KR102779977B1 - Composition for reducing body fat of sprout oat extract - Google Patents

Abstract

The present invention relates to a composition for reducing body fat or a composition for preventing or treating obesity, comprising an extract of sprouted oats or a compound derived therefrom as an effective ingredient.

Description

{Composition for reducing body fat of sprout oat extract}

The present invention relates to a composition for reducing body fat, which comprises a sprouted oat extract or a compound derived therefrom as an effective ingredient, and more particularly, to confirm the body fat reducing and obesity preventing or treating effects of a sprouted oat extract or a compound derived therefrom, and to provide the same as a food composition, a pharmaceutical composition, and a feed composition.

In modern society, due to the rapid development of medical technology, awareness of health is spreading, and the trend of pursuing a high quality of life is a phenomenon that is not only in our country but also worldwide, and it is expected that this trend will accelerate further in the future. Therefore, the development of new biomedical materials, functional food materials, and wellness materials that can meet the rapidly changing social phenomenon and the increasingly diverse expectations of consumers is required.

The above-ground parts of cereal crops, including young leaves after sowing, such as cereal sprouts, such as oat sprouts, have long been known as nutritionally excellent food sources with high vitamin and mineral content, and contain functional secondary metabolites that exhibit antioxidant effects, increasing their potential for industrial use as health functional foods and pharmaceutical materials.

Meanwhile, AMPK (AMP-activated protein kinase) is an enzyme that acts on cellular energy homeostasis. It is a heterotrimer with three subunits, a, b, and g, and is a functional enzyme that is conserved from yeast to humans, and is known to induce inhibition of neutral fat synthesis in the body and lipolysis of fat cells. When the energy homeostasis in the body that AMPK is involved in is broken and energy imbalance occurs, dysfunction of muscles, liver, and fat cells occurs, and various metabolic diseases occur as a result.

Accordingly, the inventors of the present invention have conducted extensive research efforts to find a substance effective in reducing body fat and preventing or treating obesity from a natural product whose safety in the human body is ensured, and as a result, have confirmed that sprouted oat extract exhibits excellent AMPK activity in reducing body fat and preventing or treating obesity, thereby completing the present invention.

One object of the present invention is to provide a food composition for reducing body fat comprising a sprouted oat extract or a compound derived therefrom as an effective ingredient.

Another object of the present invention is to provide a method for inhibiting differentiation into adipocytes, comprising the step of treating isolated preadipocytes or isolated stem cells capable of differentiating into adipocytes with an extract of oat sprouts or a compound derived therefrom.

Another object of the present invention is to provide a pharmaceutical composition for reducing body fat comprising an extract of sprouted oats or a compound derived therefrom as an effective ingredient.

Another object of the present invention is to provide a method for reducing body fat, comprising a step of administering the pharmaceutical composition to a subject other than a human.

Another object of the present invention is to provide a feed composition for reducing body fat comprising a sprouted oat extract or a compound derived therefrom as an effective ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating obesity, comprising an extract of sprouted oats or a compound derived therefrom as an effective ingredient.

Another object of the invention is to provide a food composition for preventing or improving obesity, comprising a sprouted oat extract or a compound derived therefrom as an effective ingredient.

Another object of the invention is to provide a feed composition for preventing or improving obesity, comprising a sprouted oat extract or a compound derived therefrom as an effective ingredient.

This is explained specifically as follows. Meanwhile, each description and embodiment disclosed in the present invention can also be applied to each other description and embodiment. That is, all combinations of various elements disclosed in the present invention fall within the scope of the present invention. In addition, the scope of the present invention cannot be considered limited by the specific description described below.

In order to achieve the above-mentioned purpose, the first aspect of the present invention provides a food composition for reducing body fat comprising a sprouted oat extract or a compound derived therefrom as an effective ingredient.

In addition, the second aspect of the present invention for achieving the aforementioned purpose provides a method for inhibiting differentiation into adipocytes, comprising the step of treating an extract of sprouted oats or a compound derived therefrom to isolated preadipocytes or isolated stem cells capable of differentiating into adipocytes.

In addition, the third aspect of the present invention for achieving the aforementioned purpose provides a pharmaceutical composition for reducing body fat comprising a sprouted oat extract or a compound derived therefrom as an effective ingredient.

In addition, the fourth aspect of the present invention for achieving the aforementioned purpose provides a method for reducing body fat, comprising a step of administering the pharmaceutical composition to a subject other than a human.

In addition, the fifth aspect of the present invention for achieving the aforementioned purpose provides a feed composition for reducing body fat comprising a sprouted oat extract or a compound derived therefrom as an effective ingredient.

In addition, the sixth aspect of the present invention for achieving the aforementioned purpose provides a pharmaceutical composition for preventing or treating obesity comprising a sprout extract or a compound derived therefrom as an effective ingredient.

In addition, the seventh aspect of the present invention for achieving the aforementioned purpose provides a food composition for preventing or improving obesity, comprising a sprouted oat extract or a compound derived therefrom as an effective ingredient.

In addition, the eighth aspect of the present invention for achieving the aforementioned purpose provides a feed composition for preventing or improving obesity, comprising a sprouted oat extract or a compound derived therefrom as an effective ingredient.

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

The term "sprout oats" of the present invention refers to sprouts of oats that have grown to about 10 cm. Specifically, it may contain Avenacoside, a saponin-based functional substance, or Flavone-Glycoside.

The term "extract" of the present invention means a substance separated from a liquid or solid mixture by a solvent. Specifically, it may be obtained by extraction with a solvent selected from the group consisting of water, alcohol, lower alcohols having 1 to 4 carbon atoms, and mixed solvents thereof.

The term "compound" in the present invention means a compound isolated or purified from a sprouted oat extract. Specifically, Avenafuranol, Diosgenoside, Avenacoside A, Avenacoside B, 26-deglucoavenacoside B, Isoorientin-6-β-D-xylopyranoside, Vitexin 2``-O-rhamnoside, Isoswertisin 2``-O-rhamnoside, Isovitexin 2``-O-arabinoside, 3-O-Feruloylquinic acid, and It may be any one selected from the group consisting of 26-deglucoavenacoside A.

The term "body fat reduction" of the present invention means the effect of reducing body fat and thus reducing body fat percentage even without a change in body weight, and the effect of suppressing an increase in body fat and thus suppressing an increase in body fat percentage.

The term "obesity" of the present invention refers to a state of energy imbalance in the body caused by consuming more energy than the body needs or consuming less energy than consumed, resulting in excessive adipose tissue. Specifically, obesity is mainly caused by a rapid increase in the amount of fat precursor cells in the body differentiating into fat cells due to fat accumulation.

The term "obesity-related disease" of the present invention refers to a disease caused by various abnormalities due to excessive accumulation of fat cells caused by obesity. Substances called adipokines secreted from fat cells regulate metabolism in the body, and obesity causes an imbalance of these substances, resulting in abnormalities in fat and glucose metabolism. Specifically, obesity-related diseases may include hypertension, diabetes, hyperlipidemia, fatty liver, gallstones, obstructive sleep apnea, irregular menstruation, polycystic ovary disease, infertility, decreased libido, depression, degenerative arthritis, gout, colon cancer, pancreatic cancer, prostate cancer, breast cancer, and various other cancers.

The term "prevention" of the present invention means any act of inhibiting or delaying the onset of obesity by administering a pharmaceutical composition according to the present invention.

The term "improvement" in the present invention means any action that at least reduces a parameter related to a condition being treated by administering a composition according to the present invention, for example, the severity of a symptom.

The term "treatment" of the present invention means any action whereby the symptoms of a subject suspected of obesity or suffering from obesity are improved or beneficially changed by administration of the pharmaceutical composition.

A food composition comprising the sprouted oat extract of the present invention or a compound derived therefrom as an effective ingredient is characterized as being for reducing body fat or preventing or treating obesity.

It can be included in a food composition for the purpose of reducing body fat, preventing or improving obesity, and since the food composition can be consumed on a daily basis, a high effect can be expected for reducing body fat, or preventing or improving obesity.

At this time, the descriptions of the above “oat sprouts”, “extract”, “compound”, “body fat reduction”, “obesity”, “treatment”, “improvement” and “prevention” are as described above.

The term "health functional food" of the present invention means a food manufactured and processed using raw materials or ingredients having functionality useful to the human body according to Act No. 6727 on Health Functional Food, and "functionality" means obtaining a useful effect for health purposes such as regulating nutrients for the structure and function of the human body or physiological effects. Meanwhile, health food means a food that has a more active health maintenance or promotion effect than general food, and health supplement food means a food for the purpose of health supplementation. In some cases, the terms health functional food, health food, and health supplement food may be used interchangeably.

The sprouted oat extract of the present invention or a compound derived therefrom may be added as is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method.

The food of the present invention can be manufactured by a method commonly used in the art, and can be manufactured by adding raw materials and ingredients commonly added in the art during the manufacturing process. Specifically, the food composition may additionally contain a physiologically acceptable carrier, and the type of the carrier is not particularly limited, and any carrier commonly used in the art may be used. In addition, the food composition may contain food additives such as preservatives, sterilizers, antioxidants, colorants, coloring agents, bleaching agents, seasonings, sweeteners, flavorings, leavening agents, reinforcing agents, emulsifiers, thickeners, film agents, gum bases, antifoaming agents, solvents, and improvers. The additives may be selected depending on the type of food and used in an appropriate amount.

In addition, the formulation of the food can be manufactured without limitation as long as it is a formulation recognized as a food. The food composition of the present invention can be manufactured in various forms of formulations, and unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long time by using food as a raw material, and is highly portable, so the food of the present invention can be consumed as a supplement to enhance the effect of reducing body fat and preventing or improving obesity.

The sprouted oat extract of the present invention or a compound derived therefrom may be included in a food composition in various weight % if it can exhibit the effect of reducing body fat or preventing or improving obesity. Specifically, it may be included in an amount of 0.00001 to 100 wt % or 0.01 to 80 wt % relative to the total weight of the food composition, but is not limited thereto. In case of long-term intake for health and hygiene purposes, the content below the above range may be included, and since there is no problem in terms of safety, the active ingredient may also be used in an amount above the above range.

In addition, the pharmaceutical composition of the present invention is characterized by having a use for reducing body fat and preventing or treating obesity-related diseases.

At this time, the descriptions of the above “oat sprouts”, “extract”, “compound”, “body fat reduction”, “obesity”, “treatment”, “improvement” and “prevention” are as described above.

The pharmaceutical composition of the present invention may additionally contain a pharmaceutically acceptable salt.

The term "pharmaceutically acceptable salt" of the present invention means a salt that can be used pharmaceutically among salts which are substances in which cations and anions are combined by electrostatic attraction, and can typically be a metal salt, a salt with an organic base, a salt with an inorganic acid, a salt with an organic acid, a salt with a basic or acidic amino acid, etc. For example, the metal salt can be an alkali metal salt (sodium salt, potassium salt, etc.), an alkaline earth metal salt (calcium salt, magnesium salt, barium salt, etc.), an aluminum salt, etc., and the salt with an organic base can be a salt with triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N,N-dibenzylethylenediamine, etc., and the salt with an inorganic acid can be a salt with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, etc. Salts with organic acids include salts with formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc. Salts with basic amino acids include salts with arginine, lysine, ornithine, etc. Salts with acidic amino acids include salts with aspartic acid, glutamic acid, etc.

The pharmaceutical composition of the present invention may additionally comprise a pharmaceutically acceptable carrier, excipient or diluent, wherein the carrier may comprise a non-naturallyoccuring carrier.

Specifically, carriers, excipients and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, polycaprolactone, poly lactic acid, poly-L-lactic acid, mineral oil, and the like. The above pharmaceutical composition can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods, and the carrier may include various irregular carriers, microspheres, nanofibers, etc. When formulating, it can be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations can be prepared by mixing the extract and its fractions with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups, and may include various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives in addition to commonly used simple diluents such as water and liquid paraffin. Preparations for parenteral administration may include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, and the like. Non-aqueous solvents and suspending agents may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. The content of the sprouted oat extract or a compound derived therefrom included in the pharmaceutical composition of the present invention is not particularly limited. Another embodiment of the present invention for achieving the above object provides a pharmaceutical composition for reducing body fat, preventing or treating obesity, comprising the sprouted oat extract or a compound derived therefrom.

Another aspect of the present invention for achieving the above object provides a method for inhibiting differentiation into adipocytes, comprising the step of treating an extract of sprouted oats or a compound derived therefrom to isolated preadipocytes or isolated stem cells capable of differentiating into adipocytes.

At this time, the description of the “oat sprouts”, “extract”, and “compound” are as described above.

The term "adipocyte precursor cell" of the present invention means that fat accumulates and differentiates into adipocytes. Specifically, when differentiation of adipocyte precursor cells is induced, the cell cycle is arrested, cell proliferation is stopped, and differentiation into mature adipocytes is induced, and a rapid increase in this amount is the cause of obesity.

The term "adipocyte" of the present invention refers to a cell that accounts for 20 to 40% of adipose tissue and controls food intake energy consumption by transmitting hormones.

The term "stem cell" of the present invention refers to a cell that is relatively undifferentiated and has the ability to differentiate into a specific tissue cell.

In the above method, the method of treating the sprouted oat extract or a compound derived therefrom to the separated preadipocytes or stem cells can be applied to various known methods without limitation.

The sprouted oat extract of the present invention or a compound derived therefrom inhibits differentiation of preadipocytes into mature adipocytes, and thus, as a natural compound, does not exhibit cytotoxicity and can be used for preventing or treating obesity and reducing body fat without affecting organs or causing side effects.

Another aspect of the present invention for achieving the above object provides a method for reducing body fat, comprising a step of administering the pharmaceutical composition to a subject.

At this time, the description of the “pharmaceutical composition” and “body fat reduction” are as described above.

The term "subject" of the present invention means any animal, including a human, such as a rat, mouse, or livestock, that has developed or may develop increased body fat or obesity. As a specific example, it may be a mammal, including a human.

The term "administration" of the present invention means introducing a given substance into a subject by any suitable method, and the route of administration of the pharmaceutical composition of the present invention may be oral or parenteral administration through any common route as long as it can reach the target tissue. In addition, the pharmaceutical composition may be administered by any device through which the active substance can move to the target cell.

The term "dosage" of the present invention may vary depending on the age, sex, and weight of the subject, and may be easily determined by those skilled in the art. Specifically, the pharmaceutical composition of the present invention may be administered orally or parenterally. In addition, the dosage of the pharmaceutical composition may be determined by those skilled in the art according to the type of the composition, which is an active ingredient, along with various related factors such as the route of administration, the degree of the disease, sex, weight, and age, so the scope of the present invention is not limited by the beneficial dosage.

The term "administration method" of the present invention refers to various methods such as oral, intravenous, subcutaneous, intradermal, intranasal, intraperitoneal, intramuscular, and transdermal.

In addition, the feed composition comprising the sprouted oat extract of the present invention or a compound derived therefrom as an effective ingredient is characterized as being for reducing body fat or preventing or treating obesity.

At this time, the descriptions of the above “oat sprouts”, “extract”, “compound”, “body fat reduction”, “obesity”, “treatment”, “improvement” and “prevention” are as described above.

The sprouted oat extract or a compound derived therefrom according to the present invention exhibits an excellent body fat reduction effect, and therefore can be included in a feed composition for the purpose of preventing or treating obesity. Since the feed composition can be consumed by animals on a daily basis, a high effect in preventing or improving obesity can be expected.

The term "feed" of the present invention means any natural or artificial diet, meal, etc., or ingredients of said meal, which are suitable for animals to eat, ingest, and digest. The type of said feed is not particularly limited, and feed commonly used in the relevant technical field can be used. Non-limiting examples of said feed include plant feeds such as grains, roots, food processing by-products, algae, fibers, pharmaceutical by-products, fats, starches, meal, or grain by-products; animal feeds such as proteins, inorganic substances, fats, minerals, fats, single-cell proteins, zooplankton, or foodstuffs. These may be used alone or in combination of two or more.

In a specific example, the cytotoxicity evaluation, adipocyte differentiation inhibition activity, and lipolysis activity evaluation of sprouted oat extract or compounds derived from sprouted oat extract were confirmed.

Therefore, through the above specific example, it was confirmed that a composition containing a sprouted oat extract or a compound derived from a sprouted oat extract can be used as a food composition, pharmaceutical composition or feed composition for reducing body fat, or preventing, improving or treating obesity, and this further suggests that it can be used for the purpose of preventing, improving or treating obesity-related diseases.

The sprouted oat extract of the present invention can provide a composition having a body fat reduction effect by not exhibiting cytotoxicity and at the same time showing an effect of inhibiting adipocyte differentiation.

Figure 1 is a graph showing the measurement of adipocyte toxicity of the sprout oat extract of the present invention.
Figure 2 is a photograph showing the inhibition of adipocyte differentiation by the sprouted oat extract of the present invention.
Figure 3 is a graph showing the inhibition of adipocyte differentiation by the sprouted oat extract of the present invention.
Figure 4 is a photograph showing that the sprouted oat extract of the present invention shows AMPK protein expression in adipocytes.
Figure 5 is a graph showing the expression of AMPK protein in adipocytes by the sprouted oat extract of the present invention.
Figure 6 is a photograph showing the adipocyte differentiation inhibition activity of each compound in the sprout oat extract of the present invention.
Figure 7 is a graph showing the adipocyte differentiation inhibition activity of each compound in the sprout oat extract of the present invention.

Hereinafter, the present invention will be described in more detail through examples. These examples are intended to explain the present invention more specifically, and the scope of the present invention is not limited by these examples.

Example 1: Preparation of sprouted oat extract

Oats were sown and grown in a greenhouse at a temperature of 18 to 20℃ for about 7 to 8 days to harvest sprouted oats. The harvested sprouted oats were dried and cut, then soaked in 50 to 100% fermented alcohol to extract, filtered to obtain a liquid component, and freeze-dried to prepare a sprouted oat extract.

Example 2: Purification of compounds derived from sprouted oat extract

In the above Example 1, the sprouted oat extract obtained was subjected to column chromatography such as silica gel, reversed-phase C18 resin, and Sephadex to separate and purify the active ingredients contained therein. More specifically, 5 kg of sprouted oat powder was placed in an extraction vessel at room temperature, and 20 L of 70% ethanol (ethanol) was used to extract by shaking and rotating three times at room temperature for 48 hours. The extracted sample was filtered through a filter paper and concentrated under reduced pressure to obtain about 540 g of a crude extract. 10 L of purified water was added thereto to suspend the extract, and fractionated into n-hexane, ethyl acetate, and butanol in that order to obtain an n-hexane fraction (140 g), an ethyl acetate fraction (150 g), and a butanol fraction (220 g), respectively. The obtained ethyl acetate fraction (150 g) was subjected to reverse-phase C18 silica gel column chromatography (1 kg, Merk), and 10 fractions (Fr1 to Fr10) were obtained by sequentially using a mixed solvent of water and methanol (40:1 to 1:10 mixing ratio) as a mobile phase solvent. Fr. E8 (5.5 g) was adsorbed onto reverse-phase C18 silica gel (250 g, Merk), and then column chromatography was performed. At this time, a mixed solvent of methanol was used as a mobile phase solvent, and a pale yellow color was obtained, which was purified to obtain compounds 1 (40 mg), compound 2 (14 mg), and compound 3 (45 mg). Fr9 (1.5 g) was adsorbed onto silica gel (20 g, Merk), then onto reversed-phase C18 silica gel (250 g, Merk), and column chromatography was performed. A methanol mixed solvent was used as the mobile phase solvent to obtain a pale yellow compound, which was purified to obtain compound 4 (35 mg), compound 5 (18 mg), and compound 6 (28 m,g). The butanol fraction was obtained using reversed-phase C18 silica gel column chromatography (1,500 g, 70-230 mesh, Merk). A mixed solvent of water, methanol, and acetonitrile (mixing ratio of 50:5:1 to 1:5:10) was sequentially used as the mobile phase solvent to obtain eight fractions (Fr. B1 to Fr. B6). Fr. B3 (15.8 g) was adsorbed onto reversed-phase C18 silica gel (120 g), and column chromatography was performed. At this time, a mixed solvent of water, methanol, and acetonitrile (mixing ratio of 30:5:1 to 1:5:20) was used as the mobile phase solvent. A yellow compound was obtained, which was purified to obtain compound 7 (50 mg) and compound 8 (80 mg). Fr. B4 (8.5 g) was adsorbed onto reversed-phase C18 silica gel (56 g), and column chromatography was performed. At this time, a mixed solvent of water, methanol, and acetonitrile (mixing ratio of 10:5:1 to 1:10:30) was used as the mobile phase solvent. Another pale yellow precipitate was obtained, which was purified to obtain compound 9 (65 mg), compound 10 (25 mg), and compound 11 (35 mg). The total separated compounds are shown in Table 1 below.

Compound number Compound name Chemical structure 1 Avenafuranol 2 Diosgenoside 3 Avenacoside A 4 Avenacoside B 5 26-deglucoavenacoside B 6 Isoorientin-6-β-D-xylopyranoside 7 Vitexin 2``-O-rhamnoside 8 Isoswertisin 2``-O-rhamnoside 9 Isovitexin 2``-O-arabinoside 10 3-O-Feruloylquinic acid 11 26-degluco avenacoside A

Experimental Example 1: Evaluation of Adipocyte Toxicity of Sprout Oat Extract

3T3-L1 cells, which are preadipocytes, were purchased and cultured in DMEM (Dulbecco's modified eagle medium) medium supplemented with 10% FBS (fetal bovine serum) and 1% antibiotics (penicillin 100 U/ml and streptomycin 0.1 mg/ml) at 37°C under 5% _CO2 (95% air) conditions. The above cultured 3T3-L1 cells were treated with the extract of Example 1 and the compound of Example 2 at 10, 50, and 100 μg/mL, and cultured for 72 hours in a medium containing differentiation-inducing substances (FBS, IBMX, dexanmethasone, and insulin) to differentiate into adipocytes. MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) solution was added, and the cells were reacted at 37°C for 4 hours, then DMSO was added to dissolve the formazan produced in the cells, and the cell viability was measured by measuring the absorbance at 570 nm (Fig. 1). At this time, adipocytes that were not treated with the substances corresponding to the extract of Example 1 and the compound of Example 2 were used as a control.

Figure 1 is a graph showing changes in adipocyte viability according to the treatment concentration of the extract of Example 1 and the compound of Example 2. As can be seen in Figure 1, the extract of Example 1 and the compound of Example 2 did not reduce cell viability during the differentiation process into adipocytes, confirming that they were not cytotoxic.

Experimental Example 2: Evaluation of Adipocyte Differentiation Inhibitory Activity of Sprout Oat Extract and Sprout Oat Extract-Derived Compounds

3T3-L1 cells, which are preadipocytes, were treated with the extract of Example 1 at concentrations of 10, 50, and 100 μg/mL and the compounds of Example 2 (compounds 1, 2, 4, 5, 8, and 9) at 50 μM to differentiate into adipocytes, and the differentiated cells were stained with oil-red staining (an experimental method for staining fat globules accumulated in adipocytes) to stain fat within the adipocytes, and the levels of stained fat were compared under a microscope (Figs. 2 and 6). At this time, undifferentiated preadipocytes (normal group) were used as a negative control group.

FIG. 2 is a micrograph showing the level of fat included in differentiated adipocytes according to the treatment concentration of the extract of Example 1 and the compound of Example 2, FIG. 3 is a graph showing the results of measuring the level of fat included in adipocytes according to the treatment concentration of the extract of Example 1, and FIG. 7 is a graph showing the results of measuring the level of fat included in adipocytes by treating with compounds 1, 2, 4, 6, 8, and 9 of Example 2 at a treatment concentration of 50 μM. As can be seen in FIGS. 2 and 6, the level of fat decreased in cells treated with the extract of Example 1 and the compound of Example 2, and it was confirmed that the level of fat decreased as the concentration of the extract of Example 1 increased. In addition, as can be seen in FIGS. 3 and 7, the OD (Oil Red Optical density) value of cells treated with the sprouted oat extract decreased according to the concentration, and it was confirmed that cells treated with the sprouted oat-derived compound reduced fat compared to the adipocyte-induced group. Therefore, it was found that the extract of Example 1 could reduce fat in a concentration-dependent manner, and the compound of Example 2 also reduced fat, and among them, it was confirmed that compound 6 reduced fat levels the best.

Experimental Example 3: Evaluation of lipolytic activity of sprouted oat extract

3T3-L1 adipocytes treated with sprouted oat extract were lysed with lysis buffer (10 mM Tris-HCl [pH 7.4], 0.1 M EDTA, 10 mM NaCl, and 0.5% Triton X-100, proteinase inhibitor), centrifuged at 13,000 rpm, 4℃ for 20 minutes, and the supernatant was obtained. The protein concentration was quantified from the supernatant using the Bradford method, and 40 μg of protein was extracted. 1× SDS sample buffer (50 mM Tris, pH 6.8, 2% SDS, 10% glycerol, 0.01% bromophenol blue, 5% β-mercaptoethanol) was added to 40 μg of the extracted protein, and the mixture was heated at 100℃ for 5 minutes. The prepared sample was electrophoresed on 10% SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) and transferred to a PVDF membrane. The membrane was blocked with 5% nonfat dry milk dissolved in TBST buffer (10 mM Tris-HCl, 150 mM NaCl, 0.1% Tween 20). The primary and secondary antibodies were reacted on the membrane for 1 hour at room temperature. After washing with TBST buffer, protein bands were detected and visualized by chemiluminescence using ECL reagent. The protein bands were quantified and numerically analyzed using software (Gel-Pro Analyzer 4.0), and actin was used as a reference gene for standardization.

As shown in FIGS. 3 to 5, it can be confirmed that the activation of AMPK by the extract of Example 1 phosphorylates each protein and induces β-oxidation of adipocytes. In addition, it can be determined that the extract of Example 1 has an anti-obesity effect because β-oxidation increases when p-AMPK is formed. The expression level of AMPK did not differ depending on the sample treatment, but the expression of p-AMPK, the phosphorylated form of AMPK, increased in a concentration-dependent manner, indicating that the extract of Example 1 has a β-oxidation activation effect.

In summary of Experimental Examples 1 to 3 of the present invention, it was confirmed that sprouted oat extract and compounds derived from sprouted oat extract exhibited no cytotoxicity and at the same time showed the ability to inhibit adipocyte differentiation, thereby exhibiting body fat reduction efficacy.

From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without changing the technical idea or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are exemplary in all respects and not restrictive. The scope of the present invention should be interpreted as including all changes or modifications derived from the meaning and scope of the patent claims described below rather than the above detailed description and their equivalent concepts within the scope of the present invention.

Claims (11)

A food composition for reducing body fat, comprising isoorientin-6-β-D-xylopyranoside as an effective ingredient.
A food composition in claim 1, wherein the composition further comprises at least one selected from the group consisting of avenafuranol, diosgenoside, avenacoside B, isoswertisin 2``-O-rhamnoside, and isovitexin 2``-O-arabinoside.
delete A method for inhibiting differentiation into adipocytes, comprising the step of treating isolated preadipocytes or isolated stem cells capable of differentiating into adipocytes with isoorientin-6-β-D-xylopyranoside.
delete delete delete A feed composition for reducing body fat, comprising isoorientin-6-β-D-xylopyranoside as an effective ingredient.
A food composition for preventing or improving obesity, comprising isoorientin-6-β-D-xylopyranoside as an active ingredient.
A food composition in claim 9, wherein the composition further comprises at least one selected from the group consisting of avenafuranol, diosgenoside, avenacoside B, isoswertisin 2``-O-rhamnoside, and isovitexin 2``-O-arabinoside.
A feed composition for preventing or improving obesity, comprising isoorientin-6-β-D-xylopyranoside as an effective ingredient.