Egg allergy inhibiting composition

Abstract

To provide a composition capable of inhibiting the onset of an egg allergy with a specific effect of cannabidiol on which attention is focused.SOLUTION: An egg allergy inhibiting composition contains cannabidiol. The egg allergy inhibiting composition is preferably a health food, a special-use food, a specified health food, a food with nutrient function claims, a functional food, a nutritional supplement, a health supplement, a supplement, or a pharmaceutical. The cannabidiol preferably suppresses production of mast cell protease-1 by an intestinal mucosal mast cell. The cannabidiol preferably suppresses expression of interleukin-4 in a small intestine and a large intestine. The cannabidiol preferably suppresses expression of mRNA of SOCS-3 (suppressor of cytokine signaling 3) in the small intestine and the large intestine.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] The present invention relates to a composition for suppressing egg allergy. [Background technology]

[0002] Food allergy, which is one of the allergic diseases, is caused by foods such as chicken eggs and milk, and various symptoms appear throughout the body, such as skin symptoms such as urticaria and itching, gastrointestinal symptoms such as vomiting and diarrhea, and respiratory symptoms such as coughing and difficulty breathing. Food allergies often occur at a young age, and infants up to about 5 years old account for about 80% of all cases (Non-Patent Documents 1 and 2). It has been reported that if a person develops food allergy in infancy, the risk of developing bronchial asthma, allergic rhinitis, allergic dermatitis, etc. as they grow up, which is called the "allergic march," increases (Non-Patent Document 3). Therefore, preventing the onset of food allergy is one of the important clinical issues.

[0003] So far, various efforts have been made to prevent food allergies. For example, in order to analyze the possibility of developing food allergies due to sensitization by antigens transferred from the mother during fetal development or lactation, a test was conducted in which pregnant or lactating mothers avoided ingesting allergens such as chicken eggs and milk. However, even if pregnant or lactating mothers avoid allergen inoculation, no effect of preventing allergies in the newborn child was observed (Non-Patent Document 4). It has also been reported that delaying the intake of chicken eggs by three months during infancy increases the risk of developing chicken egg allergy (odds ratio 4.4) (Non-Patent Document 5). Furthermore, it has also been reported that infants who first ingested artificial milk after two weeks of age have a higher risk of developing milk allergy (odds ratio 19.30) compared to infants who ingested artificial milk within two weeks of birth (Non-Patent Document 6).

[0004] On the other hand, for example, Patent Document 1 discloses a therapeutic agent for allergies that contains a cannabinoid receptor modulator as an active ingredient, and Patent Document 4 discloses a therapeutic agent for non-immediate type allergies that contains a cannabinoid peripheral cell type receptor inverse agonist as an active ingredient. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Sugita K (Akdis CA.), “Recent developments and advances in atopic dermatitis and food allergy.”, Allergol.int., 69, 204-214 (2020). [Non-Patent Document 2] Peters RL, Krawiec M, Koplin JJ, “Update on food allergy.”, Pediatr. allergy Immunol., 32, 647-657 (2021). [Non-Patent Document 3] Yang L, Fu J, Zhou, “Research progress in atopic march.”, Front. Immunol., 11, 1907 (2020). [Non-Patent Document 4] Kramer MS, Kakuma R, “Optimal duration of exclusive breastfeeding.”, Cochrane Database Syst.Rev., 2012, CD003517(2012) [Non-Patent Document 5] Koplin JJ, et al., “Can early introduction of egg allergy in infants? A population-based study.”, J. allergy Clin. Immunol., 126, 807-813 (2010). [Non-Patent Document 6] Katz Y, et al. 6 others, “Early exposure to cow's milk protein is protective against IgE-mediated cow's milk protein allergy.”, J. allergy Clin. Immunol., 126, 77-82 e71 (2010). [Patent documents]

[0006] [Patent Document 1] JP 2003-252796 A [Patent Document 2] JP 2005-15422 A Summary of the Invention [Problem to be solved by the invention]

[0007] However, the techniques described in Patent Documents 1 and 2 relate to allergies such as asthma and atopic dermatitis, and no consideration was given to allergies derived from chicken eggs. Although much research has been conducted on the prevention of food allergies, food allergies, particularly allergies derived from chicken eggs, have not been sufficiently suppressed.

[0008] Therefore, an object of the present invention is to solve the above-mentioned problems of the conventional technology and to provide a composition capable of suppressing the onset of hen's egg allergy by focusing on the specific effect of cannabidiol. [Means for solving the problem]

[0009] As a result of intensive research aimed at solving the above problems, the present inventors discovered that the above object can be achieved by utilizing the specific functions of cannabidiol itself, and thus completed the present invention.

[0010] That is, the composition for suppressing egg allergy of the present invention is characterized by containing cannabidiol.

[0011] In addition, in the chicken egg allergy suppressing composition of the present invention, it is preferable that the cannabidiol suppresses the production of mast cell protease-1 from intestinal mucosal mast cells, and it is preferable that the cannabidiol suppresses the expression of interleukin-4 in the small intestine and large intestine.

[0012] Furthermore, in the chicken egg allergy suppressing composition of the present invention, it is preferable that the cannabidiol suppresses the expression of SOCS-3 (suppressor of cytokine signaling 3) mRNA in the small intestine and large intestine, and it is preferable that the cannabidiol suppresses the infiltration of Th2 cells into the large intestine mucosa.

[0013] Furthermore, the composition for suppressing egg allergy of the present invention is preferably a health food, a food for special dietary uses, a food for specified health uses, a food with nutrient function claims, a functional food, a nutritional supplement, a health supplement, a supplement or a medicine. Effect of the Invention

[0014] According to the present invention, a composition capable of suppressing the onset of hen's egg allergy can be provided. [Brief description of the drawings]

[0015] [Figure 1] FIG. 1 shows the administration schedule for Experimental Example 1-1. [Diagram 2] 1 is a graph showing the results of fecal moisture content in Experimental Example 1-1. [Diagram 3] 1 is a graph showing the results of body weight in Experimental Example 1-1. [Figure 4] FIG. 1 shows the administration schedule for Experimental Example 1-2. [Diagram 5] 1 is a graph showing the results of the effect of administration period in Experimental Example 1-2. [Figure 6] 1 is a graph showing the results of Mcpt-1 concentration in Experimental Example 2-1. [Figure 7] 1 is a graph showing the results of IgE antibody concentration in Experimental Example 2-1. [Figure 8] 1 is a graph showing the results of the expression level of IL-4 mRNA in the small intestine in Experimental Example 2-2. [Figure 9] 1 is a graph showing the results of the expression level of IL-4 mRNA in the large intestine in Experimental Example 2-2. [Figure 10] 1 is a graph showing the results of the expression level of SOCS-3 mRNA in the small intestine in Experimental Example 2-3. [Figure 11] 1 is a graph showing the results of the expression level of SOCS-3 mRNA in the large intestine in Experimental Example 2-3. [Figure 12] 1 is a graph showing the results of blood CCL2 concentration in the large intestine in Experimental Example 2-3. [Figure 13] 1 is a graph showing the effect of CBD on the infiltration of CD4-positive cells in the large intestine in Experimental Example 2-4. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] The composition for suppressing egg allergy of the present invention will be specifically described below. The composition for suppressing chicken egg allergy of the present invention is characterized by containing cannabidiol (hereinafter also referred to as "CBD"). Foods causing food allergies include chicken eggs, milk, wheat, nuts, etc., and among these, chicken egg allergy is the most common. Chicken egg allergens are proteins contained in egg white, and representative ones include ovalbumin (OVA) (hereinafter also referred to as "OVA"), ovotransferrin, ovomucoid, lysozyme, etc. Among them, ovalbumin (OVA) accounts for 54% of the proteins contained in egg white and contributes greatly to the onset of allergy. The composition for suppressing chicken egg allergy of the present invention contains cannabidiol, which can suppress the onset of chicken egg allergy.

[0017] Here, cannabidiol is a type of cannabinoid, a polyphenol contained in hemp, and is expressed as "CBD." In the present invention, there is no limitation as long as it can be used for normal consumption, but for example, "CBD crystalline powder" from PharmaHemp Laboratories (Ljubljana, Slovenia) can be used.

[0018] In addition, in the composition for suppressing egg allergy of the present invention, the cannabidiol preferably suppresses the production of mast cell protease-1 (hereinafter also referred to as "Mcpt-1") from intestinal mucosal mast cells. Such mast cell protease-1 is the only chymase expressed in intestinal mucosal mast cells found in the intestinal epithelium.

[0019] Furthermore, in the composition for suppressing egg allergy of the present invention, the cannabidiol preferably suppresses the expression of interleukin-4 in the small intestine and large intestine. Interleukin-4 is a type of cytokine, a protein composed of 129 amino acids, and is produced by mast cells and the like.

[0020] Furthermore, in the composition for suppressing egg allergy of the present invention, the cannabidiol preferably suppresses the expression of mRNA of SOCS-3 (suppressor of cytokine signaling 3) in the small intestine and large intestine. SOCS-3 is a negative feedback factor of the JAK-STAT pathway, which is essential for maintaining homeostasis in the body, such as immune response.

[0021] In addition, in the composition for suppressing egg allergy of the present invention, the cannabidiol preferably suppresses the infiltration of Th2 cells into the mucosa of the large intestine. Such Th2 cells are a type of helper T (Th) cells, which provide helper functions to antigen-presenting cells (APCs) such as macrophages, dendritic cells, and B cells that are responsible for the immune system, and play an important role in their activation and maturation processes.

[0022] In addition, the composition for suppressing egg allergy of the present invention is preferably a health food, a food for special dietary uses, a food for specified health uses, a food with nutrient functions, a functional food, a nutritional supplement, a dietary supplement, a supplement, or a medicine. By orally taking a composition containing cannabidiol, the onset of egg allergy can be suppressed.

[0023] In the present invention, health foods refer to all foods that are advertised as being good for health, and special purpose foods (excluding foods for specified health uses) are foods that are labeled with a special purpose, such as being suitable for the development of infants, or for maintaining or restoring the health of pregnant women, lactating women, people with swallowing difficulties, and sick people, and foods for specified health uses are foods that contain components with health benefits (involving components) that affect the physiological functions of the body, and are labeled (labeled for health uses) to indicate that the intake of such foods is expected to achieve a specific health purpose, and functional foods are foods that are used to supplement specific nutritional components. This refers to foods that are used for a variety of purposes and display the functions of their nutritional ingredients. The target foods are general-purpose processed foods and general-purpose fresh foods that are packed in containers and sold to consumers. Functional foods are foods that focus on the tertiary function of foods (physical conditioning) and tout their functionality. Dietary supplements are foods that supply nutritional ingredients or contribute to specific health uses. Dietary supplements are foods that supply nutritional ingredients or are suitable for specific health uses, or other foods taken to maintain or improve health and for health management. Supplements are health foods that are mainly in the form of tablets or capsules in which specific ingredients are concentrated, such as dietary supplements in the United States, and also include snack foods and beverages.

[0024] The egg allergy suppressant composition of the present invention is mainly in the form of tablets or capsules, but it may also be in the form of various beverages, various dairy products such as yogurt, cheese, butter, and lactic acid bacteria fermentation products, liquid foods, jellies, candies, retort foods, tablet sweets, cookies, castella cakes, bread, biscuits, etc.

[0025] Furthermore, in the present invention, other ingredients can be appropriately added within a range that does not impair the effects of the present invention. Any ingredient other than those mentioned above can be blended within a qualitative and quantitative range, and ingredients that are usually blended in foods, such as food additives, oily ingredients, emulsifiers, antioxidants, flavorings, various vitamins, coloring agents, medicinal ingredients, mineral ingredients, etc., can be blended.

[0026] In the present invention, the composition for suppressing egg allergy can be produced by a method for producing a general food product.

[0027] The present invention will be described in more detail below using examples, but the present invention is not limited to these examples. In the following "Examples", the blending percentage indicates mass %. In the figure, "CBD" indicates cannabidiol, % indicates blending mass %, and "control" and "cont" indicate controls. EXAMPLES

[0028] (Experimental Example 1) (reagent) Cannabidiol (CBD) was used as CBD crystalline powder (Batch No. CB099020171A) purchased from PharmaHemp Laboratories (Ljubljana, Slovenia). This powder contains 99.34% by mass of cannabidiol. Ovalbumin (OVA) and aluminum gel were purchased from Sigma Aldrich (St. Louis, Missouri, USA). The other reagents used were the most pure commercially available products.

[0029] (Animals used) Six-week-old male BALB / c mice were purchased from Japan SLC Co., Ltd. (Hamamatsu, Shizuoka Prefecture). The mice were kept in a facility with a temperature of 24±1°C and humidity of 55±5%, and were kept under the following light / dark conditions: lights on at 8:00 a.m., lights off at 8:00 p.m. In conducting this experiment, we followed the Hoshi University Animal Experiment Guidelines, which were established with the aim of promoting the implementation of appropriate animal experiments from the standpoint of animal welfare as well as science, and obtained approval from the Hoshi University Animal Experiment Committee, and gave full consideration to the ethical aspects of animals.

[0030] (Mouse treatment) Food allergy model mice exhibiting gastrointestinal symptoms were prepared according to the method of Yamamoto et al. (Yamamoto T, et al., “Oral tolerance induced by transfer of food antigens via breast milk of allergic mothers prevents offspring from developing allergic symptoms in a mouse food allergy model.”, Clin. Dev. Immunol., 2012, 721085 (2012).).

[0031] (Experimental Example 1-1) Effect of cannabidiol (CBD) on ovalbumin (OVA)-induced gastrointestinal allergy model mice (hereinafter referred to as "OVA-induced gastrointestinal allergy model mice") FIG. 1 shows the administration schedule of Experimental Example 1-1. Mice were divided into four groups: a control group (hereinafter referred to as the "control group"), an ovalbumin (OVA) group (hereinafter referred to as the "OVA group"), an OVA + cannabidiol (CBD) low-dose group (hereinafter referred to as the "CBD low-dose group"), and an OVA + cannabidiol (CBD) high-dose group (hereinafter referred to as the "CBD high-dose group"). 0.2 mL of OVA (50 μg / mL)·phosphate-buffered saline (PBS) solution containing aluminum hydroxide gel (final concentration: 1 mg / mL) as an adjuvant was intraperitoneally administered to the OVA group, the CBD low-dose group, and the CBD high-dose group. The same treatment was repeated two weeks later, and sensitization was performed twice in total. Two weeks after the final sensitization, the OVA group, the CBD low-dose group, and the CBD high-dose group were orally administered 0.25 mL of OVA (200 mg / mL) aqueous solution once every two days for a total of four times. The control group was administered PBS during the sensitization period, and purified water during the elicitation period. The control group and OVA group were administered a normal sample, the CBD low-dose group was administered a sample containing 0.01% CBD (25 mg / kg / day), and the CBD high-dose group was administered a sample containing 0.02% CBD (50 mg / kg / day) in their feed from the first day of sensitization to the last day of the experiment. The effect of CBD on the OVA-induced gastrointestinal allergy model mice was evaluated by measuring the amount of water in the feces during the experimental period and the body weight on the last day of the experiment.

[0032] (Calculation of water content in feces) The feces excreted within 1 hour of oral administration of OVA were collected and dried for 24 hours using a freeze-dryer FDM-1000 (product name) manufactured by Tokyo Rikakikai Co., Ltd., and then the fecal mass was calculated using the following formula: Moisture content in feces (mass%) = (mass of feces before drying - mass of feces after drying) / mass of feces after drying × 100 The water content (mass%) in the feces was calculated based on this.

[0033] (Statistical significance test) Experimental values ​​were expressed as mean ± standard error (SE). Statistical significance was tested using Tukey's test, and differences were considered significant when the risk level was less than 5% (p<0.05).

[0034] CBD (25mg / kg / day or 50mg / kg / day) was administered to OVA-induced gastrointestinal allergy model mice in the feed from the first day of sensitization, and the degree of diarrhea was evaluated. Figure 2 is a graph showing the results of fecal moisture content in Experimental Example 1-1, and Figure 3 is a graph showing the results of body weight in Experimental Example 1-1. The fecal moisture content of the OVA group began to increase from the fourth day of the induction phase, and on the sixth day it was significantly higher than the control group by about two times, and the onset of diarrhea was observed. The fecal moisture content when treated with a low dose of CBD (25mg / kg / day) was similar to that of the OVA group, and no improvement in diarrhea was observed. On the other hand, the fecal moisture content when treated with a high dose of CBD (50mg / kg / day) was significantly lower than that of the OVA group, diarrhea was improved, and OVA-induced gastrointestinal allergy symptoms could be suppressed. There was no significant difference in body weight when OVA and CBD were administered.

[0035] (Experimental Example 1-2) A study of the effect of cannabidiol (CBD) administration duration on ovalbumin (OVA)-induced gastrointestinal allergy model mice Figure 4 shows the administration schedule of Experimental Example 1-2. Mice were divided into 4 groups: a control group (hereinafter referred to as the "control group"), an ovalbumin (OVA) group (hereinafter referred to as the "OVA group"), an OVA + cannabidiol (CBD) entire period group (hereinafter referred to as the "CBD entire period group"), and an OVA + cannabidiol (CBD) induction phase group (hereinafter referred to as the "CBD induction phase group"). 0.2 mL of OVA (50 μg / mL)·phosphate-buffered saline (PBS) solution containing aluminum hydroxide gel (final concentration: 1 mg / mL) as an adjuvant was intraperitoneally administered to the OVA group, the CBD entire period group, and the CBD induction phase group. Two weeks later, the same treatment was repeated for a total of two sensitizations. Two weeks after the final sensitization, 0.25 mL of OVA (200 mg / mL) aqueous solution was orally administered once every two days for a total of four times to the OVA group, the CBD entire period group, and the CBD induction phase group. The control group was administered PBS during the sensitization period, and purified water during the induction period. The control group and OVA group were given normal food, and the CBD entire period group was administered food containing 0.02% CBD (50 mg / kg / day) from the first day of sensitization, and the CBD induction phase group was administered food from the end of sensitization (oral administration of OVA). Feces were collected up to 1 hour after the final OVA administration, and the amount of water in the feces was calculated to evaluate the effect of the administration period of CBD on the OVA-induced gastrointestinal allergy model mice.

[0036] Figure 5 is a graph showing the results of the effect of the administration period in Experimental Example 1-2. When CBD (50 mg / kg / day) was administered from the first day of sensitization, the amount of water in the feces was significantly lower than that in the OVA group, and the onset of diarrhea was suppressed. In contrast, even when CBD (50 mg / kg / day) was administered from the end of sensitization, no inhibitory effect on the onset of diarrhea was observed. From this, it was found that CBD could suppress ovalbumin (OVA)-induced gastrointestinal allergy symptoms by continuously administering it from the first day of sensitization.

[0037] In the present invention, the CBD dose (50 mg / kg / day) administered to mice is equivalent to about 4 mg / kg in humans weighing 60 kg. It has been reported that administration of 600 mg of CBD to healthy humans does not affect blood pressure or heart rate, and no dependence or abuse was observed (Martin-Santos R, et al., “Acute effects of a single, oral dose of d9-tetrahydrocannabinol (THC) and cannabidiol (CBD) administration in healthy volunteers,” Curr. Pharm. Des., 18, 4966-4979, 2012).

[0038] (Experimental Example 2) (reagent) Cannabidiol (CBD) was used as CBD crystalline powder (Batch No. CB099020171A) purchased from PharmaHemp Laboratories (Ljubljana, Slovenia). This powder contains 99.34% by mass of cannabidiol. TRI Reagent, ovalbumin (OVA) and aluminum gel were purchased from Sigma Aldrich (St. Louis, Missouri, USA). High Capacuty cDNA synthesis kit was purchased from Applied Biosystems (Foster City, California, USA). SsoAdvanced® Universal SYSR® Green Supermix was purchased from Bio-Rad Laboratries (Hercules, California, USA). Various primers were purchased from Hokkaido System Science Co., Ltd. FSC22 Frozen Section Media was purchased from Lesica Biosystems (Nussloch, Germany). Rat anti-mouse CD4 antibody was purchased from Biolegend Inc. (San Diego, CA, USA). Alexa Flour 488 donkey anti-rat IgG (H+L) was purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). VECTASHIELD mounting medium with 4',6-diamidino-2-phenylindole (DAPI) was purchased from VECTOR Laboratories Inc. (Newark, NJ, USA). Other reagents were of the highest purity available from commercial sources.

[0039] (Animals used) Six-week-old male BALB / c mice were purchased from Japan SLC Co., Ltd. (Hamamatsu, Shizuoka Prefecture). The mice were kept in a facility with a temperature of 24±1°C and humidity of 55±5%, and were kept under the following light / dark conditions: lights on at 8:00 a.m., lights off at 8:00 p.m. In conducting this experiment, we followed the Hoshi University Animal Experiment Guidelines, which were established with the aim of promoting the implementation of appropriate animal experiments from the standpoint of animal welfare as well as science, and obtained approval from the Hoshi University Animal Experiment Committee, and gave full consideration to the ethical aspects of animals.

[0040] (Mouse treatment) Mice were divided into three groups: a control group (hereafter referred to as the "control group"), an ovalbumin (OVA) group (hereafter referred to as the "OVA group"), and an OVA + cannabidiol (CBD) group (hereafter referred to as the "CBD combination group"). The OVA group and the CBD combination group were intraperitoneally administered 0.2 mL of OVA (50 μg / mL)·phosphate-buffered saline (PBS) solution containing aluminum hydroxide gel (final concentration: 1 mg / mL) as an adjuvant. Two weeks later, the same treatment was repeated, and sensitization was performed twice in total. Two weeks after the final sensitization, the OVA group and the CBD combination group were orally administered 0.25 mL of OVA (200 mg / mL) aqueous solution once every two days, a total of four times. The control group was administered PBS during the sensitization period and purified water during the elicitation period. The control and OVA groups were fed a normal diet, while the CBD-combined group was fed a diet containing 0.02% CBD (50mg / kg / day) from the first day of sensitization to the last day of the experiment. One hour after the final oral administration of OVA, 5mm pieces of the central part of the small intestine and large intestine (corresponding to the jejunum and colon) were collected under isoflurane anesthesia and flash frozen in liquid nitrogen. Blood was also collected and centrifuged at 3,000×g for 15 minutes at 4℃, and the supernatant was stored as a plasma sample.

[0041] (Measurement of mast cell protease-1 (Mcpt-1) concentration) The concentration of Mcpt-1 in blood was measured by enzyme-linked immunosorbent assay (ELISA). Specifically, a 96-well plate was coated with capture antibody according to the protocol of mouse Mcpt-1 (mMcpt-1) Uncoated ELISA kit (Thermo Fisher Scientific Inc.). Mouse plasma samples were added to the plate and incubated with biotin-conjugated anti-mouse Mcpt-1 antibody, then reacted with avidin-horseradish peroxidase (HRP), and the absorbance at 450 nm and 570 nm was measured.

[0042] (Measurement of OVA-specific IgE antibody concentration) The concentration of OVA-specific IgE antibody in the blood was measured by ELSIA. Specifically, according to the protocol of the LEGEND MAX (registered trademark) Mouse OVA specific IgE ELISA kit (Biolegend Inc.), mouse plasma samples were added to a 96-well plate, incubated with mouse OVA specific antibody, reacted with avidin-HRP, and the absorbance at 450 nm and 570 nm was measured.

[0043] (Measurement of CC motif chemokine ligand (CCL)2 concentration) The blood CCL2 concentration was measured by ELISA. Specifically, mouse plasma samples were added to a 96-well plate and reacted with a polyclonal antibody specific for mouse MCP-1 conjugated to HRP according to the protocol of Quantikine ELISA Mouse CCL2 / JE / MCP-1 Immunoassay (R&D systems Inc., Minneapolis, MN, USA), and the absorbance at 450 nm and 570 nm was measured.

[0044] (Total RNA purification from small and large intestine) Total RNA was extracted from approximately 20 mg of small or large intestine using TRI Reagent. The absorbance of the resulting solution was measured at 260 nm and 280 nm using a nanodrop-liete (Thermo Fisher Scientific Inc.) to confirm the purity and calculate the RNA concentration (ng / μL).

[0045] (Real-Time RT-PCR) cDNA was synthesized from 1 μg of RNA using a high capacity cDNA synthesis kit. Various primers shown in Table 1 below were created, and Real-Time PCR was performed to detect the expression of each gene. That is, 5 μL of SsoAdvanced (registered trademark) Universal SYBR (registered trademark) Green Supermix, 0.6 μL of the forward primer (5 pmol / μL) for the target gene, 0.6 μL of the reverse primer (5 pmol / μL), 2 μL of cDNA solution, and 2.8 μL of RNase free water were added to each well of the PCR plate. The temperature conditions were a denaturation temperature of 72°C for 30 seconds. The fluorescence intensity during the amplification process was monitored using a CFX Connect Real-Time System (Bio-Rad Laboratories). The mRNA expression level was normalized using 18SrRNA. In Table 1, IL stands for interleukin, and SOCS stands for suppressor of cytokine signaling.

[0046] [Table 1]

[0047] (Fluorescent immunohistochemical staining) One hour after the final OVA administration, a 10 mm section from the center of the large intestine (corresponding to the colon) was taken and immersed in a 4% by mass paraformaldehyde-PBS solution to fix the tissue. The tissue was embedded in FSC22 Frozen Section Media and 10 μm thick sections were prepared on slides. The slides were treated with a blocking solution (0.001% by mass Triton-X, 0.03% by mass fetal bovine serum in PBS) and then reacted with rat anti-mouse CD4 antibody and then Alexa Flour 488 donkey anti-rat IgG (H+L). The sections were mounted in VECTASHIELD mounting medium with DAPI and then viewed under a microscope.

[0048] (Statistical significance test) Experimental values ​​were expressed as mean ± standard error (SE). Statistical significance was tested using Tukey's test, and differences were considered significant when the risk level was less than 5% (p<0.05).

[0049] (Experimental Example 2-1) (Effect of CBD on Mcpt-1 and OVA-specific IgE antibody concentrations) FIG. 6 is a graph showing the results of the Mcpt-1 concentration (Plasma concentration) in Experimental Example 2-1, and FIG. 7 is a graph showing the results of the IgE antibody concentration (Plasma concentration) in Experimental Example 2-1. From FIG. 6, the blood concentration of serine protease (Mcpt)-1 produced from intestinal mucosal mast cells was significantly increased in the OVA group by administration of OVA compared to the control group. In contrast, the CBD combination group in which CBD was used in combination had a lower Mcpt-1 concentration due to OVA, suppressing the production of Mcpt-1 by OVA. Furthermore, as shown in FIG. 7, there was no significant difference in the blood concentration of OVA-specific IgE antibodies between the OVA group and the CBD combination group. From this, CBD suppressed the activation of mast cells without affecting the production amount of OVA-specific IgE antibodies.

[0050] (Experimental Example 2-2) (Effect of CBD on IL-4 expression in the small and large intestine) FIG. 8 is a graph showing the results of the expression level (Relactive expression) of IL-4 mRNA in the small intestine in Experimental Example 2-2, and FIG. 9 is a graph showing the results of the expression level (Relactive expression) of IL-4 mRNA in the large intestine in Experimental Example 2-2. When the expression level (Relactive expression) of IL-4 mRNA in the small intestine was examined, it was increased by about 30 times in the OVA group compared to the control group. When the expression level (Relactive expression) of IL-4 mRNA in the large intestine was examined, it was increased by about 150 times in the OVA group compared to the control group. In contrast, in the CBD combination group in which CBD was used in combination, the expression level (Relactive expression) of IL-4 mRNA by OVA was significantly suppressed in both the small intestine and the large intestine. From this, CBD suppressed the increase in IL-4 expression by OVA in the small intestine and the large intestine.

[0051] (Experimental Example 2-3) (Effect of CBD on Th2 cytokine production in the small and large intestine) FIG. 10 is a graph showing the results of the expression level (Relactive expression) of SOCS-3 mRNA in the small intestine in Experimental Example 2-3, and FIG. 11 is a graph showing the results of the expression level (Relactive expression) of SOCS-3 mRNA in the large intestine in Experimental Example 2-3. When the expression level (Relactive expression) of SOCS-3 mRNA in the small intestine was examined, it was increased by about 4.5 times in the OVA group compared to the control group. When the expression level (Relactive expression) of SOCS-3 mRNA in the large intestine was examined, it was increased by about 10 times in the OVA group compared to the control group. In contrast, in the CBD combination group in which CBD was used in combination, the expression level (Relactive expression) of SOCS-3 mRNA by OVA was significantly suppressed in both the small intestine and large intestine. In addition, FIG. 12 is a graph showing the results of the CCL2 blood concentration (Plasma concentration) in the large intestine in Experimental Example 2-3. The plasma concentration of CCL2, which is involved in the regulation of Th2 cell differentiation, was significantly increased in the OVA group, whereas this increase was suppressed in the CBD-combined group. Based on the results of the expression level of SOCS-3 mRNA, which correlates with the production of IL-4 in Th2 cells, and the plasma concentration of CCL2, which is involved in the regulation of Th2 cell differentiation, CBD suppressed the activation of Th2 cells in the small and large intestines.

[0052] (Experimental Example 2-4) (Effect of CBD on infiltration of CD4 positive cells in the large intestine) Figure 13 is a graph showing the effect of CBD on the infiltration of CD4 positive cells in the large intestine in Experimental Example 2-4, and shows the number of Th2 cells in the large intestine mucosa by staining the CD4 antigen found on the membrane surface of Th2 cells. A large number of CD4 positive cells were observed in the OVA group, whereas the number of CD4 positive cells in the CBD-combined group was about the same as in the control group. This shows that CBD suppresses the infiltration of Th2 cells into the large intestine mucosa.

[0053] (Example 1, Comparative Example 1) A composition for suppressing egg allergies of Example 1 and a composition of Comparative Example 1 were prepared.

[0054] The composition for suppressing egg allergy in Example 1 had a better effect of suppressing OVA-induced gastrointestinal allergy symptoms than the composition in Comparative Example 1.

Claims

1. A composition for suppressing egg allergy, comprising cannabidiol.

2. 2. The composition for suppressing egg allergy according to claim 1, wherein the cannabidiol suppresses the production of mast cell protease-1 from intestinal mucosal mast cells.

3. The composition for suppressing egg allergy according to claim 1, wherein the cannabidiol suppresses the expression of interleukin-4 in the small intestine and large intestine.

4. 2. The composition for suppressing egg allergy according to claim 1, wherein the cannabidiol suppresses the expression of SOCS-3 (suppressor of cytokine signaling 3) mRNA in the small intestine and large intestine.

5. The composition for suppressing egg allergy according to claim 1, wherein the cannabidiol suppresses the infiltration of Th2 cells into the large intestine mucosa.

6. 2. The composition for suppressing egg allergy according to claim 1, which is a health food, a food for special dietary uses, a food for specified health uses, a food with nutrient functions, a functional food, a nutritional supplement, a health supplement, a supplement or a medicine.

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