KR20230040032A - Composition for improving, preventing or treating inflammatory bowel diseases comprising Agastachis Herba extract - Google Patents

Abstract

The present invention relates to a composition for preventing or treating inflammatory bowel disease (IBD) containing an extract of Agastache rugosa, wherein the composition exhibits intestinal immunity enhancing activity and promotes the recovery of damage to the colon tissue and the recovery of cell necrosis, thereby being able to be effectively used to manufacture pharmaceuticals or food for alleviating, preventing or treating IBD.

Description

Composition for improving, preventing or treating inflammatory bowel diseases comprising Agastachis Herba extract}

The present invention relates to a composition for improving, preventing or treating inflammatory bowel disease (IBD) comprising an extract of Baechohyang ( Agastache rugosa O. Kuntze ), and more particularly, intestinal immunity comprising a water extract of Baechohyang outpost It relates to a composition for promotion and a composition for improving, preventing or treating inflammatory bowel disease using the same.

Inflammatory bowel disease (IBD) is a disease that causes chronic inflammation in the gastrointestinal tract.

Generally, it is classified into two types: Ulcerative colitis (UC) and Crohn's disease (CD). Ulcerative colitis mainly invades the mucous membrane and frequently forms rashes or ulcers, which is an unknown cause of the large intestine. As a kind of diffuse nonspecific inflammation, it is accompanied by various systemic symptoms including bloody diarrhea. Crohn's disease is discontinuous in the entire digestive tract from the mouth to the anus, and ulcers and fibrosis occur in all layers of the intestinal tract from the mucous membrane. It is a granulomatous inflammatory lesion of unknown etiology in which strictures and lesions develop, accompanied by systemic symptoms such as abdominal pain, chronic diarrhea, fever, and malnutrition.

The cause of the inflammatory bowel disease has not yet been specifically identified, but it is known that abnormalities in immune function are involved. Immunological factors involved in this include innate immunity, production of cytokines, activation of CD4, etc. this is known In particular, it is known that cytokines play an important role, and the production of tumor necrosis factor (TNF-α), interleukin (IL)-1, IL-6, and IL-8 at the site of inflammation is It was confirmed that it is significantly increased in patients with colitis and Crohn's disease.

TNF-α, a major proinflammatory cytokine, exacerbates the inflammatory response through the NF-κnuclear factor kappa-B pathway. Anti-TNF-α antibodies such as infliximab and adalimumab are effective in regulating intestinal immunity by preventing apoptosis of mucosal T lymphocytes. has been approved to treat

IL-1β causes colonic inflammation by recruiting granulocytes and activates CD4+ Th17 cells to secrete IL-17. Infiltration of neutrophils is known to be fundamentally necessary for the progression of chronic inflammation. IL-6 promotes neutrophil migration to areas of colitis (colitis).

Drugs used to treat existing inflammatory bowel disease include steroidal immunosuppressants and 5-aminosalicylic acid (5-ASA) drugs that block the production of prostaglandins (e.g., sulfasalazine, etc.) , mesalazine, etc. are used. As exemplified above, anti-inflammatory drugs and immunosuppressants are prescribed to alleviate colitis, but it is well known that they have undesirable side effects such as nausea, vomiting, headache, hemolytic anemia and male infertility.

On the other hand, irritable bowel syndrome is a condition in which the function of the large intestine has changed. It refers to a syndrome in which there are no ulcers but changes in bowel habits such as diarrhea, abdominal pain, and constipation. Although this does not belong to a serious life-threatening disease, it is necessary to treat it because changes in bowel habits can cause changes in the body, such as making daily life uncomfortable or physically weak, and reducing the quality of life. Accordingly, there is a continuous demand for the development of safer and more effective inflammatory bowel disease and irritable bowel syndrome treatments.

Accordingly, the inventors of the present invention have made intensive research efforts to develop a composition for improving, preventing or treating safe and effective inflammatory bowel disease (IBD), and as a result, the present invention Baechohyang ( Agastache rugosa O. Kuntze ) extract is far superior. confirmed to complete the present invention.

Accordingly, an object of the present invention is to provide a health functional food composition for improving inflammatory bowel disease, which includes an extract of baechohyang.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of inflammatory bowel disease containing the extract of Baechohyang.

Another object of the present invention is to provide a composition for enhancing the intestinal immunity containing the baechohyang extract.

Another object of the present invention relates to the use of the extract of baechohyang for enhancing intestinal immunity or improving, preventing or treating inflammatory bowel disease.

The present invention relates to a composition for improving, preventing or treating inflammatory bowel disease (IBD) containing an extract of Baechohyang ( Agastache rugosa O. Kuntze ). By promoting the recovery of damage to colon tissue and the recovery of cell necrosis, it exhibits an effect of improving, preventing or treating inflammatory bowel disease.

Hereinafter, the present invention will be described in more detail.

One aspect of the present invention is a health functional food composition for improving inflammatory bowel disease comprising a blackberry extract.

The term "baechohyang" in the present specification is a perennial plant of the Lamiaceae family, and the stem of the above-ground part is square and has a diameter of about 5 mm. The outer surface is dark brown, with vertical patterns and nodes, and the interval between nodes is 3 to 10 cm. The peel is light and soft, and the folded surface is uneven, fibrous, and hollow inside. The upper side of the leaf is green to dark green, and the lower side is yellowish green to grayish brown, and when the leaf is spread out, it has a shallow core shape. Sometimes the flower stalk is attached. This herb has a unique fragrance and the taste is plain and slightly cool.

The part used in the preparation of the baechohyang extract may be the whole plant, but may be an aerial part or part, but is not limited thereto.

In the present invention, the inflammatory bowel disease is selected from the group consisting of ulcerative colitis (UC), Crohn's disease (CD), irritable bowel syndrome, intestinal Behcet's disease, infectious enteritis, ischemic bowel disease and radiation enteritis It may be one or more species.

The 'extract' includes a crude solvent extract, a specific solvent-soluble extract (solvent fraction), and a solvent fraction of the crude solvent extract, and the Baechohyang extract may be in a solution, concentrate or powder state.

The baechohyang extract may be a crude extract obtained by extracting baechohyang with water and one or more solvents selected from the group consisting of straight-chain or branched alcohols having 1 to 4 carbon atoms.

The baechohyang extract according to the present invention may be a solvent fraction obtained by fractionating the crude solvent extract with an additional solvent, for example, using one or more solvents selected from the group consisting of ethyl ether, ethyl acetate, and butanol in the crude solvent extract. It may be a solvent fraction.

For example, the baechohyang extract is a solvent crude extract obtained by extracting baechohyang with one or more solvents selected from the group consisting of water and a straight chain or branched alcohol having 1 to 4 carbon atoms selected from the group consisting of ethyl ether, ethyl acetate, and butanol. It may be a solvent fraction using one or more solvents.

One aspect of the present invention relates to a method for preparing an extract of baechohyang.

The baechohyang extract includes a solvent crude extract and a solvent fraction of baechohyang, and is as described above.

The baechohyang extract is a crude extract obtained by extracting at least one selected from the group consisting of the outpost, aerial parts, or parts of baechohyang with one or more solvents selected from the group consisting of water and a straight chain or branched alcohol having 1 to 4 carbon atoms. can

The manufacturing process of the baechohyang extract according to the present invention in more detail is as follows:

A washing step of cutting baechohyang and washing with water to remove constrictions;

Extraction step of extracting at room temperature with an extraction solvent of 10 to 30 times the weight, 10 to 20 times the weight, or 15 to 18 times the weight of the baechohyang;

A concentration step of preparing a concentrate by filtering after extraction; and

A drying step of spray drying the concentrate.

The extraction step may be performed at a temperature condition of 30 to 150 °C, preferably 50 to 120 °C.

The extraction step may use room temperature extraction, cold needle extraction, hot water extraction, ultrasonic extraction, steam extraction, reflux cooling extraction, reduced pressure or pressure extraction, and may preferably be performed using pressure extraction. The pressurized extraction is performed by raising the boiling point of the solvent by applying pressure in the container, and can be extracted at a higher temperature than conventional extraction methods.

The extraction step may be to additionally perform a conventional fractionation process, but is not limited thereto.

The extraction process can be repeated once or several times, and in one preferred example of the present invention, a method of re-extracting again after the first extraction can be adopted, which means that even if effective filtration is performed when herbal extracts are mass-produced, the herbal medicine itself This is to prevent loss due to the high moisture content, and the extraction efficiency is reduced by only the first extraction. In addition, as a result of verifying the extraction efficiency at each stage, it was found that about 80 to 90% of the total extraction amount was extracted by the secondary extraction.

In one example of the present invention, when the extraction process is repeated twice, the obtained residue is extracted with reflux again with about 5 to 15 times the volume of the extraction solvent, preferably 8 to 12 times the volume. After extraction, it is filtered and combined with the previously obtained filtrate and concentrated under reduced pressure to prepare an extract of Baechohyang. Although the extraction efficiency can be increased by mixing the filtrate obtained after the second extraction and each extraction, the extract of the present invention is not limited to the number of extractions.

If the amount of the solvent used in the preparation of the baechohyang extract is too small, it is difficult to stir, the solubility of the extract is lowered, and the extraction efficiency is lowered. Since handling problems may occur, the amount of the solvent used is preferably set within the above range.

The concentration step may be to additionally perform purification of the extract using a conventional purification method, but is not limited thereto. For example, a further purified fraction can be obtained using various chromatography methods such as silica gel column chromatography, thin layer chromatography, and high performance liquid chromatography. there is.

The concentration step is to increase the concentration of solids by removing the liquid of the liquid material having a high liquid content, and may be to use a vacuum concentration method. The vacuum concentration method is a method of increasing the solute concentration of a solution by rapidly evaporating the solvent by lowering the boiling point of the solvent under reduced pressure.

The drying step may be performed by air drying (AD), cold air drying, vacuum drying (Vacuum Dry, VD), spray drying (SD) and freeze drying (Freeze Dry, FD), preferably may use spray drying. Spray drying is a method of spraying slurry or liquid material with fine liquid particles (10 to 200 υ) in a drying room (around 100 to 180 ° C) and contacting the fine liquid particles with hot air to dry them within a short time (1 to 10 seconds) am.

The filtered extract obtained in this way is about 10 to 30 times by weight, preferably 15 to 25 times by weight, more preferably about 18 times by weight of the total amount of the concentrate in order to adjust the content of the remaining lower alcohol to be suitable for use as a raw material. It can be azeotropically concentrated 1 to 5 times, preferably 2 to 3 times with water in an amount of 20 to 20 times by weight, and then homogeneously suspended by adding the same amount of water again, and then freeze-dried to prepare a powdered Baechohyang extract.

In the present invention, "extraction" is a method of dissolving and separating useful soluble components contained in liquid or solid raw materials in a solvent, and may be any method commonly used, for example, hot water extraction method, cold extraction method, reflux cooling It may be an extraction method, a solvent extraction method, a steam distillation method, an ultrasonic extraction method, an elution method, or a compression method, but is not limited thereto.

When using the health functional food composition of the present invention as a food additive, the health functional food composition may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. In general, when preparing food or beverage, the health functional food composition of the present invention may be added in an amount of 15% by weight or less, preferably 10% by weight or less, based on the raw material.

There is no particular limitation on the type of food. Examples of foods to which the substance can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages, vitamin complexes, and the like, and includes all foods in a conventional sense.

The beverage may contain various flavoring agents or natural carbohydrates as additional ingredients. The aforementioned natural carbohydrates may include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, and synthetic sweeteners such as saccharin and aspartame. . The ratio of the natural carbohydrates may be appropriately determined by a person skilled in the art.

In addition to the above, the health functional food composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, and glycerin. , alcohol, a carbonating agent used in carbonated beverages, and the like. In addition, the health functional food composition of the present invention may contain fruit flesh for preparing natural fruit juice, fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The ratio of these additives can also be appropriately selected by those skilled in the art.

Another aspect of the present invention is a pharmaceutical composition for preventing or treating inflammatory bowel disease comprising an extract of Baechohyang.

In the present invention, the inflammatory bowel disease may be at least one selected from the group consisting of ulcerative colitis, Crohn's disease, irritable bowel syndrome, intestinal Behçet's disease, infectious enteritis, ischemic bowel disease, and radiation enteritis.

In the present invention, the baechohyang extract may be a crude extract obtained by extracting baechohyang with water and at least one solvent selected from the group consisting of straight-chain or branched alcohols having 1 to 4 carbon atoms.

The pharmaceutical composition of the present invention may be used as a pharmaceutical composition comprising a pharmaceutically effective amount of the extract of Baechohyang and / or a pharmaceutically acceptable carrier.

As used herein, the term "pharmaceutically effective amount" refers to an amount sufficient to achieve the efficacy or activity of the above-described Baechohyang extract.

Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used in formulation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, including, but not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil; it is not going to be The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like in addition to the above components.

The pharmaceutical composition according to the present invention may be administered to mammals including humans by various routes. The administration method may be any method commonly used, and may be administered by, for example, oral, dermal, intravenous, intramuscular, subcutaneous, etc. routes, preferably orally.

The suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, administration method, patient's age, weight, sex, morbid condition, food, administration time, administration route, excretion rate and reaction sensitivity, A ordinarily skilled physician can readily determine and prescribe dosages effective for the desired treatment or prophylaxis.

The daily dosage may be different depending on the subject of administration.

According to one embodiment of the present invention, the daily dose (weight/body weight of the subject) of the pharmaceutical composition of the present invention may be 50 to 500 mg/kg, 2 to 5 times a week, that is, 2 to 5 times a week It may be administered daily, for example, 3 to 4 times a week, that is, it may be administered 3 to 4 days a week, but is not limited thereto.

Preferably, for mice, the daily dose is 50 to 300 mg/kg, 100 to 200 mg/kg, 100 to 500 mg/kg, 100 to 300 mg/kg, or 100 to 200 mg/kg. It may be kg, for example, it may be 200 mg / kg, but is not limited thereto.

According to another embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention may be 500 to 3000 mg.

Preferably, when targeting human adults, the daily dosage may be 500 to 2500 mg, 1000 to 5000 mg, 1000 to 3000 mg, or 1000 to 2500 mg, but is not limited thereto.

The pharmaceutical composition of the present invention is prepared in unit dosage form by formulation using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily performed by those skilled in the art. or it may be prepared by incorporating into a multi-dose container. At this time, the formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet, capsule or gel (eg, hydrogel), and may additionally contain a dispersing agent or stabilizer. .

Another aspect of the present invention is a composition for enhancing intestinal immunity comprising an extract of Baechohyang.

The term "enhancement of intestinal immunity" as used herein refers to a function of promoting an immune response to an antigen non-specifically in the initial activation process of immune cells present in the intestinal tract or a function of enhancing intestinal immunity by increasing the activity of cells of the immune system. do. Diseases caused by decreased intestinal immune function may be any one or more selected from the group consisting of infectious diseases caused by pathogenic microorganisms, inflammatory diseases, and cancer. The composition for enhancing intestinal immunity has an application effect of improving or preventing seasonal flu in humans, improving or preventing bacterial diarrhea, improving or preventing bacterial infections, and enhancing post-wound treatment effects.

In the present invention, the baechohyang extract may be a crude extract obtained by extracting baechohyang with water and at least one solvent selected from the group consisting of straight-chain or branched alcohols having 1 to 4 carbon atoms.

The daily dosage of the composition for enhancing intestinal immunity may vary depending on the subject of administration.

Preferably, when targeting mice, the daily dose (weight/body weight of the subject) is 50 to 500 mg/kg, 50 to 300 mg/kg, 100 to 200 mg/kg, 100 to 500 mg/kg , 100 to 300 mg/kg, or 100 to 200 mg/kg, for example, 200 mg/kg, but is not limited thereto.

Preferably, when targeting human adults, the daily dosage may be 500 to 3000 mg, 500 to 2500 mg, 1000 to 5000 mg, 1000 to 3000 mg, or 1000 to 2500 mg, but is limited thereto It is not.

In addition, according to a preferred embodiment of the present invention, the composition may be administered 2 to 5 times a week, that is, 2 to 5 days a week, for example, 3 to 4 times a week, that is, 3 to 4 days a week. It may, but is not limited thereto.

The present invention relates to a composition for preventing or treating inflammatory bowel disease (IBD) containing an extract of Agastache rugosa O. Kuntze , wherein the composition exhibits intestinal immunity enhancing activity and recovers damage to colon tissue And since it promotes the recovery of cell necrosis, it can be effectively used in the manufacture of medicines or foods for the improvement, prevention or treatment of inflammatory bowel disease.

Figure 1a is a graph showing the survival rate in HT29 cell line upon treatment with baechohyang (Agastache rugosa O. Kuntze) extract according to an embodiment of the present invention.
Figure 1b is a graph showing the survival rate in the HCT116 cell line upon treatment with a black pepper extract according to an embodiment of the present invention.
Figure 2a is a graph showing the degree of inhibition of NO in macrophages upon treatment with the baechohyang extract according to an embodiment of the present invention.
Figure 2b is a graph showing the survival rate in macrophages when treated with baechohyang extract according to an embodiment of the present invention.
Figure 3a is a graph showing the expression level of TNF-α in macrophages upon treatment with baechohyang extract according to an embodiment of the present invention.
Figure 3b is a graph showing the expression level of IL-6 in macrophages upon treatment with the baechohyang extract according to an embodiment of the present invention.
Figure 4 is a schematic diagram showing the process of inducing ulcerative colitis (UC) in mice by DSS (Dextran sodium sulfate) stimulation according to an embodiment of the present invention.
Figure 5a is a graph showing the change in body weight by the administration of baechohyang extract in the ulcerative colitis mouse model according to an embodiment of the present invention.
Figure 5b is a photograph and a graph showing the change in the length of the large intestine by the administration of baechohyang extract in the ulcerative colitis mouse model according to an embodiment of the present invention.
Figure 6 is a photograph and a graph of the results of H&E staining observing the recovery pattern of damage to colon tissue by administration of baechohyang extract in an ulcerative colitis mouse model according to an embodiment of the present invention.
Figure 7 is a photograph and a graph of the result of tunnel analysis (TUNEL assay) observing the cell necrosis recovery pattern of colon tissue by the administration of baechohyang extract in an ulcerative colitis mouse model according to an embodiment of the present invention.
Figure 8a is a graph showing the change in the amount of COX-2 gene mRNA expression level by administration of Baechohyang extract in an ulcerative colitis mouse model according to an embodiment of the present invention.
Figure 8b is a graph showing the change in iNOS gene mRNA expression level by administration of the extract of Baechohyang in an ulcerative colitis mouse model according to an embodiment of the present invention.
Figure 8c is a graph showing the change in TNF-α gene mRNA expression level by administration of baechohyang extract in an ulcerative colitis mouse model according to an embodiment of the present invention.
Figure 8d is a graph showing changes in the expression level of IL-6 gene mRNA by the administration of the baechohyang extract in the ulcerative colitis mouse model according to an embodiment of the present invention.
Figure 8e is a graph showing changes in the expression level of IL-10 gene mRNA by administration of the baechohyang extract in an ulcerative colitis mouse model according to an embodiment of the present invention.
Figure 9a is a graph showing changes in phosphorylation of p38 by administration of the extract of Baecho-hyang in an ulcerative colitis mouse model according to an embodiment of the present invention.
Figure 9b is a graph showing the expression level of Bcl-2 by administration of the extract of pear extract in an ulcerative colitis mouse model according to an embodiment of the present invention.

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

Throughout this specification, "%" used to indicate the concentration of a particular substance is (weight/weight)% for solids/solids, (weight/volume)% for solids/liquids, and liquid/liquid is (volume/volume) %.

Preparation Example: Preparation of baechohyang water extract

Baechohyang ( Agastache rugosa O. Kuntze ) Cut the outpost and wash it with water to remove the constriction, add water corresponding to its weight 16 times (Example 1) and 20 times (Example 2) at 100 ℃ for 6 hours By extraction, a water extract of baechohyang was prepared. The extract was filtered through a 60 mesh filter, then concentrated to prepare a concentrate, and the concentrate was spray-dried to finally prepare an extract in powder form. The manufacturing process and extract information for the extract are shown in Table 1.

extract extraction temperature extraction time extraction solvent solids content Sugar content (Brix) recovery rate Example 1 100℃ 6h multiple of 16 0.90% 1.1 9.32% Example 2 100℃ 6h multiple of 20 0.79% 0.9 11.28%

Experimental Example 1: Cytotoxicity evaluation of Baechohyang extract

In order to confirm the cytotoxicity of the extract of Baechohyang in the intestinal tract, it was evaluated using HT29 and HCT116 cell lines, which are colorectal cancer cell lines.

Specifically, after seeding 2×10 ⁴ cells in a 96-well plate, the extracts of Examples 1 and 2 were dissolved in tertiary distilled water (DW, Distilled Water), and then diluted step by step. Cells were treated with various concentrations, and cell viability was measured after 24 hours.

Cell viability (HT29, %) Concentration (mg/ml) 0 0.001 0.01 0.05 0.1 0.5 One 2 Example 1 One One 1.08 1.05 1.03 1.15 1.13 1.17 Example 2 One 1.2 1.25 1.05 1.01 1.12 1.04 1.18

As can be seen in Table 2 and FIG. 1a, cytotoxicity was not observed when the extracts of Examples 1 and 2 were administered to HT29.

Cell viability (HCT116, %) Concentration (mg/ml) 0 0.001 0.01 0.05 0.1 0.5 One 2 Example 1 One 1.04 0.85 0.9 1.02 One 0.95 0.99 Example 2 One 1.12 1.26 1.20 1.27 1.26 1.19 1.30

As can be seen in Table 3 and FIG. 1b, when the extracts of Examples 1 and 2 were administered to HCT116, cell viability tended to decrease at some concentrations, but the number was low, so it is difficult to see that it is toxic. .

Experimental Example 2: Nitric oxide measurement

In order to determine whether the reduction in the anti-inflammatory effect and the level of nitric oxide (NO) of the extract of baechohyang was due to cytotoxicity, the NO level was measured using the Griess test method in the cell experiment. RAW 264.7 cells (macrophages) were inoculated into each well of 2×10 ⁴ in a 96-well plate, and then the extracts of Examples 1 and 2 were added at concentrations of 0.001, 0.01, 0.05, 0.1, 0.5, 1, and 2 mg/ml. After treatment, 500 ng/ml of LPS was treated 1 hour later. After 23 hours of LPS treatment, Griess reagent (0.1% naphthylethylenediamine + 1% sulfanilamide dissolved in 5% phosphoric acid) was added and reacted, and NO levels were measured by measuring absorbance at a wavelength of 549 nm. In addition, the cytotoxicity of the extracts of Examples 1 and 2 and LPS was confirmed in RAW 264.7 cells.

NO level (μg/ml) LPS 500 ng/ml - + + + + + + + + Concentration (mg/ml) 0 0 0.001 0.01 0.05 0.1 0.5 One 2 Example 1 3 64.5 60.0 57.0 57.0 48.2 36.0 33.8 24.1 Example 2 5.5 65.5 65.1 61.0 60.0 58.6 56.6 45.8 26.2

As can be seen in Table 4 and FIG. 2a, when the extracts of Examples 1 and 2 were administered to RAW 264.7 cells, the NO level tended to decrease as the treatment concentration of the extract increased. NO is increased when an inflammatory reaction occurs in vivo, and the decrease in this value is a result confirming that the extracts of Examples 1 and 2 have anti-inflammatory effects.

Cell viability (RAW264.7, %) LPS 500 ng/ml - + + + + + + + + Concentration (mg/ml) 0 0 0.001 0.01 0.05 0.1 0.5 One 2 Example 1 100.0 108.0 108.8 107.2 108.8 106.4 105.6 107.2 105.6 Example 2 100.0 114.6 118.8 109.3 107.8 105.4 100.7 104.6 101.5

As can be seen in Table 5 and FIG. 2b, as a result of measuring cell viability, the extracts of Examples 1 and 2 did not affect cell viability.

Experimental Example 3. Cytokine measurement

In order to confirm the anti-inflammatory effect of the extract of Baechohyang, ELISA (Enzyme-Linked Immunosorbent Assay) was used to measure the cytokine expression level. RAW 264.7 cells were inoculated into each well of 2×10 ⁴ in a 96-well plate, and the extracts of Examples 1 and 2 were treated at concentrations of 1 and 2 mg/ml, and then 500 ng/ml of LPS was added after 1 hour. processed. After 23 hours of LPS treatment, the supernatant of the cell-inoculated well in a 96-well plate was measured using an ELISA kit (TNF-α, IL-6) according to the manufacturer's instructions.

TNF-α (pg/ml) LPS 500 ng/ml - + + + Concentration (mg/ml) 0 0 One 2 Example 1 11 298 89 5 Example 2 20 401 22 35

IL-6 (pg/ml) LPS 500 ng/ml - + + + Concentration (mg/ml) 0 0 One 2 Example 1 46 397 66 57 Example 2 28 411 30 38

As can be seen in Tables 6 and 7, and FIGS. 3a and 3b, when the extracts of Examples 1 and 2 were treated, the expression levels of TNF-α and IL-6 were reduced. TNF-α and IL-6 are factors that induce an inflammatory response, and the decrease in the expression of these cytokines means that the extracts of Examples 1 and 2 have excellent anti-inflammatory effects.

When comparing the recovery rate of the extract and the anti-inflammatory effect in the cell experiment, the anti-inflammatory effect was almost similar in the two extracts, and the extract of Example 2 was higher in the recovery rate, so animal experiments were conducted using the extract of Example 2. proceeded.

Experimental Example 4: DSS-induced ulcerative colitis mouse model

Animal experiments were conducted as follows to confirm whether the Baechohyang extract had a therapeutic effect on mice induced with ulcerative colitis. DSS was administered to induce ulcerative colitis in mice. When DSS is administered to mice, it induces an inflammatory response by adding toxicity to mucosal epithelial cells that serve as an immune barrier in the large intestine, thereby causing loss of the cell layer. Depending on the process, intestinal immunity is damaged and ulcerative colitis is induced.

Experimental groups of mouse model animals were divided into three groups: a normal group (untreated group), a control group (DSS administration), and an experimental group (DSS + administration of the extract of Example 2). The total experimental period was 3 weeks. For the first 2 weeks, the extract of Example 2 was dissolved in 200 ul of purified water so that 200 mg/kg of sample was injected at one time and orally administered three times a week, and in the last week, 3% DSS (Dextran sodium sulfate) solution was orally administered in combination with 200 ul. A schematic diagram of the process for the experiment is shown in FIG. 4 .

Experimental Example 5: Observation of morphological changes

Changes in body weight and the length of the large intestine were measured during the period of administration of DSS to see the therapeutic effect of the extract of Baechohyang in an ulcerative colitis mouse model.

As can be seen in Figure 5a, in the case of mice whose ulcerative colitis was induced by DSS, they generally tended to lose weight, and it was confirmed that the decrease in weight was reduced in the experimental group compared to the control group.

As can be seen in Figure 5b, the length of the large intestine is also shorter than that of normal mice in the case of ulcerative colitis-induced mice, and the length of the large intestine in the experimental group showed a recovery compared to the control group. From the results, it was confirmed that the extract of Example 2 exerts a therapeutic effect in the ulcerative colitis mouse model.

Experimental Example 6: Observation of histological changes

In order to confirm the therapeutic effect of the extract of Baechohyang in the ulcerative colitis mouse model, the colon tissue was post-fixed and sectioned to produce a block. The fabricated block was subjected to H&E staining (stsining) and tunnel analysis (TUNEL assay) to observe histological changes by administration of the extract of Example 2.

As can be seen in Figure 6, in the H & E staining, it was confirmed through a microscope that the damage was recovered by administration of the extract of Example 2 in the state of tissue damage caused by the induction of ulcerative colitis through a microscope. As a result, as shown in the control group, ulcerative colitis induced Tissue damage due to infiltration of the mucosal and submucosa was observed in the colon tissue.

As can be seen in Figure 7, as a result of confirming the degree of cell necrosis caused by ulcerative colitis by tunnel analysis, it was confirmed that the experimental group showed a decrease in cell necrosis when compared to the control group. This is a result confirming that baechohyang exhibits a therapeutic effect in ulcerative colitis-induced mice through histological observation.

Experimental Example 7: Gene analysis of colon tissue

In order to confirm by which mechanism the therapeutic effect of Baechohyang extract proceeds in the ulcerative colitis mouse model, a gene analysis was performed by taking a colon sample. Gene analysis was performed by using the qRT-PCR technique to confirm the mRNA expression of COX-2, iNOS, TNF-α, IL-6, and IL-10, which are genes related to the inflammatory response.

As can be seen in Figures 8a to 8e, the mRNA expression of genes related to the inflammatory response was increased by the induction of ulcerative colitis, but in the experimental group, this expression was suppressed compared to the control group. This means that the inflammatory response overexpressed by ulcerative colitis is controlled by the Baechohyang extract by suppressing the expression of mRNA related to the inflammatory response.

Experimental Example 8: Analysis of protein expression in colon tissue

In order to confirm by which mechanism the therapeutic effect of Baechohyang extract proceeds in the ulcerative colitis mouse model, a protein analysis was performed by taking a sample of the large intestine. Protein analysis was performed using a Western blotting technique, and the phosphorylation of p38, a protein related to inflammatory response and cell necrosis, and the expression of Bcl-2, a protein involved in apoptosis inhibition, were confirmed.

9a and 9b, the phosphorylation of p38 was increased and the expression of Bcl-2 was suppressed by the induction of ulcerative colitis. However, it can be confirmed that the phosphorylation of p38 was decreased in the experimental group compared to the control group, and the expression of Bcl-2 was increased close to the normal group. This means that the baechohyang extract inhibits the inflammatory response overexpressed by ulcerative colitis and inhibits the progress of apoptosis.

Claims (8)

A health functional food composition for improving inflammatory bowel disease (Inflammatory bowel disease, IBD) containing an extract of Baechohyang ( Agastache rugosa O. Kuntze ). The method of claim 1, wherein the inflammatory bowel disease is a group consisting of ulcerative colitis (UC), Crohn's disease (CD), irritable bowel syndrome, intestinal Behcet's disease, infectious enteritis, ischemic bowel disease, and radiation enteritis. At least one member selected from, health functional food composition for improving inflammatory bowel disease. The method of claim 1, wherein the baechohyang extract is a crude extract obtained by extracting baechohyang with water and at least one solvent selected from the group consisting of straight-chain or branched alcohols having 1 to 4 carbon atoms. Nutraceutical composition. Baechohyang ( Agastache rugosa O. Kuntze ) Inflammatory bowel disease (Inflammatory bowel disease, IBD) prevention or treatment pharmaceutical composition comprising an extract. The method of claim 4, wherein the inflammatory bowel disease is a group consisting of ulcerative colitis (UC), Crohn's disease (CD), irritable bowel syndrome, intestinal Behcet's disease, infectious enteritis, ischemic bowel disease, and radiation enteritis. A pharmaceutical composition for preventing or treating inflammatory bowel disease, which is one or more selected from. The method of claim 4, wherein the baechohyang extract is a crude extract obtained by extracting baechohyang with water and at least one solvent selected from the group consisting of straight-chain or branched alcohols having 1 to 4 carbon atoms, preventing or treating inflammatory bowel disease. pharmaceutical composition for use. Baechohyang ( Agastache rugosa O. Kuntze ) Composition for enhancing intestinal immunity containing an extract. According to claim 7, wherein the baechohyang extract is a crude extract obtained by extracting baechohyang with water and one or more solvents selected from the group consisting of straight-chain or branched alcohols having 1 to 4 carbon atoms.

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