Composition for preventing or treating inflammatory bowel disease comprising germinated oat extract

Abstract

This invention provides a means of improving inflammatory bowel diseases, such as ulcerative colitis (UC), Crohn's disease (CD), and Behçet's disease, in which inflammation and ulcers occur in the intestines due to unknown causes. [Solution] A composition for the prevention, improvement, or treatment of inflammatory bowel disease is provided, comprising germinated oat (Avena sativa L.) extract. Since the composition improves intestinal permeability and the integrity of tight connections within the inflamed intestinal environment, it can be effectively used in the manufacture of pharmaceuticals, health functional foods, or foods for the prevention, improvement, or treatment of inflammatory bowel disease.

Description

[Technical Field]

[0001] The present invention relates to a composition for the prevention or treatment of inflammatory bowel disease, comprising a germinated oat extract with increased avenanthramide content. [Background technology]

[0002] Inflammatory bowel disease refers to chronic inflammatory diseases in which inflammation or ulcers occur in the intestines due to unknown causes, such as ulcerative colitis (UC), Crohn's disease (CD), and Behçet's disease.

[0003] Inflammatory bowel disease (IBD) is prevalent worldwide, and recently, there has been a noticeable increase in its incidence in South Korea. However, its causes and mechanisms are relatively complex and still not fully understood.

[0004] Drugs currently used to treat inflammatory bowel disease include steroidal immunosuppressants, 5-aminosalicylic acid (5-ASA) drugs that block prostaglandin production (e.g., sulfasalazine), and mesalazine. As mentioned above, anti-inflammatory and immunosuppressant drugs are prescribed, but they cause side effects such as nausea, vomiting, headache, hemolytic anemia, and male infertility. A considerable number of patients do not respond to treatment or experience reduced efficacy, so there is a continuous need for the development of safer and more effective therapeutic agents.

[0005] Inflammatory bowel disease (IBD) is characterized by damage to the epithelial barrier due to inflammation. Damaged barrier integrity and increased permeability of epithelial tectonic connections lead to a weakened barrier, known as 'leaky gut'. Recent research has revealed that the barrier is crucial not only in IBD but also in a variety of other diseases, including autoimmune diseases associated with barrier integrity. Maintaining and strengthening barrier integrity is important in the prevention and treatment of IBD and various other diseases. Recently, natural bioactive substances known for their anti-inflammatory effects have attracted attention as important potential therapeutic agents for IBD.

[0006] Oats (Avena sativa L.) contain abundant biologically active components, exhibiting a variety of health-promoting effects, including antioxidant, anti-diabetic, antibacterial, anti-cancer, anti-hypertensive, immunomodulatory, anti-hyperlipidemia, anti-obesity, cardioprotective, and anti-inflammatory effects. Previous studies have shown that germinated oats regulate the expression of skin junctional proteins, improve skin barrier function, and treat skin barrier dysfunction, making them effective against atopic dermatitis.

[0007] Oats contain various phenolic components, among which avenanthramides (AVNs), a type of phenolic alkaloid represented by the following chemical formula I, are specifically found in oats. The content of avenanthramides and phenolic compounds in germinated oats is far higher than that of ungerminated oat seeds.

[0008] [Chemical formula I] JPEG2026079813000001.jpg3576

[0009] Abenanthramide is known as a potent antioxidant and is known to have effects such as blood pressure regulation, suppression of skin irritation, and anti-atherosclerotic effects. Recently, it has also been discovered that it can be used to treat hearing loss and degenerative neurological diseases. Thus, research on the uses of avenanthramide is being actively conducted, and it is attracting attention as a useful ingredient that can be used to treat various diseases.

[0010] However, in the context of inflammatory bowel disease (IBD), the specific effects of the active ingredients of germinated oats on intestinal permeability and tight junction (TJ) integrity have not been clarified.

[0011] Therefore, the inventors defined inflammatory bowel disease (IBD) as a phenotype associated with the interaction between the bioactive compounds of germinated oats and the genetic and proteomic environment that determines close connectivity integrity. They adopted an integrated approach to initial silico evaluation using a Combination-Oriented Natural Product Database with Unified Terminology (COONUT), combining bioinformatics analysis and chemical profiling. Furthermore, focusing on efficacy evaluation through in silico prediction validated in a test-tube (in vitro) model using a Caco-2 cell model, they elucidated the fundamental mechanism of the effect of germinated oats on intestinal permeability and confirmed that germinated oat extract (GOE) improves close connectivity integrity in the inflammatory intestinal environment and has an effect against inflammatory bowel disease, thus completing the present invention. [Prior art documents] [Patent Documents]

[0012] [Patent Document 1] Registered Patent No. 10-1745734 of the Republic of Korea [Patent Document 2] Registered Patent No. 10-1935500 of the Republic of Korea [Overview of the project] [Problems that the invention aims to solve]

[0013] The object of the present invention is devised to solve the above problems, and it is to provide a pharmaceutical composition for preventing or treating inflammatory bowel disease containing an extract of germinated oat (Avena sativa L.).

[0014] Another object of the present invention is to provide a health functional food composition for improving inflammatory bowel disease containing an extract of germinated oat.

[0015] Still another object of the present invention is to provide a probiotic composition for improving intestinal flora containing an extract of germinated oat as an active ingredient.

Means for Solving the Problems

[0016] The present invention relates to a composition for preventing, improving or treating inflammatory bowel disease (IBD) containing an extract of germinated oat (Avena sativa L.). The composition according to the present invention shows a preventive, improving or therapeutic effect on inflammatory bowel disease by improving the tight junction integrity in the intestinal environment with intestinal permeability and inflammation.

[0017] In one embodiment of the present invention, a pharmaceutical composition for preventing or treating inflammatory bowel disease containing an extract of germinated oat is provided. [[ID=XX]]

[0018] [[ID=XX]] In another embodiment of the present invention, a health functional food composition for preventing or improving inflammatory bowel disease containing an extract of germinated oat is provided.

[0019] In still another embodiment of the present invention, a probiotic composition for improving intestinal flora containing an extract of germinated oat as an active ingredient is provided.

Effects of the Invention

[0020] The present invention relates to a composition for preventing, ameliorating or treating inflammatory bowel disease containing an extract of germinated oat (Avena sativa L.). Since the composition improves intestinal permeability and improves the tight junction integrity in an inflamed intestinal environment, it can be effectively used for the production of pharmaceuticals, health functional foods or probiotics for preventing, ameliorating or treating inflammatory bowel disease.

Brief Description of Drawings

[0021] [Figure 1] Figure 1 shows the qualitative analysis chromatography of phenols and alkaloids in the extract of germinated oat, (A) positive mode; (B) negative mode. [Figure 2a] Figure 2a shows the results of predictive analysis of potential target genes and biological pathways, and is a diagram showing the complex gene expression phenotype (inflammatory bowel disease) network of germinated oat. [Figure 2b] Figure 2b shows the results of predictive analysis of potential target genes and biological pathways, and is a diagram showing genes related to tight junction (GO:CC). [Figure 2c] Figure 2c shows the results of predictive analysis of potential target genes and biological pathways. In the PPI analysis, it is a diagram showing the protein-protein interaction (PPI) analysis of two gene sets (complex genes and tight junction genes) of the red network of the top 15 genes. [Figure 2d] Figure 2d shows the results of predictive analysis of potential target genes and biological pathways, and is a diagram showing 15 genes analyzed through PPI analysis and related biological processes. [Figure 3a] Figure 3a shows the results of intestinal integrity evaluation, showing the change in epithelial electrical resistance (TEER) in the Caco-2 cell monolayer 24 hours after treatment with germinated oat. p < 0.05. [Figure 3b] Figure 3b shows the results of intestinal integrity evaluation, showing the comparative TEER values after 24 hours. p < 0.05. [Figure 3c]Figure 3c shows the results of the intestinal integrity assessment, measured by FITC-dextran permeability. The error bars represent the standard error (SE) values, and values ​​expressed with other letters in the graph indicate meaningful differences based on Duncan's multiple range test. p<0.05. [Figure 4a] Figure 4a shows the effect of germinated oats on potential inflammatory bowel disease (IBD) biomarkers, specifically focusing on tight junction proteins. Error bars represent standard error (SE) values, while other letters in the graph indicate meaningful differences based on Duncan's multiple range test. p<0.05. [Figure 4b] Figure 4b shows the effect of germinated oats on potential inflammatory bowel disease (IBD) biomarkers, specifically inflammatory proteins. The error bars represent standard error (SE) values, and other letters in the graph indicate meaningful differences based on Duncan's multiple range test. p<0.05. [Figure 4c] Figure 4c shows the effect of germinated oats on potential inflammatory bowel disease (IBD) biomarkers, specifically on cell signaling genes. Error bars represent standard error (SE) values, while other letters in the graph indicate meaningful differences based on Duncan's multiple range test. p<0.05. [Figure 4d] Figure 4d shows the effects of germinated oats on potential inflammatory bowel disease (IBD) biomarkers, specifically cell cycle-related genes. Error bars represent standard error (SE) values, and other letters in the graph indicate meaningful differences based on Duncan's multiple range test. p<0.05. [Figure 4e] Figure 4e shows the effects of germinated oats on potential inflammatory bowel disease (IBD) biomarkers, illustrating cellular structure and function. Error bars represent standard error (SE) values, and other letters in the graph indicate meaningful differences based on Duncan's multiple range test. p<0.05. [Figure 4f]Figure 4f shows the effect of germinated autoclaved wheat on potential inflammatory bowel disease (IBD) biomarkers and is a figure showing cell death molecules. Error bars represent standard error (SE) values, and values represented by other letters in the graph indicate significant differences by Duncan's multiple range test. p < 0.05. [Figure 5a] Figure 5a shows the correlation between metabolites and IBD-related biomarkers, and each square represents the Pearson correlation coefficient value (r). Red indicates a positive correlation (0 < r < 1), blue indicates a negative correlation (-1 < r < 0), and asterisks indicate significant differences (p < 0.05). It is a figure showing the correlation analysis for the negative mode of the 25 μg / mL germinated autoclaved wheat treatment group. [Figure 5b] Figure 5b shows the correlation between metabolites and IBD-related biomarkers, and each square represents the Pearson correlation coefficient value (r). Red indicates a positive correlation (0 < r < 1), blue indicates a negative correlation (-1 < r < 0), and asterisks indicate significant differences (p < 0.05). It is a figure showing the correlation analysis for the positive mode of the 25 μg / mL germinated autoclaved wheat treatment group. [Figure 6] Figure 6 is a graph illustrating the experimental results confirming that the germinated autoclaved wheat extract promotes the growth of lactic acid bacteria according to an embodiment of the present invention.

Mode for Carrying Out the Invention

[0022] Unless otherwise defined, all terms (including technical and scientific terms) used herein can be used in a meaning commonly understood by those of ordinary skill in the technical field to which this invention belongs. Also, terms defined in commonly used dictionaries are not to be interpreted ideally or overly unless specifically defined otherwise.

[0023] As used herein, the stepwise form can include multiple forms unless it clearly indicates different cases in the context.

[0024] In this specification, when a part is described as "containing" a component, this means, unless otherwise stated, that it may include other components rather than excluding them.

[0025] Furthermore, all numbers and expressions describing the quantities of components, reaction conditions, etc., as described herein should be understood to be modified in all cases by the term "drug" unless otherwise specified.

[0026] In this specification, “pharmaceutically acceptable” means that, when used in normal medicinal doses, it is approved by a government or equivalent regulatory body for use in animals, more specifically in humans, by avoiding significant toxic effects, is listed in a pharmacopoeia, or is otherwise recognized as a general pharmacopoeia.

[0027] In this invention, the term “prevention” means suppressing the development of a disease or illness in an individual who has never been diagnosed with having a disease or illness but is susceptible to such diseases or illnesses. In this specification, the term “treatment” means suppressing the development of a disease or illness; alleviating a disease or illness; and / or eliminating a disease or illness. In this specification, the term “improvement” includes alleviating symptoms, suppressing the onset of such symptoms, delaying the onset, and eliminating the onset of such symptoms.

[0028] Furthermore, unless otherwise specified, the experimental procedures described herein are the same as those commonly performed in the art.

[0029] The present invention will be described in detail below.

[0030] One aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of inflammatory bowel disease (IBD) comprising a germinated oat (Avena sativa L.) extract.

[0031] As used herein, the term "sprouted oats" may, but is not limited to, oats with a sprout length of an average of 3 to 6 mm, specifically 4 to 5 mm, and more specifically 4.2 to 4.7 mm.

[0032] In one specific example of the present invention, the germinated oats are obtained by germinating oat seeds at room temperature for two days in a 2:1 light-dark cycle, and then treating them with an inducer under the same light conditions for three days. The inducer may be, but is not limited to, abscis acid, methyl jasmonate, or a combination thereof.

[0033] The germinated oat extract, which is the active ingredient of the present invention, may be produced by a method comprising the following steps, but is not limited thereto: 1) The step of adding an extraction solvent to germinated oats and extracting the contents; and 2) The step of filtering the extract from step 1).

[0034] In the present invention, the germinated oats in step 1) can be cultivated or commercially available without limitation.

[0035] In the present invention, the extraction solvent in step 1) is preferably one or more solvents selected from the group consisting of water and organic solvents, and the organic solvent is more preferably one or more selected from the group consisting of C1 to C5 alcohols, ethyl acetate, acetone, ether, chloroform, benzene, hexane, and dichloromethane. The alcohol can be selected from the group consisting of methanol, ethanol, propanol, butanol, and isopropanol, and is preferably ethanol. As for the extraction method, hot water extraction, ultrasonic extraction, shaking extraction, Soxhelt extraction, or reflux extraction can be used, and specifically hot water extraction can be used, but is not limited thereto. The extraction time may be 1 to 24 hours, specifically 2 to 10 hours, 2 to 9 hours, or 2 to 8 hours, but is not limited thereto. The number of extractions may be 1 to 5, but is not limited thereto.

[0036] Furthermore, the germinated oat extract of the present invention may include fractions obtained by further fractionating a primary extract extracted using the aforementioned extraction solvent with an extraction solvent of a different polarity. For example, the germinated oat extract may be a fraction obtained by extracting the active ingredients contained in germinated oats with an alcohol having 1 to 5 carbon atoms, and then further fractionating it with solvents of different polarity such as ether, benzene, or hexane.

[0037] During the fractionation process, two or more solvents can be used. Each solvent extract can be produced by using them sequentially or by mixing them according to their polarity, but the method is not limited to this.

[0038] The extract produced through the process described above, or the fraction obtained by the fractionation process, can then be filtered, concentrated, or dried to remove the solvent. Both filtering, concentration, and drying are possible. Specifically, filtration can be performed using filter paper or a vacuum filter, concentration can be performed using a vacuum concentrator, for example a rotary evaporator, and drying can be performed, for example, by spray drying.

[0039] In the present invention, the germinated oat extract may be contained at a concentration of 5 to 500 μg / ml, specifically 10 to 400 μg / ml, more specifically 15 to 300 μg / ml, more specifically 20 to 200 μg / ml, and more specifically 25 to 100 μg / ml, but is not limited thereto.

[0040] As used herein, the term "avenanthramides (AVNs)" refers to alkaloid compounds that are mainly known to be produced from oats. The avenanthramide may be one or more selected from the group consisting of avenanthramide A, avenanthramide B, avenanthramide C, avenanthramide O, and avenanthramide P. Specifically, it can mean one or more components selected from the group consisting of avenanthramide A, avenanthramide B, and avenanthramide C, but is not limited thereto.

[0041] The avenanthramide can also be named anthranilic acidamides and can be represented by the following [Chemical Formula I]:

[0042] [Chemical Formula I] JPEG2026079813000002.jpg3576

[0043] In the above chemical formula, n = 1, and R 2 , 3 , 1 , 2 , 3 , 1 , 2 , 3 is H, R 2 is OH, R 3 is H, and the compound is "avenanthramide A". When n = 1, R 1 is OCH3, R 2 is OH, R 3 is H, and the compound is "avenanthramide B". When n = 1, R [[ID=3​​​​​​​ In this specification, the term "inflammatory bowel disease" may refer to one or more conditions selected from the group consisting of ulcerative colitis (UC), Crohn's disease (CD), irritable bowel syndrome, intestinal Behcet's disease, indeterminate colitis, infectious enteritis, bacterial enteritis, viral enteritis, amoebic enteritis, hemorrhagic rectal ulcer, ischemic bowel disease, radiation enteritis, tuberculous enteritis, and leaky bowel syndrome.

[0045] In one specific example of the present invention, the inventors have confirmed that the germinated oat extract improves intestinal permeability and tight junction (TJ) integrity in inflammatory bowel disease, and therefore the extract can be very usefully used as an active ingredient in pharmaceutical compositions for the prevention or treatment of inflammatory bowel disease.

[0046] Furthermore, according to one specific example of the present invention, the germinated oat extract can promote the growth of beneficial intestinal bacteria, specifically, it has been confirmed that it promotes the growth of one or more lactic acid bacteria selected from the group consisting of Lactobacillus plantarum and Lactobacillus delbrueckii, thereby improving the intestinal environment. Therefore, the germinated oat extract can be usefully used as an active ingredient in pharmaceutical compositions for the prevention or treatment of inflammatory bowel disease.

[0047] The pharmaceutical composition according to the present invention may further contain a pharmaceutically acceptable carrier in addition to an active ingredient such as germinated oat extract.

[0048] The aforementioned carriers are those commonly used in the formulation of pharmaceutically acceptable components and include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, arunate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. The pharmaceutical compositions of the present invention may additionally contain lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0049] Furthermore, the appropriate dosage of the pharmaceutical composition according to the present invention may be prescribed in various ways depending on factors such as the formulation method, administration method, the patient's age, weight, sex, medical condition, diet, administration time, administration route, excretion rate, and response sensitivity.

[0050] Furthermore, the pharmaceutical composition of the present invention can be administered orally or parenterally. When administered parenterally, it can be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, or transdermal administration. However, it is desirable that the route of administration be determined according to the type of disease to which it is applied.

[0051] Furthermore, the pharmaceutical compositions according to the present invention may be manufactured in unit volume form or by being contained in multi-volume containers by formulation using pharmaceutically acceptable carriers and / or excipients in a method readily available to a person with ordinary skill in the art to which the invention pertains. In this case, the dosage form may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or in the form of an X-agent, powder, granules, tablet or capsule, and may additionally contain dispersants or stabilizers.

[0052] Furthermore, the pharmaceutical compositions according to the present invention may additionally include carriers and vehicles commonly used in the pharmaceutical field. Specifically, these may include, but are not limited to, ion exchange resins, alumina, aluminum stearate, lecithin, serum proteins (e.g., human serum albumin), buffering substances (e.g., various phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids), water, salts or electrolytes (e.g., protamate phosphate, yttrium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride and zinc salts), colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose substrates, polyethylene glycol, sodium carboxymethyl cellulose, polyarylate, wax or lanolin, etc.

[0053] Furthermore, the pharmaceutical composition according to the present invention may be in the form of granules, powders, coated tablets, tablets, capsules, suppositories, syrups, juices, suspensions, emulsions, drips, injections, or sustained-release formulations of the active compound, and can be administered orally or parenterally in various dosage forms. When compounding, it can be prepared using fillers, bulking agents, binders, wetting agents, disintegrants, surfactants, and other diluents or excipients commonly used in the pharmaceutical field.

[0054] Another aspect of the present invention provides a composition of a health functional food for the prevention or improvement of inflammatory bowel disease (IBD) comprising germinated oat (Avena sativa L.) extract.

[0055] In the present invention, the health functional food composition may contain, but is not limited to, 0.0001 to 100% by weight of germinated oat extract.

[0056] Furthermore, since the extraction method of the germinated oat extract and the types of inflammatory bowel disease described are the same as those described in the pharmaceutical composition for the prevention or treatment of inflammatory bowel disease containing the germinated oat extract as an active ingredient, the specific explanation will refer to the above description, and below we will only explain the unique composition of the health functional food.

[0057] In one specific example of the present invention, the inventors have confirmed that the germinated oat extract improves intestinal permeability and tight junction (TJ) integrity in inflammatory bowel disease, and therefore the extract can be very useful as an active ingredient in a health functional food composition for the prevention or improvement of inflammatory bowel disease.

[0058] Furthermore, according to one specific example of the present invention, the germinated oat extract can promote the growth of beneficial bacteria in the intestines, specifically promoting the growth of one or more lactic acid bacteria selected from the group consisting of Lactobacillus plantarum and Lactobacillus delbrueckii, and improving the balance of intestinal microorganisms. Therefore, the germinated oat extract can be usefully used as an active ingredient in health functional food compositions for the prevention or improvement of inflammatory bowel disease.

[0059] The health functional food composition of the present invention can be manufactured by methods commonly used in the ordinary art, and during such manufacturing, raw materials and components commonly added in the ordinary art can be added.

[0060] Furthermore, the health functional food of the present invention can be used by adding the extract as is or by using it together with other foods or food ingredients, and can be used appropriately according to conventional methods.

[0061] The aforementioned health functional food may be, but is not limited to, one or more dosage forms selected from the group consisting of health functional food preparations such as tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, bars, beverages, gums, and candies.

[0062] Another aspect of the present invention provides a food composition for the prevention or improvement of inflammatory bowel disease (IBD) comprising germinated oat (Avena sativa L.) extract.

[0063] There are no particular restrictions on the types of food mentioned above. Examples of such food include beverages, meats, sausages, bread, biscuits, mochi, chocolate, candies, snacks, sweets, pizza, ramen, other noodle products, gums, dairy products including ice cream, various soups, drinking water, alcoholic beverages, vitamin complexes, and all other health foods in the usual sense.

[0064] The composition may contain food-grade food additives in addition to the active ingredient, and the amount of the active ingredient may be appropriately determined depending on the intended use (prevention, health, or therapeutic treatment).

[0065] The content of the extract according to the present invention may be appropriately determined depending on its intended use (prevention or improvement). Generally, the amount of the extract in a health food can be added to 0.01 to 15% by weight of the total food weight. However, in the case of long-term intake for health and hygiene purposes, or for health regulation purposes, the aforementioned amounts may be below the aforementioned range, and since there are no safety concerns, the active ingredients may also be used in amounts exceeding the aforementioned range.

[0066] The health functional beverage composition of the present invention contains the extract as an essential component in the indicated proportion, but other than that, there are no special restrictions on other components, and like ordinary beverages, it can contain various flavorings or natural carbohydrates as additional components. Examples of the natural carbohydrates mentioned above include monosacaryls, e.g., glucose, fructose; disacaryls, e.g., maltos, sucrose; and polysacaryls, e.g., dextrin, cyclodextrin and other common sugars, and sugar alcohols such as xylitol, sorbitol, and erythritol. In addition to the above, natural flavorings (thaumatin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavorings (sacaryl, aspartum, etc.) can be advantageously used.

[0067] In addition to the foregoing, the food of the present invention may contain various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring agents and medialing agents (for cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonating agents used in carbonated beverages. Furthermore, the extract of the present invention may contain fruit pulp for the production of natural fruit juices, fruit juice beverages, and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not particularly important, but it is generally selected in the range of more than 0 to about 20 parts by weight per 100 parts by weight of the extract of the present invention.

[0068] Another aspect of the present invention is to provide a prebiotic composition for improving the intestinal flora, which contains germinated oat (Avena sativa L.) extract as an active ingredient.

[0069] The term "for improving the gut microbiota" means that the product promotes the growth or proliferation of beneficial bacteria in the gut and suppresses the growth or proliferation of harmful bacteria in the gut, while maintaining a balance between beneficial and harmful bacteria in the gut.

[0070] The aforementioned "beneficial gut bacteria" can be broadly defined as microorganisms that reside in the intestines and exert beneficial effects on the human body. For example, beneficial gut bacteria may include probiotics.

[0071] The aforementioned "harmful intestinal bacteria" is a general term for microorganisms that inhabit the intestines and cause harmful effects on the human body, such as enteritis, and may include Escherichia coli, Clostridium sp., and Staphylococcus sp.

[0072] In this invention, "probiotics" can mean microorganisms that have a beneficial effect on health in the body, and may include genera such as Lactobacillus sp., Lactococcus sp., Streptococcus sp., Enterococcus sp., Pediococcus sp., and Bifidobacterium sp., and more specifically, one or more selected from the group consisting of Lactobacillus plantarum and Lactobacillus delbruecki, and more specifically, Lactobacillus plantarum subsp. Plantarum and Lactobacillus delbrueckii subsp. Bulagaricus.

[0073] In this invention, "prebiotics" can refer to components that are utilized by microorganisms, including beneficial bacteria, and that promote the growth and activity of these microorganisms, thereby exhibiting beneficial effects on the host's health.

[0074] This invention uses network biology and cheminformatics approaches to predict target biomarkers and molecular mechanisms in order to confirm the therapeutic effect of germinated oat extract on inflammatory bowel disease, and the effects of the predicted biomarkers were confirmed using a cellular model of colitis.

[0075] The effects of germinated oat extract were verified through in vitro studies, revealing a significant improvement in epithelial electrical resistance (TEER) and a decrease in fluorescent isothiocyanate (FITC) permeability. Analysis of mRNA expression of IBD-related biomarkers in Caco-2 cells showed a significant increase in mRNA levels of TJ proteins, including TJP1, TJP2, occludin, and claudin-1, compared to the inflamed group. Furthermore, germinated oat extract significantly reduced mRNA expression levels of inflammatory cytokines such as TNF-α, IL-6, and CXCL8. COCONUT and chemical profiling analysis were combined to confirm the underlying molecular mechanisms of germinated oat extract. Through these results, systematically using big data-based network biology to confirm the effects of food components, the germinated oat extract of this invention can be utilized for the prevention or treatment of IBD.

[0076] The present invention will be described in more detail below with reference to examples and experimental cases. However, the following examples and experimental cases are for illustrative purposes only, and the scope of the present invention is not limited to these.

[0077] <Example 1> Production of germinated oat extract The germinated oat extract powder was obtained from Imagine the Next Green Revolution, Inc. (INGR, Yongin, South Korea), and germinated oats rich in avenanthramide were cultivated using molecular switching technology at a smart farm.

[0078] Specifically, oat seeds (Avena sativa cv Daeyang) were germinated at 25°C for 2 days under a 16-hour:8-hour light-dark cycle. They were then treated with the inducers abscisic acid (ABA) and methyl jasmonate (MJ) at 22°C under the same light conditions for 3 days. After harvesting and washing the germinated oats, they were hot-air dried at 40°C for 1 day. The dried germinated oats were then subjected to a primary hot water extraction at 120°C for 5 hours. Following the primary extraction, the residue was rehydrated with distilled water and a secondary extraction was performed at 105°C for 3 hours. Both the primary and secondary extracts were treated with 0.2% (v / v) 1,4-alpha-D-glucan glucanohydrolase (BAN 480LS, Novozymes) at 75°C for 1 hour, after which 0.2% (v / v) amylase (Termamyl 2X, Novozymes) was added and the reaction was continued at 95°C for 1 hour. The mixture was heated at 105°C for 1 hour to inactivate the enzymes, filtered through a 1 μm filter, concentrated to over 20 Brix at 65°C, and finally spray-dried using a spray dryer (SPRAY DRYER FS-2.0D, Fine ST Co.). The spray-dried germinated oat flour was dissolved in a 30% ethanol solution, and the following in-vitro experiments were performed.

[0079] <Experimental Example 1> Phenolic compound profiling and qualitative analysis of germinated oats and quantitative evaluation of avenanthramides Analysis method Combination of high-performance liquid chromatography and Q Exactive Orbitrap mass spectrometry (HPLC / Q Exactive Orbitrap MS) Qualitative analysis of the germinated oat extract obtained in Example 1 was performed using a Thermo Vanquisher instrument (Thermo Fisher, Waltham, Massachusetts, USA). A Waters Cortects T3 column (2.1 mm × 150 mm, 1.6 μm particle size; water) was used for chromatographic analysis.

[0080] Chromatographic analysis was performed using mobile phase A, which consisted of water with 0.1% formic acid added, and mobile phase B, which consisted of acetonitrile with 0.1% formic acid added, at a flow rate of 0.25 mL / min using gradient elution.

[0081] The mass spectrometry conditions are as follows: The ion source type is heated electro-nebulized ionization (H-ESI); the cation mode voltage is 3.5kV; the anion mode voltage is 3kV; the cover gas flow rate is 50 Arb; the auxiliary gas flow rate is 10 Arb; the sweep gas flow rate is 1 Arb; and the ion transfer tube temperature is 320°C.

[0082] The mass spectrometer operates within a scan range of 100–1500 m / z, with a full width at half maximum (FWHM) of 70,000 for MS1 and 17,500 for MS2. It is configured to select the top 10 precursor ions for fragmentation using stepwise normalized collision energy (NCE) settings of 10, 30, and 50, and to perform data-dependent acquisition in topN mode. These settings, leveraging advanced mass spectral libraries and analytical software, facilitate detailed analysis and identification of compounds based on isotopic distribution patterns, ion fragmentation patterns, and accurate mass measurements.

[0083] Analysis results Analysis of germinated oats for phenols, alkaloids, and avenanthramides using HPLC-MS / MS revealed a total of 23 compounds in the spray-dried oats (Figure 1).

[0084] Table 1 shows detailed information such as retention time, ionization mode, precursor and generated ions, as well as chromatograms of the results of quantitative analysis of the major components of germinated oat compounds via HPLC-MS / MS. Table 2 shows the results of quantitative analysis of avenanthramide contained in spray-dried germinated oat extract, with each value representing the mean ± standard deviation.

[0085] [Table 1] JPEG2026079813000003.jpg225169JPEG2026079813000004.jpg224164JPEG2026079813000005.jpg223164JPEG2026079813000006.jpg99170

[0086] [Table 2] JPEG2026079813000007.jpg62128

[0087] In negative mode, gentisic acid, protocatechuic aldehyde, vanillin, 4-coumarate, 4-feruloylquinic acid, 6-hydroxy-7-methoxycoumarin, ferulate, procyanidin B2, and nedocromil were detected. In positive mode, dianthoside, romucosine D, lyxoflavin, avenanthramide 1c, and zanthodioline were identified. Additionally, AVN A, AVN B, AVN C, AVN E, AVN G, AVN 2, AVN1s, AVN L, AVN 2fd-1, AVN 2fd-2, and (S)-Annocherine A were detected in both negative and positive modes.

[0088] <Experimental Example 2> Insilico: Gene-Phenomenon Network Approach Silico experiments were used to predict potentially important biomarkers that may be controlled by germinated oats.

[0089] 2-1. Comprehensive analysis of tight tunic genes regulated by germinated oat compounds Based on the compound profiling in Experimental Example 1, the effects of germinated oats on intestinal health were analyzed.

[0090] Selection of a test gene set Network analysis was performed using the COCONUT database and QuickGeneOntology (GO) to accurately identify closely linked and germinated oat plant-related genes. Compound-gene and gene-phenotypic association data were collected using COCONUT data. COCONUT provides a standardized, structured, and integrated database for natural products and is used to explore component genes, associated efficacy, and target genes, and to investigate diseases that may be affected by genetic alterations from medicinal plants.

[0091] Close tangency-related genes were collected using Gene Ontology annotations, and the genes were collected from 'cellular components' specifically focused on genes associated with close tangency.

[0092] Of the 25 compounds found in germinated oats, 14 were identified as major compounds associated with IBD.

[0093] The two-layered network consists of 404 nodes and 1,220 edges (Figure 2a). We focused on genes potentially regulated by compounds found in germinated oats, with particular emphasis on identifying genes associated with gut health. A total of 389 genes were mapped to the compound-gene-phenotype network scaffold (Figure 2a). To identify specific functions related to gut barrier health, we investigated close tethering-related genes in Homo sapiens species. Genes associated with gut permeability and close tethering were selected into the category of fast GO cell components (CCs), and although a total of 147 Homo sapiens genes associated with close tethering function were specifically identified, they were integrated into a gene set as shown in 2b.

[0094] 2-2. Confirmation of interactions between target genes and closely coupled genes derived from germinated oats. We focused on identifying genes potentially regulated by compounds in germinated oats, particularly those related to the maintenance and function of close tangency, which is crucial for gut health. To confirm the relationship between close tangency-related genes and compound-driven genes, we joined two gene datasets and performed protein-protein interaction (PPI) network analysis on these datasets.

[0095] Analysis method PPI analysis was performed using the aforementioned dataset to identify potential biomarkers associated with both compound-related genes and tightly coupled genes. Compound-driven phenotype-related genes and tightly coupled genes were coupled for PPI analysis, and the PPI analysis was performed using the coupled genes. The STRING database (version 12.0) was used for protein-protein network analysis. All genes were mapped to Homosapiens genes, and Cystoscope (version 3.10.0) was used for visualization.

[0096] Analysis results Fifteen genes were identified in the following order: JP1, CTNNB1, OCLN, AKT1, IL6, ACTB, TNF, BCL2, TJP3, TJP2, CXCL8, CCND1, CLDN1, CLDN3, and ALB (Figure 2c).

[0097] PPI analysis classified the genes shown in Table 3 by function, and as a result, six functions were confirmed for the classified genes: tight tethering, cell signaling, cell cycle, inflammatory response, cell death, and cell structure and function.

[0098] [Table 3] JPEG2026079813000008.jpg223159JPEG2026079813000009.jpg231150JPEG2026079813000010.jpg32160

[0099] 2-3. Prediction of biological pathways in IBD inflammatory responses and barrier integrity genes through gene ontology analysis. Analysis method To confirm the mechanism by which germinated oat extract promotes gut health in Caco-2 cells, we analyzed biological pathways using DAVID (https: / / david.ncifcrf.gov / tools.jsp). The top 15 genes from the PPI analysis were used for pathway prediction analysis, which was represented by gene ontology biological processes. To identify biological pathways associated with inflammatory bowel disease (IBD), we selected only IBD-related pathways. To select biological pathways related to IBD, we collected IBD-related keywords through the Human Phenotype Ontology (https: / / hpo.jax.org / app / , (Gargano et al., 2023)).

[0100] A phenotypic keyword search using Quick GO identified biological pathways associated with the aforementioned terms. Subsequently, sequential analysis was performed focusing on the top 15 genes and biological pathway lists derived from Quick GO, utilizing Human Phenotype Ontology (HPO) keywords as a reference. Only biological pathways that showed overlap between analyses were included in the results, and Tableau (version 2022.2.0) was used for visualization. All pathway results show p<0.05.

[0101] Analysis results Enrichment analysis of the gene ontology related to IBD identified key biological processes, including inflammatory responses, intracellular signaling, positive regulation of gene expression, negative regulation of cell death processes, and positive and negative control of cell proliferation. Processes such as "inflammatory response," "cell-cell junction tissue," "positive regulation of gene expression," "positive regulation of wound healing," "maintenance of blood-brain barrier permeability," and "establishment of the endothelial gut barrier" are among those ranked (Figure 2d).

[0102] The associations between genes identified by PPI analysis and specific biological processes, along with detailed statistical significance values, are shown in Table 4. All biological processes listed in Table 4 have a statistical significance level of less than 0.05 p-value.

[0103] [Table 4] JPEG2026079813000011.jpg228168JPEG2026079813000012.jpg226163JPEG20260798130000 13.jpg211163JPEG2026079813000014.jpg226163JPEG2026079813000015.jpg226163JPEG202 6079813000016.jpg219163JPEG2026079813000017.jpg226163JPEG2026079813000018.jpg2 18163JPEG2026079813000019.jpg226163JPEG2026079813000020.jpg226163JPEG2026079813 000021.jpg211163JPEG2026079813000022.jpg226163JPEG2026079813000023.jpg226163JP EG2026079813000024.jpg211163JPEG2026079813000025.jpg226163JPEG2026079813000026. jpg226163JPEG2026079813000027.jpg225163JPEG2026079813000028.jpg218163JPEG202607 9813000029.jpg212163JPEG2026079813000030.jpg226163JPEG2026079813000031.jpg74169

[0104] <Experiment Example 3> In vitro experiment In vitro experiments were conducted to confirm that the genes predicted through the aforementioned silico analysis are regulated by germinated oat treatment, thereby improving gut health.

[0105] cell culture Caco-2 cells (passages 33-37) were purchased from ATCC (Manassas, Virginia, USA). The cells were supplemented with 10% Utea serum (FBS) (Gibco BRL, New York, USA), 1% penicillin-streptomycin (P / S; Corning Inc., New York, USA), and 1% non-essential amino acids (NEAAs; Gibco, Rockville, Maryland, USA) in Dulbecco's modified Eagle's Medium (DMEM) (Biowest, Cells were cultured in a 37°C, 5% CO2 incubator (JPEG2026079813000032.jpg427, Cholet, France). For the experiment, cells with a 0.4 μm pore size were cultured for differentiation for 21 days at a density of 3.75 × 10⁵ cells / well in polyester 6-well transwell Costar® (Kennebunk, ME, USA). The culture medium was changed every 2-3 days, and experimental concentrations without cytotoxic effects were determined and applied to the study, based on the results obtained by CCK8 analysis.

[0106] Cell viability Cell viability was evaluated using the Cell Counting Kit-8 (CCK8) test (Sigma-Aldrich), a cell proliferation test based on the cleavage of 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium (WST-8).

[0107] Specifically, cells were divided into 24-well plates at a cell density of 1 × 10⁵ cells per well and cultured for 21 days to induce differentiation. After 21 days, the cultured cells were replaced with serum-free medium. The cells were treated with various GOE concentrations (1, 2.5, 5, 10, 25, 50, and 100 μg / mL) for 24 hours. After 24 hours of culture, the cells were supplemented with 100 μL CCK-8 and cultured for 4 hours in a 5% CO₂ incubator at 37°C.

[0108] 3-1. Assessment of intestinal health status using TEER and FITC-dextran This invention primarily focuses on the integrity of close junctions, which are crucial in the pathophysiology of inflammatory bowel disease (IBD). An in vitro model reflecting colitis observed in IBD was established using an inflammatory cytokine cocktail, and the integrity and permeability of the epithelial barrier were evaluated using epithelial electrical resistance (TEER) and FITC-dextran permeability measurements.

[0109] Trans-epithelial electrical resistance (TEER) A combined therapeutic approach was used to investigate the effects of germinated oat extract on intestinal barrier integrity in an inflammatory state. Initially, a cytokine cocktail consisting of 25 ng / mL interleukin (IL)-1β, 10 ng / mL lipopolysaccharide (LPS), 50 ng / mL tumor necrosis factor (TNF)-α, and 50 ng / mL interferon (IFN)-γ was administered basally to the extrabasal transwell to simulate the colitis environment. Simultaneously, germinated oat extract at concentrations of 2.5 and 25 μg / mL and LPS (10 μg / mL) were introduced into serum-free medium from the apical side. After simultaneous application of germinated oat extract and cytokine cocktail, TEER values ​​were measured at 0, 3, 6, 12, and 24-hour intervals using the Millicell® ERS (Millipore) system.

[0110] Fluorescein isothiocyanate (FITC)-dextran flux High intensity values ​​indicate high transmittance. FITC-dextran solution (100 μg / mL) and germinated oat extract were added to the apical side, and after 24 hours of incubation, fluorescence was collected on the basal outer side of the transwell. The collected fluorescence solution was measured at 485 nm and 535 nm (excitation and emission wavelengths, respectively) using a SpectraMax i3x Multi-Mode Microplate Reader.

[0111] Analysis results To confirm the effects on the integrity and permeability of Caco-2 epithelial cells, epithelial electrical resistance (TEER) and FITC-dextran permeability were measured. The results showed that the TEER value was significantly lower in the cocktail treatment group than in the control group.

[0112] Samples were treated with 25 or 2.5 μl / mL, and a significant effect in the germinated oat extract treatment group appeared 12 hours after treatment (p<0.05). In the germinated oat extract treatment group, TEER values ​​improved in a concentration-dependent manner compared to the cocktail group (Figures 3a, 3b). In the cocktail-administered group, FITC-dextran flux increased approximately 2.5 times compared to the control group, and germinated oat treatment significantly reduced FITC flux (Figure 3c).

[0113] Treatment with germinated oat extract significantly improved barrier function, as evidenced by improved TEER values ​​and reduced FITC-dextran flux. These results suggest that the biologically active components in germinated oat extract can exert a protective effect against cytokine-induced barrier disruption, thus confirming their potential role in regulating intestinal permeability in the context of IBD.

[0114] 3-2. PCR validation of gene regulation for germinated oats Subsequently, we analyzed the potential genetic causes of these effects and predicted genes associated with gut integrity and potentially affected by germinated oat compounds. This led to the identification of 15 candidate genes thought to mediate the protective effect of oats on gut integrity. To validate the predicted genes, we used PCR in a Caco-2 cell monolayer model focused on various biomarkers.

[0115] RNA isolation and real-time reverse transcription polymerase chain reaction (qRT-PCR) Following the manufacturer's guidelines, samples were treated with TRIZOR® reagent (LifeTechnologies, Rockville, Maryland, USA) for 24 hours, after which total RNA was isolated from Caco2 cells. After treating the cells with TRIZOR and chloroform, the resulting mixture was centrifuged at 12,000 rpm at 4°C for 20 minutes. The upper layer was carefully transferred, and the mixture, to which the same amount of isopropanol was added, was centrifuged again under the same conditions to precipitate mRNA. NanoDrop was used to evaluate the amount of total RNA. TMA LiteSpectrophotometer was used. cDNA was synthesized from total RNA using the Transcriptor First Strand cDNA Synthesis Kit (Roche).

[0116] The top 14 genes analyzed via protein-protein interaction (PPI) analysis were validated using qRT-PCR. mRNA expression levels were normalized relative to the expression of the housekeeping gene GAPDH. ΔΔCT It was quantified using the method.

[0117] Analysis results Of the 15 genes analyzed by PPI, TJP1, TJP2, TJP3, OCLN, CLDN1, and CLDN3 are associated with tight junction function. TJP1, TJP2, and OCLN, which are important for maintaining epithelial barrier integrity, were significantly reduced under cocktail conditions compared to the control group. TJP1, TJP2, and OCLN are important for maintaining epithelial barrier integrity and were significantly improved when treated with germinated oat extract compared to the cocktail group (p=0.0027, p=0.0025, p<0.0001).

[0118] On the other hand, the expression of TJP3 and CLDN3, members of the tightly coupled protein system, did not change significantly in the germinated oat extract treatment group, but showed an increasing trend (Figure 4a). TNF-α, IL-6, and CXCL8 are inflammatory cytokines that are upmodified in IBD, and mRNA expression of TNF-α, CXCL-8, and IL-6 increased significantly by about four times in the cocktail group, but was significantly decreased in the germinated oat extract group (p<0.0001) (Figure 4b). In addition, CTNNB1 decreased significantly in response to germinated oat treatment (p<0.0001), and AKT1 showed a similar trend, although it did not show statistical significance, and both were regulated by germinated oat extract treatment (Figure 4c). In the case of CCND1, different responses were observed depending on the extract concentration.

[0119] In this invention, two concentrations of germinated oat extract, 2.5 μg / mL and 25 μg / mL, were used. The results showed that CCND1 expression decreased at a concentration of 2.5 μg / mL of germinated oat extract, while CCND1 expression increased at the higher concentration of 25 μg / mL (Figure 4d), tightly linked protein expression increased significantly, and inflammatory cytokine levels decreased remarkably. Through these results, it was confirmed that germinated oat extract has a dose-dependent effect on improving intestinal health.

[0120] ACTB, a β-actin gene related to cell structure and function, was significantly decreased in the group administered the inflammatory cocktail, but significantly increased in the group administered germinated oat extract (Figure 4e). On the other hand, cyclin D (CCND1), which is involved in cell cycle regulation, and BCL-2, which is related to cell death, were both significantly increased in the cocktail administration group. Bcl-2 expression was significantly decreased in the group administered germinated oat extract (Figure 4f). Fourteen out of fifteen genes were examined (Figure 4). Through these results, we confirmed that the germinated oat extract of the present invention contributes to maintaining barrier integrity by regulating the ACTB gene.

[0121] 3-3. Correlation analysis of germinated oat compounds abundant in avenanthramides and IBD-related markers at various treatment concentrations Figure 5 shows the strength of the correlations between the analyzed factors, with red, blue, and white indicating positive, negative, and no correlation, respectively. Characteristically, avenanthramide compounds in germinated oats showed stronger correlations in the negative mode than in the positive mode. TEER values ​​generally showed positive correlations with avenanthramide compounds, while FITC-dextran permeability test values ​​showed a fairly negative correlation with avenanthramide compounds. Among tightly coupled genes, TJP1 and CLDN3 showed the strongest correlations with avenanthramide compounds, while TJP2 and TJP3 showed positive correlations not with avenanthramides, but with other phenolic compounds such as 4-coumarate and nedocromil. In the context of inflammation-related genes, TNF-α and IL-6 showed significant negative correlations with avenanthramide compounds, protocatexanaldehyde, and phenolic compounds including romucosin D. The effects on CXCL8 showed a negative correlation with non-avenanthramide phenol compounds such as gentisic acid and 4-coumarate. Cellular signaling genes such as akt-1 and CTNNB1 generally showed a negative correlation with avenanthramide compounds, while CCND1 showed a positive correlation with non-avenanthramide alkaloids and phenol compounds in a negative mode. The actin cytoskeleton gene ACTB mainly showed a positive correlation with avenanthramide compounds, while the BCL-2 gene generally showed a significant negative correlation with avenanthramide compounds. These results confirm specific interactions between germinated oat metabolites and key biological markers, which imply potential therapeutic effects on inflammatory bowel disease.

[0122] <Experimental Example 4> Effect of germinated oat extract on promoting the growth of lactic acid bacteria Intestinal lactic acid bacteria are said to improve the balance of intestinal microorganisms by producing organic acids, which lower the intestinal pH and suppress the growth of harmful bacteria. Therefore, in this invention, in order to confirm the effect of germinated oat extract on the growth of beneficial intestinal bacteria, changes in the proliferation of lactic acid bacteria were measured after treatment with germinated oat extract. As a result, it was confirmed that the germinated oat extract of this invention promotes the growth of lactic acid bacteria and can contribute to the improvement of the intestinal environment.

[0123] badge MRSA + Germinated Oat Extract (0%, 1%, 5%) MRS broth agar (MRSA) Raw batch 50 plate: MB-P1040-P50 Germinated oat extract (0%, 1%, 5%)

[0124] Test method - Standard plate method 1. Dilute 10 to 25 g (mL) of the sample with the diluent in a 1:9 ratio (dilute only as needed). 2. Dispense 1 ml of diluent into two Petri dishes (KS-P0101). 3. Dispense the prepared MRSA, cooled to 43 or 45°C, mix well, and let it solidify (once the badge has solidified, add another 3 to 5 ml and overlap). 4. Incubate at 4.30±1℃ for 24 hours.

[0125] Specifically, Lactobacillus plantarum subsp. Plantarum and Lactobacillus delbrueckii subsp. Bulagaricus were added to MRS medium containing 2% and 5% germinated oat extract prepared in Example 1, respectively. 9 1% Lactobacillus CFU / mL (10%) 7 After inoculation at a CFU / mL concentration, the samples were cultured in a 30°C incubator under aerobic conditions for 24 hours, and the change in the total number of cultured lactic acid bacteria was measured. The lactic acid bacteria count was measured using the standard plate method based on Section 8, General Test Methods - 4.5.1, General Bacterial Count Measurement Method, of the Food Safety Standards Act. 10 to 25 g (mL) of the sample was diluted with the diluent in a 1:9 ratio, and 1 mL was hastily placed on two Petri dishes (KS-P0101), and the lactic acid bacteria count (CFU / mL) was calculated. For the control group, MRS medium (0% germinated oat extract) was used, and the lactic acid bacteria count was calculated using the same method. The results are shown in Table 5 and Figure 6 below.

[0126] [Table 5] JPEG2026079813000033.jpg85163Data are expressed as the mean±SD(n=2).Values ​​with different uppercase letters within the same column are significantly different(P<0.05)according to Tukey's range test.CFU,colony-forming unit.

[0127] As a result, it was revealed that both Lactobacillus plantarum and Lactobacillus delbuer lactic acid bacteria significantly increased in the germinated oat extract treatment groups (2%, 5%) of the present invention compared to the control group (0%).

[0128] statistical analysis Data are expressed as mean ± standard error (SE). Statistical evaluation was performed using one-way analysis of variance (one-way ANOVA) with SAS 9.4 (SAS Institute, Carrey, North Carolina, USA). For post-hoc analyses, Duncan's multiple range test was used for both TEER and FITC-dextran measurements and PCR data analysis. Statistical significance in comparisons with a p-value less than 0.05 is expressed using different letters for each group.

[0129] In conclusion, experimental validation of germinated oat extract demonstrated positive regulation of permeability and effective regulation of 9 out of 14 genes tested (TJP1, TJP2, OCLN, CLDN1, TNF, IL-6, CXCL8, CTNNB1, CCND1). In particular, germinated oats effectively regulated genes related to tight junctions and inflammation. Furthermore, germinated oat extract was found to promote the growth of beneficial gut bacteria (especially lactic acid bacteria) and improve the balance of gut microbiota. Therefore, the germinated oat extract of this invention can be effectively used for the prevention, improvement, or treatment of inflammatory bowel disease by maintaining the integrity of tight junctions between intestinal epithelial cells, regulating intestinal permeability and inflammatory responses, and simultaneously promoting the growth of beneficial gut bacteria.

[0130] The above description of the present invention is illustrative, and a person with ordinary skill in the art will understand that it can be easily modified into other specific forms without altering the technical idea or essential features of the present invention. Therefore, the embodiments described above should be understood in all respects as illustrative and not limiting.

Claims

1. A pharmaceutical composition for the prevention or treatment of inflammatory bowel disease (IBD), comprising germinated oat (Avena sativa L.) extract and a pharmaceutically acceptable carrier.

2. The inflammatory bowel disease is one or more selected from the group consisting of ulcerative colitis (UC), Crohn's disease (CD), irritable bowel syndrome, intestinal Behcet's disease, indeterminate colitis, infectious enteritis, bacterial enteritis, viral enteritis, amoebic enteritis, hemorrhagic rectal ulcer, ischemic bowel disease, radiation enteritis, tuberculous enteritis, and leaky enter syndrome, according to claim 1.

3. The germinated oats are obtained by germinating oat seeds in a 2:1 light-dark cycle for 1 to 3 days, and then treating them with an inducer in a 2:1 light-dark cycle for 2 to 5 days, as described in claim 1.

4. The pharmaceutical composition according to claim 3, wherein the inducer is abscis acid, methyl jasmonate, or a combination thereof.

5. The aforementioned germinated oat extract is 1) The step of adding an extraction solvent to germinated oats and extracting the oats; and 2) Produced by a method comprising filtering the extract from step 1) and then concentrating or drying it, Here, the extraction solvent in step 1) is one or more solvents selected from the group consisting of water and organic solvents. The pharmaceutical composition according to claim 1, wherein the drying in step 2) is performed by spray drying.

6. The pharmaceutical composition according to claim 5, wherein the organic solvent is one or more selected from the group consisting of C1 to C5 alcohols, ethyl acetate, acetone, ether, chloroform, benzene, hexane, and dichloromethane.

7. The pharmaceutical composition according to claim 1, characterized in that the germinated oat extract improves intestinal permeability and tight junction (TJ) integrity that occur due to inflammatory bowel disease.

8. The pharmaceutical composition according to claim 1, characterized in that the germinated oat extract promotes the growth of beneficial bacteria in the intestines.

9. The pharmaceutical composition according to claim 8, characterized in that the beneficial bacteria are one or more lactic acid bacteria selected from the group consisting of Lactobacillus plantarum and Lactobacillus delbruecki.

10. A functional food composition for the prevention or improvement of inflammatory bowel disease (IBD), containing germinated oat (Avena sativa L.) extract.

11. The inflammatory bowel disease is one or more selected from the group consisting of ulcerative colitis (UC), Crohn's disease (CD), irritable bowel syndrome, intestinal Behcet's disease, indeterminate colitis, infectious enteritis, bacterial enteritis, viral enteritis, amoebic enteritis, hemorrhagic rectal ulcer, ischemic bowel disease, radiation enteritis, tuberculous enteritis, and leaky enter syndrome, as described in claim 10.

12. The health functional food composition according to claim 10, characterized in that the germinated oat extract improves intestinal permeability and tight junction (TJ) integrity that occur in inflammatory bowel disease.

13. The health functional food composition according to claim 10, wherein the health functional food is one or more formulations selected from the group consisting of health functional food formulations such as tablets, capsules, pills, granules, liquids, powders, flattened forms, pastes, syrups, gels, jellies, bars, beverages, gums, and candies.

14. A food composition for the prevention or improvement of inflammatory bowel disease (IBD), containing germinated oat (Avena sativa L.) extract.

15. The food composition according to claim 14, wherein the inflammatory bowel disease is one or more selected from the group consisting of ulcerative colitis (UC), Crohn's disease (CD), irritable bowel syndrome, intestinal Behcet's disease, indeterminate colitis, infectious enteritis, bacterial enteritis, viral enteritis, amoebic enteritis, hemorrhagic rectal ulcer, ischemic bowel disease, radiation enteritis, tuberculous enteritis, and leaky enter syndrome.

16. The food composition according to claim 14, wherein the food is one or more selected from the group consisting of various drinks, meats, sausages, bread, biscuits, mochi, chocolates, candies, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, soups, drinking water, alcoholic beverages, and vitamin complexes.

17. A prebiotic composition for improving the gut microbiota, containing germinated oat (Avena sativa L.) extract as an active ingredient.

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