KR20200081267A - Composition for Anti-obesity Using an Extract of Spiraea prunifolia - Google Patents

Abstract

The present invention discloses a composition for anti-obesity using a bridal wreath extract. The composition for anti-obesity using the bridal wreath extract of the present invention inhibits differentiation of 3T3-L1, which is a mouse pro-adipocyte, into adipocytes in a concentration-dependent manner, and also significantly suppresses weight gain in an obese animal model. The composition for anti-obesity using the bridal wreath extract is active not only to significantly reduce the amount of visceral fat and subcutaneous fat, but also to reduce accumulation of liver fat, subcutaneous fat, epididymal fat, and subperitoneal fat, and reduce blood triglycerides.

Description

Composition for anti-obesity using an extract of Spiraea prunifolia}

The present invention relates to a composition for anti-obesity using an extract of Spiraea prunifolia .

In modern society, the obesity population is increasing due to the convenient living environment due to rapid automation, excessive nutrition and reduced physical activity due to the increase in processed foods and eating out. In the United States, the country with the highest obesity rate among developed countries, about 60% of adults are known to be overweight, and according to the National Health and Nutrition Survey, the proportion of adults with a body mass index (BMI) of 25 or higher was 31.7 in 2007. %, which is steadily increasing compared to 26.0% in 1998, and this rate is increasing by 5% every 10 years, which is recognized as a serious problem that threatens public health (JR Soc. Med.99:250-257 2006). The World Health Organization (WHO) classifies obesity as a disease rather than a symptom or symptom, and reported that in 2015, the global obesity population would increase to 1.5 billion, which would be a serious health problem (Ann Epidemiol. 2005. 15:87-97; J Life Sci. 2013. 23:69-78). Although the cause of obesity is not clearly known, it is not caused by a single condition, but by various causes such as improper eating and lifestyle, genetic factors, and decreased physical activity (.Korean J. Food Science Nutrition 22(1 )110-12.2009; Korean J. Food Science Nutrition 23(5):363-367.1990).

Obesity is an increase in the number and size of fat cells due to excessive accumulation of body fat due to an imbalance between energy intake and consumption (Cell. 104:531-543 2001), and energy in the body is in the form of triglyceride in fat cells. When stored and depleted energy sources, the stored fat is decomposed into free fatty acids and glycerol to be used as energy sources, but excessive intake of energy promotes differentiation of fat cells and increases the amount of stored fat in the body, which is a direct cause of obesity. (Metabolism. 30: 425-427 1981; Biochem J. 1;435(3):723-32 2011). Obesity acts as a risk factor to increase the incidence of various diseases as well as changes in body shape according to fat accumulation in the intestines and abdomen (Korean J. Pediatr.48:126-137. 2005) When excessive accumulation of visceral fat causes sugar in the body Problems with metabolism occur, and symptoms such as hormone secretion abnormalities and cytokine secretion abnormalities occur. Increased triglycerides, LDL-cholesterol, and HDL-cholesterol due to obesity cause fat metabolism in the body, reduce insulin receptors present in tissues, and also reduce insulin sensitivity, thereby inhibiting the transport of glucose to the cells. It also causes high blood sugar and diabetes. In addition, obesity is known to have a deep relationship with the occurrence of metabolic diseases such as hyperlipidemia, cardiovascular disease, cancer, respiratory disorders, stroke, osteoarthritis (Med. Int. 22 385-388 1994;.Ann.Intern.Med 103:1024-1029 1985).

Control of the physiological action of adipocytes can be largely divided into induction of adipocyte differentiation, fat biosynthesis, and lipid breakdown. In the differentiation stage of adipocytes, various transcription factors such as peroxisome proliferator activated receptor (PPAR), CCAAT/enhancerbinding proteins (C/EBP), and sterol regulatory element binding protein (SREBP) are induced step by step in adipocytes undergoing differentiation. , It regulates the expression of various adipocyte specific genes through the interaction of each factor (Genes Dev. 14:1293-1307. 2000). Differentiated adipocytes undergo triglyceride biosynthesis using fatty acids and glucose introduced into them, and the enzymes involved in this are FAS (fatty acid synthase), ACC (acetyl-CoA carboxylase), SCD (stearoyl-CoAdesaturase), and ACS (acyl-CoA synthetase), DGAT (diacylglycerol acyltransferase), etc. (Trends Endocrinol. Metab., 23, 56-64. 2011; Biochem. Biophys. Res. Commun., 420,805-810. 2012). Triglycerides synthesized by the action of these enzymes are used as energy sources. Lipid breakdown is controlled by catecholamines and insulin according to nutritional status. The enzymes involved in lipid breakdown include adipose triglyceride lipase (ATGL), hormone sensitive lipase (HSL), and monoacyl glycerol lipase (MGL). When the triglyceride of a molecule is hydrolyzed by the action of these enzymes, a molecule of glycerol and a fatty acid It is divided into three molecules (Trends Endocrinol. Metab., 23, 56-64. 2011; Biochem. Biophys. Res. Commun., 420,805-810. 2012). Fatty acids produced by hydrolysis are oxidized in the mitochondria and consumed as energy or esterified and synthesized as triglycerides and stored in cells.

Adipose fat cells 3T3-L1 were first isolated from 3T3 cells by Green and Meuth (Cell. 3(2):127-33. 1974), and the properties of differentiation into adipocytes under biological characteristics and proper culture conditions were revealed. It has been widely used to study the differentiation process of fat cells and decomposition of accumulated fat. Adipose progenitor cells, 3T3-L1 cells, differentiate into mature adipocytes by various hormones, PPAR, C/EBP, and SREBP, accumulating triglycerides in cells by increasing the expression of related genes and related enzyme activity ( J Nutr 130: 3116S-3121S, 2000; J Nutr 130: 3122S-3126S, 2000). For the study of functional material development, a method of suppressing the differentiation process or exploring a material that promotes fat breakdown using 3T3-L1 cells as described above has been widely used in order to inhibit excessive accumulation of triglycerides in adipose tissue.

Conventional anti-obesity drugs, such as orlistat and sibutramine, are known to have serious side effects such as vomiting, constipation, gastrointestinal disorders, and cardiovascular disease (Int J Obes Relat Metab Disord. Jul;25(7)) :1095-9.2001; Obes Res. 8(6):431-7. 2000; Lancet. 6;369(9555):71-7. 2007; Front Physiol. 2014 Jun 24;5:228. 2014), Efforts to develop effective and safe materials continue. Retinol, vitamin E, vitamin U, and herbal extracts have been reported as substances that inhibit fat cell differentiation (Mol Cell Biol. 16:15671575. 1996; Ann Dermatol. Feb;24(1):39- 44 2012; J Nutr. 139(1):51-7 2009; Ann Dermatol. 24(1):39-44 2012) Research on the development of anti-obesity agents based on natural substances that can be safely and continuously consumed is actively being conducted. .

Spiraea prunifolia for. simpliciflora Nakai is a medicinal plant belonging to the rose family, and is widely distributed throughout the country in Korea and grows well in sunny places such as forests and fields. In general, in traditional medicine, the root is used as a medicine, and it can be used for fever, contraction of vascular tissue, and convergence of branches. It can also be used for fever, neuralgia, swelling and soreness caused by a cold, school quality (a symptom of chills and fever at a certain time every day), and diarrhea.

Nevertheless, there are not many studies on the pharmacological components and uses for Chopop trees, so the applications for Chopop trees are not wide.

Specifically, Korean Patent Registration No. 10-1176526 has known about the anti-aging effect of skin, anti-wrinkle effect, skin whitening effect and skin damage suppression effect through the antioxidant effect of Jopop tree extract, but studies on other uses Is not known.

The present invention discloses the use of anti-obesity of the barberry extract.

An object of the present invention is to provide a composition for anti-obesity using a Jopop tree extract.

Other or specific objects of the present invention will be presented below.

The present invention, as can be seen in the Examples and Experimental Examples below, inhibits the differentiation of mouse progenitor cells 3T3-L1 into adipocytes in a concentration-dependent manner, and clearly suppresses weight gain even in obese animal models. Completed by confirming that it not only significantly reduces the amount of fat in visceral fat and subcutaneous fat, but also reduces the accumulation of liver fat, subcutaneous fat, epididymal fat, and subperitoneal fat and reduces triglyceride in the blood.

When considering the experimental results of the above, the composition for anti-obesity of the present invention is characterized in that it comprises a crude poplar extract as its active ingredient.

In the present specification, the term "Jopop tree extract" refers to Jopop tree leaves, stems, ground parts, roots, roots, underground parts, or mixtures thereof, which are extracted, water, lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.) and methylene chloride. , Ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or mixed solvents thereof The extract obtained by leaching by means of an extract obtained using a supercritical extraction solvent such as carbon dioxide, pentane, or a fraction obtained by fractionating the extract, and the extraction method is cold immersion in consideration of the polarity, extraction degree, and storage degree of the active substance. , Reflux, warming, ultrasonic radiation, supercritical extraction, and other methods can be applied. In the case of fractionated extracts, the fractions obtained by suspending the extract in a specific solvent and mixing and policing with a solvent having a different polarity, adsorb the crude extract on a column filled with silica gel, etc., and then add a hydrophobic solvent, a hydrophilic solvent or a mixed solvent thereof. It means that it contains the fraction obtained as a mobile phase. In addition, the meaning of the extract includes a concentrated liquid extract or a solid extract in which the extraction solvent is removed by a method such as freeze drying, vacuum drying, hot air drying, spray drying, and the like. Preferably, an extract obtained by using water, an alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, and more preferably an extract obtained by using a mixed solvent of water and an alcohol having 1 to 4 carbon atoms as an extraction solvent. do.

In addition, in the present specification, "active ingredient" refers to a component that can exhibit a desired activity alone or itself can exhibit activity together with an inactive carrier.

Also, in the present specification, "anti-obesity" means a reduction in body fat, inhibition of body fat accumulation, and/or a decrease in body weight. Therefore, it is meant to include prevention of obesity, treatment of obesity, and further includes reducing body weight/body fat for cosmetic or health purposes (aka diet purposes), although the weight is not classified as obesity or overweight.

Also, in the present specification, the term “obesity” refers to a state in which adipose tissue is abnormally increased, whether it is obesity due to genetic factors or obesity due to environmental factors, and is high when the body mass index (BMI) is classified. It includes obesity (if BMI is over 30.0), obesity (if BMI is 25 to 30), and overweight (if BMI is between 23 and 25).

The composition for anti-obesity of the present invention can be included in any amount (effective amount) according to the use, formulation, blending purpose, etc., as long as the active ingredient can exhibit anti-obesity activity, the typical effective amount is based on the total weight of the composition Will be determined within the range of 0.001% to 20.0% by weight. Here, the "effective amount" refers to the intended functional and pharmacological effects, such as anti-obesity effects, when the composition of the present invention is administered to a mammal, preferably a person, to which the object is applied, according to the recommendation of a medical expert, Refers to the amount of active ingredient contained in the composition of the present invention. Such effective amount can be determined empirically within the range of ordinary skill in the art.

The anti-obesity composition of the present invention, in addition to the active ingredient, in order to increase or strengthen the anti-obesity effect or to improve the convenience of taking or ingesting through the addition of similar activity such as blood pressure control activity, safety has already been verified in the art Any compound known to have this activity or natural extracts may be further included. These compounds or extracts include compounds or extracts, medicines, etc. that are listed in the Pharmacopoeia of each country ("Korea Republic of Korea Pharmacopoeia"), and health functional food exhibitions of each country ("Korea Food and Drug Administration" "Health functional food standards and standards") Laws governing the manufacture and sale of compounds or extracts and health functional foods that have been granted an item in accordance with the laws of each country governing the manufacture and sale of foods (the "Pharmaceutical Law" in Korea). ”) includes compounds or extracts whose functionality is recognized.

For example, according to the Korean Act on Health Functional Food, Garcinia cambogia bark extract, which has been recognized as functional with'reduction of body fat', conjugated linolenic acid (free fatty acid), conjugated linolenic acid (triglyceride), green tea extract, chitosan, lactobacillus gaseri BNR17 ( Lactobacillus gasseri BNR17), L-carnitine tartrate, green mate extract, green coffee bean extract, sesame leaf extract, soybean germ extract, etc., sol extract, alcohol extract powder, lactoferrin (milk purified protein), lemon balm extract mixed powder , Mate hydrothermal extract, seaweed and other complex extracts (Xantien), fermented vinegar pomegranate complex, boy tea extract, Seomok Tae (Rice Bean) peptide complex, vegetable oil and fat diglyceride, wild mango seed extract, medium-chain fatty acid (MCFA) L-glutamic acid-derived GABA-containing powder, which has been recognized as functional by controlling blood pressure, such as choleus forskolin extract, chito-oligosaccharide, fingerroot extract powder, hibiscus, etc., Katsuobushi oligopeptide, NATO bacteria culture powder, Seomok Tae Soybean) Peptide Complex, Salmon Peptide, Olive Leaf Extract, Sardine Peptide, Casein Hydrolyzate, Coenzyme Q10, Grapeseed Enzyme Decomposition Extract Powder, Haitai Oligopeptide, etc. , Indigestible maltodextrin, bamboo leaf extract, vegetable fat diglyceride, sardine refined fish oil, refined squid oil, etc., L-arabinose, nopal extract, cinnamon extract powder, guava leaf extract, ovary recognized as'glycemic control' Hwaseong maltodextrin, freeze-dried silkworm powder, hemp alcohol extract, banaba leaf extract, upper leaf extract, etc., functional fermented amino acid complexes recognized as functional with'fatigue improvement', hut fruit extract, honggyeongcheon extract, etc., functional as'anti-stress' The recognized L-theanine, ash aganda extract, milk protein hydrolyzate, extra-leaves leaf extract, etc. may correspond to these compounds or extracts.

In another aspect, the present invention relates to a composition for improving hyperlipidemia, which includes a crude poplar extract as an active ingredient.

In the present specification, "hyperlipidemia" refers to a condition in which fat metabolism such as triglycerides and cholesterol is not properly performed, and thus the amount of fat in the blood is increased more than necessary, and includes hypertriglyceridemia and hypercholesterolemia.

In addition, in the present specification, "improving hyperlipidemia" means lowering the concentration of triglycerides in the blood or lowering the cholesterol level in the blood.

In the anti-hyperlipidemic composition of the present invention, the above-mentioned with respect to the anti-obesity composition of the present invention is effective as it relates to the meaning of the barberry extract and its effective amount.

The composition for anti-obesity and the composition for improving hyperlipidemia of the present invention can be identified as a food composition in a specific embodiment.

The food composition of the present invention can be produced in any form, such as tea, juice, carbonated beverages, beverages such as ionic beverages, processed oils such as milk, yogurt, gums, rice cakes, sweets, bread, cookies, noodles, etc. Food, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, bars, etc. can be prepared as health functional food formulations.

In addition, the food composition of the present invention can be classified into any product as long as it complies with the enforcement regulations at the time of manufacture and distribution in terms of legal and functional classification. For example, it is a health functional food pursuant to the Korean Act on Health Functional Food, or a confectionary, soybean, tea, or beverage according to each food type in the Food Fair of the Korean Food Sanitation Act (「Food Standards and Standards」) , Special-purpose food.

The food composition of the present invention may include a food additive in addition to the active ingredient. Food additives can be generally understood as substances added to foods to be mixed or infiltrated in manufacturing, processing, or preserving foods, and their safety must be ensured because they are consumed daily and for a long time with foods. Food additives in accordance with national laws governing the manufacture and distribution of food ("Food Sanitation Act" in Korea) are limited in terms of ingredients or functions in terms of food additives with guaranteed safety. In the Korean Food Additives Fair (「Food Additive Standards and Standards」 published by the Ministry of Food and Drug Safety), food additives are classified into chemical synthetic products, natural additives, and mixed preparations in terms of ingredients, and these food additives are sweeteners and flavors in terms of functionality. It is divided into agents, preservatives, emulsifiers, acidulants, and thickeners.

Sweeteners are used to impart a moderate sweetness to food, and both natural and synthetic can be used in the food composition of the present invention. Preferably, a natural sweetener is used. Examples of the natural sweetener include sugar syrup such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavoring agents are used to improve taste or aroma, and both natural and synthetic ones can be used. Preferably, it is the case of using a natural thing. In addition to flavor, when using natural ones, the purpose of enhancing nutrition can also be combined. As a natural flavoring agent, it may be obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, or the like, or may be obtained from green tea leaves, perilla, large leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo can be used. Natural flavoring agents may be liquid concentrates or solid extracts. In some cases, synthetic flavoring agents may be used. As the synthetic flavoring agent, esters, alcohols, aldehydes, terpenes, and the like may be used.

As a preservative, calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. can be used, and as an emulsifier, acacia gum, carboxymethylcellulose, xanthan gum, pectin Etc. can be used, and as acidulant, arithmetic, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid and the like can be used. The acidulant may be added so that the food composition has an appropriate acidity for the purpose of suppressing the growth of microorganisms in addition to the purpose of enhancing taste. As a thickener, a suspending agent, sedimentation agent, gel forming agent, swelling agent, etc. may be used.

The food composition of the present invention, in addition to the food additives as described above, may include bioactive substances or minerals known in the art for the purpose of supplementing and reinforcing functional and nutritional properties and guaranteed stability as food additives.

Examples of such bioactive substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoyl thiamine, etc., and minerals include calcium preparations such as calcium citrate, magnesium stearate Magnesium preparations such as iron, iron preparations such as iron citrate, chromium chloride, potassium iodine, selenium, germanium, vanadium, zinc, and the like.

The food composition of the present invention may include the food additives as described above in an appropriate amount to achieve the purpose of addition according to the product type.

With regard to other food additives that may be included in the food composition of the present invention, it is possible to refer to national foods or food additives.

The composition of the present invention may be identified as a pharmaceutical composition in other specific embodiments.

The pharmaceutical composition of the present invention may be prepared in an oral dosage form or a parenteral dosage form according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. Here, the route of administration may be any suitable route including a local route, an oral route, an intravenous route, an intramuscular route, and direct absorption through mucosal tissue, and two or more routes may be used in combination. An example of a combination of two or more routes is when two or more formulations of the drug are combined according to the route of administration, for example, when one drug is administered by the intravenous route and the second drug is administered by the local route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or formulation, and specifically refer to the pharmacopeia of each country, including "Korea Pharmacopoeia".

When the pharmaceutical composition of the present invention is prepared in an oral dosage form, powders, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers according to methods known in the art with suitable carriers And the like. At this time, examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, xylitol, starches such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Celluloses such as hydroxypropyl methylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, green Serol, etc. are mentioned. In the case of formulation, if necessary, suitable binders, lubricants, disintegrants, colorants, diluents, etc. may be included. Suitable binders include starch, magnesium aluminum silicate, starch ferist, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax, etc., and lubricants include oleic acid Sodium, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, magnesium salts and calcium salts thereof, polydetylene glycol, and the like, and starch and methyl cellulose as disintegrants , Agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt. Moreover, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine etc. are mentioned as a diluent.

When the pharmaceutical composition of the present invention is prepared in a parenteral dosage form, it may be formulated in the form of injections, transdermal administrations, nasal inhalants and suppositories according to methods known in the art with suitable carriers. When formulated as an injectable agent, an aqueous isotonic solution or suspension may be used as a suitable carrier. Specifically, an isotonic solution such as PBS (phosphate buffered saline) containing triethanol amine, sterile water for injection, or 5% dextrose may be used. . When formulated as a transdermal dosage form, it can be formulated in the form of ointments, creams, lotions, gels, external solutions, pasta, linen agents, aerosols, and the like. For nasal inhalants, dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc. can be formulated in the form of an aerosol spray using a suitable propellant. witepsol), tween 61, polyethylene glycols, cacao butter, laurin, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, sorbitan fatty acid esters, and the like.

Regarding the specific formulation of the pharmaceutical composition, it is known in the art, and for example, see Remington's Pharmaceutical Sciences (19th ed., 1995) and the like. The above documents are regarded as part of this specification.

Preferred dosages of the pharmaceutical compositions of the present invention range from 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g per day, depending on the patient's condition, weight, sex, age, patient severity, and route of administration. /kg range. Administration can be made once a day or divided into several times. Such dosage should not be construed as limiting the scope of the invention in any aspect.

As described above, according to the present invention, it is possible to provide an anti-obesity composition and an anti-hyperlipidemic composition using a camphor tree extract. The composition for anti-obesity of the present invention can be commercialized as a functional food, drug, or the like.

1 is a result of quantifying the fat content of the cells and micrographs stained with Oil Red O reagent after treating the mouse fat precursor cells 3T3-L1 according to the concentration of the camphor tree extract.
2 is a block diagram of an obese animal model experiment.
3 is a result of measuring body weight and feed intake.
4 is a result of measuring visceral fat and subcutaneous fat.
5 is a result of measuring triglycerides in the blood.
6 is a photograph of staining of liver adipose tissue.
7 is a photograph of the size of adipocytes in liver tissue and the results of quantifying them graphically.
8 is a result of measuring leptin concentration in the blood.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these examples and experimental examples.

< Example > Preparation of Chopop tree extract

Chopop trees (extractions, leaves) were washed 2-3 times with distilled water, then removed, dried with hot air, and then ground and crushed. Thereafter, distilled water was added to the pulverized product at a ratio of 1:10 (w:v), stirred and extracted at 80° C. for 3 hours, the extract was filtered, and the filtrate was concentrated under reduced pressure and lyophilized to obtain a crude poplar extract.

< Experimental Example > Experiment on the improvement of obesity of Jopopus extract

<Experiment 1> adipocyte differentiation inhibitory activity experiment

3T3-L1 (ATCC CL-173), a mouse progenitor cell, was added with 10% BCS DMEM medium and cultured at 37°C and 5% CO ₂ . After dispensing 3T3-L1 progenitor adipocytes into a cell number of 5×10 ⁴ /well in a 24 well plate, it was maintained for 2 days at the time of 100% confluency. Progenitor cells differentiate into adipocytes with 10% FBS DMEM medium containing MDI (0.5 mM 3-isobutyl-1-methylxanthine (Calbiochem 410957), 1 uM dexamethasone (Calbiochem 265005), 1 ug/ml insulin (Sigma I9278)). Was induced for 2 days, and after 48 hours of culture, the cells were cultured for 2 days with 10% FBS DMEM containing 1 ug/ml insulin. Then, every 2 days, the cells were replaced with 10% FBS DMEM culture for 4 days. During the adipocyte differentiation induction, the extracts of the examples were treated in each culture at a concentration of 50, 100 µg/ml, and the degree of adipocyte differentiation was observed on the 8th day when the differentiation was completed. The degree of adipocyte differentiation was primarily confirmed through a microscope through Oil Red O staining, and the adipocyte staining degree was measured by using an ELISA Reader (SPECTRAmax 340PC, Molecular Devices, USA) at 510 nm absorbance. Did. The results were calculated through 3 repeated experiments for each concentration.

The results are shown in FIG. 1, and referring to FIG. 1, it can be seen that the camphor tree extract has an activity of inhibiting the differentiation of fat precursor cells into adipocytes and a fat accumulation inhibitory activity depending on concentration.

<Experimental Example 2> Obese animal model experiment

1. Experimental method

1.1 Production of obese animal models by high-fat diet

After purchasing a 6-week-old C57BL/6 mouse, the groups were arranged in order of high weight, and the groups (n=10) were separated so that the mean and standard deviation between the groups were equal, and as shown in the table below and FIG. 2, the control group (HFD) was The high-fat diet (containing 60% fat) was fed, and the experimental group (50, 100, 200, 400 MPK) was also fed with the high-fat diet and the samples of the examples. The high-fat diet was freely ingested, and the example samples were orally administered for 6 weeks at a concentration of 50, 100, 200, 400 mg/kg using sonde daily. The sample was not administered to the normal group (ND), and the feed was fed as a normal feed.

1.2 Measurement of changes in body weight and diet

The experiment was conducted for 6 weeks, and body weight and feed intake were measured twice a week.

1.3 Visceral fat, subcutaneous fat shooting, fat mass measurement

At the 3rd and 6th week of the 6-week experiment period, all animals were photographed using animal micro-CT (Inveon, Simens). Quantitative analysis of the visceral fat and subcutaneous fat in animals was performed using the captured image files.

1.4 Quantitative analysis of triglycerides in blood

After oral administration for 6 weeks, animals were sacrificed and blood was collected from the abdominal aorta. The collected blood was centrifuged at 3,000 rpm using a centrifuge for analysis, and plasma was separated, and triglyceride in the blood was measured using a blood biochemical analyzer (Hitachi 7020, Japan).

1.5 Adipose tissue staining for each part such as liver, adipose tissue change, and size of fat tax

After sacrificing the experimental animals, liver, subcutaneous fat, epididymal fat, and peritoneal fat were extracted for each site. After fixation with 4% formaldehyde for 7 days, the fixed tissue is subdivided again into a size of 1×0.5 cm and placed in a tissue cassette (tissue processor) (Thermo scientific, USA) After dehydration, transparent, and paraffinization, store at -20℃, cut the paraffin block to a thickness of 4~5㎛ with a microtome (Thermo scientific, USA), and then dry it on a 60℃ heating plate. Ordered. To measure lesions of skin tissue, deparaffinization of skin tissue fragments with xylene, followed by deparaffinization of the skin tissues, hematoxylin/eosin, toludine blue O solution ( toluidine blue O solution), stained each tissue cell, sealed with mount regent, and observed with an optical microscope. Next, the slide of adipose tissue stained with toludine blue O was compared with an optical microscope to measure the size of adipose tissue by region and calculate the average and percentage.

1.6 Blood Leptin Analysis

Leptin is a type of adipocaine that increases insulin sensitivity in muscles and liver and affects glucose utilization in peripheral blood. In obesity, leptin resistance increases blood levels (Nature 1998;392:398). -401). Therefore, substances exhibiting anti-obesity activity reduce the concentration of leptin in the blood.

In this experiment, leptin in blood was measured using ELISA. Specifically, the anti-leptin/adiponectin coated plate was reacted with biotin-conjugated anti-leptin and plasma for 2 hours as a primary antibody. After washing 3 times, the secondary antibody, HRP-conjugated streptavidin, was added and reacted for 1 hour. After washing 4 times, the substrate was reacted for 30 minutes, the reaction was stopped by adding a stop buffer, and absorbance was measured at 450 nm at the ELISA reader. Leptin was quantified against a standard curve.

2. Experimental results

1.1 Measurement results of weight and dietary changes

The experiment was conducted for 6 weeks, and the weight and feed intake were measured twice a week and the results are shown in FIG. 3. Body weight increased the highest in the control group, and the degree of increase was significantly lower in the experimental group than the control group. Table 2 below shows the percentage of weight suppression in the experimental group compared to the control group for 6 weeks. The amount of diet did not show a special difference between the control group and the experimental group.

1.2 Visceral fat, subcutaneous fat shooting, fat mass measurement results

4 shows the results of measuring visceral fat and subcutaneous fat during the 6-week trial period. Both the amount of fat in visceral fat and subcutaneous fat was significantly decreased in proportion to the dose in the experimental group compared to the control group. The lower picture in FIG. 4 is a picture of visceral fat.

1.3 Quantitative analysis of triglycerides in the blood

The results of measuring blood triglycerides during the 6-week test period are shown in FIG. 5. Referring to Figure 5, triglyceride in the blood was significantly lower in all experimental groups compared to the control group.

1.4 Fat tissue staining for each part such as liver, fat tissue change amount and fat tax size measurement result

Fat accumulation inhibition rate and body fat percentage compared to the control group of subcutaneous fat, epididymal fat, and peritoneal fat are shown in Tables 3 to 5 below. It can be seen that subcutaneous fat, epididymal fat, and peritoneal fat decrease in proportion to the dose.

In addition, a photo of staining of adipose tissue of the liver is shown in FIG. In the liver tissue, it shows that the adipose tissue decreases in proportion to the dose in the experimental group compared to the control group.

In addition, a photograph of the size of adipocytes in liver tissue and the result of quantification by graph are shown in FIG. 7. The size of adipocytes also showed a decrease in proportion to the dose in the experimental group compared to the control group.

1.5 Blood Leptin Analysis

The results are shown in FIG. 8. Figure 8 shows that the concentration of leptin in the blood decreased in proportion to the dose in the experimental group compared to the control group.

Claims (8)

A food composition for anti-obesity, which contains Jopop tree extract as an active ingredient.
According to claim 1,
The composition of the anti-obesity is obtained by extracting the leaves of Jopop tree as an object of extraction and extracting with water, ethanol or a mixed solvent thereof.
A pharmaceutical composition for anti-obesity, comprising a jopop tree extract as an active ingredient.
According to claim 3,
The crude poplar extract is a pharmaceutical composition for anti-obesity, characterized in that obtained by extracting the leaves of poplar and extracted with water, ethanol or a mixed solvent thereof.
A food composition for improving hyperlipidemia, which includes a Jopop tree extract as an active ingredient.
The method of claim 5,
The composition for the improvement of hyperlipidemia, characterized in that the crude poplar extract is obtained by extracting the leaves of the poplar and extracted with water, ethanol or a mixed solvent thereof.
A pharmaceutical composition for improving hyperlipidemia, which includes a Jopop tree extract as an active ingredient.
The method of claim 7,
The crude tree extract is a pharmaceutical composition for improving hyperlipidemia, characterized in that it is obtained by extracting a tree branch tree leaf with water, ethanol or a mixed solvent thereof.

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