KR101539289B1 - A composition comprising extract of Artemisia annua L. for preventing or treating fatty liver or obesity - Google Patents

Abstract

The present invention relates to a composition for prevention or treatment of fatty liver or obesity which contains a ragweed extract. The above-mentioned milk powder extract has the effect of inhibiting the activity of SREBP-1 and FAS, which are transcription factors of fatty acid synthesis, and enhances the activity of AMPK, which is involved in the inhibition of lipogenesis. Thus, weight, liver weight, subcutaneous fat, blood glucose concentration And is effective in inhibiting the synthesis of fatty acids, cholesterol and triglycerides, and can be easily used as a therapeutic agent for fatty liver or obesity.

Description

[0001] The present invention relates to a composition for preventing or treating fatty liver or obesity,

The present invention relates to a composition for prevention or treatment of fatty liver or obesity which contains a ragweed extract.

In recent years, the population of obesity has been rapidly increasing due to improvements in living standards due to economic development, lack of exercise due to busy living environment, and excessive intake of nutrition. The proportion of obesity (BMI> 25) increased from 11.7% in 1995 to 30.7% in 2008 and the proportion of obesity (BMI> 30) increased from 2.3% in 1998 to 4.1% Has been growing rapidly. Obesity is a condition in which energy is excessively accumulated in the body due to unbalanced energy intake and consumption, resulting in an abnormally increased adipose tissue. The World Health Organization (WHO) considers Oriental BMI (body mass index) 23-25 as overweight, 25-30 obese, and over 30 as high obesity.

The causes of obesity include environmental factors such as high fat, high calorie diet, lack of exercise due to busy social environment, endocrine abnormality, and genetic factors. Of these, about 50 to 70% of obesity is due to environmental factors And the rest is known to be caused by genetic factors.

Currently, typical examples of obesity remedies include Orlistat (Roche), which is the main ingredient of Roche, and Reductil (Abbott), which is a main ingredient of sibutramine. Nausea, diarrhea, abdominal pain, insomnia, And fibrates, which are represented by clofibrate, have been used in clinical practice, but side effects such as hepatic dysfunction have been reported.

Fatty liver (steatosis, fatty liver) is a disease of accumulation of triglyceride in hepatocyte, non-alcoholic fatty liver and alcoholic fatty liver disease can be divided into the symptoms, fat accumulation due to obesity can be caused by fatty liver is mainly nonalcoholic fatty liver . Fatty liver is a condition in which fat lipids, especially triglycerides, accumulate excessively and fat vacuoles are observed in more than half of the hepatocytes due to the failure of normal lipid metabolism. Clinically, when the fat is observed in 5% or more of hepatocytes or when the fat is more than 5 mg per 100 mg of liver, it is classified as fatty liver. In recent years, patients with fatty liver have been increasing rapidly due to high fat, high calorie diet and lack of exercise due to the development of civilization, and the age group has been on the whole from the teens to the 50s and 60s. When left untreated, liver can be progressed to hepatitis, liver cirrhosis, liver cancer and the like. The causes of fatty liver include alcohol, diabetes, obesity, malnutrition (long-term low-fat diet, high fat diet, starvation, starvation, vitamin deficiency, excess carbohydrate intake), drug abuse Journal of Medicine 175 (1995.12, pp.785-794).

Along with obesity, fatty liver is currently suffering from a number of people, but due to the complexity of the disease with various symptoms, there is no definitive diagnosis and treatment method.

On the other hand, AMP-activated protein kinase (AMPK) is activated by LKB1 (liver kinase B1) or CaMKKβ (calcium / calmodulin-dependent protein kinase kinase beta) in conditions such as glucose deficiency, body stress, hypoxia, (Fryer, Parbu-Patel, & Carling, 2002; Hurley et al., 2005; Hardie, 2007).

AMPK inhibits the production of de novo fatty acids through inactivation of ACC (acetyl-CoA carboxylase) (increased phosphorylation of ACC), while CPT-1 (carnitine palmitoyltransferase- 1), PPAR-α (peroxisome proliferator-activated receptor-α), and UCP (uncoupling protein).

AMPK already plays an important role in glucose and fat metabolism in diabetes, obesity and cancer (Carling, 2004) and is known to be a major target for treating such metabolic diseases (Zhang, Zhou & Li, 2009 ). SREBP is a transcription factor that regulates the expression of fat-related enzymes such as ACC (acetyl-CoA carboxylase), FAS (fatty acid synthase) and HMGCR (3-hydroxy-3-methylglutaryl CoA reductase) Goldstein, Rawson, & Brown, 2002), increased expression of SREBP has been shown to be strongly associated with fatty liver in diabetic animal models (Horton, Goldstein, & Brown, 2002; Yahagi et al., 2002).

In addition, the expression of SREBP-1c is regulated by the nuclear receptors LXR-α and LXR-β, which are involved in the metabolism of glucose, fat and cholesterol. -Phosphoric acid has been reported to be direct agonists of the LXR-alpha and LXR-beta.

In recent years, it has been known that AMPK inactivates SREBP-1 in high fat diet induced animal models to inhibit fatty liver or obesity. Thus, AMPK and SREBP are targets for treatment of fatty liver and obesity It is proving that it is being used.

Artemisia annua L. is an annual herbaceous plant belonging to the Asteraceae family. It is distributed worldwide in tropical Asia. In Korea, it is wild in the streets and fields all over the country. In the oriental medicine, the top part of the mugwort is used as an antipyretic agent, a hemostatic agent, a treatment for skin diseases, and an insecticide. It is also known as a source of the anti-malaria agent artemisinin. In addition, the various pharmacological effects of white mugwort are well known in various literature. However, it is difficult to obtain and it is difficult to cultivate it, so it is not commercialized so that it can be consumed as a healthy food of the general public.

The inventors of the present invention conducted studies on the treatment of obesity or fatty liver, and found that the extract of dog gall mugwort had an effect of inhibiting weight gain and fat accumulation in mouse liver tissue and activating AMP-activated protein kinase (AMPK) Thereby completing the present invention.

Meanwhile, although it has been disclosed that the extract of Artemisia officinalis has an obesity-suppressing effect (Choi, Y, et al., 2013), the inventors of the present invention have found that Artemisia sp. Extract has an effect of inhibiting obesity or fatty liver.

Choi, Y, et at., Artemisia iwayomogi Extract Attenuates High-Fat Diet-Induced Obesity by Decreasing the Expression of Genes Associated with Adipogenesis in Mice. Fryer, L. G., Parbu-Patel, A., & Carling, D. (2002). The anti-diabetic drugs rosiglitazone and metformin stimulate AMP-activated protein kinase through distinct signaling pathways. The Journal of Biological Chemistry, 277, 25226-25232. Hardie, D. G. (2007). AMP-activated / SNR1 protein kinases: conserved guardians of cellular energy. Nature reviews. Molecular cell biology, 8, 774-785. Horton, J. D., Goldstein, J. L., & Brown, M. S. (2002). SREBPs: activators of the complete program of cholesterol and fatty acid synthesis in the liver. The Journal of Clinical Investigation, 109, 1125-1131. Hurley, R. L., Anderson, K. A., Franzone, J. M., Kemp, B. E., Means, A. R., & Witters, L. A. (2005). The Ca2 + / calmodulin-dependent protein kinase kinases are AMP-activated protein kinase kinases. The Journal of Biological Chemistry, 280, 29060-29066. T., Ide, T., Yoshikawa, T., Amemiya-Kudo, M., Tomita, S., Okazaki, H., Tamura, Y. , Iizuka, Y., Ohashi, K., Osuga, J., Harada, K., Gotoda, T., Nagai, R., Ishibashi, S., & Yamada, N. (2002). (SREBP-1) ameliorates fatty livers but not obesity or insulin resistance in Lep (ob) / Lep (ob) mice. The Journal of Biological Chemistry, 277, 19353-19357.

It is an object of the present invention to provide a composition for prevention or treatment of fatty liver or obesity which contains a ragweed extract.

The present invention relates to a composition for prevention or treatment of fatty liver or obesity which contains a ragweed extract.

The present invention relates to a composition for preventing or treating fatty liver or obesity containing Artemisia annua L. extract.

The rabbit extract can be prepared by extracting ragweed with C1 to C4 lower alcohol, water or a mixed solvent thereof.

The rabbit extract has an effect of increasing the activity of AMPK (AMP-activated protein kinase).

In another aspect, the present invention provides a health functional food for prevention or amelioration of fatty liver or obesity which contains a ragweed extract.

Hereinafter, the present invention will be described in detail.

The rabbit extract can be prepared by using a lower alcohol of C1 to C4, water or a mixed solvent thereof, and can be prepared by extraction with an organic solvent (alcohol, ether, acetone, etc.), distribution of hexane and water, A known method used for the separation and extraction of plant components, such as a method based on a synthetic method, or a method based on a synthetic method, or a method using a synthetic method.

The extraction temperature, extraction time and weight of the extraction solvent can be appropriately adjusted according to the extraction conditions, but the extraction temperature is preferably 40 to 130 ° C, more preferably 50 to 100 ° C, most preferably 60 Lt; 0 > C. In addition, the extraction time of the ragweed extract may be 0.5 to 12 hours, more preferably 0.5 to 6 hours, and most preferably 0.5 to 2 hours. In order to prepare the ragweed extract, May be added in an amount of 1 to 20 times.

The chromatography can be carried out using silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, medium pressure liquid chromatography chromatography, thin layer chromatography (TLC), silica gel vacuum liquid chromatography, and high performance liquid chromatography.

The rabbit extract according to the present invention can be used as an additive for foods and medicines because it is prepared using ragweed which has been used as a traditional medicinal material and is easy to manufacture and has high stability.

In addition, the present invention provides a pharmaceutical composition for prevention or treatment of fatty liver or obesity, which comprises a ragweed extract and a pharmaceutical excipient. The pharmaceutical composition may contain 0.001 to 99.9% by weight of Lauroginae Extract. The pharmaceutical excipient may be a carrier, excipient or diluent. The pharmaceutical composition containing the ragweed extract may be formulated in the form of powders, granules, tablets, capsules, oral preparations such as suspensions, emulsions, syrups and aerosols, external preparations, suppositories and sterilized injection solutions, Can be used. Examples of the carrier, excipient or diluent which can be contained in the pharmaceutical composition 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 and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may contain at least one excipient, such as starch, calcium carbonate, sucrose, Lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The dosage of the pharmaceutical composition for prevention or treatment of fatty liver or obesity containing the ragweed extract of the present invention depends on the age, sex, and weight of the subject to be treated, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, And the prescriber's judgment. Dosage determinations based on these factors are within the level of ordinary skill in the art and generally the dosage ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferable dosage is 1 mg / kg / day to 500 mg / kg / day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

The pharmaceutical composition for prevention or treatment of fatty liver or obesity containing the ragweed extract of the present invention can be administered to mammals such as rats, livestock, and humans in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection. Since the compound of the present invention has little toxicity and side effects, it can be safely used even for long-term administration for preventive purposes.

The present invention provides a health functional food for preventing or ameliorating fatty liver or obesity containing the dog's milk worm extract and a food acceptable food supplementary additive. The health functional food of the present invention includes forms such as tablets, capsules, pills, and liquids. Examples of the foods to which the extract of the present invention can be added include various foods, beverages, gums, tea, vitamins , And health functional foods.

The present invention relates to a composition for prevention or treatment of fatty liver or obesity which contains a ragweed extract. The above-mentioned milk powder extract has the effect of inhibiting the activity of SREBP-1 and FAS, which are transcription factors of fatty acid synthesis, and enhances the activity of AMPK, which is involved in the inhibition of lipogenesis. Thus, weight, liver weight, subcutaneous fat, blood glucose concentration And is effective in inhibiting the synthesis of fatty acids, cholesterol and triglycerides, and can be easily used as a therapeutic agent for fatty liver or obesity.

1 is a schematic diagram of an experimental procedure for confirming the anti-obesity effect in a high fat diet (HFD) mouse of the present invention.
FIG. 2 is a graph showing the results of measuring the body weight of mice administered with the extract of Example 1, the high fat diet + the extract of Comparative Example 1, and the non-treated group, the high fat diet group, and the high fat diet group.
FIG. 3 is a graph showing the liver weight of mice treated with the extract of Example 1, the high fat diet + the extract of Comparative Example 1, the non-treated group, the high fat diet group, and the high fat diet group.
FIG. 4 is a photograph showing the subcutaneous fat of mice administered with the extract of Example 1-1.
FIG. 5 shows the results of microscopic observation of liver sections of mice treated with the extract of Example 1-1, without treatment, high fat diet, high fat diet.
FIG. 6 shows the results of AST (aspartate aminotransferase) and ALT (alanine) liberated in serum from hepatocytes of the mice treated with the extract of Example 1, the extract of Example 1 and the extract of Comparative Example 1, aminotransferase) activity of the present invention.
FIG. 7 is a graph showing the triglyceride content in liver tissues of the mice to which the extract of Example 1, the high fat diet system + the extract of Comparative Example 1 was administered, the untreated group, the high fat diet group, the high fat diet group +
FIG. 8 is a graph showing the total cholesterol content in blood of the mice treated with the extract of Example 1, the high-fat diet + and the extract of Comparative Example 1, the non-treated group, the high fat diet group, and the high fat diet group.
FIG. 9 is a graph showing blood glucose levels of mice treated with the extract of Example 1, the high fat diet + group, and the extract of Comparative Example 1, without treatment group, high fat diet group, and high fat diet group.
FIG. 10 shows the results of Western blotting of expression of p-AMPK, p-ACC and β-actin in the mice to which the extract of Example 1-1 was administered in the untreated group, the high fat diet group and the high fat diet group.
FIG. 11 shows the expression of FAS (Fatty acid synthase) and SREBP-1c (sterol regulatory element-binding protein-1c) in mice treated with the extract of Example 1-1, This is the result of checking with blot.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, the intention is to provide an exhaustive, complete, and complete disclosure of the principles of the invention to those skilled in the art.

< Example  1. Preparation of Dogwood Mugwort Extract>

Example  1-1. Water extract

300 g of distilled water was added to 100 g of dried mugwort, and the mixture was extracted at 90 ° C. for 30 minutes. The extract was cooled to 25 ° C. and filtered through a 0.45 μm membrane filter to separate the filtrate and residue. The concentrate was concentrated under reduced pressure at 35 캜 using a vacuum concentrator, and the concentrate was purified by using a freeze-dried vacuum dryer. The herb was purchased from Kyungnam Agricultural Cooperative Federation.

Example  1-2. 100% ethanol extract of dog mugwort

The same method as in Example 1-1 was used, except that 100% ethanol was used instead of water as the extraction solvent, and the extraction temperature was set at 60 ° C and the filtrate was concentrated under reduced pressure to prepare a 100% ethanol extract powder of Artemisia princeps.

Example  1-3. Extract of 70% ethanol aqueous solution of mugwort

Using the same method as in Example 1-2, the extraction solvent was concentrated under reduced pressure using 70% aqueous ethanol instead of 100% ethanol and lyophilized to prepare a powder of aqueous methanol solution of 70% ethanol.

< Comparative Example  One. Inosugi  Preparation of Extract>

Comparative Example  1-1. Inosugi  Water extract

The same method as in Example 1-1 was used except that Artemisia syrup extract was prepared using Artemisia sp.

Comparative Example  1-2. Inosugi  100% ethanol extract

Using the same method as in Example 1-2, instead of dog wormwood, 100% ethanol extract of Artemisia sp.

Comparative Example  1-3. Inosugi  70% ethanol aqueous solution extract

The same method as in Example 1-3 was used except that an aqueous solution of 70% ethanolic aqueous solution of arspookus was prepared using Artemisia sp.

Experimental Example 1. Anti-obesity effect in high-fat diet mice [

Experimental Example 1-1. Administration of high-dose mouse and mouse powder extract to mice

After 4 weeks (28 days) administration of high fat diet to male ICR mice of 4 weeks of age, mice weighing increasing were weighed and selected mice were divided into the untreated group (negative control, normal diet, normal diet, (28 mg / kg body weight) was divided into the experimental group (50 mg / kg of the crude extract of Example 1 or 50 mg / kg of the extract of Artemisia princeps of Comparative Example 1) ). Five mice were used in each experimental group. The mice were maintained at a temperature of 23 ° C, humidity of 50 ± 5%, and night and day for 12 hours. In the above, 8% kcal dietary diets (Research Diets, USA) were used for the normal control diet, and 45% kcal dietary diets (Research Diets, USA) were used. Each sample was dissolved in 100 μl of physiological saline per mouse and administered orally. Neutral saline was administered only to the negative control group or the positive control group. The effect of the mugwort extract on body weight was measured on the 56th day when the experiment was completed using the animal scale, and the result of the experiment was shown in the graph in FIG. The results of the experiment are shown in FIG. 3. Referring to FIGS. 2 and 3, in the case of the group to which the ragweed extract was administered, And liver weight were remarkably decreased, and it was found that the reduction effect was superior to that of Artemisia sp.

In addition, in order to confirm the effect of the hot water extract of Laetmugwara on the weight of the abdominal fat (white fat) in Example 1-1, the abdominal fat was extracted and the weight was measured. The results of the experiment are shown in FIG. 4, 1-1 showed a significant decrease in abdominal fat weight compared with those of high fat diet group, and it was almost similar to that of untreated group.

Experimental Example 1-2. Liver biopsy

The effect of the mugwort extract on hepatocyte injury was examined by microscopic observation after tissue sectioning after the anesthetization with ether after 56 days of experiment in Experimental Example 1-1 and the liver was extracted from each group. 5 in each group. Hematoxylin and eosin and masson's trichrome (Vyberg et al., 1987) were used for hematoxylin-eosin and masen trichrome staining.

Referring to FIG. 5, it can be seen that the liver condition similar to that of the untreated control group can be confirmed in the group treated with the high-fat control method and in Example 1-1, -1 in the treated group was higher than that in the treated group.

Experimental Examples 1-3. Experiment to measure hepatic damage index enzyme activity

The levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), which are used as indicator enzymes for the degree of liver damage, were measured after 56 days of experiment in Experimental Example 1-1, (ASAN PHARMACEUTICAL, Korea), and the results of the experiment are shown in FIG.

6, the AST and ALT values of the group administered with the ragweed extract of Example 1 were significantly lower than those of the non-ragweed group, Could know.

Experimental Examples 1-4. Measurement of serum triglyceride and total cholesterol in blood

After 56 days of experiment in Experimental Example 1-1, the effect of the milk curd extract on blood total cholesterol and blood triglyceride content was examined and the serum of each mouse was separated and analyzed using an assay kit (Asan Pharm, Korea) The results of the experiment are shown in FIG. 7 and FIG. 8. Triglyceride (Triglyceride) (FIG. 7) and total cholesterol (total cholesterol Total cholesterol, Fig. 8) was low, and the effect was more excellent than that of the group treated with Artemisia sp.

Experimental Examples 1-5. Blood glucose measurement experiment

After 56 days of experiment in Experimental Example 1-1, the effect of the crude glutinous rice extract on the blood glucose content was measured using a blood glucose analyzer (MediSense, ABBOTT, USA) in each group of mice. Is shown in Fig.

Referring to FIG. 9, it was found that the blood glucose (Glucose) content was lower in the mice treated with the dog gall bladder extract than in the mice maintained only in the high-fat feeding method, and the effect was more excellent than those of the mice treated with the Artaxanthin extract .

Experimental Example 1-6. Measurement of expression levels of FAS, SREBP-1c, p-AMPK and p-ACC by Western blotting

In order to confirm the effect of the water extract of P. japonicus on the AMPK activation of the abdominal fat (white fat) after 56 days of experiment in Experimental Example 1-1, the animal was anesthetized with ether, The results were shown in FIGS. 10 and 11. FIG.

For Western blot analysis, the adipose tissues treated with the sample were washed twice with PBS and incubated with 1 mM PMSF, 1 mM Na ₃ VO ₄ , 1 mM NaF, 1 μg / ml aprotinin, 1 μg / ml pepstatin, 1 μg / ml leupeptin and 10% The supernatant was obtained by centrifugation (15000 rpm, 4 ° C, 20 minutes) by vortexing and digesting with protein separating reagent containing RIPA lysis buffer (Upstate Biotechnology, USA) for 1 hour. Protein concentrations were measured and quantified at 595 nm in a microreader using a Bio-rad protein assay kit (Bio-rad, USA). 40 μg of the protein was separated by electrophoresis on a 10% SDS-polyacrylamide gel and then transferred (100V, 120 min) to a polyvinylidene difluoride membrane (Milipore, USA). Protein-transferred PVDF membranes were blocked with 0.1% TTBS (Tris buffered saline with 0.1% Tween 20) solution containing 5% skim milk or 5% BSA (bovine serum albumin) for 1 hour at room temperature. And then reacted with primary antibodies (p-AMPK, p-ACC, FAS, SREBP-1c, and β-actin). (1: 1,000, Santa Cruz Biotechnology, USA), SREBP-1c antibody (1: 1,000, Santa Cruz Biotechnology, USA), p-AMPK antibody (1: 1,000, Cell Signaling, USA) and β-actin antibody (1: 2,000, Santa Cruz Biotechnology, USA) overnight at 4 ° C. The membranes were washed with 0.1% TTBS, and the peroxidase-conjugated secondary antibody (anti-rabbit IgG-HRP, Santa Cruz Biotechnology, USA) was diluted to 1: 2,000, 1: 5,000 or 1: 10,000 Diluted, reacted for 1 hour, and washed with TTBS. The expression of each protein was measured using a chemiluminescence measuring device (Chemi-doc, Bio-Rad, USA) after reacting with a WEST-ZOL western blot detection system (iNtRON Biotechology, Korea) Respectively.

10, the AMPK and ACC phosphorylation (p-AMPK, p-ACC) was significantly reduced in the high-fat diet group compared to the untreated group, whereas the high fat diet group 1-1, the phosphorylation of AMPK and ACC (p-AMPK, p-ACC) was recovered to a similar level to that of the control group.

11, the expression of FAS and SREBP-1c was increased in the high-fat diet group compared to the untreated group. However, in the high fat diet group, the FAS and SREBP- And SREBP-1c were restored to similar levels to those of the control group.

Therefore, through the above results, it was confirmed that the milk curcuma extract of the present invention activates AMPK and inhibits signal transduction of FAS, thereby inhibiting lipid synthesis and having therapeutic effect of obesity or fatty liver.

<Experimental Example 2> Toxicity test>

Experimental Example 2-1. Acute toxicity

This study was conducted to investigate the toxicity of the extract of Example 1-1 of the present invention to an animal body acutely (within 24 hours) when an excessive amount of the extract of the present invention was consumed in a short period of time and to determine the mortality rate. Twenty ICR mice were injected into each of the control and experimental groups. 400 / tween-80 / EtOH (8/1/1, v / v / v) was administered to the control group and the extract of Example 1-1 of the present invention was applied to the PEG- EtOH (8/1/1, v / v / v) for oral administration. After 24 hours of administration, the respective mortality rates were examined. As a result, it was confirmed that all the mice survived in the control group and the experimental group administered with the herbal mixture extract of Example 1-10 of the present invention at a concentration of 2 g / kg / day.

Experimental Example 2-2. Organ organs toxicity test in experimental group and control group

In the long-term toxicity test, the extract of Example 1-1 of the present invention was administered to C57BL / 6J mice (10 mice per group) for 8 weeks at each concentration. In order to investigate the effect on the organs (tissues) of the animals, an experimental group to which the extract of the crude drug mixture of Example 10 of the present invention was administered and PEG-400 / tween-80 / EtOH (8/1/1, v / v / v), the blood concentration of glutamate-pyruvate transferase (GPT) and blood urea nitrogen (BUN) was measured using a Select E (Vital Scientific NV, Netherland) instrument Respectively. As a result, GPT, which is known to be related to hepatotoxicity, and BUN, which is known to be related to renal toxicity, showed no significant difference compared to the control group. In addition, liver and kidney were cut from each animal, followed by a general tissue section preparation, histological observation with an optical microscope, and no abnormalities were observed in all tissues.

&Lt; Formulation Example 1 >

Formulation Example 1-1. Manufacture of tablets

200 g of the extract of Example 1-1 of the present invention was mixed with 175.9 g of lactose, 180 g of potato starch and 32 g of colloidal silicic acid. To this mixture was added a 10% gelatin solution, which was pulverized and passed through a 14-mesh sieve. This was dried, and a mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate was made into tablets.

Formulation Example 1-2. Injection preparation

1 g of the extract of Example 1-1 of the present invention, 0.6 g of sodium chloride and 0.1 g of ascorbic acid were dissolved in distilled water to make 100 ml. This solution was placed in a bottle and sterilized by heating at 20 DEG C for 30 minutes.

<Formulation Example 2: Food Preparation>

Formulation Example 2-1. Manufacture of cooking seasonings

The extract of Example 1-1 of the present invention was added to the cooking seasoning in an amount of 1 wt% to prepare a cooking sauce for health promotion.

Formulation Example 2-2. Manufacture of flour food products

The extract of Example 1-1 of the present invention was added to wheat flour at 0.1% by weight, and bread, cake, cookies, crackers and noodles were prepared using the mixture to prepare a food for health promotion.

Preparation Example 2-3. Manufacture of soups and gravies

Health enhancing soup and juice were prepared by adding the extract of Example 1-1 of the present invention to the soup and juice at 0.1 wt%.

Formulation Example 2-4. Manufacture of dairy products

Various dairy products such as butter and ice cream were prepared from the extract of Example 1-1 of the present invention using the milk.

Formulation Example 2-5. Vegetable juice manufacturing

0.5 g of the extract of Example 1-1 of the present invention was added to 1,000 ml of tomato juice or carrot juice to prepare vegetable juice for health promotion.

Formulation example  2-6. Fruit juice  Produce

0.1 g of the extract of Example 1-1 of the present invention was added to 1,000 ml of apple juice or grape juice to prepare fruit juice for health promotion.

Claims (5)

A composition for the prevention or treatment of fatty liver or obesity, which comprises Artemisia annua L., a C. elegans extract prepared by extracting C1 to C4 lower alcohol, water or a mixed solvent thereof. delete The method according to claim 1,
Wherein the rabbit extract has an effect of increasing the activity of AMP-activated protein kinase (AMPK). A health functional food for preventing or ameliorating fatty liver or obesity containing Artemisia annua L. by extracting from Artemisia annua L. with lower alcohol of C1 to C4, water or a mixed solvent thereof. delete

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