KR102038887B1 - Pharmaceutical composition for the prevention or treatment of obesity, Hyperlipidemia or fatty liver comprising extract of Lonicera caerulea L. var. edulis fruits and method for manufacturing thereof - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of obesity, hyperlipidemia or fatty liver, including a powder of the oleander fruit, and a method of preparing the same. When the pharmaceutical composition of the present invention is used, it can be used for the treatment of obesity, hyperlipidemia or fatty liver by reducing the amount of body fat, reducing total cholesterol, and decreasing the rate of change of liver fat.

Description

Pharmaceutical composition for the prevention or treatment of obesity, Hyperlipidemia or fatty liver comprising extract of Lonicera caerulea L. var. edulis fruits and method for manufacturing knowledge}

The present invention relates to a pharmaceutical composition for the prevention or treatment of obesity, hyperlipidemia or fatty liver, including a powder of the oleander fruit, and a method of preparing the same.

Obesity is the cause of various lethal metabolic diseases such as cardiovascular disease and hypertension. When obesity, fat cells are a long-term energy source that stores lipids as well as various adipocaines that are involved in the metabolism and inflammation of fat and non-fat cells. secretion of adipokine causes chronic inflammation and causes related metabolic diseases such as insulin resistance.

Recently, the incidence of metabolic syndrome is increasing due to obesity due to high calorie diet and lack of exercise worldwide, and it is predicted that by 2025, metabolic syndrome patients will nearly double the present number, or about 300 million people.

Obesity due to abnormal local fat accumulation, such as abdominal obesity, provides a variety of causes that can seriously cause atherosclerosis, such as hypertension, hyperlipidemia, blood clotting disorders, vascular inflammation, insulin resistance with abnormal insulin secretion, Finally, it is a factor that increases the prevalence of fatal cardiovascular disease.

Significant obesity is also induced in rodent mice by feeding high fat diet (HFD) and significantly higher blood sugar, insulin resistance, and nonalcoholic steatohepatitis (NASH) compared to mice fed normal pellet diet (NFD). Mild mild diabetic nephropathy and hyperlipidemia have been identified in HFD induced obese mice. In other words, HFD feeding caused diabetic obesity similar to human metabolic syndrome in ICR mice.

Metformin is a representative oral antihyperglycemic anti-diabetic agent orally known as an AMPK activator. Metformin is the first drug used to treat patients with normal overweight and obese type 2 diabetes, which can reduce the incidence of cardiovascular disease, one of the serious side effects of diabetes, and is directly involved in the digestion of fat. It is known to be the only drug that can regulate the secretion of enzyme source granules of the pancreas and the activity of enzymes related to liver glucose metabolism. Metformin is currently the world's most prescribed antidiabetic drug, with approximately 48 million people prescribed metformin in 2010 in the United States alone. Metformin is primarily used to treat type 2 diabetes, but prescriptions for polycystic ovary syndrome, NAFLD and precocious puberty are also on the rise.

Currently, various metabolic syndrome treatments have been developed, but various side effects have been considered, and the use thereof has been limited. Therefore, the development of alternative medicines derived from natural products with lower side effects and more effective have been actively attempted. In particular, with adequate blood sugar control and adequate control of oxidative stress, which has become a major etiology for diabetes and related complications, has emerged as the most important method for treating diabetes, resulting in lower side effects and more effective a-glucosidase blockers. Or the situation is trying to develop antioxidants.

The present inventors made diligent research efforts to develop a pharmaceutical composition for preventing or treating obesity, hyperlipidemia or fatty liver. As a result, the present invention was completed by identifying that the extract of L. larvae (Lonicera caerulea L. var.edulis) reduced body fat mass, decreased total cholesterol, and decreased liver fat change.

Accordingly, it is an object of the present invention to provide a pharmaceutical composition for the prevention or treatment of obesity, hyperlipidemia or fatty liver, which includes the extract of Acacia.

It is another object of the present invention to provide a food composition for improving obesity, hyperlipidemia or fatty liver, which includes an extract from Acacia.

Another object of the present invention is to provide a method for preparing a pharmaceutical composition for the prevention or treatment of obesity, hyperlipidemia or fatty liver, which includes the extract of the oleander fruit.

According to one aspect of the present invention, the present invention provides a pharmaceutical composition for the prevention or treatment of obesity, hyperlipidemia or fatty liver, comprising an extract of Lonicera caerulea L. var.edulis fruit.

The present inventors made diligent research efforts to develop a pharmaceutical composition for preventing or treating obesity, hyperlipidemia or fatty liver. As a result, it was found that the extract of L. larvae (Lonicera caerulea L. var.edulis) decreased body fat mass, total cholesterol and decreased liver fat change.

As used herein, the term 'dangle' refers to Lonicera caerulea var. Belonging to the genus Caprifoliaceae. It is a mature fruit of edulis , which has been used as a traditional medicinal plant in North Russia, China, and Japan, but relatively unfamiliar in North America and Europe. Taraxacum contains abundant ascorbic acid and phenolic components, and is particularly rich in anthocyanins, flavonoids and low-molecular phenolic acids, which are known to have relatively good antioxidant effects.

Taraxacum fruit extract according to the present invention may be prepared by treating an enzyme, for example, may be an extract treated with proteolytic enzymes on the mulberry fruit. The Taraxacum fruit extract may be in solution, concentrate or powder state.

The pharmaceutical composition may further comprise chitosan and / or guar gum. The chitosan and / or guar gum have the effect of removing proteins and foreign substances.

The pharmaceutical composition may be to reduce the weight gain, body fat amount or abdominal fat amount increased by a high fat diet.

The pharmaceutical composition may be to reduce the thickness of fat tissue or the diameter of fat cells increased by high fat diet.

The pharmaceutical composition comprises one or more blood contents selected from the group consisting of total cholesterol (TC), triglyceride (TG), and low density lipoprotein (LDL), which are increased by a high fat diet. It may be to reduce.

The pharmaceutical composition may be to increase the blood content of high density lipoprotein (HDL) reduced by a high fat diet.

The pharmaceutical composition is aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (ALP) increased by high fat diet. Lactate dehydrogenase (LDH), gamma-glutamyl transpeptidase (GGT), blood urea nitrogen (BUN) and creatinine to reduce the blood content of one or more selected from the group consisting of It may be.

The pharmaceutical composition may be to reduce the rate of hepatic fat change, hepatocellular diameter or number of degenerative urinary tract increased by high fat diet.

The pharmaceutical composition may be to reduce the expression of ACC1 mRNA increased by high fat diet in liver tissue.

The pharmaceutical composition is 1 selected from leptin, leptin, C / EBPa (CCAAT-enhancer binding protein-a), C / EBPβ (CCAAT-enhancer binding protein-β) and SREBP1c increased by high fat diet in adipose tissue. It may be to reduce species abnormalities.

The pharmaceutical composition of the present invention can be used as a pharmaceutical composition comprising a pharmaceutically effective amount of a powder of the oleander fruit extract and / or a pharmaceutically acceptable carrier.

As used herein, the term “pharmaceutically effective amount” means an amount sufficient to achieve the efficacy or activity of the above-mentioned Rootberry extract.

Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like.

The pharmaceutical compositions according to the invention can be administered to a mammal, including humans, by various routes. The mode of administration may be any of the methods commonly used, and may be administered by, for example, oral, skin, vein, muscle, subcutaneous, and the like, and preferably, orally.

Suitable dosages of the pharmaceutical compositions of the invention vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion and response to the patient, Usually a skilled practitioner can easily determine and prescribe a dosage effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dose of the pharmaceutical composition of the present invention is 100 to 400 mg / kg.

The pharmaceutical compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporating into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of extracts, powders, granules, tablets, capsules, or gels (eg, hydrogels), and may further include dispersants or stabilizers. .

According to another aspect of the present invention, the present invention provides a food composition for improving obesity, hyperlipidemia or fatty liver, which includes an extract of Lonicera caerulea L. var.edulis fruit.

Taraxacum fruit extract according to the present invention may be prepared by treating an enzyme, for example, may be an extract treated with proteolytic enzymes on the mulberry fruit. The Taraxacum fruit extract may be in solution, concentrate or powder state.

The food composition may further include chitosan or guar gum.

The content of the powder of the oleander tree extract as an active ingredient contained in the food composition is not particularly limited according to the form of the food, desired use, and the like, for example, it may be added to 0.01 to 15% by weight of the total food weight, The health beverage composition may be added at a ratio of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml.

When the food is a beverage, there is no particular limitation on the liquid components, besides containing the extract of the dainty berry as an essential ingredient in the indicated ratio, and it may contain various flavors or natural carbohydrates as additional ingredients, like ordinary drinks. Can be.

The natural carbohydrates are monosaccharides such as disaccharides such as glucose, fructose and the like, and conventional sugars and xylitol such as maltose, sucrose and the like, and polysaccharides such as dextrin, cyclodextrin and the like. Sugar alcohols such as sorbitol and erythritol.

As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, alginic acid and Salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like.

In addition, the food composition of the present invention may contain a fruit flesh for producing natural fruit juice and fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

According to another aspect of the present invention, the present invention is a method for producing a pharmaceutical composition for the prevention or treatment of obesity, hyperlipidemia or fatty liver, comprising an enzyme treatment step of mixing proteolytic enzymes and processing for 1 to 5 hours into a tadberry fruit. To provide.

The protease may be pectinase.

The pectinase may be 0.01 to 1.0% by weight relative to the dagger.

The enzyme treatment step may be to be treated for 2 to 2.5 hours.

The method of preparing the pharmaceutical composition may further include adding chitosan and / or guar gum.

According to another aspect of the present invention, the present invention provides a method for preparing a pharmaceutical composition for preventing or treating obesity, hyperlipidemia or fatty liver, comprising the following steps:

Pulverizing step of heating the linden tree (Lonicera caerulea L. var. Edulis) fruit to 40 to 60 ℃;

An enzyme treatment step of treating proteolytic enzymes for 1 to 5 hours by mixing the powdered wood fruit;

A sediment removal step of removing the sediment from the enzyme-treated daimberry fruit;

A heating step of heating the off-tree fruit from which the precipitate is removed to 60 to 80 ° C;

An additive input step of adding chitosan and / or guar gum to 0.001 to 0.1% by weight, respectively, in the heated daimberry fruit;

A sterilization step of filtering and concentrating the powdered dendrite fruit into which the additive is added and sterilizing at 80 to 100 ° C; And

Lyophilization step of lyophilizing by mixing the sterilized Daimyeo fruit and dextrin in a weight ratio of 1: 2 to 2: 1.

The first heating step may be to heat the wood off 45 to 55.

The protease may be pectinase.

The pectinase may be 0.01 to 1.0% by weight relative to the dagger.

The enzyme treatment step may be to be treated for 2 to 2.5 hours.

The precipitate removal step may be performed by centrifugation, but is not limited thereto.

The chitosan or guar gum may be 0.001 to 0.1 wt%, 0.001 to 0.01 wt%, 0.001 to 0.005 wt%, or 0.005 to 0.1 wt%, for example, 0.005 to 0.01 wt%, but is not limited thereto.

The weight ratio of the dagger and dextrin may be 1: 1.

The features and advantages of the present invention are summarized as follows:

(a) The present invention provides a pharmaceutical composition for the prevention or treatment of obesity, hyperlipidemia or fatty liver, which includes an extract of Lonicera caerulea L. var.edulis.

(B) The present invention provides a method for preparing a pharmaceutical composition for the prevention or treatment of obesity, hyperlipidemia or fatty liver, comprising the step of treating proteolytic enzymes on the oleander tree fruit.

(c) When using the pharmaceutical composition for the prevention or treatment of obesity, hyperlipidemia or fatty liver comprising the extract of the present invention, the weight loss of obese, hyperlipidemia by reducing the body fat mass, total cholesterol, and the rate of change of liver fat Or for treating fatty liver.

Figure 1 shows a graph comparing the change in weight according to the treatment of the Daengberry fruit extract in high fat diet mice.
Figure 2 shows a graph comparing the changes in body fat and abdominal fat according to the treatment of the Daengberry fruit extract in a high fat diet mouse.
Figure 3 shows the decrease in mean diameter of pancreatic islets and the increase in zymogen granules according to the treatment of the extract of R. olecus in high fat diet mice.
Figure 4 shows the content of fat in the feces of the total cholesterol and triglycerides according to the treatment of the extract from the barberry fruit in a high fat diet mouse.
Figure 5 shows a decrease in liver fat change rate, a decrease in hepatic cell diameter, and a degeneration of the number of denatured tubules according to the treatment of the Daengberry fruit extract in a high-fat diet mouse.
Figure 6 shows the histological changes of kidneys according to the treatment of the extracts of the barberry fruit in high-fat diet mice.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example 1: Taraxacum Extract Preparation

The wood fruit was heated at 45 to 55 ° C. for 3 minutes and pulverized with a grinder to treat pectinase (Weectbiotch, Weissbiotch, Germany) which was 0.1% by weight of the wood fruit for 2 to 2.5 hours. The precipitate was removed by continuous centrifugation at 4000 rpm / min, heated at 80 ° C. for 15-30 seconds, and chitosan and guar gum were added at a rate of 0.005% by weight of the daggers, respectively, and filtered at 1500 rpm / min. Press filtration. Thereafter, the mixture was concentrated at 50 ° C. for 1 minute, sterilized at 90 to 95 and mixed with dextrin in a weight ratio of 1: 1, and then lyophilized to obtain a powder of dried barberry fruit (BHx).

Example 2: Preparation of Experimental Animals

A total of 85 6-week-old female SPF ICR mice (Oriental Bio, Seongnam, Gyeonggi-do) were acclimated to the laboratory environment for 7 days, then adapted to high-fat diet for 1 week, and then 8 animals per group. Divided into groups and used for the experiment.

The classification of the experimental group is as follows:

(1) normal control group: normal diet intake, oral administration of 10 ml / kg of distilled water,

(2) Type 2 diabetes (NIDDM) control group: high fat feed intake, oral administration of 10 ml / kg of distilled water,

(3) metformin control: high fat feed, metformin 250 mg / kg oral administration,

(4) BHx 400 mg / kg administration group: high fat feed, BHx 400 mg / kg oral administration,

(5) BHx 200 mg / kg administration group: high fat feed, BHx 200 mg / kg oral administration,

(6) BHx 100 mg / kg administration group: high fat feed, BHx 100 mg / kg oral administration group.

High fat feed (Cat. No. D12451; Research Diet, New Brunswick, NJ, USA) was supplied from 1 week before the start of the test substance administration, and normal high fat feed (Cat. No. 38057; Purinafeed) in the normal control group. , Seungnam, Korea).

After 1 week of high-fat diet, the Rootberry Extract was dissolved in sterile distilled water at concentrations of 40, 20 and 10 mg / ml (the Rootberry Extract itself except for dextrin), and the volume of 10 ml / kg ( 400, 200 and 100 mg / kg) was forcedly administered orally using a 1 ml syringe with metal sonde. Metformin was also dissolved in sterile distilled water at a concentration of 25 mg / ml, followed by a volume of 10 ml / kg (250 mm / kg). kg) orally.

In the normal and high fat diet control group, only sterile distilled water was forced orally administered for the same period of time in the same manner in order to apply the same administration and correction stress.

Example 3: Determination of body weight and feed intake

Body weight was measured on the day before the administration of high fat feed (Day-8), the day before the test substance (day-1), the test substance administration (day0), and once a week during the test substance administration period using electronic scales. . Fasting (12 hours) was performed on the day before and the day of sacrifice.

Weight gain (g) was calculated in the following manner.

Adaptation period (7 days) = start date of body weight-start date of high fat feed (Day -8 ~ Day 0)

Duration of Test Substance Administration (84 Days) = Day of Sacrifice Weight-Start Date of Test Substance Administration (Day 0 ~ Day)

Feeding 150 grams per breeding cage was measured by electronic balance after 24 hours. The amount of feed measured was divided by the number of animals in the breeding cage and calculated as the daily feed intake per individual (g / day / mice). Daily feed intake was examined weekly during the 84-day experimental period.

Metformin 250 mg / kg, Dandelion Fruit Extract (BHx) 400 and 200 mg / kg, respectively, from 28 days after the start of the administration, and Dandelion Berry Extract from 100 mg / kg, 42 days after the start of administration, significantly more weight than the control group. The decrease of was started to be observed, and all the test substance administration group including metformin 250 mg / kg showed a weight loss decrease during the 84 days of the administration of the test substance significantly compared to the obese control group is shown in Table 1 and Figure 1, respectively.

division 8 days ago
[A]
(g) 1 day before
[B]
(g) 0 day *
[C]
(g) 84 days *
[D]
(g) Adaptation period
[BA]
(g) Duration of administration
[DC]
(g) Average Feed Intake
(g) Normal control 27.86
± 0.52 28.34
± 0.57 25.36
± 0.67 30.13
± 1.59 0.48
± 0.15 4.76
± 1.30 4.63
± 0.32 Obesity Control 27.88
± 0.84 31.03
± 0.90 28.25
± 0.73 51.75
± 1.85 3.15
± 0.26 23.50
± 1.68 3.87
± 0.21 Metformin 27.91
± 0.45 31.09
± 0.58 28.21
± 0.45 40.66
± 1.30 3.18
± 0.21 12.45
± 1.00 3.90
± 0.25 BHx 400 27.89
± 0.60 31.03
± 0.63 28.15
± 0.69 40.81
± 0.92 3.14
± 0.05 12.66
± 1.03 3.90
± 0.22 BHx 200 27.85
± 0.74 31.03
± 0.75 28.36
± 0.62 44.51
± 1.79 3.18
± 0.20 16.15
± 1.81 3.86
± 0.25 BHx 100 27.89
± 0.82 31.01
± 0.77 28.20
± 1.10 46.54
± 3.39 3.13
± 0.15 18.34
± 2.71 3.89
± 0.23

As can be seen from Table 1 and Figure 1, the weight gain during the 84 days administration period showed a change of 393.44% in the obese control group, compared to the normal control group, metformin 250 mg / kg, Dandelion fruit 400, 200 and 100 The mg / kg group showed changes of -47.02, -46.12, -31.28 and -21.97%, respectively, compared to the obese control group.

Significant decreases in mean feed intake were observed in all HFD feeders compared to the normal control group, but significant changes in mean feed intake were observed in all experimental groups, including Daengberry Fruit 400 mg / kg. Did.

The average feed intake was -16.37% in the obese control group compared to the normal control group, but metformin 250 mg / kg and dagger extract 400, 200 and 100 mg / kg were 0.83 and 0.85, respectively, compared to the obese control group. , -0.26 and 0.50%.

Therefore, all doses of the Daimberry Fruit extract showed dose-dependent body weight reduction and weight gain during the 84-day period of test substance administration, and HFD-induced body weight similar to metformin 250 mg / kg It was confirmed that the effect of increasing weight gain was shown.

Example 4 Confirmation of Changes in Body Fat and Abdominal Fat

Body fat and abdominal fat of each experimental animal were measured on the day of sacrifice using equipment ( in live DEXA, InAlyzer; Medikors, Seungnam, Korea).

As can be seen in Figure 2, the body fat amount in the obese control group showed a change of 295.14%, respectively, compared to the normal control group, metformin 250 mg / kg, dagger fruit extract 400, 200 and 100 mg / kg administration group in the obesity control group Compared with the changes in body fat mass of -58.03, -56.10, -43.12 and -26.29%, respectively.

Abdominal fat mass was 287.14% in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger extract 400, 200, and 100 mg / kg were -55.96,-compared to the control group, respectively. 54.12, -38.78 and -24.78% change, respectively.

Therefore, the dose-dependent body fat and abdominal fat decreased in all the Daengberry fruit extract groups, and the high fat diet, similar to metformin 250 mg / kg, suppressed the increase of body fat and abdominal fat in doses. The effect was confirmed

Example 5: Serum Biochemistry

Blood collected from the posterior vena cava on the day of sacrifice was placed in a clotting activated serum tube, followed by centrifugation at 15,000 rpm for 10 minutes to separate serum, followed by an automated blood analyzer (Dri-Chem NX500i; Fuji Medical System). Co., Ltd., Tokyo, Japan).

Total Cholesterol (TC), Triglyceride (TG), Low Density Lipoprotein (LDL) and High Density Lipoprotein (HDL) content in blood are shown in Table 2.

division Total cholesterol
(mg / dl) Triglycerides
(mg / dl) Low density lipoprotein
(mg / dl) High density lipoprotein
(mg / dl) Normal control 103.88 ± 19.58 61.25 ± 12.73 15.63 ± 3.38 96.13 ± 19.90 Obesity Control 282.50 ± 29.32 212.50 ± 29.77 66.25 ± 11.54 21.50 ± 10.99 Metformin 167.00 ± 15.98 117.75 ± 22.58 30.75 ± 10.11 63.88 ± 19.21 BHx 400 168.25 ± 34.08 119.38 ± 26.50 31.13 ± 10.58 60.50 ± 15.73 BHx 200 195.00 ± 24.88 138.00 ± 25.53 39.38 ± 10.25 52.00 ± 10.39 BHx 100 210.38 ± 23.77 167.63 ± 21.25 49.50 ± 10.20 40.38 ± 10.47

As can be seen from Table 2, the TC content in the blood was 171.96% in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger fruit extract 400, 200 and 100 mg / kg in the obese control group The changes were -40.88, -40.44, -30.97 and -25.53%, respectively.

The TG content of the control group was 246.94% in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger fruit extract 400, 200 and 100 mg / kg were -44.59 and-, respectively. 43.82, -35.06, and -21.12%.

The blood LDL content of the obese control group was 324.00% higher than that of the normal control group, but metformin 250 mg / kg, dagger fruit extract 400, 200 and 100 mg / kg were -53.58 and-, respectively. 53.02, -40.57 and -25.28% change.

Serum HDL content was -77.63% in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger fruit extract 400, 200 and 100 mg / kg were 197.09 and 181.40, respectively. , 141.86 and 87.79%.

Therefore, the dose-dependent decreases in TC, TG, LDL and HDL contents were observed in all Daengberry fruit extract groups, and the high fat diet similar to metformin 250 mg / kg was induced in the Dandelion fruit extract 400 mg / kg group. Inhibitory effect of increasing TC, TG, LDL and HDL content in blood was shown.

Blood aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), gamma glutamyl trans Peptides (Gamma-glutamyl transpeptidase (GGT), blood urea nitrogen (BUN) and creatinine content is shown in Table 3.

division AST
(IU / L) ALT
(IU / L) ALP
(IU / L) LDH
(IU / L) GGT
(IU / L) BUN
(mg / dl) Creatinine
(mg / dl) Normal control 65.38
± 12.93 32.25
± 10.71 72.50
± 17.06 600.63
± 258.42 1.88
± 0.83 30.36
± 12.28 0.46
± 0.18 Obesity Control 198.63
± 18.10 126.63
± 18.32 228.00
± 33.72 3239.63
± 912.22 9.88
± 2.03 96.00
± 22.23 2.05
± 0.24 Metformin Control 116.13
± 26.18 70.25
± 21.63 162.63
± 25.91 1513.50
± 462.28 4.25
± 1.67 51.00
± 13.14 0.86
± 0.18 BHx 400 111.63
± 17.18 69.63
± 14.17 155.25
± 21.37 1375.50
± 363.23 4.13
± 1.36 49.25
± 11.85 0.80
± 0.26 BHx 200 132.75
± 17.73 81.63
± 13.67 173.25
± 17.19 1660.50
± 283.42 5.63
± 1.51 58.00
± 11.38 1.03
± 0.27 BHx 100 162.88
± 13.60 102.13
± 14.23 186.25
± 18.98 2169.25
± 342.83 6.88
± 1.73 68.25
± 13.18 1.54
± 0.24

As can be seen in Table 3, the blood AST content of the obese control group showed a 203.82% change compared to the normal control group, but metformin 250 mg / kg, Daggerberry extract 400, 200 and 100 mg / kg high-fat diet group The changes of -41.54, -43.80, -33.17 and -18.00%, respectively, were compared with the control group.

The ALT content of blood was 292.64% in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger fruit extract 400, 200 and 100 mg / kg were -44.52 and-, respectively. Changes of 45.01, -35.54 and -19.35%.

The blood ALP content of the obese control group was 214.48% higher than that of the normal control group, but metformin 250 mg / kg, dagger fruit extract 400, 200 and 100 mg / kg were -28.67 and-, respectively. 31.91, -24.01, and -18.31%.

The LDH content in the control group was 439.38% in the obese control group compared with the normal control group, but metformin 250 mg / kg, dagger fruit extract 400, 200, and 100 mg / kg were -53.28,-compared to the obese control group, respectively. 57.54, -48.74 and -33.04%.

The blood GGT content of the obese control group was 426.67% higher than that of the normal control group, but metformin 250 mg / kg, dagger fruit extract 400, 200 and 100 mg / kg were -56.96 and-, respectively. 58.23, -43.04, and -30.38%.

The BUN content of the control group was 213.47% in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger extract 400, 200 and 100 mg / kg were -46.88,-compared to the HFD control group, respectively. Changes of 48.70, -39.58 and -28.91% were shown.

Serum creatinine content was 252.83% in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger extract 400, 200, and 100 mg / kg group -48.74,- 46.45, -37.62 and -23.53%.

Therefore, the dose-dependent decrease in blood AST, ALT, ALP, LDH, GGT, BUN and creatinine contents were found in all the Daengberry fruit extract groups, and metformin 250 mg / kg Increasing levels of AST, ALT, ALP, LDH, GGT, BUN and creatinine in blood induced by similar high fat diets were inhibited.

Example 6: Determination of organ weight

On the day of sacrifice, the pancreas, left ovarian fat mass, left abdominal wall fat mass attached to the muscularis quadratus lumborum, liver and left kidney were extracted.

(Relative weight (%) = extraction weight / day of sacrifice weight * 100).

The ovarian fat mass and abdominal wall fat mass were measured and shown in Table 4.

division Fat mass around the ovary
(Absolute weight (g)) Abdominal wall
(Absolute weight (g)) Fat mass around the ovary
(Relative weight (%)) Abdominal wall
(Relative weight (%)) Normal control 0.089 ± 0.027 0.076 ± 0.031 0.297 ± 0.094 0.252 ± 0.104 Obesity Control 0.775 ± 0.110 0.564 ± 0.105 1.500 ± 0.226 1.091 ± 0.208 Metformin Control 0.220 ± 0.069 0.194 ± 0.052 0.541 ± 0.172 0.479 ± 0.134 BHx400 0.236 ± 0.054 0.217 ± 0.092 0.577 ± 0.129 0.533 ± 0.235 BHx200 0.350 ± 0.097 0.307 ± 0.053 0.784 ± 0.211 0.690 ± 0.121 BHx100 0.498 ± 0.130 0.377 ± 0.073 1.077 ± 0.314 0.813 ± 0.162

As can be seen in Table 4, the absolute and relative weights of the fat accumulated around the ovary were 772.29 and 405.35%, respectively, in the obese control group, but metformin 250 mg / kg and Taraxacum berry extract 400 and 200, respectively. And the 100 mg / kg administration group showed changes in absolute weight of -71.65, -69.61, -54.92 and -35.75%, respectively, compared to the obesity control group, and relative weight changes of -63.94, -61.53, -47.72 and -28.18%, respectively. Indicated.

Absolute and relative weights of abdominal wall accumulation fat were 647.19 and 333.39% in the obese control group, respectively, compared to the normal control group, but metformin 250 mg / kg and dagger extract 400, 200 and 100 mg / kg were overweight. The relative weights of -65.57, -61.56, -45.62 and -33.17% were compared with the control, respectively, and the relative weights of -56.12, -51.12, -36.76 and -25.49%, respectively.

Therefore, dose-dependent ovarian peritoneal and abdominal wall accumulation fat relative and absolute weight reduction were observed in all groups of daphnia fruit extracts, and high fat diets similar to metformin 250 mg / kg were induced in dagger extract 400 mg / kg groups. It showed the effect of suppressing the increase in the ovary and abdominal wall accumulated fat weight.

In addition, by dividing the weight of the extracted organ by the weight of the relative organs are shown in Table 5 and FIG.

division Liver (g) Kidney (g) Liver (%) Kidney (%) Normal control 1.073 ± 0.096 0.176 ± 0.007 3.563 ± 0.300 0.585 ± 0.044 Obesity Control 1.774 ± 0.085 0.301 ± 0.019 3.429 ± 0.140 0.581 ± 0.038 Metformin Control 1.415 ± 0.090 0.231 ± 0.020 3.483 ± 0.246 0.569 ± 0.059 BHx 400 1.413 ± 0.028 0.230 ± 0.017 3.464 ± 0.098 0.564 ± 0.047 BHx 200 1.509 ± 0.092 0.249 ± 0.008 3.398 ± 0.288 0.559 ± 0.023 BHx 100 1.596 ± 0.061 0.262 ± 0.011 3.446 ± 0.291 0.566 ± 0.044

As can be seen in Table 5, the absolute and relative weights of the liver showed changes of 65.35 and -3.77%, respectively, in the obese control group, but metformin 250 mg / kg and dagger extract 400, 200 and 100 mg, respectively. The / kg administration group showed changes in absolute weights of -20.23, -20.33, -14.93 and -10.01%, respectively, and relative weight changes of 1.58, 1.03, -0.91 and 0.52%, respectively, compared to the obese control group.

The absolute and relative weights of the kidneys were 71.05 and -0.65%, respectively, in the obese control group compared to the normal control group, but metformin 250 mg / kg and dagger extract 400, 200 and 100 mg / kg in the obese control group. The relative weight changes were -23.12, -23.53, -17.26 and -12.77%, respectively, compared to -2.04, -3.02, -3.78 and -2.66%, respectively.

Therefore, dose-dependent liver and kidney weight reductions were observed in all Daengberry fruit extract groups, respectively, and in the 400mg / kg dose of Dandelion fruit extract, livers induced by high-fat diets similar to metformin 250 mg / kg. And suppressing an increase in the absolute weight of the kidney.

On the other hand, no significant changes in absolute and relative weights of liver and kidney were found in all high fat diets compared to the normal control group.

Example 7 Analysis of Thickness of Adipose Tissue and Diameter of Adipocyte

The thickness of the adipose tissue and the diameter of the adipose cell were measured from the fat extracted in Example 6 and are shown in Table 6.

division Thickness (mm, ovary) Fat cell diameter (um, ovary) Thickness (mm, abdominal wall) Fat cell diameter (um, abdominal wall) Normal control 2.20 ± 0.66 29.81 ± 5.94 2.03 ± 0.67 37.24 ± 4.87 Obesity Control 4.50 ± 0.46 120.44 ± 12.38 5.32 ± 1.14 132.47 ± 14.58 Metformin 2.76 ± 0.48 46.02 ± 10.22 3.03 ± 0.85 66.17 ± 12.75 BHx 400 mg / kg 2.81 ± 0.54 43.03 ± 13.29 3.14 ± 0.58 60.96 ± 21.41 BHx 200 mg / kg 3.46 ± 0.50 55.99 ± 11.89 3.60 ± 0.41 75.16 ± 18.32 BHx 100 mg / kg 3.78 ± 0.49 64.77 ± 15.83 3.79 ± 0.70 83.82 ± 18.97

As can be seen in Table 6, the thickness and fat cell mean diameter of the ovarian accumulated adipose tissue showed changes of 104.72 and 304.03%, respectively, in the obese control group, but metformin 250 mg / kg and the Taraxacum berry extract. The 400, 200, and 100 mg / kg dose groups showed -38.57, -37.65, -23.09, and -15.98% changes in accumulated adipose tissue thickness, respectively, compared to the obese control group, and -61.79, -64.27, -53.51, and -46.22%. Changes in the average diameter of adipocytes were shown.

The thickness of abdominal wall adipose tissue and the mean diameter of adipocytes showed changes of 161.95 and 255.71% in the obese control group, respectively, but metformin 250 mg / kg and dandelion fruit extract 400, 200 and 100 mg / kg. Compared with the obese control group, the administration group showed changes in accumulated fat tissue thickness of -43.09, -40.97, -32.28 and -28.73%, respectively, and changed the average diameter of fat cells of -50.05, -53.98, -43.27 and -36.73%. Respectively.

Therefore, the dose-dependent ovarian peritoneal and abdominal wall-accumulating adipose tissue thickness and adipocyte diameter were decreased in all the groups of daphnia fruit extracts, respectively, and similar to metformin 250 mg / kg in the dagger fruit 400 mg / kg group. The high fat diet suppressed the increase in the thickness and fat cell diameter of the induced adipose tissue.

Example 8: Fat Analysis in Feces

Fecal samples were collected for 8 hours after the last administration of the test substance (TC colorimetric assay kit, Cat. No. 100102303, Cayman, Ann Arbor, MI, USA; Total Cholesterol Assay Kit (Colorimetric), Cat.No. STA-384 , Cell Biolabs, San Diego, Calif., USA) were used to measure fecal total cholesterol (TC) and triglyceride (TG) with a microplate reader.

As can be seen in Figure 4, the TC content in the feces showed a change of 13.04% compared to the normal control group in the obesity control group, metformin 250 mg / kg, dagger fruit 400, 200 and 100 mg / kg administration group obesity Changes of 131.20, 130.34, 110.26 and 74.57%, respectively, were compared with the control group.

TG content in the feces was 16.25% in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger extract 400, 200 and 100 mg / kg were 174.65 and 177.57, respectively. , 127.04 and 81.57%.

Therefore, the dose and dose-dependent fecal TC and TG contents were observed to increase in all the Taraxacum berry extract groups, and the TC and TG contents in fecal similar to metformin 250 mg / kg were increased in the 400mg / kg dose It showed an increasing effect.

Example 9: Real-Time RT-PCR Analysis

ACC1 (Acetyl-CoA carboxylase), AMPKa1 and AMPKa2 mRNA expression in liver tissues were analyzed by real-time RT-PCR, and leptin, uncoupling proteins (UCP2), adiponectin, MRNA expression of CCAAT-enhancer binding protein-a (C / EBPa), CCAAT-enhancer binding protein-β (C / EBPβ) and sterol regulatory element-binding protein 1 (SREBP-1) mRNA was analyzed. . In brief, RNA was extracted with Trizol reagent (Invitrogen, Carlsbad, Calif., USA), and RNA concentration and quality were determined with the CFX96 ^™ Real-Time System (Bio-Rad, Hercules, Calif., USA). Treated with recombinant DNase I (DNA-free; Ambion, Austin, TX, USA) to remove contaminated DNA and using a kit (High-Capacity cDNA Reverse Transcription Kit, Applied Biosystems, Foster City, CA, USA) Reverse transcribe. Subsequently, it was analyzed by a detection device (ABI Step One Plus Sequence Detection System, Applied Biosystems, Foster City, CA, USA), and the expression pattern was calculated by comparison with the control group. Analytical values were corrected by GAPDH mRNA expression.

The results of ACC1, AMPKa1 and AMPKa2 mRNA expression analysis in adipose tissue are shown in Table 7.

division Normal control Obesity Control Metformin Control 400 BHx 200 BHx 100 BHx ACC1 1.01 ± 0.13 4.38 ± 1.22 1.73 ± 0.33 1.76 ± 0.47 2.29 ± 0.51 2.98 ± 0.23 AMPKa1 1.00 ± 0.09 0.48 ± 0.10 0.83 ± 0.12 0.84 ± 0.08 0.73 ± 0.13 0.66 ± 0.13 AMPKa2 1.01 ± 0.12 0.54 ± 0.09 0.87 ± 0.12 0.83 ± 0.11 0.73 ± 0.11 0.66 ± 0.04

As can be seen from Table 7, ACC1 mRNA expression in the liver tissues showed a change of 334.86% in the obese control group compared to the normal control group, but metformin 250 mg / kg, Dandelion fruit extract 400, 200 and 100 mg / kg group Showed a change of -60.43, -59.80, -47.67 and -32.01%, respectively, compared to the obese control group.

The expression of AMPKa1 mRNA in the liver tissues was -51.75% in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger fruit extract 400, 200 and 100 mg / kg compared to the obese control group, respectively. 71.43, 74.03, 50.65 and 36.36% change.

The expression of AMPKa2 mRNA in the liver tissues was -46.34% in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger extract 400, 200, and 100 mg / kg were compared with the obese control group, respectively. 61.43, 53.35, 35.57 and 21.71%.

Therefore, all doses of ANC extract showed a decrease in ACC1, AMPKa1, and AMPKa2 mRNA expression in dose-dependent liver tissue, respectively, and 400 mg / kg of ALP extract in liver tissues similar to metformin 250 mg / kg. An increase in the expression of ACC1, AMPKa1, and AMPKa2 mRNA was suppressed.

The results of C / EBPa, C / EBPβ and SREBP1c mRNA expression analysis in adipose tissue are shown in Table 8.

division Normal control Obesity Control Metformin 400 BHx 200 BHx 100 BHx Leptin 0.96 ± 0.08 5.94 ± 1.08 1.89 ± 0.76 1.95 ± 0.50 2.79 ± 0.72 3.64 ± 0.89 UCP2 0.99 ± 0.06 0.24 ± 0.07 0.61 ± 0.12 0.58 ± 0.18 0.47 ± 0.12 0.39 ± 0.08 Adiponectin 1.00 ± 0.12 0.15 ± 0.08 0.67 ± 0.20 0.63 ± 0.13 0.47 ± 0.10 0.36 ± 0.10 C / EBPa 1.00 ± 0.08 1.89 ± 0.23 1.27 ± 0.14 1.28 ± 0.12 1.35 ± 0.11 1.46 ± 0.15 C / EBPβ 0.98 ± 0.06 3.23 ± 0.72 1.62 ± 0.33 1.52 ± 0.33 1.89 ± 0.26 2.28 ± 0.37 SREBP 1c 1.03 ± 0.16 2.29 ± 0.42 1.44 ± 0.13 1.37 ± 0.21 1.53 ± 0.26 1.77 ± 0.24

As can be seen from Table 8, leptin mRNA expression in the adipose tissue showed a change of 517.01% in the obese control group, compared to the normal control group, but metformin 250 mg / kg, Dandelion fruit extract 400, 200 and 100 mg / kg administration group Showed a change of -68.13, -67.14, -53.04, and -38.69%, respectively, compared to the control group.

UCP2 mRNA expression in adipose tissues was -76.29% in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger extract 400, 200, and 100 mg / kg were compared with the obese control group, respectively. 160.11, 145.21, 100.00 and 63.83% change.

Adiponectin mRNA expression in adipose tissue showed -85.50% change in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger fruit extract 400, 200 and 100 mg / kg compared to the obese control group, respectively. 362.93, 336.31, 222.41 and 145.69%.

C / EBPa mRNA expression in adipose tissue showed 88.67% change in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger fruit extract 400, 200 and 100 mg / kg compared to the obese control group. The changes were -33.14, -32.61, -28.71 and -23.17%, respectively.

The expression of C / EBPβ mRNA in adipose tissue was 228.66% in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger extract 400, 200 and 100 mg / kg compared to the obese control group. The changes were -49.65, -52.83, -41.47 and -29.42%, respectively.

Therefore, all doses of A. extracts were recognized to reduce mRNA expression of leptin, C / EBPa, C / EBPβ and SREBP1c in dose-dependent adipose tissue, respectively. Increasing mRNA expression of leptin, C / EBPa, C / EBPβ and SREBP1c in adipose tissue similar to kg was suppressed.

Example 10 Histopathological Examination

After measuring the tissue weight, a part of the left lobe, left kidney, pancreas, ovarian fat and abdominal wall fat were fixed in 10% neutral formalin solution, embedded with paraffin, and cut into 3 to 4 um in succession, and then part of hematoxylin & eosin. (Hematoxylin &eosin; HE) staining was performed. Tissue samples were observed under an optical microscope. Some tissues were dehydrated in 30% sucrose solution and subjected to oil red staining after freezing sections.

Observations included steatohepatitis, mean hepatocyte diameter (HE staining), and automated image analysis process (Model i Solution FL ver 9.1; IMT i -solution Inc., Vancouver, Quebec, Canada). It measured using.

Specifically, the steatohepatitis area was calculated as the percentage of lipopolysaccharide in the hepatic parenchyma area (% / mm ² of hepatic parenchyma, oil red stain), and the hepatocyte diameter was at least 10 using an automated image analysis process. Hepatic cells were measured in dogs.

In addition, the number of fatty droplet deposition (fear) was measured in 100 tubules of the new tubule. The mean diameter of adipocytes was measured for at least 10 adipocytes using an automated image analysis process to determine Liver steatosis, mean hepatocyte diameter, and degenerative renal tubule numbers in Tables 9 and 5. Indicated.

division Hepatic steatosis
(% / mm ² of hepatic tissues) Average hepatocyte diameter (um / cell) Degenerative Kidney Urine Count (%) Normal control 7.20 ± 2.87 13.36 ± 1.04 4.25 ± 2.38 Obesity Control 78.84 ± 10.03 34.30 ± 2.65 77.38 ± 10.31 Metformin Control 45.65 ± 10.12 22.64 ± 3.75 31.88 ± 5.41 BHx 400 42.87 ± 10.51 22.71 ± 4.25 27.75 ± 8.58 BHx 200 53.03 ± 10.05 24.04 ± 4.66 39.63 ± 10.70 BHx 100 60.33 ± 12.81 27.31 ± 2.34 49.88 ± 12.72

As can be seen in Table 9 and Figure 5, the rate of change in liver fat (the ratio of the real body showing a change in fat) showed a change of 995.16% compared to the normal control group in the obese control group, metformin 250 mg / kg, dagger fruit extract The 400, 200 and 100 mg / kg administration groups showed changes of -42.09, -45.62, -32.74 and -23.48%, respectively, compared to the obesity control group.

The hepatocellular diameter showed a change of 156.79% in the obese control group compared to the normal control group, but metformin 250 mg / kg, dagger extract 400, 200, and 100 mg / kg were -33.99 and -33.80, respectively. , -29.91 and -20.38%.

The number of denatured tubules characterized by phobia caused by fat accumulation was 1720.59% in the obese control group compared to the normal control group, but metformin 250 mg / kg, dandelion fruit extract 400, 200 and 100 mg / kg Showed a change of -58.80, -64.14, -48.79 and -35.56%, respectively, compared to the obese control group.

As can be seen in FIG. 6, a significant increase in degenerative panic renal tubules was observed in the HFD control group as compared to the intact control group as a result of diabetic nephropathy-induced lipid droplets, but this diabetic nephropathy was metformin 250 mg / Significant normalization was achieved by treatment of the extract of Damascus fruit containing kg. Compared to the HFD control group, the dose-dependent decrease in the number of proliferative kidney tubules was shown, especially in the Dandelion fruit extract 400, 200 and 100 mg / kg treatment groups.

Therefore, dose-dependent liver fat reduction rate, hepatic cell diameter reduction, and degeneration of tubule number were confirmed in all Daengberry fruit extract groups, and similar to metformin 250 mg / kg in Dandelion fruit extract 400 mg / kg group. It showed an inhibitory effect on increasing the localization rate of the liver matrix, indicating a change.

Example 11: Histological Immunology

Tissue sections were subjected to ABO (Avidin-biotin peroxidase) immunostaining, insulin was the guinea pig polyclonal insulin antiserum kit (Abcam, Cat no.ab7842), and glucagon was the rabbit polyclonal glucagon antiserum kit (Abcam, cat No. ab133195) and immunohistochemical analysis was performed.

Specifically, the endogenous peroxidase activity was stopped by incubation for 30 minutes with methanol and 0.3% hydrogen peroxide solution, and the nonspecific binding immunoglobulin was blocked for 1 hour in a wet chamber with a vector lab serum (Vector Lab). . Treated with primary antisera and left overnight in a wet chamber at 4 ° C. and wet chamber with a secondary antibody (biotinylated universal secondary antibody, vector lab) and ABC reagent (Cat no. PK-6200, Vectastain Elite ABC kit, Vector Lab). It was left to stand at room temperature for 1 minute. Finally, the mixture was left at room temperature for 3 minutes in a peroxide substrate kit (Vector lab, cat No. SK-4100).

Cells showing more than 20% immunoreactivity compared with each of the insulin and glucagon inactive cells were determined as positive, and the number of insulin- and glucagon-immunoreactive cells per mm ^{2 of} pancreatic matrix was analyzed by an automated image analysis process. Analyzed.

Automated image analysis process for occupying area (% / mm ² ) of mean zymogen granules in pancreatic matrix, number of pancreatic islets (number / mm ² ) and diameter of pancreatic islet (um) Table 4 and shown in Figure 3 (Scale bars = 80 um) (A: normal control group, B: type 2 diabetes (NIDDM) control group, C: metformin control group, D: dagger fruit extract 400 mg / kg administration group, E: BHx 200 mg / kg administration group, F: BHx 100 mg / kg administration group).

Insulin / glucagon cells (ratio) = (average insulin immunoreactive cells / average glucagon immune active cells)

division Zymogen Granules
(% / mm ² ofexocrine) Average islet cell count
(No./10 mm ² ) Average pancreatic islet diameter
(um / islet) Insulin immune response cells
(cells / mm ² ) [A] Glucagon immune response cells
(cells / mm ² ) [B] Insulin / Glucagon Rate
[A / B] Normal control 44.93 ± 5.19 5.88 ± 1.25 104.23 ± 20.72 471.50 ± 109.88 149.88 ± 34.01 3.18 ± 0.46 Obesity Control 13.99 ± 2.95 22.25 ± 3.11 274.28 ± 51.83 2618.25 ± 493.27 347.13 ± 57.27 7.58 ± 1.15 Metformin Control 32.44 ± 5.92 10.50 ± 2.33 145.52 ± 28.73 1143.75 ± 205.40 227.75 ± 51.94 5.11 ± 0.79 BHx 400 35.82 ± 5.46 11.00 ± 1.60 137.79
± 25.86 1131.63 ± 254.92 215.75 ± 52.91 5.27 ± 0.24 BHx 200 28.44 ± 7.38 15.38 ± 3.02 164.25 ± 11.81 1411.00 ± 219.82 245.88 ± 30.36 5.76 ± 0.75 BHx 100 24.77 ± 5.02 17.38 ± 2.77 186.78 ± 22.27 1640.88 ± 242.61 268.13 ± 36.05 6.13 ± 0.59

As can be seen in Table 10 and Figure 3, the ratio of Zymogen granules in the pancreatic exocrine showed a change of -68.87% in the case of the high-fat diet control group, compared to the normal control group, metformin 250 mg / kg, dandelion fruit In the 400, 200 and 100 mg / kg administration group, the high-fat diet showed a change of 131.93, 156.04, 103.28 and 77.07%, respectively, compared to the control group.

Therefore, the dose-dependent increase in the ratio of Zymogen granules was observed in all the Daengberry fruit extract groups, and the high fat diet-induced pancreatic diet similar to metformin 250 mg / kg in the Danangu fruit extract 400 mg / kg group. It showed an inhibitory effect on the proportion of imogen granules.

Claims (12)

A step S1 of grinding a linden tree (Lonicera caerulea L. var.edulis) fruit at 45 to 55 ° C. for heating;
S2 step of contacting the pung tree fruit of the step S1 to 0.1% by weight of pectinase of the dagger fruit of the step S1 for 2 to 2.5 hours;
S3 step of removing the precipitate from the daengyeok fruit that the step S2;
A step S4 of heating the laden tree fruit which has been subjected to the step S3 to 60 to 80 ° C. for 15 to 30 seconds;
S5 step of adding chitosan and guar gum to 0.005% by weight of each of the daggers of the step S1 to the daggers of the step S4;
S6 step of filtering and concentrating a dendrite tree fruit performing the S5 step; including, obesity, hyperlipidemia or fatty liver (Lonicera caerulea L. var. Edulis) fruit extract manufacturing method for the prevention, treatment or improvement. A pharmaceutical composition for the prevention or treatment of obesity, hyperlipidemia or fatty liver, comprising an extract of Lonicera caerulea L. var.edulis fruit prepared by the method of claim 1. A food composition for improving obesity, hyperlipidemia, or fatty liver, comprising an extract of Lonicera caerulea L. var. Edulis fruit prepared by the method of claim 1. delete delete delete delete delete delete delete delete delete

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