sterile injectable solutions. A food composition for the alleviation of allergic diseases, which comprises an extract of Sadomasu sp. 10. The method of claim 9,
A thickener, a thickener, a preservative, a bactericide, an antioxidant, a coloring agent, a bleaching agent, a seasoning, a sweetener, an flavoring agent, an expanding agent, an enhancer, an emulsifier, a thickener, a stabilizer, a coagulant, a gum base agent, a defoaming agent, &Lt; / RTI &gt; and at least one food additive selected from the group consisting of at least one food additive.

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