WO2019093739A1 - Composition comprising mealworm beetle extract as effective ingredient for prevention, alleviation, or treatment of fatty liver disease - Google Patents

Composition comprising mealworm beetle extract as effective ingredient for prevention, alleviation, or treatment of fatty liver disease Download PDF

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Publication number
WO2019093739A1
WO2019093739A1 PCT/KR2018/013386 KR2018013386W WO2019093739A1 WO 2019093739 A1 WO2019093739 A1 WO 2019093739A1 KR 2018013386 W KR2018013386 W KR 2018013386W WO 2019093739 A1 WO2019093739 A1 WO 2019093739A1
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Prior art keywords
extract
fatty liver
liver disease
alcoholic
brown
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PCT/KR2018/013386
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French (fr)
Korean (ko)
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채성욱
임아랑
이주영
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한국 한의학 연구원
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/20—Animal feeding-stuffs from material of animal origin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L35/00—Food or foodstuffs not provided for in groups A23L5/00 – A23L33/00; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/56—Materials from animals other than mammals
    • A61K35/63—Arthropods
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/20—Natural extracts
    • A23V2250/204—Animal extracts

Definitions

  • the present invention relates to a composition for preventing, ameliorating or treating a fatty liver disease, which comprises an extract of brown gruel as an active ingredient.
  • the death rate of domestic liver disease is very high, 23.5 per 100,000 population.
  • the number of deaths in the 40s is the first (41.1 / 100,000), the second in the 50s (72.4 / 100,000) (10 people / 100,000 people), including liver disease is the leading cause of death in the Korean middle-aged population.
  • liver diseases fatty liver suggests that abnormal fat, which is not present in normal cells, is abnormally deposited in liver cells.
  • abnormal fat which is not present in normal cells, is abnormally deposited in liver cells.
  • triglyceride fatty acid, phospholipid, cholesterol and cholesterol ester are the main components of fat.
  • triglyceride fatty acid, phospholipid, cholesterol and cholesterol ester are the main components of fat.
  • triglyceride fatty acid, phospholipid, cholesterol and cholesterol ester
  • hepatocytes When the fatty liver becomes worse and fat mass in hepatocyte becomes larger, important components of the cells including nucleus are pushed to one side, the function of the hepatocyte is lowered, and the expanded hepatocytes due to the accumulated fat in the cells pressurize the microvascular and lymphatic vessels between the hepatocytes Resulting in obstacles to the circulation of blood and lymph in the liver. In this way, hepatocytes can not properly receive oxygen and nutrient supply, and liver function is deteriorated.
  • Non-alcoholic fatty liver disease is defined as the accumulation of fatty acids in the form of triglycerides in the parenchymal cells of the liver in excess of 5%, not by liver damage caused by alcohol. Pathologically, it is classified as simple steatosis and inflammatory fatty liver disease (steatohepatitis). It can be converted to severe liver disease such as hepatitis, liver fibrosis and cirrhosis when left for a long time. In Korea, the frequency of nonalcoholic liver disease is increasing due to changes in lifestyle.
  • Alcoholic fatty liver disease is caused by excessive alcohol consumption. Individuals have different genetic characteristics and gender. However, alcohol intake of 80g or more per day is very likely to cause liver disease such as alcoholic fatty liver disease. Women are more likely to develop alcoholic liver disease in lesser amounts. Generally, it can be calculated that one cup of shochu, one cup of beer, one cup of wine, and one cup of makkolli contain about 10 g of alcohol. The most mild form of alcoholic fatty liver disease is asymptomatic, but mild hepatic enlargement (greater than normal liver) may cause mild tenderness in the right upper abdomen.
  • the brown goose is 30 ⁇ 38cm in length
  • body color is yellow-green
  • head and chest are yellowish brown.
  • the compound eyes are egg-shaped and lustrous grayish brown.
  • the wings are yellowish brown and longer than the end of the boat but do not fly much. Inhabits grassy areas near rice fields and cultivated areas. It occurs once a year, and it drifts with a bunch of ground in the ground.
  • the alum is wrapped in a thin film of collagen.
  • the full-grown nymphs have a wing crest at the fourth node and a body length of about 30 mm.
  • the use of pesticides as a major pest of rice has decreased, but it is gradually increasing. In 1930, it originated in Hwanghae Province and bought eggs from the government. It is distributed in Korea, Japan, and China.
  • Korean Patent Laid-Open No. 2016-0041115 discloses a composition for preventing or treating diabetes comprising an effective amount of a larva or extract thereof in Korean Patent Application No. 1424125, A composition for treating inflammatory diseases including larva is known, but a composition for preventing, ameliorating or treating a fatty liver disease containing the brown gruel extract of the present invention as an active ingredient is not known.
  • the present invention provides a composition for preventing, ameliorating or treating a fatty liver disease comprising an extract of brown gruel as an active ingredient, wherein the brown gruel extract is a mixture of triglyceride And confirming that there is an effect of inhibiting accumulation, thereby completing the present invention.
  • the present invention provides a health functional food composition for preventing or ameliorating fatty liver disease, which comprises an extract of brown gruel as an active ingredient.
  • the present invention also provides a pharmaceutical composition for the prevention or treatment of fatty liver disease, which comprises an extract of brown gruel as an active ingredient.
  • the present invention also provides a veterinary composition for the prevention or treatment of fatty liver disease in an animal other than a human, which comprises an extract of brown gruel as an active ingredient.
  • the present invention provides a feed additive for preventing or ameliorating fatty liver disease in an animal other than humans, which comprises an extract of brown gruel as an active ingredient.
  • the present invention provides a method for preventing or treating fatty liver disease comprising administering a brown gruel extract to a subject other than a human.
  • the present invention relates to a composition for preventing, ameliorating or treating a fatty liver disease comprising an extract of brown gruel as an active ingredient. Since the brown gruy extract as an active ingredient of the present invention is a raw material known to be edible, it has no side effects and toxicity , because liver cancer cell line (HepG 2) reduction in the accumulation of triglycerides effectively, the composition of the present invention may be useful as a functional food, a drug, or a feed additive for the prevention, improvement or treatment of fatty liver disease.
  • HepG 2 liver cancer cell line
  • liver cancer cell line (HepG 2) on oleate and palmitate is 2: 1 concentration ratio after the process the free fatty acids (FFA) and mealworm extract (Cu) of a mixture of 1.0mM, fat absorption (lipid uptake) .
  • FFA free fatty acids
  • Cu mealworm extract
  • #### showed a statistically significant increase in the fat absorption rate in the group treated with free fatty acids compared with the control group (p ⁇ 0.0001), while * and ** indicate that the brown of the present invention
  • the absorption of free fatty acids in the sorghum extract-treated group was statistically significantly decreased.
  • the * indicates p ⁇ 0.05 and ** indicates p ⁇ 0.01.
  • FIG. 2 shows the results of (A), (B), (C), (C) and (C) (HF + Mi), which was administered with milk fat extract (100 mg / kg / day) as a positive control with high fat diets, and the brown gruel extract administered group (HF + Cu) And (B) the above tissues are shown as oil droplet staining results (above) and their graphs (below).
  • P ⁇ 0.01 was found to be a statistically significant increase in the liver weight of the high-fat diet group compared to the normal group, and ** was higher in the high fat diet group compared to the control group of the brown goat extract- The statistically significant decrease in weight was p ⁇ 0.01.
  • FIG. 3 shows the cell viability (%) after treatment of hepatocellular carcinoma cell line (HepG 2 ) with ethanol and brown goat extract.
  • the cell survival rate of the group was statistically significantly increased, ** is p ⁇ 0.01 and *** is p ⁇ 0.001.
  • FIG. 4 shows the results of examining the expression level of Caspase-3 after treatment of hepatocellular carcinoma cell line (HepG 2 ) with ethanol and brown goat extract.
  • the expression level of Caspase-3 in the ethanol-treated group was significantly higher than that in the control (p ⁇ 0.0001), and **** was significantly higher than that of the control group
  • the expression level of caspase-3 in the treatment group was statistically significantly decreased, and p ⁇ 0.0001.
  • FIG. 5 shows the result of confirming whether the brown goat's extract (Cu) of the present invention reduces ALT increase in the blood by the high fat diet.
  • Mi was a positive control group, which received milk-thirst extract (100 mg / kg / day) with high fat diet. (P ⁇ 0.001) and *** was significantly higher than that of the control group (control group) compared to that of the control group (control group), the brown goat extract treatment group of the present invention or the positive control group , which is a statistically significant decrease in the ALT value of the in-milk-thistle extract-treated group, is p ⁇ 0.001.
  • FIG. 6 shows the result of confirming whether the brown gruel extract (Cu) of the present invention reduces the increase of total cholesterol (TC) in the blood by the high fat diet.
  • Mi was a positive control group, which received milk-thistle extract (100 mg / kg / day) with high fat diet.
  • P ⁇ 0.001, ** and **** were higher than those of the control group (control group), and those of the control group (control group) were significantly higher than those of the control group
  • the total cholesterol in the blood of the extract-treated group or the milk-treated cholesterol extract group, which is a positive control group was statistically significantly decreased, ** is p ⁇ 0.01 and **** is p ⁇ 0.0001.
  • FIG. 7 shows the result of confirming whether the brown gruel extract (Cu) of the present invention reduces the glucose increase in the blood due to the high fat diet.
  • Mi was a positive control group, which received milk-thistle extract (100 mg / kg / day) with high fat diet.
  • ### showed a statistically significant increase in the glucose content of the high-fat diet group compared to the control group (p ⁇ 0.001), ** indicates that the glucose content of the brown goat extract-treated group of the present invention Statistically significant decrease, p ⁇ 0.01.
  • FIG. 8 shows the results of confirming whether the brown gruel extract (Cu) of the present invention reduces the fat droplets of liver tissue formed by high fat diets (HFD).
  • Mi was a positive control group, which received milk-thistle extract (100 mg / kg / day) with high fat diet.
  • A is a liver tissue of a normal control group
  • B is a liver tissue of a high fat diet group
  • C is a liver tissue of a high fat fat diet group and a positive control milk group
  • the liver tissue of the brown gruel extract administration group of the present invention is a positive control group, which received milk-thistle extract (100 mg / kg / day) with high fat diet.
  • the present invention relates to a health functional food composition for preventing or ameliorating fatty liver disease, which comprises an extract of brown gruel as an active ingredient.
  • the brown gutter extract may be prepared by a method including, but not limited to, the following steps:
  • step (3) The step of extracting the filtered extract of step (2) by concentration under reduced pressure and drying.
  • the extraction solvent is preferably selected from water, a C 1 -C 4 lower alcohol or a mixture thereof, more preferably ethanol, even more preferably 70% (v / v) ethanol But is not limited thereto.
  • any conventional method known in the art such as filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction may be used.
  • the extraction solvent is preferably added in an amount of 1 to 20 times, preferably 5 to 15 times, the weight of the dried brown gruel.
  • the extraction temperature is preferably 4 to 50 DEG C, but is not limited thereto.
  • the extraction time is preferably 0.5 to 10 hours, more preferably 0.5 to 5 hours, most preferably 1 hour, but not always limited thereto.
  • the drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.
  • the fatty liver disease is non-alcoholic or alcoholic simple fatty liver; Non-alcoholic or alcoholic fatty liver disease; And non-alcoholic or alcoholic cirrhosis, but is not limited thereto.
  • the brown gruel extract is characterized by reducing the content of triglyceride in liver tissue, reducing the content of total triglyceride in liver tissue and decreasing lipid uptake.
  • the health functional food may be prepared by adding the composition of the present invention to a food for the purpose of preventing or ameliorating fatty liver disease.
  • the health functional food is not particularly limited, and may be a health functional food, a nutritional supplement, but it can be any type of food such as pharmafood, health food, nutraceutical, designer food, food additive, etc.
  • the food can be meat, sausage, bread, chocolate, candy, snack, confectionery, , Other noodles, gums, dairy products including ice cream, various soups, drinks, tea, drinks, alcoholic beverages and vitamin complexes.
  • the method of adding is appropriately used according to a conventional method, and the amount to be added can be suitably determined according to the intended use (prevention, health or therapeutic treatment).
  • composition of the present invention may further include various components other than the active component.
  • the various components may be various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, Salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks and the like. It may also contain flesh for the production of natural fruit juices, fruit juice beverages and vegetable beverages to add palatability and / or functionality, and these ingredients may be used independently or in combination.
  • the present invention relates to a pharmaceutical composition for preventing or treating fatty liver disease, which comprises an extract of brown gruel as an active ingredient.
  • the fatty liver disease is a non-alcoholic or alcoholic simple fatty liver; Nonalcoholic or alcoholic fatty liver disease; And non-alcoholic or alcoholic liver cirrhosis, but is not limited thereto.
  • the pharmaceutical compositions of the present invention may comprise a pharmaceutically acceptable carrier.
  • the pharmacologically acceptable carrier to be contained in the pharmaceutical composition of the present invention is one which is usually used at the time of formulation and includes saline, sterilized water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, But are not limited to, lactose, sucrose, sucrose, sorbitol, mannitol, starch, acacia, calcium phosphate, alginate, gelatin, calcium calcium, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methylcellulose, methylhydroxybenzoate, Hydroxybenzoate, talc, magnesium stearate, and mineral oil, but the present invention is not limited thereto.
  • the pharmaceutical composition of the present invention may further comprise at least one auxiliary selected from an antioxidant, a buffer, a bacteriostat, a diluent, a surfactant, a binder, a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, .
  • at least one auxiliary selected from an antioxidant, a buffer, a bacteriostat, a diluent, a surfactant, a binder, a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, .
  • the pharmaceutical composition of the present invention can be administered orally or parenterally, and is preferably administered orally.
  • the appropriate dosage of the pharmaceutical composition of the present invention may vary depending on such factors as formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, route of administration, excretion rate, .
  • the dosage of the pharmaceutical composition of the present invention is preferably 0.001-100 mg / kg on an adult basis.
  • the pharmaceutical composition of the present invention may be formulated into a unit dosage form by using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by those having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container.
  • the formulations may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of excipients, powders, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.
  • the present invention also relates to a veterinary composition for the prevention or treatment of fatty liver disease in an animal other than humans, which comprises an extract of brown gruel as an active ingredient.
  • the veterinary composition comprising the brown gruel extract of the present invention may further comprise suitable excipients and diluents according to conventional methods.
  • excipients and diluents that can be included in the veterinary composition containing the brown gutter extract of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin , Calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, cetanol, stearyl alcohol, Paraffin, sorbitan monostearate, polysorbate 60, methylparaben, propylparaben, and mineral oil.
  • the veterinary composition comprising the brown gruel extract according to the present invention may further contain a filler, an anti-coagulant, a lubricant, a wetting agent, a spice, an emulsifier, an antiseptic, etc.
  • the veterinary composition according to the present invention may further comprise May be formulated using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient, and the formulations may be formulated as powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, Soft or hard gelatine capsules, suppositories, sterile injectable solutions, sterile external preparations, and the like.
  • the veterinary composition according to the present invention may vary depending on the age, sex, and body weight of the animal, but may be administered in an amount of 0.1 to 100 mg / kg once or several times a day.
  • the dosage may vary depending on the route of administration, Sex, weight, age, and the like. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.
  • the present invention also relates to a feed additive for preventing or ameliorating fatty liver disease in an animal other than a human, which comprises an extract of brown gruel as an active ingredient.
  • the feed additive may be in the form of a high concentrate, powder or granulate containing from 20 to 90% by weight of brown gruel extract.
  • the feed additive of the present invention can be used as a feed additive containing a phosphate such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, or organic acids such as sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polymerized phosphate), polyphenol, catechin, Natural antioxidants such as tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid may be further included.
  • a phosphate such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, or organic acids such as sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polymerized phosphate), polyphenol, catechin, Natural antioxidants such as tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid may be further included
  • the animal feed additive containing the brown goat's extract of the present invention and the feed comprising the animal feed additive can be supplemented with amino acids, inorganic salts, vitamins, antibiotics, antimicrobials, antioxidants, antifungal enzymes, digestion and absorption enhancers, growth promoters, And the like.
  • the feed additive may be administered alone to the animal in combination with other feed additives in the edible carrier.
  • the feed additives can also be easily administered as top dressing or they can be mixed directly with the animal feed or separately from the feed, in separate oral formulations, by injection or transdermal, or in combination with other ingredients.
  • a single daily dose or a divided daily dose can be used as is well known in the art.
  • the dosage form of the extract may be prepared in an immediate release or sustained release formulation in combination with a non-toxic pharmaceutically acceptable food carrier.
  • Such edible carriers may be solid or liquid, for example, corn starch, lactose, sucrose, soy flakes, peanut oil, olive oil, sesame oil and propylene glycol.
  • the dosage form of the extract may be a tablet, capsule, powder, troche or emulsion or top-dressing in finely divided form.
  • a liquid carrier it may be in the form of a soft gelatin capsule, or a syrup or liquid suspension, emulsion or solution.
  • the dosage form may contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, solution promoting agents and the like.
  • the animal feed in which the brown gruel extract is included as a feed additive can be any protein-containing organic gut that is commonly used to meet the dietary needs of an animal. These protein-containing flours usually consist mainly of corn, soy flour or corn / soy flour mix.
  • the feed additive may be added to the animal feed by immersion, spraying or mixing.
  • the veterinary composition or feed additive of the present invention can be applied to the companion animal's diet.
  • the present invention also relates to a method for preventing or treating fatty liver disease comprising administering a brown gruel extract to a subject other than a human.
  • Such an individual includes all animals, including those with a possibility of fatty liver disease, including those suffering from fatty liver disease.
  • the dried brown powdery mildew was refluxed for 3 hours using 70% (v / v) ethanol.
  • a 70% (v / v) ethanol extract was obtained and distilled under reduced pressure to prepare a brown goat ethanol extract.
  • Liver cancer cell line (HepG 2) were purchased from ATCC (American Type Culture Collection). Were cultured in the DMEM-F12 containing the streptomycin (Dulbecco's modified Eagle's medium) medium-liver cancer cell line (HepG 2) is 10% FBS, 1% penicillin. The cells were cultured in a 5% CO 2 incubator at 37 ° C, and the cultured cells were tested by dividing a certain number of cells (3 ⁇ 10 4 cells / 500 ⁇ l well) into 24-well plates.
  • streptomycin Dulbecco's modified Eagle's medium
  • a 0.1 M NaOH solution was used as a solvent at 70 ⁇ C, filtered using a 0.2 ⁇ filter, and then sterilized.
  • the concentration of each fatty acid prepared in (1) above was measured using a 5% (w / v) BSA solution containing no free fatty acid dissolved in the tertiary distilled water prepared in (2) 10 ml of a fatty acid storage solution was prepared so as to be 5 mM.
  • 100 ⁇ l of palmitate and 100 mM of the olate storage solution prepared in (1) above were mixed with 165 ⁇ l (1.65 mM) of palmitate and 330 ⁇ l (3.3 mM) of oleate and 9505 ⁇ l of 5% w / v) dropwise dropwise to the BSA solution (vortexing). Thereafter, the mixture was cooled to room temperature and then filtered with a 0.2 ⁇ m filter (the mixed solution was stable at -20 ° C. for 3 to 4 weeks).
  • Oil red O staining was performed to determine whether the brown goat extract (Cu) reduced the lipid accumulation in liver tissues due to high fat diets. Lipids are stained red by Oil red O staining method.
  • the lipid accumulation increased by the high fat diet was reduced by the brown goat extract (Cu), and the liver tissue protective effect was confirmed.
  • liver weight increase due to fat accumulation of liver tissue was statistically decreased by brown goat extract (Cu).
  • brown gruel extract (Cu) of the present invention reduced the content of ALT, total cholesterol (TC) and glucose in the blood increased by the high fat diet.
  • H & E staining was performed to determine if the brown goat extract (Cu) of the present invention reduced lipid accumulation in the liver by high fat diet.

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Abstract

The present invention relates to a composition comprising a mealworm beetle extract as an effective ingredient for prevention, alleviation, or treatment of fatty liver disease. A mealworm beetle extract, which is an effective ingredient of the present invention, was found to effectively induce fat absorption from a liver cancer cell line (HepG2) and to have the effect of reducing lipids in liver tissues. Therefore, the composition of the present invention can be useful as a health functional food, a medicine product, or a feed additive for prevention, alleviation, or treatment of fatty liver disease.

Description

갈색거저리 추출물을 유효성분으로 포함하는 지방간 질환의 예방, 개선 또는 치료용 조성물A composition for preventing, improving or treating a fatty liver disease comprising an extract of brown gruel as an active ingredient
본 발명은 갈색거저리 추출물을 유효성분으로 포함하는 지방간 질환의 예방, 개선 또는 치료용 조성물에 관한 것이다.The present invention relates to a composition for preventing, ameliorating or treating a fatty liver disease, which comprises an extract of brown gruel as an active ingredient.
국내 간질환 사망률은 인구 십만명 당 23.5명으로 매우 높으며, 40대 사망원인 1위(41.1명/10만 명), 50대 사망원인 2위(72.4명/10만 명), 30대 사망원인 3위(10명/10만 명)를 차지하는 등 간질환은 한국 중년층 인구의 주요 사망원인이다. The death rate of domestic liver disease is very high, 23.5 per 100,000 population. The number of deaths in the 40s is the first (41.1 / 100,000), the second in the 50s (72.4 / 100,000) (10 people / 100,000 people), including liver disease is the leading cause of death in the Korean middle-aged population.
상기 간질환 중에서 지방간은 정상세포 내에는 존재하지 않는 중성지방이 간 세포 내에 비정상적으로 침착되어 보이는 현상이 나타난 것을 말한다. 정상 간은 약 5%가 지방조직으로 구성되어 있으며 중성지방, 지방산, 인지질, 콜레스테롤 및 콜레스테롤 에스터가 지방의 주요 성분이나, 일단 지방간이 발생하면 대부분의 성분이 중성지방으로 대체되며 중성지방의 양이 간 중량의 5% 이상이면 지방간으로 진단된다. 지방간이 악화되어 간세포 속의 지방 덩어리가 커지면 핵을 포함한 세포의 중요한 구성성분이 한쪽으로 밀려 간세포의 기능이 저하되며, 세포 내에 축적된 지방으로 인하여 팽창된 간세포들이 간세포 사이에 있는 미세 혈관과 임파선을 압박하여 간 내의 혈액과 임파액의 순환에 장애가 생기게 된다. 이렇게 되면 간세포는 산소와 영양공급을 적절히 받을 수 없어 간기능이 저하되는 것이다. Among the liver diseases, fatty liver suggests that abnormal fat, which is not present in normal cells, is abnormally deposited in liver cells. In normal liver, about 5% is composed of adipose tissue, and triglyceride, fatty acid, phospholipid, cholesterol and cholesterol ester are the main components of fat. However, once fatty liver occurs, most components are replaced with triglyceride. If the liver weight is 5% or more, it is diagnosed as fatty liver. When the fatty liver becomes worse and fat mass in hepatocyte becomes larger, important components of the cells including nucleus are pushed to one side, the function of the hepatocyte is lowered, and the expanded hepatocytes due to the accumulated fat in the cells pressurize the microvascular and lymphatic vessels between the hepatocytes Resulting in obstacles to the circulation of blood and lymph in the liver. In this way, hepatocytes can not properly receive oxygen and nutrient supply, and liver function is deteriorated.
비알콜성 지방간 질환(non-alcoholic fatty liver disease, NAFLD)은 알코올에 의한 간 손상이 아니라, 지방산이 중성지방의 형태로 간의 실질세포 내에 5% 이상 축적된 경우로 정의한다. 병리학적으로는 단순 지방간(simple steatosis)과 염증을 동반한 지방간염(steatohepatitis)으로 분류되는데, 장기간 방치 시, 간염, 간 섬유, 간경변 등의 심각한 간 질환으로 이행될 수 있다. 국내에서도 생활양식의 변화로 인해 비알코올성 간질환 발생빈도가 증가하는 추세이다.Non-alcoholic fatty liver disease (NAFLD) is defined as the accumulation of fatty acids in the form of triglycerides in the parenchymal cells of the liver in excess of 5%, not by liver damage caused by alcohol. Pathologically, it is classified as simple steatosis and inflammatory fatty liver disease (steatohepatitis). It can be converted to severe liver disease such as hepatitis, liver fibrosis and cirrhosis when left for a long time. In Korea, the frequency of nonalcoholic liver disease is increasing due to changes in lifestyle.
또한, 알콜성 지방간은 과다한 음주로 인해 발생하는데, 개인별로 유전적인 특징, 성별에 따라 차이가 있지만, 하루 80g 이상의 알코올의 섭취를 하는 경우 알콜성 지방간과 같은 간질환이 발생할 가능성이 크다. 여성의 경우 이보다 적은 양으로도 알코올성 간질환이 발생할 가능성이 커진다. 일반적으로 소주 1잔, 맥주 1잔, 양주 1잔, 막걸리 1홉에 약 10g의 알코올이 포함된 것으로 계산할 수 있다. 증상으로는 가장 경미한 형태인 알코올성 지방간 환자들의 경우 대부분 무증상이지만, 경미한 간비대(간이 정상보다 크게 된 상태)가 나타난 경우 오른쪽 상복부에 가벼운 압통을 호소할 수 있다.Alcoholic fatty liver disease is caused by excessive alcohol consumption. Individuals have different genetic characteristics and gender. However, alcohol intake of 80g or more per day is very likely to cause liver disease such as alcoholic fatty liver disease. Women are more likely to develop alcoholic liver disease in lesser amounts. Generally, it can be calculated that one cup of shochu, one cup of beer, one cup of wine, and one cup of makkolli contain about 10 g of alcohol. The most mild form of alcoholic fatty liver disease is asymptomatic, but mild hepatic enlargement (greater than normal liver) may cause mild tenderness in the right upper abdomen.
한편, 갈색거저리는 몸길이 30~38cm으로, 몸빛깔은 황록색이나 머리와 가슴은 황갈색이다. 겹눈은 달걀 모양이고 광택이 있는 회갈색이다. 앞가슴등판에 가느다란 3개의 가로 홈이 있고 양쪽에 갈색의 세로줄이 있다. 날개는 황갈색이고 배 끝보다 길지만 별로 날아다니지 않는다. 논이나 경작지 근처의 풀밭에 서식한다. 연 1회 발생하며, 땅 속에서 알무더기로 월동한다. 알무더기는 아교질의 엷은 막으로 싸여 있다. 다 자란 약충은 날개딱지가 제4배 마디에 이르며 몸길이는 약 30mm이다. 벼의 주요 해충으로 농약을 사용하면서 그 수가 줄어들었으나 차차 늘어나고 있다. 1930년 황해도에서 발생해 알 덩어리를 정부에서 사들인 일도 있다. 한국·일본·중국 등지에 분포한다. On the other hand, the brown goose is 30 ~ 38cm in length, body color is yellow-green, and head and chest are yellowish brown. The compound eyes are egg-shaped and lustrous grayish brown. There are three thin lateral grooves on the pronotum, with brown vertical stripes on both sides. The wings are yellowish brown and longer than the end of the boat but do not fly much. Inhabits grassy areas near rice fields and cultivated areas. It occurs once a year, and it drifts with a bunch of ground in the ground. The alum is wrapped in a thin film of collagen. The full-grown nymphs have a wing crest at the fourth node and a body length of about 30 mm. The use of pesticides as a major pest of rice has decreased, but it is gradually increasing. In 1930, it originated in Hwanghae Province and bought eggs from the government. It is distributed in Korea, Japan, and China.
갈색거저리를 이용하는 의약용도 관련 기술로는 한국공개특허 제2016-0041115호에 갈색거저리 유충 또는 이의 추출물을 유효성분으로 포함하는 당뇨 예방 또는 치료용 조성물이 알려져 있고, 한국등록특허 제1424125호에 갈색거저리 유충을 포함하는 염증성 질환 치료용 조성물이 알려져 있으나, 본 발명의 갈색거저리 추출물을 유효성분으로 함유하는 지방간 질환의 예방, 개선 또는 치료용 조성물에 대해서는 알려진 바 없다.Korean Patent Laid-Open No. 2016-0041115 discloses a composition for preventing or treating diabetes comprising an effective amount of a larva or extract thereof in Korean Patent Application No. 1424125, A composition for treating inflammatory diseases including larva is known, but a composition for preventing, ameliorating or treating a fatty liver disease containing the brown gruel extract of the present invention as an active ingredient is not known.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명은 갈색거저리 추출물을 유효성분으로 포함하는 지방간 질환의 예방, 개선 또는 치료용 조성물을 제공하고, 상기 갈색거저리 추출물은 간조직 내의 중성지방의 축적을 억제하는 효과가 있다는 것을 확인함으로써, 본 발명을 완성하였다.The present invention provides a composition for preventing, ameliorating or treating a fatty liver disease comprising an extract of brown gruel as an active ingredient, wherein the brown gruel extract is a mixture of triglyceride And confirming that there is an effect of inhibiting accumulation, thereby completing the present invention.
상기 목적을 달성하기 위하여, 본 발명은 갈색거저리 추출물을 유효성분으로 함유하는 지방간 질환의 예방 또는 개선용 건강기능식품 조성물을 제공한다.In order to accomplish the above object, the present invention provides a health functional food composition for preventing or ameliorating fatty liver disease, which comprises an extract of brown gruel as an active ingredient.
또한, 본 발명은 갈색거저리 추출물을 유효성분으로 포함하는 지방간 질환의 예방 또는 치료용 약학 조성물을 제공한다.The present invention also provides a pharmaceutical composition for the prevention or treatment of fatty liver disease, which comprises an extract of brown gruel as an active ingredient.
또한, 본 발명은 갈색거저리 추출물을 유효성분으로 포함하는 인간을 제외한 동물의 지방간 질환의 예방 또는 치료용 수의학적 조성물을 제공한다.The present invention also provides a veterinary composition for the prevention or treatment of fatty liver disease in an animal other than a human, which comprises an extract of brown gruel as an active ingredient.
또한, 본 발명은 갈색거저리 추출물을 유효성분으로 포함하는 인간을 제외한 동물의 지방간 질환의 예방 또는 개선용 사료 첨가제를 제공한다.In addition, the present invention provides a feed additive for preventing or ameliorating fatty liver disease in an animal other than humans, which comprises an extract of brown gruel as an active ingredient.
또한, 본 발명은 갈색거저리 추출물을 인간을 제외한 개체에게 투여하는 단계를 포함하는 지방간 질환의 예방 또는 치료방법을 제공한다.In addition, the present invention provides a method for preventing or treating fatty liver disease comprising administering a brown gruel extract to a subject other than a human.
본 발명은 갈색거저리 추출물을 유효성분으로 포함하는 지방간 질환의 예방, 개선 또는 치료용 조성물에 관한 것으로, 본 발명의 유효성분인 갈색거저리 추출물은 식용 가능한 것으로 알려진 원료이므로, 부작용 및 독성이 없을 뿐만 아니라, 간암 세포주(HepG2)에서 중성지방의 축적을 효과적으로 감소시키므로, 본 발명의 조성물은 지방간 질환의 예방, 개선 또는 치료를 위한 건강기능식품, 의약품 또는 사료첨가제로 유용하게 사용될 수 있다.The present invention relates to a composition for preventing, ameliorating or treating a fatty liver disease comprising an extract of brown gruel as an active ingredient. Since the brown gruy extract as an active ingredient of the present invention is a raw material known to be edible, it has no side effects and toxicity , because liver cancer cell line (HepG 2) reduction in the accumulation of triglycerides effectively, the composition of the present invention may be useful as a functional food, a drug, or a feed additive for the prevention, improvement or treatment of fatty liver disease.
도 1은 간암 세포주(HepG2)에 올레이트 및 팔미테이트가 2:1의 농도비로 혼합된 1.0mM의 유리지방산(FFA) 및 갈색거저리 추출물(Cu)을 처리한 후, 지방 흡수율(lipid uptake)을 측정한 결과이다. ####은 대조군(Control)에 비해 유리지방산을 처리한 군에서의 지방 흡수율이 통계적으로 유의미하게 증가했다는 것으로, p<0.0001이고, *, **은 유리지방산 처리군에 비해 본 발명의 갈색거저리 추출물 처리군의 유리지방산의 흡수율이 통계적으로 유의미하게 감소했다는 것으로, *은 p<0.05이며, **은 p<0.01이다.1 is a liver cancer cell line (HepG 2) on oleate and palmitate is 2: 1 concentration ratio after the process the free fatty acids (FFA) and mealworm extract (Cu) of a mixture of 1.0mM, fat absorption (lipid uptake) . #### showed a statistically significant increase in the fat absorption rate in the group treated with free fatty acids compared with the control group (p <0.0001), while * and ** indicate that the brown of the present invention The absorption of free fatty acids in the sorghum extract-treated group was statistically significantly decreased. The * indicates p <0.05 and ** indicates p <0.01.
도 2는 갈색거저리 추출물(Cu)이 고지방 식이에 의한 간 무게와 간조직 내 지질 축적 증가를 감소시키는지 확인한 결과로, (A)는 조건별(정상군(N), 고지방식이군(HF), 고지방 식이와 함께 양성대조군으로 밀크시슬 추출물(100mg/kg/day)을 투여한 군(HF+Mi) 및 고지방 식이와 함께 및 본 발명의 갈색거저리 추출물 투여군(HF+Cu)) 간조직의 사진이고, (B) 상기 조직을 오일레드 오 염색 결과 사진(위) 및 이를 그래프(아래)로 나타낸 것이다. ##은 정상군 대비 고지방식이군의 간 무게가 통계적으로 유의미하게 증가하였다는 것으로 p<0.01이며, **는 고지방식이군 대비 본 발명의 갈색거저리 추출물 투여군 또는 양성대조군인 밀크시슬 추출물 투여군의 간 무게가 통계적으로 유의미하게 감소했다는 것으로, p<0.01이다.FIG. 2 shows the results of (A), (B), (C), (C) and (C) (HF + Mi), which was administered with milk fat extract (100 mg / kg / day) as a positive control with high fat diets, and the brown gruel extract administered group (HF + Cu) And (B) the above tissues are shown as oil droplet staining results (above) and their graphs (below). P <0.01 was found to be a statistically significant increase in the liver weight of the high-fat diet group compared to the normal group, and ** was higher in the high fat diet group compared to the control group of the brown goat extract- The statistically significant decrease in weight was p <0.01.
도 3은 간암 세포주(HepG2)에 에탄올 및 갈색거저리 추출물을 처리한 후 세포생존률(%)을 확인한 결과이다. ####은 대조군(Control)에 비해 에탄올 처리군의 세포 생존률이 통계적으로 유의미하게 감소했다는 것으로, p<0.0001이고, **, ***은 에탄올 처리군에 비해 본 발명의 갈색거저리 추출물 처리군의 세포 생존률이 통계적으로 유의미하게 증가했다는 것으로, **은 p<0.01이고, ***은 p<0.001이다.FIG. 3 shows the cell viability (%) after treatment of hepatocellular carcinoma cell line (HepG 2 ) with ethanol and brown goat extract. P <0.0001, ** and *** were significantly higher than those of the ethanol-treated group compared to the control group (Control) The cell survival rate of the group was statistically significantly increased, ** is p <0.01 and *** is p <0.001.
도 4는 간암 세포주(HepG2)에 에탄올 및 갈색거저리 추출물을 처리한 후 Caspase-3의 발현량 변화를 확인한 결과이다. ####은 대조군(Control)에 비해 에탄올 처리군의 Caspase-3의 발현량이 통계적으로 유의미하게 증가했다는 것으로, p<0.0001이고, ****은 에탄올 처리군에 비해 본 발명의 갈색거저리 추출물 처리군의 Caspase-3의 발현량이 통계적으로 유의미하게 감소했다는 것으로, p<0.0001이다.FIG. 4 shows the results of examining the expression level of Caspase-3 after treatment of hepatocellular carcinoma cell line (HepG 2 ) with ethanol and brown goat extract. The expression level of Caspase-3 in the ethanol-treated group was significantly higher than that in the control (p <0.0001), and **** was significantly higher than that of the control group The expression level of caspase-3 in the treatment group was statistically significantly decreased, and p <0.0001.
도 5는 본 발명의 갈색거저리 추출물(Cu)이 고지방 식이에 의한 혈액 내 ALT 증가를 감소시키는지 여부를 확인한 결과이다. Mi는 양성 대조군으로, 고지방 식이와 함께 밀크시슬 추출물(100mg/kg/day)을 투여한 군이다. ###은 대조군(Control)에 비해 고지방식이군의 ALT 값이 통계적으로 유의미하게 증가했다는 것으로, p<0.001이고, ***은 고지방식이군에 비해 본 발명의 갈색거저리 추출물 처리군 또는 양성대조군인 밀크시슬 추출물 처리군의 ALT 값이 통계적으로 유의미하게 감소했다는 것으로, p<0.001이다.FIG. 5 shows the result of confirming whether the brown goat's extract (Cu) of the present invention reduces ALT increase in the blood by the high fat diet. Mi was a positive control group, which received milk-thirst extract (100 mg / kg / day) with high fat diet. (P <0.001) and *** was significantly higher than that of the control group (control group) compared to that of the control group (control group), the brown goat extract treatment group of the present invention or the positive control group , Which is a statistically significant decrease in the ALT value of the in-milk-thistle extract-treated group, is p <0.001.
도 6은 본 발명의 갈색거저리 추출물(Cu)이 고지방 식이에 의한 혈액 내 총콜레스테롤(TC) 증가를 감소시키는지 여부를 확인한 결과이다. Mi은 양성 대조군으로, 고지방 식이와 함께 밀크시슬 추출물(100mg/kg/day)을 투여한 군이다. ###은 대조군(Control)에 비해 고지방식이군의 혈액 내 총콜레스테롤이 통계적으로 유의미하게 증가했다는 것으로, p<0.001이고, **, ****은 고지방식이군에 비해 본 발명의 갈색거저리 추출물 처리군 또는 양성대조군인 밀크시슬 추출물 처리군의 혈액 내 총콜레스테롤이 통계적으로 유의미하게 감소했다는 것으로, **은 p<0.01이고, ****은 p<0.0001이다.FIG. 6 shows the result of confirming whether the brown gruel extract (Cu) of the present invention reduces the increase of total cholesterol (TC) in the blood by the high fat diet. Mi was a positive control group, which received milk-thistle extract (100 mg / kg / day) with high fat diet. P <0.001, ** and **** were higher than those of the control group (control group), and those of the control group (control group) were significantly higher than those of the control group The total cholesterol in the blood of the extract-treated group or the milk-treated cholesterol extract group, which is a positive control group, was statistically significantly decreased, ** is p <0.01 and **** is p <0.0001.
도 7은 본 발명의 갈색거저리 추출물(Cu)이 고지방 식이에 의한 혈액 내 글루코스(glucose) 증가를 감소시키는지 여부를 확인한 결과이다. Mi은 양성 대조군으로, 고지방 식이와 함께 밀크시슬 추출물(100mg/kg/day)을 투여한 군이다. ###은 대조군(Control)에 비해 고지방식이군의 글루코스 함량이 통계적으로 유의미하게 증가했다는 것으로, p<0.001이고, **은 고지방식이군에 비해 본 발명의 갈색거저리 추출물 처리군의 글루코스 함량이 통계적으로 유의미하게 감소했다는 것으로, p<0.01이다.FIG. 7 shows the result of confirming whether the brown gruel extract (Cu) of the present invention reduces the glucose increase in the blood due to the high fat diet. Mi was a positive control group, which received milk-thistle extract (100 mg / kg / day) with high fat diet. ### showed a statistically significant increase in the glucose content of the high-fat diet group compared to the control group (p <0.001), ** indicates that the glucose content of the brown goat extract-treated group of the present invention Statistically significant decrease, p <0.01.
도 8은 본 발명의 갈색거저리 추출물(Cu)이 고지방 식이(HFD)에 의해 형성된 간조직의 지방방울을 감소시키는지 여부를 확인한 결과이다. Mi은 양성 대조군으로, 고지방 식이와 함께 밀크시슬 추출물(100mg/kg/day)을 투여한 군이다. (A)는 정상대조군의 간조직이고, (B)는 고지방식이군의 간조직이며, (C)는 고지방식이 및 양성대조군인 밀크시슬 추출물의 투여군의 간조직이고, (D)는 고지방식이 및 본 발명의 갈색거저리 추출물 투여군의 간조직이다.FIG. 8 shows the results of confirming whether the brown gruel extract (Cu) of the present invention reduces the fat droplets of liver tissue formed by high fat diets (HFD). Mi was a positive control group, which received milk-thistle extract (100 mg / kg / day) with high fat diet. (A) is a liver tissue of a normal control group, (B) is a liver tissue of a high fat diet group, (C) is a liver tissue of a high fat fat diet group and a positive control milk group, And the liver tissue of the brown gruel extract administration group of the present invention.
본 발명은 갈색거저리 추출물을 유효성분으로 함유하는 지방간 질환의 예방 또는 개선용 건강기능식품 조성물에 관한 것이다.The present invention relates to a health functional food composition for preventing or ameliorating fatty liver disease, which comprises an extract of brown gruel as an active ingredient.
상기 갈색거저리 추출물은 하기의 단계를 포함하는 방법에 의해 제조할 수 있으나, 이에 한정하지 않는다:The brown gutter extract may be prepared by a method including, but not limited to, the following steps:
(1) 갈색거저리 건조 분말에 추출용매를 가하여 추출하는 단계;(1) extracting a brown gummy dry powder with an extraction solvent;
(2) 단계 (1)의 추출물을 여과하는 단계; 및 (2) filtering the extract of step (1); And
(3) 단계 (2)의 여과한 추출물을 감압 농축하고 건조하여 추출물을 제조하는 단계. (3) The step of extracting the filtered extract of step (2) by concentration under reduced pressure and drying.
상기 단계 (1)에서 추출용매는 물, C1~C4의 저급 알코올 또는 이들의 혼합물 중에서 선택하는 것이 바람직하며, 더 바람직하게는 에탄올이고, 더욱더 바람직하게는 70%(v/v) 에탄올이지만 이에 한정하지 않는다.In step (1), the extraction solvent is preferably selected from water, a C 1 -C 4 lower alcohol or a mixture thereof, more preferably ethanol, even more preferably 70% (v / v) ethanol But is not limited thereto.
상기 제조방법에 있어서, 추출방법은 여과법, 열수 추출, 침지 추출, 환류 냉각 추출 및 초음파 추출 등의 당 업계에 공지된 모든 통상적인 방법을 이용할 수 있다. 상기 추출용매는 건조된 갈색거저리 중량의 1~20배 첨가하여 추출하는 것이 바람직하며, 더 바람직하게는 5~15배 첨가하는 것이다. 추출온도는 4 내지 50℃인 것이 바람직하나 이에 한정하지 않는다. 또한, 추출시간은 0.5~10시간인 것이 바람직하며, 0.5~5시간이 더욱 바람직하고, 1시간이 가장 바람직하나 이에 한정하지 않는다. 상기 방법에 있어서, 단계 (3)의 감압농축은 진공 감압 농축기 또는 진공회전증발기를 이용하는 것이 바람직하나 이에 한정하지 않는다. 또한, 건조는 감압건조, 진공건조, 비등건조, 분무 건조 또는 동결 건조하는 것이 바람직하나 이에 한정하지 않는다.In the above production method, any conventional method known in the art such as filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction may be used. The extraction solvent is preferably added in an amount of 1 to 20 times, preferably 5 to 15 times, the weight of the dried brown gruel. The extraction temperature is preferably 4 to 50 DEG C, but is not limited thereto. The extraction time is preferably 0.5 to 10 hours, more preferably 0.5 to 5 hours, most preferably 1 hour, but not always limited thereto. In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in the step (3), but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.
상기 지방간 질환은 비알콜성 또는 알콜성 단순 지방간; 비알콜성 또는 알콜성 지방간염; 및 비알콜성 또는 알콜성 간경변 중에서 선택된 어느 하나인 것이 바람직하지만 이에 한정하는 것은 아니다. The fatty liver disease is non-alcoholic or alcoholic simple fatty liver; Non-alcoholic or alcoholic fatty liver disease; And non-alcoholic or alcoholic cirrhosis, but is not limited thereto.
상기 갈색거저리 추출물은 간 조직 내 중성지방의 함량을 감소시키는 것이 특징이며, 간 조직 내 총 중성지방의 함량을 감소시키며, 지방 흡수율(lipid uptake)을 감소시키는 것이 특징이다.The brown gruel extract is characterized by reducing the content of triglyceride in liver tissue, reducing the content of total triglyceride in liver tissue and decreasing lipid uptake.
상기 본 발명의 조성물을 지방간 질환의 예방 또는 개선을 목적으로 식품에 첨가하여 건강기능성 식품을 제조할 수 있는데, 상기 건강기능 식품은 특별히 이에 제한되지 않으며, 건강 기능성 식품, 영양 보조제, 영양제, 파머푸드(pharmafood), 건강 식품, 뉴트라슈티칼(nutraceutical), 디자이너 푸드, 식품 첨가제 등의 모든 형태의 식품이 될 수 있는데, 바람직하게는 육류, 소세지, 빵, 쵸코렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 될 수 있다. 이때, 첨가하는 방법은 통상적인 방법에 따라 적절하게 사용되고, 첨가량은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. The health functional food may be prepared by adding the composition of the present invention to a food for the purpose of preventing or ameliorating fatty liver disease. The health functional food is not particularly limited, and may be a health functional food, a nutritional supplement, but it can be any type of food such as pharmafood, health food, nutraceutical, designer food, food additive, etc. Preferably, the food can be meat, sausage, bread, chocolate, candy, snack, confectionery, , Other noodles, gums, dairy products including ice cream, various soups, drinks, tea, drinks, alcoholic beverages and vitamin complexes. At this time, the method of adding is appropriately used according to a conventional method, and the amount to be added can be suitably determined according to the intended use (prevention, health or therapeutic treatment).
상기 본 발명의 조성물은 유효성분 이외의 다양한 성분을 추가로 포함할 수도 있는데, 이들 다양한 성분은 특별히 이에 제한되지 않으나, 바람직하게는 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등이 될 수 있다. 또한, 기호성 및/또는 기능성을 부가하기 위하여 천연 과일 주스, 과일 주스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있으며, 이들 성분들은 독립적으로 또는 조합하여 사용할 수 있다.The composition of the present invention may further include various components other than the active component. The various components may be various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, Salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks and the like. It may also contain flesh for the production of natural fruit juices, fruit juice beverages and vegetable beverages to add palatability and / or functionality, and these ingredients may be used independently or in combination.
본 발명은 갈색거저리 추출물을 유효성분으로 포함하는 지방간 질환의 예방 또는 치료용 약학 조성물에 관한 것이다.The present invention relates to a pharmaceutical composition for preventing or treating fatty liver disease, which comprises an extract of brown gruel as an active ingredient.
상기 지방간 질환은 비알코올성 또는 알코올성 단순 지방간; 비알코올성 또는 알코올성 지방간염; 및 비알코올성 또는 알코올성 간경변 중에서 선택된 어느 하나인 것이 바람직하지만 이에 한정하지 않는다.The fatty liver disease is a non-alcoholic or alcoholic simple fatty liver; Nonalcoholic or alcoholic fatty liver disease; And non-alcoholic or alcoholic liver cirrhosis, but is not limited thereto.
본 발명의 약학 조성물은 약학적으로 허용되는 담체를 포함할 수 있다. 본 발명의 약학 조성물에 포함되는 약학적으로 허용되는 담체는 제제 시에 통상적으로 이용되는 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 덱스트로즈 용액, 말토덱스트린 용액, 글리세롤, 에탄올, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 중에서 선택된 1종 이상의 담체를 더 포함할 수 있으나, 이에 한정되는 것은 아니다. The pharmaceutical compositions of the present invention may comprise a pharmaceutically acceptable carrier. The pharmacologically acceptable carrier to be contained in the pharmaceutical composition of the present invention is one which is usually used at the time of formulation and includes saline, sterilized water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, But are not limited to, lactose, sucrose, sucrose, sorbitol, mannitol, starch, acacia, calcium phosphate, alginate, gelatin, calcium calcium, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methylcellulose, methylhydroxybenzoate, Hydroxybenzoate, talc, magnesium stearate, and mineral oil, but the present invention is not limited thereto.
본 발명의 약학적 조성물은 상기 성분들 이외에 항산화제, 완충액, 정균제, 희석제, 계면활성제, 결합제, 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제 및 보존제 중에서 선택된 1종 이상의 보조제 등을 추가로 포함할 수 있다. The pharmaceutical composition of the present invention may further comprise at least one auxiliary selected from an antioxidant, a buffer, a bacteriostat, a diluent, a surfactant, a binder, a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, .
본 발명의 약학적 조성물은 경구 또는 비경구 투여할 수 있으며, 바람직하게는 경구 투여 방식으로 적용된다. 본 발명의 약학적 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다. 본 발명의 약학적 조성물의 투여량은 바람직하게는 성인 기준으로 0.001-100 ㎎/kg이다.The pharmaceutical composition of the present invention can be administered orally or parenterally, and is preferably administered orally. The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on such factors as formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, route of administration, excretion rate, . The dosage of the pharmaceutical composition of the present invention is preferably 0.001-100 mg / kg on an adult basis.
본 발명의 약학적 조성물은 당해 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다. 이때 제형은 오일 또는 수성 매질중의 용액, 현탁액, 시럽제 또는 유화액 형태이거나 엑스제, 산제, 분말제, 과립제, 정제 또는 캅셀제 형태일 수도 있으며, 분산제 또는 안정화제를 추가적으로 포함할 수 있다.The pharmaceutical composition of the present invention may be formulated into a unit dosage form by using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by those having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container. The formulations may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of excipients, powders, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.
또한, 본 발명은 갈색거저리 추출물을 유효성분으로 포함하는 인간을 제외한 동물의 지방간 질환의 예방 또는 치료용 수의학적 조성물에 관한 것이다.The present invention also relates to a veterinary composition for the prevention or treatment of fatty liver disease in an animal other than humans, which comprises an extract of brown gruel as an active ingredient.
본 발명의 갈색거저리 추출물을 포함하는 수의학적 조성물은 통상의 방법에 따른 적절한 부형제 및 희석제를 더 포함할 수 있다. 본 발명의 갈색거저리 추출물을 포함하는 수의학적 조성물에 포함될 수 있는 부형제 및 희석제로는, 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트, 세탄올, 스테아릴알콜, 유동파라핀, 솔비탄모노스테아레이트, 폴리소르베이트 60, 메칠파라벤, 프로필파라벤 및 광물유를 들 수 있다. The veterinary composition comprising the brown gruel extract of the present invention may further comprise suitable excipients and diluents according to conventional methods. Examples of excipients and diluents that can be included in the veterinary composition containing the brown gutter extract of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin , Calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, cetanol, stearyl alcohol, Paraffin, sorbitan monostearate, polysorbate 60, methylparaben, propylparaben, and mineral oil.
본 발명에 따른 갈색거저리 추출물을 포함하는 수의학적 조성물은 충진제, 항응집제, 윤활제, 습윤제, 향신료, 유화제, 방부제 등을 추가로 포함할 수 있는데, 본 발명에 의한 수의학적 조성물은 동물에 투여된 후 활성 성분의 신속, 지속 또는 지연된 방출을 제공할 수 있도록 당업계에 잘 알려진 방법을 사용하여 제형화될 수 있고, 제형은 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 용액, 시럽, 에어로졸, 연질 또는 경질 젤라틴 캅셀, 좌제, 멸균 주사용액, 멸균 외용제 등의 형태일 수 있다.The veterinary composition comprising the brown gruel extract according to the present invention may further contain a filler, an anti-coagulant, a lubricant, a wetting agent, a spice, an emulsifier, an antiseptic, etc. The veterinary composition according to the present invention may further comprise May be formulated using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient, and the formulations may be formulated as powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, Soft or hard gelatine capsules, suppositories, sterile injectable solutions, sterile external preparations, and the like.
본 발명에 의한 수의학적 조성물은 동물의 나이, 성별, 체중에 따라 달라질 수 있으나, 0.1~100mg/㎏의 양을 1일 1회 내지 수회 투여할 수 있고, 투여량은 투여경로, 질병의 정도, 성별, 체중, 나이 등에 따라서 증감될 수 있다. 따라서, 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.The veterinary composition according to the present invention may vary depending on the age, sex, and body weight of the animal, but may be administered in an amount of 0.1 to 100 mg / kg once or several times a day. The dosage may vary depending on the route of administration, Sex, weight, age, and the like. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.
또한, 본 발명은 갈색거저리 추출물을 유효성분으로 포함하는 인간을 제외한 동물의 지방간 질환의 예방 또는 개선용 사료 첨가제에 관한 것이다.The present invention also relates to a feed additive for preventing or ameliorating fatty liver disease in an animal other than a human, which comprises an extract of brown gruel as an active ingredient.
상기 사료 첨가제는 20~90중량%의 갈색거저리 추출물을 함유하는 고 농축액, 분말 또는 과립형태일 수 있다. The feed additive may be in the form of a high concentrate, powder or granulate containing from 20 to 90% by weight of brown gruel extract.
본 발명의 사료 첨가제는 구연산, 후말산, 아디픽산, 젖산, 사과산 등의 유기산이나 인산나트륨, 인산칼륨, 산성피로인산염, 폴리인산염(중합인산염) 등의 인산염이나 폴리페놀, 카테킨(catechin), 알파-토코페롤, 로즈메리 추출물(rosemary extract), 비타민 C, 녹차 추출물, 감초 추출물, 키토산, 탄닌산, 피틴산 등의 천연 항산화제 중에서 선택된 하나 이상을 추가로 포함할 수 있다.The feed additive of the present invention can be used as a feed additive containing a phosphate such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, or organic acids such as sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polymerized phosphate), polyphenol, catechin, Natural antioxidants such as tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid may be further included.
본 발명의 갈색거저리 추출물을 함유하는 동물사료 첨가제 및 이를 포함하는 사료는 보조성분으로 아미노산, 무기염류, 비타민, 항생물질, 항균물질, 항산화, 항곰팡이 효소, 소화 및 흡수향상제, 성장촉진제, 질병예방제 등과 같은 물질과 함께 사용될 수 있다.The animal feed additive containing the brown goat's extract of the present invention and the feed comprising the animal feed additive can be supplemented with amino acids, inorganic salts, vitamins, antibiotics, antimicrobials, antioxidants, antifungal enzymes, digestion and absorption enhancers, growth promoters, And the like.
상기 사료 첨가제는 동물에게 단독으로 식용 담체 중에서 다른 사료 첨가제와 조합되어 투여될 수 있다. 또한, 상기 사료첨가제는 탑 드레싱으로서 또는 이들을 동물 사료에 직접 혼합하거나 또는 사료와 별도로, 별도의 경구 제형으로, 주사 또는 경피로 또는 다른 성분과 조합하여 쉽게 투여할 수 있다. 통상적으로, 당 업계에 잘 알려진 바와 같이 단독 일일 투여량 또는 분할 일일 투여량을 사용할 수 있다. 상기 사료 첨가제를 동물 사료와 별도로 투여할 경우, 당 업계에 잘 알려진 바와 같이 추출물의 투여 형태는 이들을 비-독성 제약상 허용 가능한 식용 담체와 조합하여 즉석 방출 또는 서방성 제형으로 제조할 수 있다. 이러한 식용 담체는 고체 또는 액체, 예를 들어 옥수수 전분, 락토스, 수크로스, 콩 플레이크, 땅콩유, 올리브유, 참깨유 및 프로필렌 글리콜일 수 있다. 고체 담체가 사용될 경우, 추출물의 투여형은 정제, 캡슐제, 산제, 토로키제 또는 함당정제 또는 미분산성 형태의 탑 드레싱일 수 있다. 액체 담체가 사용될 경우, 연 젤라틴 캡슐제, 또는 시럽제 또는 액체 현탁액제, 에멀젼제 또는 용액제의 투여 형태일 수 있다. 또한, 투여 형태는 보조제, 예를 들어 보존제, 안정화제, 습윤제 또는 유화제, 용액 촉진제 등을 함유할 수 있다. 또한, 갈색거저리 추출물이 사료 첨가제로 포함되는 동물사료는 동물의 식이 요구를 충족시키는데 통상적으로 사용되는 임의의 단백질-함유 유기 곡분일 수 있다. 이러한 단백질-함유 곡분은 통상적으로 옥수수, 콩 곡분 또는 옥수수/콩 곡분 믹스로 주로 구성되어 있다. 상기의 사료 첨가제는 침지, 분무 또는 혼합하여 상기 동물사료에 첨가하여 이용될 수 있다. 본 발명의 수의학적 조성물 또는 사료첨가제는 반려동물의 식이에 적용할 수 있다.The feed additive may be administered alone to the animal in combination with other feed additives in the edible carrier. The feed additives can also be easily administered as top dressing or they can be mixed directly with the animal feed or separately from the feed, in separate oral formulations, by injection or transdermal, or in combination with other ingredients. Typically, a single daily dose or a divided daily dose can be used as is well known in the art. When the feed additive is administered separately from animal feed, the dosage form of the extract, as is well known in the art, may be prepared in an immediate release or sustained release formulation in combination with a non-toxic pharmaceutically acceptable food carrier. Such edible carriers may be solid or liquid, for example, corn starch, lactose, sucrose, soy flakes, peanut oil, olive oil, sesame oil and propylene glycol. When a solid carrier is used, the dosage form of the extract may be a tablet, capsule, powder, troche or emulsion or top-dressing in finely divided form. When a liquid carrier is used, it may be in the form of a soft gelatin capsule, or a syrup or liquid suspension, emulsion or solution. In addition, the dosage form may contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, solution promoting agents and the like. In addition, the animal feed in which the brown gruel extract is included as a feed additive can be any protein-containing organic gut that is commonly used to meet the dietary needs of an animal. These protein-containing flours usually consist mainly of corn, soy flour or corn / soy flour mix. The feed additive may be added to the animal feed by immersion, spraying or mixing. The veterinary composition or feed additive of the present invention can be applied to the companion animal's diet.
또한, 본 발명은 갈색거저리 추출물을 인간을 제외한 개체에게 투여하는 단계를 포함하는 지방간 질환의 예방 또는 치료방법에 관한 것이다.The present invention also relates to a method for preventing or treating fatty liver disease comprising administering a brown gruel extract to a subject other than a human.
상기 개체는 지방간 질환을 앓는 환자를 포함하여 지방간 질환의 가능성이 있는 사람을 비롯한 동물 모두를 포함한다.Such an individual includes all animals, including those with a possibility of fatty liver disease, including those suffering from fatty liver disease.
이하, 실시예를 이용하여 본 발명을 더욱 상세하게 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로 본 발명의 범위가 이들에 의해 제한되지 않는다는 것은 당해 기술분야에서 통상의 지식을 가진 자에게 있어 자명한 것이다. Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.
실시예 1. 갈색거저리 추출물의 제조Example 1. Preparation of Brown Goat Extract
갈색거저리 건조 분말에 70%(v/v) 에탄올을 사용하여 3시간 동안 환류추출하였다. 70%(v/v) 에탄올 추출액을 수득하고 감압증류하여 갈색거저리 에탄올 추출물을 제조하였다.The dried brown powdery mildew was refluxed for 3 hours using 70% (v / v) ethanol. A 70% (v / v) ethanol extract was obtained and distilled under reduced pressure to prepare a brown goat ethanol extract.
실시예 2. 간암 세포주(HepG2)의 배양Example 2 Culture of Hepatocarcinoma Cell Line (HepG2)
간암 세포주(HepG2)는 ATCC(American Type Culture Collection)에서 구입하여 사용하였다. 간암 세포주(HepG2)는 10% FBS, 1% 페니실린-스트렙토마이신이 포함된 DMEM-F12(Dulbecco's modified Eagle's medium) 배지로 배양하였다. 상기 세포는 37℃에서, 5% CO2 배양기에서 배양하였고, 배양된 세포는 24웰 플레이트에 일정한 수의 세포(3×104 cells/ 500㎕ well)를 분주하여 실험하였다.Liver cancer cell line (HepG 2) were purchased from ATCC (American Type Culture Collection). Were cultured in the DMEM-F12 containing the streptomycin (Dulbecco's modified Eagle's medium) medium-liver cancer cell line (HepG 2) is 10% FBS, 1% penicillin. The cells were cultured in a 5% CO 2 incubator at 37 ° C, and the cultured cells were tested by dividing a certain number of cells (3 × 10 4 cells / 500 μl well) into 24-well plates.
24웰 플레이트에 세포를 분주한 후, 세포가 24웰이 완전히 붙어 세포의 모양을 갖춘 후 세포가 75%의 포화도(confluence)에 도달했을 때 사용하였다. After cells were placed in a 24-well plate, the cells were used when 24 wells were completely attached to the cells and the cells reached 75% confluence.
실시예 3. 유리 지방산(Free Fatty Acid; FFA) 제조Example 3. Preparation of Free Fatty Acid (FFA)
(1) 100mM 팔미테이트(시그마 P-0500) 및 100mM 올레이트(시그마 O-75010) 저장용액의 제조(1) Preparation of 100 mM palmitate (Sigma P-0500) and 100 mM Olate (Sigma O-75010) stock solution
70℃에서 0.1M NaOH 용액을 용매로 이용하여 제조하고, 0.2㎛의 필터를 이용하여 여과한 후, 멸균하였다.A 0.1 M NaOH solution was used as a solvent at 70 째 C, filtered using a 0.2 탆 filter, and then sterilized.
(2) 유리 지방산이 포함되지 않은 5%(w/v)의 BSA(시그마 A-6003) 용액의 제조(2) Preparation of 5% (w / v) BSA (Sigma A-6003) solution free of free fatty acids
3차 증류수를 용매로 이용하여 제조한 후 멸균하고, 4℃ 냉장보관하였다.After the third distilled water was used as a solvent, it was sterilized and stored at 4 ° C in a refrigerator.
(3) 올레이트 및 팔미테이트의 농도가 2:1이 되도록 혼합한 5mM의 혼합지방산 저장용액 제조(3) Preparation of 5 mM mixed fatty acid storage solution mixed so that the concentration of olate and palmitate was 2: 1
60℃에서, 상기 (2)에서 제조한 3차 증류수에 녹인 유리 지방산이 포함되지 않은 5%(w/v)의 BSA 용액을 용매로 이용하여 상기 (1)에서 제조한 각각의 지방산의 농도가 5mM이 되도록 지방산 저장용액 10㎖을 제조하였다. 상기 (1)에서 제조한 100mM 팔미테이트 및 100mM 올레이트 저장용액을 165㎕(1.65mM)의 팔미테이트(Palmitate) 및 330㎕(3.3mM)의 올레이트(oleate)를 취해 9,505㎕의 5%(w/v)의 BSA 용액에 방울방울 떨어뜨리면서 흔들며 섞어주었다(vortexing). 이후에 상온으로 식힌 다음, 0.2μm 필터로 여과하였다(상기 제조한 혼합용액은 -20℃에서 3~4주 동안 보관하여도 안정하다).The concentration of each fatty acid prepared in (1) above was measured using a 5% (w / v) BSA solution containing no free fatty acid dissolved in the tertiary distilled water prepared in (2) 10 ml of a fatty acid storage solution was prepared so as to be 5 mM. 100 μl of palmitate and 100 mM of the olate storage solution prepared in (1) above were mixed with 165 μl (1.65 mM) of palmitate and 330 μl (3.3 mM) of oleate and 9505 μl of 5% w / v) dropwise dropwise to the BSA solution (vortexing). Thereafter, the mixture was cooled to room temperature and then filtered with a 0.2 μm filter (the mixed solution was stable at -20 ° C. for 3 to 4 weeks).
실시예 4. 갈색거저리 추출물의 처리에 따른 지방 흡수(lipid uptake) 정도의 변화 확인 Example 4. Determination of Changes in the Level of Lipid Uptake by Treatment of Brown Goat Extract
간암 세포주(HepG2)에 상기 실시예 3에서 제조한 1mM의 올레이트 및 팔미테이트 혼합(2:1) 유리 지방산을 처리하여 증가한 지방 흡수(lipid uptake)가 갈색거저리 추출물의 처리에 의해 감소 되는지를 확인하였다.Whether by the FFA treatment increased fat absorption (lipid uptake) is reduced in the process of the mealworm extract: hepatoma cell line (HepG 2) Example oleate of 1mM prepared from 3 and palmitate mixture (1: 2) to Respectively.
그 결과 도 1에 개시한 바와 같이, 갈색거저리 추출물의 농도 의존적으로 지방 흡수(lipid uptake)가 감소한 것을 확인하였다.As a result, as shown in Fig. 1, it was confirmed that the lipid uptake decreased in a concentration-dependent manner in the brown goat extract.
실시예 5. 오일레드 오 염색(Oil Red O staining) 어세이Example 5. Oil Red O staining assay
갈색거저리 추출물(Cu)이 고지방 식이에 의한 간조직 내 지질 축적 증가를 감소시키는지 확인하기 위해 Oil red O 염색을 하였다. 지질은 Oil red O 염색법에 의해 붉은색으로 염색된다. Oil red O staining was performed to determine whether the brown goat extract (Cu) reduced the lipid accumulation in liver tissues due to high fat diets. Lipids are stained red by Oil red O staining method.
그 결과 도 2에 개시한 바와 같이, 고지방 식이에 의해 증가된 지질 축적이 갈색거저리 추출물(Cu)에 의해 감소하여 간 조직 보호효능을 확인하였다. 또한 간조직의 지방 축적으로 인한 간 무게 증가가 갈색거저리 추출물(Cu)에 의해 통계적으로 유의미하게 감소하는 것을 확인하였다.  As a result, as shown in Fig. 2, the lipid accumulation increased by the high fat diet was reduced by the brown goat extract (Cu), and the liver tissue protective effect was confirmed. In addition, it was confirmed that liver weight increase due to fat accumulation of liver tissue was statistically decreased by brown goat extract (Cu).
실시예 6. 본 발명의 갈색거저리 추출물의 처리에 따른 세포 생존율 및 Caspase-3의 발현량 변화 확인 Example 6 Confirmation of Cell Survival Rate and Caspase-3 Expression by the Treatment of Brown Goat Extract of the Present Invention
간암 세포주(HepG2)에 에탄올을 처리하여 감소한 세포 생존률 및 증진된Caspase-3 발현량이 갈색거저리 추출물의 처리에 의해 변화가 일어나는지를 확인하였다.Liver cancer cell line (HepG 2) ethanol to decreased cell viability and increase the amount of Caspase-3 expression in the identified changes are happening in the process of the mealworm extract.
그 결과 도 3 및 도 4에 개시한 바와 같이, 갈색거저리 추출물의 세포 생존율이 증진되었고, 농도 의존적으로 Caspase-3의 발현량이 현저하게 감소한 것을 확인하였다.As a result, as shown in FIG. 3 and FIG. 4, it was confirmed that the cell viability of the brown goat extract was enhanced and the expression level of Caspase-3 was remarkably decreased in a concentration-dependent manner.
실시예 7. 본 발명의 갈색거저리 추출물의 처리에 따른 혈액 내 ALT, 총콜레스테롤(TC) 및 글루코스의 함량 변화 확인Example 7 Confirmation of Changes in ALT, Total Cholesterol (TC) and Glucose Content in the Blood Following the Treatment of the Brown Goat Extract of the Present Invention
본 발명의 갈색거저리 추출물(Cu)이 고지방 식이에 의해 증가된 혈액 내 ALT, 총콜레스테롤(TC), 글루코스의 함량을 감소시키는지 여부를 확인하였다. It was confirmed whether the brown gruel extract (Cu) of the present invention reduced the content of ALT, total cholesterol (TC) and glucose in the blood increased by the high fat diet.
그 결과, 도 5~7에 개시한 바와 같이, 고지방식이에 의해 증가된 ALT, 총콜레스테롤 및 글루코스 함량이 통계적으로 유의미하게 감소한 것을 확인하였다.As a result, as shown in Figs. 5 to 7, it was confirmed that the ALT, total cholesterol and glucose contents increased by the high fat diet method statistically significantly decreased.
실시예 8. H&E (hematocylin & eosin) 염색Example 8. H & E (hematocylin & eosin) staining
본 발명의 갈색거저리 추출물(Cu)이 고지방 식이에 의한 간조직 내 지질 축적 증가를 감소시키는지 확인하기 위해 H&E 염색을 하였다. H & E staining was performed to determine if the brown goat extract (Cu) of the present invention reduced lipid accumulation in the liver by high fat diet.
그 결과 도 8에 개시한 바와 같이, 고지방 식이에 의해 증가된 지방방울의 형성을 확인할 수 있었으며, 본 발명의 갈색거저리 추출물(Cu) 또는 양성대조군인 밀크씨슬을 처리한 경우, 고지방 식이군에 비해 지방방울이 감소한 것을 확인하였다.  As a result, as shown in FIG. 8, it was confirmed that the fat droplet formation was increased by the high fat diet. When the brown goat extract (Cu) of the present invention or milkciss positive control group was treated, Compared to the control group.

Claims (11)

  1. 갈색거저리 추출물을 유효성분으로 함유하는 지방간 질환의 예방 또는 개선용 건강기능식품 조성물.A health functional food composition for prevention or amelioration of fatty liver disease containing an extract of brown goat skin as an active ingredient.
  2. 제1항에 있어서, 상기 갈색거저리 추출물의 추출용매는 물, C1~C4의 저급 알코올 또는 이들의 혼합물인 것을 특징으로 하는 지방간 질환의 예방 또는 개선용 건강기능식품 조성물.The health functional food composition according to claim 1, wherein the extraction solvent of the brown gutter extract is water, a C 1 -C 4 lower alcohol or a mixture thereof.
  3. 제1항에 있어서, 상기 지방간 질환은 비알코올성 또는 알코올성 단순 지방간; 비알코올성 또는 알코올성 지방간염; 및 비알코올성 또는 알코올성 간경변 중에서 선택된 어느 하나인 것을 특징으로 하는 지방간 질환의 예방 또는 개선용 건강기능식품 조성물.The method of claim 1, wherein the fatty liver disease is selected from the group consisting of non-alcoholic or alcoholic simple fatty liver; Nonalcoholic or alcoholic fatty liver disease; And a non-alcoholic or alcoholic liver cirrhosis.
  4. 제1항에 있어서, 상기 갈색거저리 추출물은 간 조직 내 중성지방의 함량을 감소시키는 것을 특징으로 하는 지방간 질환의 예방 또는 개선용 건강기능식품 조성물. The health functional food composition according to claim 1, wherein the brown gourd extract reduces the content of triglyceride in liver tissue.
  5. 갈색거저리 추출물을 유효성분으로 포함하는 지방간 질환의 예방 또는 치료용 약학 조성물.A pharmaceutical composition for the prevention or treatment of fatty liver disease comprising an extract of brown goat as an active ingredient.
  6. 제5항에 있어서, 상기 지방간 질환은 비알코올성 또는 알코올성 단순 지방간; 비알코올성 또는 알코올성 지방간염; 및 비알코올성 또는 알코올성 간경변 중에서 선택된 어느 하나인 것을 특징으로 하는 지방간 질환의 예방 또는 치료용 약학 조성물.6. The method of claim 5, wherein the fatty liver disease is a non-alcoholic or alcoholic simple fatty liver; Nonalcoholic or alcoholic fatty liver disease; And a non-alcoholic or alcoholic liver cirrhosis.
  7. 제5항에 있어서, 상기 유효성분 이외에 약학적으로 허용가능한 식염수, 멸균수, 링거액, 완충 식염수, 덱스트로즈 용액, 말토덱스트린 용액, 글리세롤, 에탄올, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 중에서 선택된 1종 이상의 담체를 더 포함하는 것을 특징으로 하는 지방간 질환의 예방 또는 치료용 약학 조성물.The pharmaceutical composition according to claim 5, wherein the active ingredient is selected from the group consisting of a pharmaceutically acceptable salt solution, sterilized water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, lactose, dextrose, sucrose, sorbitol, mannitol, But are not limited to, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, Magnesium, and mineral oil. &Lt; RTI ID = 0.0 &gt; 21. &lt; / RTI &gt;
  8. 제5항에 있어서, 상기 유효성분 이외에 약학적으로 허용 가능한 항산화제, 완충액, 정균제, 희석제, 계면활성제, 결합제, 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제 및 보존제 중에서 선택된 1종 이상의 보조제를 더 포함하는 것을 특징으로 하는 지방간 질환의 예방 또는 치료용 약학 조성물.The pharmaceutical composition according to claim 5, wherein at least one auxiliary selected from a pharmaceutically acceptable antioxidant, a buffer, a bacteriostat, a diluent, a surfactant, a binder, a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, Wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.
  9. 갈색거저리 추출물을 유효성분으로 포함하는 인간을 제외한 동물의 지방간 질환의 예방 또는 치료용 수의학적 조성물.A veterinary composition for the prevention or treatment of fatty liver disease in an animal other than human, which comprises an extract of brown gruel as an active ingredient.
  10. 갈색거저리 추출물을 유효성분으로 포함하는 인간을 제외한 동물의 지방간 질환의 예방 또는 개선용 사료 첨가제.A feed additive for preventing or ameliorating fatty liver disease in an animal other than humans, which contains an extract of brown goat as an active ingredient.
  11. 갈색거저리 추출물을 인간을 제외한 개체에게 투여하는 단계를 포함하는 지방간 질환의 예방 또는 치료방법.A method for preventing or treating fatty liver disease comprising administering to a subject other than a human a brown gruel extract.
PCT/KR2018/013386 2017-11-08 2018-11-06 Composition comprising mealworm beetle extract as effective ingredient for prevention, alleviation, or treatment of fatty liver disease WO2019093739A1 (en)

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