KR100662776B1 - Pharmaceutical composition comprising the crude drug extract for preventing and treating liver disease - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing and treating liver diseases containing herbal extracts, and more particularly, for preventing and treating liver diseases containing essential extracts of baekbaekpi, baekbokyeong, brown root, hyungae, safflower, shiho and windproof extract. It relates to a composition. Since the composition of the present invention exhibits excellent liver protection and is stable to the human body, the composition of the present invention can be usefully used for medicines and health functional foods for protecting liver cells and preventing or treating liver damage.

Liver disease, health functional food, baekbaekpi, baekbokyeong, brown root

Description

Pharmaceutical composition comprising the crude drug extract for preventing and treating liver disease}

The present invention relates to a pharmaceutical composition for the prevention and treatment of liver disease and health functional food containing baekbaekpi, baekbokyeong, brown root, mold dog, safflower, shiho and windproof extract.

The liver is an organ that has been known to be involved in blood storage and circulation, blood volume control and detoxification, and is also closely related to mental activity. Since our bodies are always exposed to various pollutants and toxic substances due to industrialization, the liver is also constantly suffering from detoxification. Furthermore, hepatic damage caused by mental stress is a serious problem. Damaged hepatocytes can be repaired in the event of mental rest, but in a busy modern society, they cannot afford to rest. Because of the body's defense and detoxification effect due to the immune system may cause other diseases. The liver is a large buffering organ that does not appear well in the early stages of the disease and is found only when it is significantly weakened.

Hepatotoxic agents include carbon tetrachloride and D-galactosamine. Aliphatic halogen hydrocarbons, carbon tetrachloride, are xenobiotics that cause biofilm damage and cause toxic effects on the liver and kidneys (Bruckner, JV, Fund. Appl. Toxicology , 6 , pp 16-34; Butler, TC, J. Pharmacol.Exp. Ther. , 134 , pp 311-319).

Recently, a lot of experiments have been reported on the action of inhibiting free radical generation through natural products such as edible and medicinal plants for the prevention or treatment of liver toxicity (Caragy, AB, Food Technology , 46 , pp65-68, 1992; Liang jun, Y., et al., Biochem. And Biophy.Res. Com. , 212 , pp360-366, 1995; Kim, HK, et al., Korean J. Food Sci.Technol., 27 , pp80 -85, 1995; Middleton, E., Int. J. Pharmacognosy , 34 , pp344-348, 1996). In particular, silymarin (SLM) is a substance isolated from the fruit of Silybum marianum (Carduus marianus) belonging to the Asteraceae family, and BDD (biphenyl dimethyl dicarboxylate (BDD) is similar to Schizandrin, a component of Schizandra chinensis. As an artificial synthetic material, it has been developed and used as a therapeutic agent for liver diseases derived from natural products.

Therefore, the present invention was intended to evaluate the possibility of development as a drug for improving liver function or hepatotoxicity by observing the effect of improving the liver function of herbal combinations that are expected to be effective in the prevention or treatment of liver toxicity clinically.

Mulberry root-bark ( Mori Radicis Cortex ) is the root bark of the mulberry ( Morus alba L.) or the same plant (Moraceae). 소), Haeng Haeng-su, sooyoung Cheongdam, and antipyretic antitussive expectorant, which have anti-inflammatory and diuretic effects. The main bioactive components are morusin and cyclomorucine. cyclomorusin, sanggenon AE, Kuwanone A, B, C, α-amyrin, β-amirin, fatty acids, etc. (Pharmaceutical Research Society, Modern Pharmacy, p248, 1986). It has been reported that E. coli prevents hyperglycemia by inhibiting maltase and sucrose in the intestine of mice and blocking glucose influx into the blood (Yoon Young Kim et al., Korean J. FOOD SCI.Technol . 31 ( 4) , pp1057-1064, 1999), and serum peroxidation of liver and kidney tissues of ICR mice induced by diabetes mellitus (alloxan) decreased to normal control levels (Seuk-Rin Lim, The Institute of Oriental Medicine, Daejeon University, 10 (1), p483, 2001) In rats induced diabetes with STZ (sreptozotocin), increased AST, ALT, ALP, and LDH activity was significantly restored to normal control levels due to decreased blood glucose and STZ administration. (Yoon Soo Hong et al., J. Korea Soc. Hygienic Sciences , 7 (2) , pp119-123, 2001). In addition, the hepatoprotective effect of E. coli was isolated from isolated liver cells (Ae-Kyung Kim et al., J. Korea Soc. Hygienic Sciences , 1 (1) , 1995). , ALT and MDA have been reported to restore the control level (Kim Sun-Yeung, et al., Hepatoprotective effect of extracts of hepatic bark on hepatotoxicity induced by carbon tetrachloride, International Symposium on the 100th Anniversary of the Korean Society of Radiology and Korean Symposium Competition, 2000).

Poria cocos WOLF is a fungal nucleus that parasitizes around the roots after 5 to 6 years of resection of pine trees, and includes white, white, white, red, and red. The main components are carbohydrates, water, crude fiber, minerals and trace proteins. Recently, many studies have been conducted on anticancer effects. It is reported that triterpene phase in Bokryeong has anti-soil, anti-inflammatory effects, and the fungal nucleus contains beta-patchy (β-pachyman), accounting for 93% of the dry weight. Trichypenic compounds, pachymic acid, tumulosic acid, 3beta-hydroxylanosta-7,9, and 24-triene-2 It contains acid (24-triene-2-acid), as well as plant solids, chitin, sterols, lecithin, adenine, histidine, choline, lipase, Proteinase and the like. Bokryeong acts on the digestive system such as diuretic, antimicrobial and gastric acid, and reduces blood sugar levels (Information Sup, Shin Min-duk, Doha Herbal Medicine Dictionary, Yeonglimsa, pp40-43, 1998).

Brown root ( Puerariae RADIX) is a root removed from the bark of Pueraria lobata OHWI and is widely used as a favorite food, drink, and health food in the private sector. In oriental medicine, it is used to release cold muscles, rash, anti-alcoholic pain, etc., and relieves thirst by making a essence, which increases the spleen and stomach. (Herbology, p148, 1995). Brown root contains a large amount of isoflavonoids (Chinese modern research in clinical applications, p626, 1994), and many studies have been reported on the components mainly related to isoflavonoids. Pharmacological actions related to repulsion include anti-alcoholic intoxication ( Proceeding of National Academy of Sciences of USA , 95 (5) , pp2198-203, 1998), hepatoprotective action ( Planta Medica , 64 (5) , 413-6., 1998), Proceeding of National Academy of Sciences of USA , 94 (5) , pp1675-9, 1997), and antioxidant activity ( Chem. Pharm. Bull. 40 (3) , 721-4, 1992) Known.

The elder brother (Schizonepeta tenuifolia var. Japonica KITAGAWA) uses the annual herbaceous outpost belonging to Labiatae in the sun. The oil contains 1.8% of essential oils, and d-menthone, dl-menthone and a small amount of d-limonene are included in the essential oil. Hae Pyo (解 表), Geopung (祛風), and bleeding is effective (Information, Shin Min-ryo, Dohae Hyangjedae, Yeonglimsa, pp863-4, 1998).

Safflower (Carthamus tinctorious L.) is an annual vegetation belonging to the family Compositae , originating from Afghanistan's mountainous regions or Ethiopia (Lee Chang-bok, Korea Plant Book, 1980). Safflower seed has been used for soothing and bleeding in Korea, China, Japan, etc. 1975). In particular, safflower seed oil contains a large amount of unsaturated fatty acid and tocopherol, including linolenic acid, which is not only nutritionally important, but also widely used as an antioxidant, antithrombotic, and antihypertensive new material (In-Kyu Park, Bong-je Kim, Dong Bo-gam, 1989).

Shiho ( Bupleurum falcatum L.) is a perennial herb that belongs to Umbelliferae , dried in the sun, and is used in essential oils, bupleurumol, oleic acid, and linoleic acid. acid), palmitic acid, stearic acid, lignoceric acid, glucose and saponin. Shiho has the effects of antipyretic, sedative, analgesic, anti-inflammatory, and anti-pathogenic effects (Information Sup, Shin Min Kyo, Doha Haksa Dictionary, Yeonglimsa, pp413-4, 1998).

Windbreak ( Ledebouriella seseloides WOLF.) Is a three-year-old herbaceous roots belonging to the genus Umbelliferae , dried in the sun, and the wind contains essential oils, mannitol, and high glycosides. Windproof has effects such as sweating, antipyretic, analgesic, diuretic, and antibacterial effect (Information subsidiary, Shin Minryo, Dohae medicinal metabolism dictionary, Younglimsa, p428, 1998).

As mentioned above, many researches have been conducted on the efficacy and toxicity of the baekbaekpi, bokyeong, brown root, safflower, safflower, shiho and windbreaks, which have been used in oriental medicine and folk medicine for a long time without any special toxicity or side effects. . However, there is no research literature or other data on the composition for preventing and treating liver disease containing the complex herbal extract, and the research itself has not been made.

Therefore, the present inventors focused on the research on improving liver function derived from natural products, and as a result, the composition containing the extract of Sangbaekpi, Bokryeong, brown root, safflower, safflower, shiho and windproof as essential active ingredients has excellent treatment effect of liver disease. And the present invention was completed.

An object of the present invention is to provide a pharmaceutical composition and health functional food for preventing and treating liver disease, which essentially contains a mixed extract of baekryepi, bokyeong, brown root, mold dog, safflower, shiho and windproof.

In order to achieve the above object, the present invention is a liver disease prevention, or treatment of liver disease, including baekbaekpi, bokyeong, brown root, mold opening, safflower, shiho and windproof mixed extract as an active ingredient and a pharmaceutically acceptable carrier, excipient or diluent It provides a pharmaceutical composition.

The extract includes a polar solvent selected from water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably an extract available in water.

Hereinafter, the present invention will be described in detail.

The extract is characterized in that it is included in the form of a pulverized product, extract or powder extract thereof. At this time, the composition according to the present invention includes a mixed extract comprising a single extract of each herbal medicine, or a complex extract prepared by extracting a plant containing two or more of each herbal medicine.

The extract according to the present invention may be a crude extract prepared by applying a solvent extraction method using a conventional water extract, alcohol, and the like, and may also be a fraction extract purified using column chromatography. Extraction method according to the invention can be applied to all extraction methods known to those of ordinary skill in the art, for example, extraction with water, alcohol or mixed solvents, extraction method using a bath or room temperature, etc. Including but not limited to.

In a preferred embodiment of the present invention, dried herbal medicines may be washed to remove foreign substances, etc. present on the surface of the material, and then dried in a dryer, mixed in an appropriate blending ratio, and ground to obtain the herbal powders.

In addition, 2 to 20 times, preferably 3 to 10 times the volume / weight of distilled water, lower alcohols having 1 to 4 carbon atoms or mixed solvents thereof, preferably distilled water, methanol or 70% alcohol of the powdered complex herbal medicines. Alone or a mixture of these solvents is added at a temperature of 0 ° C to room temperature, preferably 4 to 6 ° C for 12 hours to 48 hours, preferably 20 to 24 hours, or 80 to 110 ° C, preferably 1 to 24 hours, preferably 2 to 5 hours at an extraction temperature of 90 ℃ or more by extracting 2 to 5 times with water of 20 to 50 times the total amount of extraction by a hydrothermal extraction method without pressure to the complex herbal extracts available in polar solvents Extracts can be obtained, and the inventors have referred to the extracts produced by the above method as a disintegrant.

On the other hand, 3 ~ 10 times of 10% NaCl aqueous solution was added to the dry herbal pulverized powder containing baekbaekpi, bokyeong, brown root, mold opening, safflower, shiho and windproof and soaked for 30 to 52 hours and then dried, and then dried. 10-fold 15% aqueous ethanol solution was added and then immersed for 48-96 hours, followed by drying. The powder obtained was extracted by 3-10-fold water, methanol, or 70% alcohol by a similar procedure to the extraction of the above and the above-mentioned decanter. Complex herbal extracts produced by a method for preparing a pharmaceutical composition for preventing or treating liver disease can be obtained, and the present inventors referred to the extract produced by the method as poze.

The present invention provides a pharmaceutical composition for the prevention and treatment of liver disease, which comprises the extract of medicinal herb extract, leprosy or nasal extract obtained by the above-mentioned preparation and comprises a pharmaceutically acceptable carrier, excipient or diluent.

Herbal medicine extracts for preventing and treating liver diseases of the present invention is not limited thereto, but is not limited to 40 to 60% by weight of lettuce, 10 to 20% by weight, fugu 10 to 20% by weight, 3 to 8% by weight, safflower 3 to 8 It is preferable to contain the wt%, 3 to 8% by weight, and 3 to 8% by weight windproof because it shows excellent activity. The herbal extracts have been used as food for a long time as the extract of the present invention extracted therefrom also has no problems such as toxicity and side effects.

In addition, as a result of examining the effects on the carbon tetrachloride-induced chronic liver disorder of the composition obtained by the above method, the weight was increased, the number was increased in hematological tests such as leukocytes, red blood cells, etc., AST (aspartate transaminase) increased by hepatotoxicity ), Alanine transaminase (ALT), alkaline phosphatase (ALP), and total bilirubin (TB) levels have been shown to improve liver damage.

Pharmaceutical dosage forms of the extracts of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds, as well as in a suitable collection.

Pharmaceutical compositions comprising extracts according to the invention, in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterile injectable solutions, respectively, according to conventional methods. Can be formulated and used. Carriers, excipients and diluents that may be included in the composition comprising the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid preparations may include at least one excipient such as starch, calcium carbonate and sucrose in the extract. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the compositions of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably 0.001 to 100 mg / kg per day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

The extract of the present invention can be administered to mammals such as mice, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

Complex herbal extract of the present invention is a drug that can be used with confidence even when taken for a long time for the purpose of prevention because there is little toxicity and side effects.

In addition, the present invention provides a health functional food for improving liver function, including the complex herbal extract and food supplements acceptable food supplement. The health functional food of the present invention includes the form of tablets, capsules, pills, or liquids, and the food to which the compound of the present invention can be added includes, for example, various foods, beverages, gums, teas, vitamins, and the like. Complex, health functional foods, and the like.

In detail, the present invention is a functional health food for preventing and treating liver disease, including a complex herbal extract containing baekbaekpi, bokyeong, brown roots, molds, safflower, shiho and windproof as an active ingredient and a food supplement acceptable additives To provide. The health functional food of the present invention includes the complex herbal extract in the form of a ground powder, extract or powder extract.

The composition containing the extract of the complex herbal medicine of the present invention can be used in a variety of drugs, food and beverages for the prevention and treatment of liver disease. Foods to which the complex herbal extract of the present invention may be added include various foods, for example, beverages, gums, teas, vitamin complexes, health supplements, and the like, pills, powders, granules, acupuncture tablets, capsules, or the like. It can be used in the form of a drink. At this time, the amount of the extract in the food or beverage may be added in 0.01 to 15% by weight of the total food weight, the health beverage composition may be added in a ratio of 0.02 to 5 g, preferably 0.3 to 1g based on 100 ml. have.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for having the extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates, etc. as additional ingredients, as in general beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, 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 said 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 extract 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 and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the extracts of the present invention may contain flesh for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is usually selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the extract of the present invention.

Hereinafter, the present invention will be described in detail by the following Examples and Experimental Examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following Examples and Experimental Examples.

Example 1 Preparation and Preparation of Samples

1-1. Preparation of Morus bark

It was purchased from Daegu Yangnyeong Market and used as an experimental material for monodrug.

1-2. Preparation of complex herbal medicines (including moxa)

Sangbaekpi, Siho, windbreak and hyunggae were purchased at Daegu Yangnyeong Market, and Brown Root, Baekbokyeong, and Hwanghwa seeds were purchased at Gyeongdong Market, and used as a test material for mixed compound with the following weight. At the time of preparation of the above-mentioned tablets, 500g of baekbaekpi, 150g of Baekbokyeong, 150g of brown root, 50g of mold hyung, 50g of safflower seed, 50g of shiho and 50g of windbreak were mixed to prepare a total of 1kg.

1-3. Preparation of Complex Herbal Formulas Including Morus bark

3 kg of the dry matter preparation prepared in Example 1-2 was added to 10 l of 10% brine, and then immersed for 48 hours, and then dried for 48 hours in the shade, and then again immersed in 10 liters of 15% alcohol concentration for 72 hours. Next, it was dried for 48 hours in the shade to prepare a composite herbal formulation.

Example 2. Preparation of Water Extract of Samples

1 kg of each dry sample prepared in Examples 1-1, 1-2, and 1-3 was added to 10 L of distilled water, boiled for 8 hours, cooled, and filtered through a filter cloth (Hygiene extract porcelain, Hana Industrial Co., Ltd.). Then, the filtrate was concentrated under reduced pressure using a reduced pressure concentrator (NE-1, EYELA, JAPAN) to a volume of 3L, and then freeze-dried to obtain 12.5g, 13.1g, and 12.1g of each powder extract.

Example 3. Preparation of Methanol Extract of a Sample

1 kg of each dry sample prepared in Examples 1-1, 1-2, and 1-3 was added to 10 liters of methanol and deposited in a 4-liter refrigerator for 72 hours by cold extraction, followed by filtration with a filter cloth and then filtrate 3 liters. Concentrated under reduced pressure using an evaporator to the volume, and then freeze-dried to obtain 10.8g, 11.0g, 10.4g dry powder of each methanol extract.

Example 4 Preparation of 70% Alcohol Extract of a Sample

2 kg of each dry sample prepared in Examples 1-1, 1-2, and 1-3 was added to 10 l of 70% alcohol, and a cooling tube was installed, then boiled at 85 ° C. for 8 hours, extracted, and cooled again. Next, a primary extract was obtained using a filter cloth, and the residue was put into a container again, and 6 liters of 70% alcohol was added to extract and filter in the same manner as above to obtain a secondary extract. On the other hand, after repeating the above process to obtain a tertiary extract, the first, second and tertiary extracts were all mixed and concentrated to a final volume of about 3 liters under a reduced pressure concentrator below 50 ° C. The concentrate was freeze-dried in a freeze-drier square tray (40 x 30 x 4cm) 1 l each in a freeze and freeze dryer to obtain 13.5g, 12.8g, 13.9g (yield 13%) of 70% alcohol extract powder.

Reference Example 1. Preparation of experimental animals and induction of liver toxicity

In order to confirm the improvement effect on the liver damage of the composition according to the present invention using the extract prepared in Examples 2 to 4 using the following experimental method to improve the effect on liver liver damage caused by carbon tetrachloride I looked at it.

1-1. Preparation of Laboratory Animals

The experimental animals used SD (Sparague-Dawley) male rats (Samtaco Bio Korea Co., Ltd.) with a body weight of about 200 g. The feed (Sampaco Co.) and drinking water were freely ingested, and the temperature of the kennel was 23 ± 2˚C. Relative humidity was maintained at 60 ± 10%, ventilation frequency 10 ~ 15 times / hr, lighting cycle 12hr lighting / 12hr off, illuminance 150 ~ 300 Lux.

1-2. Induce liver toxicity by carbon tetrachloride treatment and sample administration

Reagents were purchased from CCl ₄ (carbon tetrachloride Sigma Co.), corn oil, alanine amino trasperase (ALT, GPT, Stanbio Co.) and aspartate amino transferase (AST, GOT, Stanbio Co.).

To induce hepatotoxicity, a single dose of CCl ₄ (carbon tetrachloride Sigma Co.) and corn oil (1: 1) (v / v) was administered to each rat at a single dose of 1.0 ml / kg per kg. Orally, it was administered orally 4 times for 2 weeks at 0.5ml / kg at 3-4 days intervals. That is, CCl ₄ was orally administered at 0, 7, 10, 14, 17, and 21 days from the start of the test, respectively, in the experimental group, the extracts of Examples 2 to 4 were administered for 2 weeks after the last exposure of CCl ₄ , and the normal group Saline is administered and positive control is administered. Carbon tetrachloride (CCL ₄ ) was administered. Experimental group settings and experimental schedules are listed in Table 1.

Extraction Method Dose (mg / kg) Composition Marisu Water extract Normal 10 Positive control 10 50 Monoclonal repellent 10 10 10 100 Monoclonal repellent 10 10 10 200 Monoclonal repellent 10 10 10 Methanol extract Normal 10 Positive control 10 50 Monoclonal repellent 10 10 10 100 Monoclonal repellent 10 10 10 200 Monoclonal repellent 10 10 10 70% spirit extract Normal 10 Positive control 10 50 Monoclonal repellent 10 10 10 100 Monoclonal repellent 10 10 10 200 Monoclonal repellent 10 10 10

1-3. Statistical processing of data

Levene's test was performed to compare the homogeneity of the variances for each test result.If the variances were homogeneous, ANOVA (one-way analysis of variance) was performed. Dunnett's t-test was performed to identify the test group with the difference. In other words, if the variance is heterogeneous, a proper data transformation was performed, and if the variance was the same, the ANOVA test was performed. On the other hand, if the variances are not homogeneous, non-parametric ANOVA tests were conducted, and if the results were significant, the appropriate t-test method was selected for statistical analysis (SPSS Inc.). USA).

Experimental Example 1. Measurement of body weight after administration of the extract of the present invention

After the administration of the extract of the present invention according to the schedule of Reference Example 1 was examined the weight change of the test animal. Body weights of the test animals used for the test were measured at the time of separation of the preliminary group, once / 7 days after the start of administration of the test substance and immediately before the autopsy. The results are shown in Table 2.

extract Dose (mg / kg) Week 0 1 week 2 weeks 3 weeks 4 Weeks Water extract Normal 215 ± 5 247 ± 14 263 ± 11 295 ± 13 335 ± 15 Positive control 211 ± 10 243 ± 12 255 ± 13 277 ± 15 289 ± 22 50 210 ± 9 ^a 239 ± 16 251 ± 16 285 ± 19 293 ± 12 216 ± 6 ^b 240 ± 8 253 ± 14 283 ± 18 301 ± 19 212 ± 6 ^c 243 ± 11 258 ± 12 288 ± 12 315 ± 13 100 222 ± 8 ^a 238 ± 13 259 ± 11 287 ± 16 305 ± 12 209 ± 6 ^b 239 ± 16 256 ± 21 285 ± 15 311 ± 7 213 ± 6 ^c 245 ± 12 257 ± 13 291 ± 12 321 ± 14 200 211 ± 13 ^a 243 ± 10 259 ± 14 285 ± 11 316 ± 17 215 ± 6 ^b 236 ± 14 255 ± 21 290 ± 13 318 ± 16 210 ± 5 ^c 241 ± 12 268 ± 11 298 ± 13 329 ± 15 Methanol extract Normal 231 ± 15 261 ± 11 297 ± 16 334 ± 19 358 ± 22 Positive control 225 ± 9 251 ± 9 273 ± 14 195 ± 15 301 ± 17 50 226 ± 10 ^a 252 ± 15 277 ± 16 291 ± 18 311 ± 13 234 ± 14 ^b 255 ± 11 279 ± 15 294 ± 14 313 ± 24 100 230 ± 12 ^a 251 ± 13 273 ± 10 296 ± 5 325 ± 22 222 ± 8 ^b 259 ± 10 285 ± 17 301 ± 21 329 ± 19 200 228 ± 6 ^a 253 ± 16 271 ± 13 299 ± 14 326 ± 21 225 ± 8 ^b 263 ± 19 282 ± 20 303 ± 18 330 ± 13 70% spirit extract Normal 223 ± 13 263 ± 17 293 ± 20 323 ± 25 353 ± 21 Positive control 221 ± 11 253 ± 18 277 ± 21 296 ± 19 313 ± 22 50 226 ± 13 ^a 259 ± 19 287 ± 15 301 ± 21 321 ± 19 220 ± 17 ^b 254 ± 13 289 ± 15 309 ± 17 339 ± 24 217 ± 13 ^c 255 ± 14 293 ± 17 315 ± 12 325 ± 16 100 219 ± 6 ^a 244 ± 12 284 ± 17 304 ± 21 334 ± 23 221 ± 7 ^b 258 ± 10 288 ± 13 314 ± 26 331 ± 21 225 ± 11 ^c 261 ± 10 289 ± 10 318 ± 15 347 ± 19 200 226 ± 11 ^a 251 ± 8 290 ± 16 319 ± 19 338 ± 25 220 ± 8 ^b 258 ± 9 283 ± 9 318 ± 15 345 ± 22 217 ± 10 ^c 252 ± 11 289 ± 13 321 ± 13 355 ± 25

1-1. Body weight change in water extract group

During the experimental period, the weight of all groups showed a tendency to increase continuously, but the positive control group showed a tendency to lose weight compared to the normal control group. Especially, after 4 weeks of experiment, the weight of the control group was significantly lower than that of the control group ( p <0.01). However, Mori Cortex water extract showed a tendency to regain the weight by the administration, the particular stage control and recovery control of weight significantly compared to the positive control group to 200 mg / kg administration group and Posay experiment after 4 weeks at 100 and 200 mg / kg group (p <0.05 or p <0.01) to increase (see Table 2).

1-2. Body weight change in the methanol extract group

During the experimental period, the weight of all groups showed a tendency to increase continuously, but the positive control group showed a tendency to lose weight compared to the normal control group. In particular, the body weight of the positive control group was significantly decreased ( p <0.01) at 4 weeks after the experiment. However, the body weights tended to recover in the monoclonal 200 mg / kg and the 100 and 200 mg / kg administration groups, and increased significantly ( p <0.05) compared to the positive control at the fourth week of the experiment (see Table 2).

1-3. Body weight change in 70% alcohol extract group

During the experimental period, the weight of all groups showed a tendency to increase continuously, but the positive control group showed a tendency to lose weight compared to the normal control group. In particular, at 4 weeks, the body weight of the positive control group was significantly decreased ( p <0.05) compared to the normal control group. However, the body weight loss tended to recover compared to the positive control group in the 200 mg / kg and 100 mg and 200 mg / kg injection groups, and increased significantly ( p <0.05) compared to the positive control group at 4 weeks. See Table 2).

Experimental Example 2 Hematological Measurements after Administration of the Extract of the Present Invention

After the administration of the extract of the present invention according to the schedule of Reference Example 1 was performed a hematological test. Specifically, after anesthetizing the animals fasted 20 to 24 hours before the autopsy, a portion of the blood obtained by pre-bleeding from the abdominal aorta is placed in an EDTA tube (BD vaccutainer, USA) and stored well, and then white blood cell count (white) within 3 hours. blood cells (WBCs), red blood cells (RBCs), red blood cell volumes (hematocrits (Hct), hemoglobin (Hb) and platelets (platelets, PLT) Measured using. The results are shown in Table 3.

extract Dose (mg / kg) WBC (10 ³ / μl) RBC (10 ⁶ / μl) Hb (g / μl) Hct (%) Plt (10 ³ / μl) Water extract Normal 7.54 ± 0.45 8.05 ± 0.37 14.8 ± 0.5 48.2 ± 1.5 765 ± 54 Positive control 9.89 ± 0.74 6.13 ± 0.53 12.6 ± 0.3 36.5 ± 1.3 713 ± 43 50 8.87 ± 0.23 ^a 6.43 ± 0.23 12.9 ± 0.4 35.3 ± 1.2 715 ± 62 7.80 ± 0.43 ^b 6.32 ± 0.61 13.3 ± 0.2 37.3 ± 0.6 721 ± 76 7.72 ± 0.61 ^c 7.03 ± 0.55 13.5 ± 0.5 38.5 ± 0.9 788 ± 54 100 7.75 ± 0.54 ^a 7.57 ± 0.41 13.7 ± 0.3 41.5 ± 1.3 695 ± 51 8.65 ± 0.66 ^b 6.54 ± 0.32 13.4 ± 0.4 39.5 ± 2.3 746 ± 68 7.78 ± 0.51 ^c 7.72 ± 0.55 13.9 ± 0.5 41.3 ± 0.6 802 ± 59 200 8.83 ± 0.73 ^a 7.87 ± 0.42 14.5 ± 0.2 44.4 ± 1.1 745 ± 91 7.65 ± 0.65 ^b 7.35 ± 0.51 13.9 ± 0.5 42.7 ± 0.8 753 ± 42 7.57 ± 0.39 ^c 7.55 ± 0.85 14.3 ± 0.4 45.6 ± 1.3 798 ± 55 Methanol extract Normal 5.65 ± 0.32 7.43 ± 0.21 13.9 ± 0.5 41.7 ± 1.3 821 ± 43 Positive control 6.32 ± 0.55 7.12 ± 0.32 12.5 ± 0.8 37.3 ± 2.6 765 ± 37 50 6.02 ± 0.65 ^a 6.98 ± 0.54 12.1 ± 0.6 36.6 ± 2.4 731 ± 25 4.15 ± 0.72 ^b 6.87 ± 0.36 12.8 ± 0.7 38.3 ± 3.7 765 ± 31 100 6.69 ± 0.39 ^a 7.34 ± 0.54 13.3 ± 0.7 40.3 ± 1.5 755 ± 34 6.98 ± 0.54 ^b 7.35 ± 0.32 13.9 ± 0.5 42.0 ± 0.9 743 ± 55 200 7.60 ± 0.67 ^a 7.44 ± 0.69 13.6 ± 0.4 41.0 ± 3.4 787 ± 47 5.54 ± 0.43 ^b 7.49 ± 0.90 13.7 ± 0.5 40.4 ± 2.7 765 ± 41 70% spirit extract Normal 8.14 ± 0.74 8.45 ± 0.42 15.3 ± 0.8 48.2 ± 3.1 735 ± 21 Positive control 12.11 ± 0.94 6.32 ± 0.77 12.3 ± 0.5 35.3 ± 3.9 541 ± 34 50 11.32 ± 0.65 ^a 7.05 ± 0.43 13.3 ± 0.6 40.2 ± 3.9 643 ± 44 10.84 ± 0.83 ^b 7.23 ± 0.37 13.7 ± 0.4 41.2 ± 3.0 657 ± 55 9.25 ± 0.45 ^c 7.88 ± 0.54 14.2 ± 0.6 42.2 ± 4.5 687 ± 65 100 9.97 ± 0.65 ^a 7.83 ± 0.92 14.1 ± 0.5 44.2 ± 1.5 683 ± 58 8.78 ± 0.76 ^b 7.43 ± 0.57 13.9 ± 0.7 42.2 ± 1.0 691 ± 48 8.87 ± 0.47 ^c 8.21 ± 0.85 14.6 ± 0.9 45.5 ± 1.7 692 ± 43 200 9.44 ± 0.71 ^a 8.21 ± 0.45 15.0 ± 0.3 46.2 ± 1.7 721 ± 43 8.84 ± 0.69 ^b 8.35 ± 0.63 15.2 ± 0.4 45.7 ± 2.8 702 ± 63 8.38 ± 0.41 ^c 8.54 ± 0.67 15.5 ± 0.9 48.6 ± 3.5 727 ± 57

2-1. Hematological test in water extract group

In the positive control group, the WBC level was significantly increased and the RBC, Heb, and Hct level were significantly decreased ( p <0.05) compared to the normal control group. However, both groups treated with 200 mg / kg of epidermis showed a general tendency to recover compared with the positive control group. In particular, WBC levels were significantly reduced to control levels by the administration of 200 mg / kg of the abdominal and sieves. Heb levels increased significantly ( p <0.05) compared to the positive control group by mg / kg administration, and Hct levels increased significantly ( p <0.05) compared to the positive control group by the administration of 200 mg / kg of foam (Table 3). Reference).

2-2. Hematological test in methanol extract group

The positive control group showed increased WBC levels and decreased RBC, Heb, Hct and Plt levels compared to the normal control group. However, RBC, Heb and Hct levels showed a tendency to recover to the normal control group by administration of 100 or 200 mg / kg of epidermis, especially WBC levels to the control group. (See Table 3).

2-3. Hematological examination in 70% alcohol extract group

In the positive control group, the WBC level was significantly increased and the RBC, Heb, Hct and Plt levels were significantly decreased ( p <0.05 or p <0.01). However, it showed a tendency to recover overall compared with the positive control group by the administration of epithelial skin. For WBC values, the monoclonal drug 100 or 200 mg / kg, the monoclonal drug 100 mg / kg and the antifoam 50 or 200 mg / kg, were positive. significantly higher than the control group (p <0.05 or p <0.01 ) decrease was, RBC numbers in stage control and recovery control 200 mg / kg administered with Posay 100 or 200 mg / by kg dose significantly higher than the positive control (p in <0.05 or p <0.01). Heb level was significantly increased ( p <0.05 or p <0.01) compared with the positive control group by monoclonal 100 or 200 mg / kg, oral remedy 200 mg / kg and sieve 50, 100 or 200 mg / kg. In the Hct level, the amount of p- 0.05 or p <0.01) was significantly increased compared to the positive control group by the administration of monoclonal and biphasic 200 mg / kg and sieve 100 or 200 mg / kg. In addition, the Plt level was significantly increased ( p <0.05 or p <0.01) compared with the positive control group by the administration of 100 or 200 mg / kg monoclonal and oral tablets and 200 mg / kg of foam (see Table 3). .

Experimental Example 3 Blood Biochemical Tests After Administration of the Extract of the Present Invention

After administering the extract of the present invention according to the schedule of Reference Example 1, the rats fasted 20 to 24 hours before the autopsy were subjected to ether anesthesia, and the blood obtained by prebleeding in the abdominal aorta was left at room temperature for 30 minutes to coagulate, followed by centrifugation ( AST (aspartate transaminase), ALT (alanine transaminase), ALP () alkaline phosphatase, TP (total protein), Alb (albumin), TB (total bilirubin), T-CHO Blood biochemical tests for total cholesterol (TG), triglyceride (TG), low density lipoprotein-cholesterol (LDL-C), and high density lipoprotein-cholesterol (HDL-C) were performed using an automated analyzer (Hitachi-747, Hitachi medical. Japan). It was measured by. The results are shown in Tables 4 and 5.

extract Dose (mg / kg) TP (g / ㎗) Albumin (g / ㎗) AST (IU / ℓ) ALT (IU / ℓ) ALP (mg / dl) TB (mg / dl) Water extract Normal 6.7 ± 0.4 3.4 ± 0.2 87 ± 14 54 ± 11 256 ± 30 0.10 ± 0.01 Positive control 6.5 ± 0.6 3.2 ± 0.3 896 ± 124 345 ± 67 578 ± 92 0.43 ± 0.17 50 6.6 ± 0.5 ^a 3.3 ± 0.2 653 ± 113 234 ± 75 435 ± 87 0.34 ± 0.04 6.5 ± 0.3 ^b 3.2 ± 0.3 726 ± 152 251 ± 43 421 ± 80 0.27 ± 0.03 6.6 ± 0.4 ^c 3.4 ± 0.3 556 ± 45 181 ± 32 337 ± 40 0.19 ± 0.02 100 6.8 ± 0.2 ^a 3.3 ± 0.3 696 ± 127 269 ± 27 441 ± 73 0.21 ± 0.02 6.4 ± 0.3 ^b 3.4 ± 0.4 542 ± 130 283 ± 43 367 ± 56 0.32 ± 0.05 6.5 ± 0.6 ^c 3.3 ± 0.2 432 ± 72 151 ± 33 293 ± 77 0.20 ± 0.05 200 6.6 ± 0.6 ^a 3.3 ± 0.3 587 ± 164 271 ± 50 378 ± 72 0.26 ± 0.04 6.6 ± 0.4 ^b 3.5 ± 0.2 496 ± 94 229 ± 38 321 ± 83 0.18 ± 0.05 6.5 ± 0.6 ^c 3.5 ± 0.3 326 ± 54 131 ± 43 276 ± 43 0.15 ± 0.05 Methanol extract Normal 6.3 ± 0.3 3.2 ± 0.2 113 ± 26 71 ± 13 186 ± 54 0.10 ± 0.02 Positive control 6.2 ± 0.5 3.0 ± 0.3 438 ± 134 235 ± 32 354 ± 57 0.41 ± 0.25 50 6.3 ± 0.2 ^a 3.1 ± 0.3 335 ± 44 212 ± 55 326 ± 45 0.24 ± 0.03 6.0 ± 0.4 ^b 3.2 ± 0.2 356 ± 87 223 ± 24 302 ± 37 0.15 ± 0.05 100 6.2 ± 0.3 ^a 3.3 ± 0.2 297 ± 57 185 ± 21 285 ± 34 0.13 ± 0.02 6.3 ± 0.5 ^b 3.0 ± 0.3 358 ± 83 205 ± 35 264 ± 51 0.20 ± 0.07 200 6.4 ± 0.3 ^a 3.0 ± 0.4 288 ± 65 135 ± 36 270 ± 28 0.21 ± 0.04 6.2 ± 0.4 ^b 3.1 ± 0.2 294 ± 54 127 ± 23 314 ± 42 0.16 ± 0.03 70% spirit extract Normal 6.9 ± 0.3 3.5 ± 0.3 85 ± 17 56 ± 11 211 ± 32 0.10 ± 0.02 Positive control 6.2 ± 0.7 3.1 ± 0.4 1125 ± 235 643 ± 113 653 ± 98 1.24 ± 0.27 50 6.6 ± 0.6 ^a 3.3 ± 0.2 874 ± 132 524 ± 122 543 ± 76 0.86 ± 0.24 6.7 ± 0.5 ^b 3.4 ± 0.3 816 ± 178 517 ± 101 512 ± 83 0.61 ± 0.17 6.6 ± 0.3 ^c 3.3 ± 0.5 596 ± 85 437 ± 85 387 ± 56 0.20 ± 0.07 100 6.6 ± 0.2 ^a 3.3 ± 0.4 764 ± 121 434 ± 95 526 ± 55 0.67 ± 0.12 6.7 ± 0.4 ^b 3.5 ± 0.3 783 ± 145 456 ± 132 492 ± 46 0.55 ± 0.15 6.5 ± 0.7 ^c 3.2 ± 0.3 436 ± 94 317 ± 43 323 ± 39 0.31 ± 0.06 200 6.6 ± 0.3 ^a 3.5 ± 0.3 774 ± 135 332 ± 86 413 ± 34 0.63 ± 0.14 6.6 ± 0.3 ^b 3.5 ± 0.3 642 ± 128 324 ± 73 463 ± 63 0.37 ± 0.05 6.5 ± 0.5 ^c 3.6 ± 0.5 212 ± 58 189 ± 33 289 ± 56 0.22 ± 0.04

extract Dose (mg / kg) T-CHO (mg / dl) TG (mg / dl) HDL-C (mg / dl) LDL-C (mg / dl) Water extract Normal 75.3 ± 5.8 48.0 ± 5.3 15.3 ± 3.5 13.3 ± 2.6 Positive control 118.6 ± 13.1 110.0 ± 11.2 29.4 ± 5.6 20.4 ± 2.2 50 85.4 ± 6.2 ^a 92.3 ± 4.2 24.4 ± 3.2 18.7 ± 2.0 96.1 ± 4.4 ^b 88.2 ± 4.2 23.3 ± 2.1 17.9 ± 3.3 84.1 ± 5.3 ^c 72.5 ± 9.4 25.2 ± 1.8 19.9 ± 3.6 100 84.5 ± 11.5 ^a 74.6 ± 4.8 21.7 ± 1.6 18.1 ± 1.9 82.3 ± 7.1 ^b 71.2 ± 3.4 24.6 ± 3.1 19.6 ± 2.7 79.5 ± 5.4 ^c 66.0 ± 5.2 23.7 ± 2.5 17.0 ± 3.4 200 78.3 ± 5.6 ^a 62.1 ± 7.2 20.5 ± 4.1 16.7 ± 1.6 65.5 ± 8.4 ^b 55.2 ± 3.8 20.2 ± 3.8 15.4 ± 2.4 68.5 ± 6.3 ^c 57.2 ± 5.8 22.6 ± 3.2 15.5 ± 3.4 Methanol extract Normal 68.2 ± 4.3 59.9 ± 4.5 13.4 ± 1.7 15.0 ± 3.8 Positive control 107.1 ± 8.0 84.6 ± 6.4 24.5 ± 2.6 35.4 ± 6.0 50 85.3 ± 4.9 ^a 76.8 ± 4.6 22.5 ± 3.3 27.3 ± 4.1 77.0 ± 7.4 ^b 77.2 ± 5.3 22.6 ± 3.1 26.3 ± 2.3 100 81.7 ± 10.2 ^a 63.4 ± 6.4 19.3 ± 2.3 21.3 ± 4.0 74.4 ± 6.6 ^b 66.6 ± 3.7 20.5 ± 3.4 18.8 ± 3.9 200 70.5 ± 6.8 ^a 59.6 ± 4.5 19.3 ± 1.7 17.5 ± 4.3 66.6 ± 4.2 ^b 64.3 ± 3.2 15.9 ± 2.1 15.6 ± 3.1 70% spirit extract Normal 37.8 ± 3.1 65.3 ± 2.3 23.3 ± 2.5 11.3 ± 1.8 Positive control 98.6 ± 5.5 118.5 ± 10.3 48.4 ± 5.3 38.9 ± 2.7 50 73.2 ± 4.9 ^a 98.5 ± 6.3 34.2 ± 2.3 33.1 ± 2.3 79.3 ± 5.3 ^b 88.6 ± 3.8 35.7 ± 5.4 31.2 ± 2.8 74.3 ± 6.9 ^c 77.6 ± 4.8 33.7 ± 2.4 29.2 ± 2.2 100 68.7 ± 4.1 ^a 74.5 ± 5.4 29.6 ± 3.4 24.4 ± 2.5 71.4 ± 3.8 ^b 81.5 ± 4.5 32.5 ± 2.6 21.6 ± 2.4 70.3 ± 6.3 ^c 76.7 ± 5.7 29.2 ± 3.7 20.5 ± 1.8 200 64.6 ± 4.8 ^a 73.4 ± 4.9 28.5 ± 3.4 18.6 ± 1.7 60.1 ± 3.3 ^b 68.4 ± 3.4 27.3 ± 2.6 16.2 ± 1.5 64.1 ± 4.4 ^c 65.2 ± 7.4 26.5 ± 1.8 14.5 ± 1.3

3-1. In case of water extract administration group

Hepatotoxicity by carbon tetrachloride showed a significant increase in AST, ALT and ALP levels in all groups compared to normal controls. In particular, in the case of AST and ALT, all groups increased significantly ( p <0.01) compared to the normal control group. However, as the extracts from the extracts were reduced compared to the positive control group, AST levels were increased by 50 or 200 mg / kg monoclonal drug, 100 or 200 mg / kg monoclonal drug, and 50, 100 or 200 mg / kg foam drug. There was a significant decrease ( p <0.05 or p <0.01) compared to the positive control. The ALT level was significantly decreased ( p <0.05 or p <0.01) in the dose group of 200 mg / kg of tablets and 50, 100 or 200 mg / kg of drug. Similarly to ALT, the concentration of ALP was significantly decreased ( p <0.01) in the group treated with sieve 50, 100 or 200 mg / kg. In the total bilirubin (TB) level, the positive control group increased significantly ( p <0.01) compared to the normal control group, but it showed a tendency to be suppressed throughout the whole epithelium-treated group. Significantly decreased ( p <0.05 or p <0.01) by mg / kg and sieve 50, 100 or 200 mg / kg administration (see Table 4).

On the other hand, the serum lipid content of TG, T-CHO and HDL-C in the positive control group was significantly increased ( p <0.01, p <0.05 or p <0.05, respectively) compared to the normal control group, and LDL-C The numbers also tended to increase. However, it showed a tendency to recover by the administration of water extracts from the epidermis. The TG level was significantly higher than that of the positive control group by 100 or 200 mg / kg monoclonal and oral tablets and 50, 100 or 200 mg / kg foam. ( p <0.05 or p <0.01) decreased. In addition, T-CHO levels decreased significantly ( p <0.05 or p <0.01) compared to the positive control group, and recovered in HDL-C and LDL-C levels compared to the positive control group. Trends are shown (see Table 5).

3-2. Methanol extract administration group

Hepatotoxicity by carbon tetrachloride showed a significant ( p <0.01) increase in AST, ALT, ALP and TB levels in the positive control group compared to the normal control group. In particular, AST increased significantly ( p <0.01) in all groups except the normal control group, but was significantly ( p <0.05) compared to the positive control group by the administration of 100 or 200 mg / kg monoclonal drug and 200 mg / kg monoclonal drug. ALT showed a significant decrease ( p <0.05) in the ALT compared to the positive control group by the administration of mono- and multi-drug 200 mg / kg. ALP levels were sporadically decreased ( p <0.05 or p <0.01) in the positive control group, 50 mg / kg monoclonal drug, and 50 or 200 mg / kg monoclonal drug compared to the normal control group. Showed a tendency to recover. In the total bilirubin (TB) level, the positive control group increased significantly ( p <0.01) compared with the normal control group, but showed a tendency to be suppressed throughout the epithelial treatment group, including the monoclonal 50 mg / kg ( p <0.05) group. Sun decreased significantly ( p <0.01) in all epithelial treated groups (see Table 4).

In serum lipid content, TG, T-CHO and LDL-C levels in the positive control group were significantly increased ( p <0.05) compared to the normal control group, and HDL-C levels also increased. However, administration of the extract of M. alba extract showed a tendency to recover compared to the positive control group. The TG value was significantly decreased ( p <0.05) compared with the positive control group by the administration of 100 or 200 mg / kg of single- or double-release drugs. The -CHO level also showed a significant decrease ( p <0.05) in the monoclonal 200 mg / kg and the multiclonal groups compared to the positive control group. In addition, LDL-C levels also showed a tendency to recover significantly ( p <0.05) in the monoclonal 200 mg / kg and the 100 or 200 mg / kg monoclonal administration groups compared to the positive control group (see Table 5).

3-3. 70% alcohol extract group

Hepatotoxicity by carbon tetrachloride showed a significant ( p <0.01) increase in AST, ALT, ALP and TB levels in the positive control group compared to the normal control group. In particular, AST showed a tendency to increase significantly ( p <0.01) in all groups compared to the normal control group. However, the AST level showed a tendency to recover significantly ( p <0.01) compared to the positive control group in all groups including monoclonal 50 mg / kg ( p <0.05). Similar trends were observed for ALT levels. ALT also showed a significant increase ( p <0.01) in all groups compared to normal controls, but with monoclonal 50, 100 or 200 mg / kg, or 100 or 200 mg / kg and sieve 100 or 200 mg / kg showed a tendency to recover significantly ( p <0.05 or p <0.01) compared to the positive control. In ALP levels, all groups showed a tendency to increase significantly ( p <0.05 or p <0.01), except for 200 mg / kg of sieves, and 200 mg / kg of monoclonal and occlusion (above). p <0.05) and all treatment groups showed a concentration-dependently ( p <0.01) recovery trend compared to the positive control group (see Table 4).

On the other hand, in the lipid content of serum, TG, T-CHO, HDL-C and LDL-C levels of the positive control group were significantly increased ( p <0.05 or p <0.01) compared to the normal control group, but 70% extract of alveolar epidermis There was a tendency to recover compared to the positive control group, and the TG values were significantly higher than those of the positive control group in both the 50 mg / kg sieves and the 100 or 200 mg / kg monoclonal and percutaneous agents ( p <0.05 or p). <0.01) and T-CHO levels also showed a significant decrease ( p <0.05) in both monoclonal 100 mg / kg and 200 mg / kg groups. In addition, LDL-C levels showed a tendency to recover significantly ( p <0.05 or p <0.01) in all 200 mg / kg administration groups, and 200 mg / kg of tablets and foams in HDL-C levels. The administration group showed a significant ( p <0.05) reduction compared to the positive control group (see Table 5).

Experimental Example 4. Determination of lipid content in liver tissue after administration of the extract of the present invention

The total lipid content in liver tissues and feces after administration of the extract of the present invention according to the schedule of Reference Example 1 was determined by Folk et al. (Folch J et al, J Bio Chem , 226, pp 497-502, 1957), Zlatkis A et al, Anal Biochem, 29 , pp143-146, 1969, and triglyceride content were measured by Bix et al. (Biggs HG et al, Clin Chem, 21 , pp437-441, 1975) (see Table 6).

extract Dose (mg / kg) T-lipid (mg / g dry weight) T-CHO (mg / g dry weight) TG (mg / g dry weight) Water extract Normal 222.5 ± 15.3 21.5 ± 3.4 65.4 ± 5.4 Positive control 484.3 ± 34.4 104.8 ± 25.7 126.0 ± 21.6 50 356.1 ± 26.7 ^a 95.7 ± 13.5 121.6 ± 19.7 332.3 ± 21.3 ^b 83.2 ± 13.4 105.1 ± 17.3 292.3 ± 17.3 ^c 77.4 ± 15.3 90.3 ± 12.3 100 342.2 ± 28.7 ^a 87.2 ± 11.4 99.3 ± 8.6 336.1 ± 15.4 ^b 73.5 ± 13.1 81.5 ± 6.2 305.3 ± 23.6 ^c 67.2 ± 17.4 77.3 ± 4.9 200 296.3 ± 21.5 ^a 86.4 ± 12.5 83.8 ± 5.5 276.3 ± 18.6 ^b 71.4 ± 9.3 79.3 ± 6.8 241.4 ± 12.6 ^c 63.5 ± 5.9 67.1 ± 4.2 Methanol extract Normal 198.5 ± 11.2 15.9 ± 5.1 46.3 ± 4.6 Positive control 321.7 ± 13.4 83.5 ± 8.4 97.5 ± 6.2 50 314.3 ± 23.6 ^a 77.4 ± 4.8 95.3 ± 8.2 329.4 ± 21.3 ^b 88.5 ± 12.5 86.3 ± 6.1 100 302.6 ± 17.4 ^a 72.6 ± 9.4 79.4 ± 5.5 296.1 ± 14.5 ^b 65.8 ± 4.3 74.3 ± 4.7 200 303.4 ± 13.7 ^a 59.4 ± 5.6 71.2 ± 4.6 267.5 ± 12.5 ^b 47.5 ± 6.2 66.5 ± 8.5 70% spirit extract Normal 168.8 ± 13.6 19.5 ± 4.5 58.5 ± 3.9 Positive control 394.0 ± 21.4 95.3 ± 8.5 112.1 ± 4.8 50 353.0 ± 17.1 ^a 92.5 ± 7.1 98.6 ± 3.9 322.0 ± 19.3 ^b 82.1 ± 5.5 88.4 ± 5.6 276.0 ± 13.7 ^c 71.1 ± 3.9 83.4 ± 3.2 100 336.5 ± 15.6 ^a 79.3 ± 5.9 85.4 ± 4.7 305.3 ± 12.1 ^b 61.5 ± 6.8 77.8 ± 5.1 289.1 ± 27.3 ^c 43.7 ± 3.5 76.0 ± 4.9 200 315.5 ± 21.6 ^a 65.8 ± 3.4 74.2 ± 4.0 256.1 ± 12.1 ^b 40.4 ± 4.1 69.2 ± 3.6 201.7 ± 13.9 ^c 33.4 ± 2.1 63.6 ± 5.1

4-1. In case of water extract administration group

Lipid content measurement in liver tissue showed a significant increase ( p <0.01) in T-lipid, T-CHO and TG values in the positive control group compared to the normal control group. In particular, T-CHO showed a tendency to increase significantly ( p <0.05 or p <0.01) in all treatment groups compared to the normal control group. However, the group treated with sieve 50, 100 or 200 mg / kg showed a significantly higher concentration-dependent recovery ( p <0.05 or p <0.01) than the positive control group, and the group receiving 200 mg / kg of tablets was significantly more than the positive control group. One ( p <0.05) decrease was shown. Significant increase ( p <0.05) was observed in T-lipid levels in the 50 and 100 mg / kg group of mono- and multi-drugs compared to the normal control group, but positive in all groups treated with mono-, multi-drugs 200 mg / kg and sieves. It showed a significant decrease ( p <0.05 or p <0.01) compared to the control. In addition, a significant increase ( p <0.05 or p <0.01) was observed in all the 50 mg / kg administration groups and the 100 mg / kg monoclonal and peritoneal administration groups compared to the normal control group. In the 100 or 200 mg / kg administration group including the overall control group showed a tendency to recover significantly ( p <0.05 or p <0.01) compared to the positive control group (see Table 6).

4-2. Methanol extract administration group

Lipid content measurement in liver tissue showed a significant increase in T-lipid, T-CHO and TG levels in the positive control group ( p <0.05, p <0.01 or p <0.01), respectively. In particular, T-CHO showed a tendency to increase significantly ( p <0.01) in all treatment groups compared to the normal control group. However, there was a significant decrease ( p <0.01) in the 100 mg / kg of the over-the-counter ( p <0.05) and 200 mg / kg of the mono- and multi-drugs. T-lipid levels In clothing control were significant (p <0.01) increase observed compared to the control group in all groups except for the 200 mg / kg administered group, repeat control 200 mg / kg administration group in is significant compared to the positive control group (p <0.05) reduction was observed. In addition, the TG level was also significantly increased ( p <0.05 or p <0.01) in all groups except the 200 mg / kg of the tablets. / kg administration significantly decreased ( p <0.05 or p <0.01) compared to the positive control group (see Table 6).

4-3. Alcohol 70% extract administration group

Lipid content measurement in liver tissue showed a significant increase ( p <0.01) in T-lipid, T-CHO and TG levels in the positive control group compared to the normal control group. In particular, T-CHO showed a tendency to increase significantly ( p <0.05 or p <0.01) in all treatment groups compared to the normal control group. However, the monoclonal 200 mg / kg, the multiclonal 100 or 200 mg / kg, and sieves showed a significant decrease ( p <0.05 or p <0.01) compared to the positive control group. T-lipid levels were significantly increased ( p <0.05 or p <0.01) in all groups except for 200 mg / kg of tablets and 100 or 200 mg / kg of tablets. In all groups treated with mg / kg and sieves, a tendency to recover significantly ( p <0.05 or p <0.01) was observed compared to the positive control group. In addition, the TG level was significantly increased ( p <0.05 or p <0.01) in all groups except the 200 mg / kg administration group, monoclonal 200 mg / kg, pill and foam 100 or 200 The mg / kg administration group showed a tendency to recover significantly ( p <0.05 or p <0.01) compared to the positive control group.

Experimental Example 5. Determination of lipid content in the stool after administration of the extract of the present invention

The amount of bile acid in the stool was measured using a method of Takahisa et al. (Takahisa T, J. Nutr Sci Vitaminol . 32 , pp111-121, 1986) and extracting the bile acid from the stool using a bile acid kit (Far East Pharmaceuticals, Japan). That is, a 1: 1 amount of acetone and ethanol were added to a predetermined amount of the mixture, mixed for 1 hour, and a predetermined amount of sample was taken from the supernatant obtained by centrifugation twice at 3000 rpm for 20 minutes each, followed by evaporation of the solvent with nitrogen gas. After dissolving with diethyl ether to extract the bile acid in the lower layer, it was measured using a bile acid analysis kit (see Table 7).

extract Dose (mg / kg) T-lipid (mg / g dry weight) T-CHO (mg / g dry weight) TG (mg / g dry weight) Water extract Normal 65.5 ± 3.7 7.32 ± 1.25 5.64 ± 1.21 Positive control 121.2 ± 21.3 21.53 ± 9.4 9.32 ± 2.54 50 130.3 ± 35.4 ^a 17.67 ± 2.61 8.37 ± 3.77 121.2 ± 22.5 ^b 16.45 ± 3.78 8.03 ± 2.54 113.4 ± 23.3 ^c 16.21 ± 7.75 7.65 ± 2.27 100 117.5 ± 36.7 ^a 18.73 ± 5.29 8.43 ± 4.65 135.5 ± 34.8 ^b 16.91 ± 7.21 7.2 ± 3.87 89.3 ± 28.5 ^c 13.44 ± 2.96 8.32 ± 3.91 200 93.6 ± 29.4 ^a 11.52 ± 4.23 7.06 ± 2.43 115.0 ± 36.3 ^b 13.43 ± 2.26 7.76 ± 1.31 88.5 ± 15.3 ^c 15.61 ± 3.98 7.98 ± 1.81 Methanol extract Normal 48.4 ± 5.9 6.85 ± 1.12 6.73 ± 1.32 Positive control 88.5 ± 13.5 17.32 ± 4.78 10.82 ± 1.37 50 89.3 ± 11.6 ^a 15.34 ± 3.72 8.33 ± 3.45 85.5 ± 9.2 ^b 15.26 ± 2.1 8.98 ± 2.32 100 76.3 ± 5.6 ^a 15.63 ± 3.56 7.26 ± 1.20 73.1 ± 12.3 ^b 14.64 ± 1.32 7.98 ± 2.05 200 69.3 ± 4.6 ^a 14.55 ± 3.54 6.43 ± 1.86 55.8 ± 6.9 ^b 13.54 ± 4.13 6.56 ± 3.32 70% spirit extract Normal 45.3 ± 3.9 5.43 ± 2.67 6.43 ± 1.13 Positive control 78.5 ± 6.3 14.94 ± 4.71 11.54 ± 2.98 50 72.1 ± 8.5 ^a 13.87 ± 3.13 11.23 ± 3.45 65.2 ± 5.6 ^b 14.21 ± 2.12 10.54 ± 2.27 63.3 ± 4.8 ^c 12.32 ± 1.92 10.98 ± 3.11 100 65.7 ± 3.7 ^a 9.32 ± 1.66 9.65 ± 1.23 59.3 ± 3.2 ^b 8.32 ± 4.32 7.45 ± 1.76 55.3 ± 6.6 ^c 7.54 ± 3.33 7.32 ± 3.54 200 67.8 ± 5.6 ^a 7.32 ± 1.26 6.97 ± 1.22 53.7 ± 7.4 ^b 7.97 ± 1.03 6.63 ± 0.87 52.6 ± 8.9 ^c 7.85 ± 1.26 6.53 ± 1.02

5-1. In case of water extract administration group

The lipid content of the feces showed that the T-lipid, T-CHO and TG levels in the positive control group increased significantly ( p <0.05) compared to the normal control group. In particular, T-lipid showed a tendency to increase significantly ( p <0.05) in all administration groups except the 200 mg / kg sieve group. However, the group treated with sieve 100 or 200 mg / kg showed a tendency to recover significantly ( p <0.05 or p <0.01) compared to the positive control group. T-CHO levels showed a tendency to increase significantly ( p <0.05) compared to the normal control group, and compared with the positive control group with 200 mg / kg monoclonal and oral tablets and 100 or 200 mg / kg foam. A significant ( p <0.05) decrease was observed. In general, TG levels showed a tendency to recover compared to the positive control group, and decreased significantly ( p <0.05 or p <0.01) in the 200 mg / kg administration group of the abdominal drug and sieve ( p <0.05 or p <0.01). Reference).

5-2. Methanol extract administration group

The lipid content of the feces showed that the T-lipid, T-CHO and TG levels in the positive control group increased significantly ( p <0.05, p <0.01 or p <0.05), respectively. In particular, T-CHO showed a tendency to increase significantly ( p <0.05 or p <0.01) in all administration groups, but decreased in comparison with the positive control group. T-lipid except the 200 mg / kg administration group in the figures, the overall significantly compared to the control group (p <0.05) for increase significantly showed a tendency, as compared to the positive control group to suit Control 200 mg / kg dose of (p <0.05) reduction was observed. In general, the TG level showed a tendency to recover compared to the positive control group, and the mono- and multi-dose 200 mg / kg administration group showed a significant decrease ( p <0.05) compared to the positive control group (see Table 7).

5-3. In case of 70% extract administration group

The lipid content of the feces showed that the T-lipid, T-CHO and TG levels in the positive control group increased significantly ( p <0.05, p <0.01 or p <0.05), respectively. T-CHO levels showed a significant increase ( p <0.05) in both monoclonal, occupantal and sieve 50 mg / kg groups compared to the normal control group. Significant recovery ( p <0.05) was observed by administration compared to the positive control group. T-lipid level was not significantly increased except for 50 mg / kg monoclonal administration ( p <0.05). A significant decrease was observed ( p <0.05) compared to the control. In the TG level, similar to T-lipid, no significant increase ( p <0.05) was observed in the monoclonal 50 mg / kg group compared with the normal control group. A 200 mg / kg dose of control and foams was observed to decrease significantly ( p <0.05) compared to the positive control (see Table 7).

Experimental Example 6. Measurement of antioxidant enzyme activity in liver fraction after administration of extract of the present invention

6-1. Preparation of Liver Fractions

30 mM Hepes containing 150 mM KCl and finely chopped the livers of the rats extracted according to the method of Bansal et al. (Bansal SK et al, Biochem Biophys Res. Commun. 117 , pp268-274, 1983). The mixture was diluted 5 times with buffer (pH 7.4), homogenized, and centrifuged at 700 g for 20 minutes to obtain a supernatant. The supernatant was centrifuged at 11,000 g for 30 minutes to remove pellets. The supernatant was further centrifuged at 105,000 g for 60 minutes to obtain a cytoplasmic fraction. The pellet was washed with 130 mM KCl-containing Hepes buffer and re- homogenized with the same buffer to obtain a microsomal fraction. The whole process of separating the microsomes and the cytoplasmic fraction was carried out in a low temperature room of 0 ~ 4 ℃, was used in the experiment while storing the prepared fraction at -70 ℃.

6-2. Catalase and Superoxide dismutase activity measurements

Catalase activity in the hepatic cytoplasm was measured using the Aebi method (Aebi H. In Method in Enzymology, Colowick SP (Kaplan NO) , pp. 121-126, 1984). Protein was quantified to prepare 0.02 mg protein per ml of sample. At the end of this process, 150 μl of 1 M Tris-HCl buffer (pH 8.0) containing 5 mM EDTA, 2.7 mL of 10 mM H ₂ O ₂ , and 90 μl of distilled water were added, followed by vortex mixing for 30 seconds. . After the reaction mixture was reacted at 37 ° C. for 10 minutes, pipetted by adding 60 μl of a sample prepared with 0.02 mg protein per ml, and the change in optical density at 240 nm was measured for 1 minute (Table 1). 8 and Table 9).

Meanwhile, the activity of SOD in liver cytoplasm was measured using Lee et al. (Lee DW et al, Aging Clin Exp Res. 2 , pp357-362, 1990). Protein was quantified to prepare 2 mg protein per ml of sample. 10 ml of 0.01 N NaOH was dissolved in 0.76 mg of xanthine reagent prepared beforehand, followed by 24.8 mg of cytochrome C, followed by 50 mM phosphate-buffered saline (pH 7.8). 50 μl of each sample prepared with 2 mg of protein per 1 ml was added to 2.9 ml of Solution A, which was made into 110 ml, and mixed by vortex for 30 seconds. Solution B (xanthine oxidase) was added to 2 M ammonium sulfate solution containing 0.1 mM EDTA in the mixed solution. After the solution B is added, the change in absorbance is set to an amount of 0.025 for 1 minute.) After adding 50 µl, pipetting for a sufficient time, the change in absorbance at 550 nm is measured for 1 minute. All lipid peroxides and enzyme activity were measured in a low temperature room at 4 ° C.

Antioxidant Activity and Lipid Peroxidation in Serum Dose (mg / kg) Catalase (IU / mg protein) SOD (IU / mg protein) TBARS (IU / mg Protein) PCOOH (IU / mg protein) Water extract Normal 3.25 ± 0.18 4.03 ± 0.13 1.46 ± 0.18 9.24 ± 1.25 Positive control 8.37 ± 1.05 9.35 ± 2.43 4.54 ± 0.78 13.56 ± 2.89 50 8.45 ± 1.01 9.57 ± 1.14 4.21 ± 0.54 13.16 ± 1.56 8.34 ± 0.97 9.16 ± 2.67 4.53 ± 0.95 13.50 ± 2.43 7.98 ± 0.83 8.99 ± 3.81 4.21 ± 1.53 13.19 ± 1.96 100 7.46 ± 2.36 8.94 ± 1.65 4.06 ± 0.76 12.87 ± 2.34 7.21 ± 1.65 8.83 ± 13.43 3.98 ± 0.84 12.96 ± 2.66 7.02 ± 1.76 8.43 ± 5.01 3.82 ± 0.43 12.54 ± 3.12 200 6.93 ± 0.54 6.42 ± 1.32 3.13 ± 0.21 11.32 ± 1.34 6.73 ± 0.88 6.26 ± 0.77 3.66 ± 0.54 11.09 ± 2.55 6.76 ± 0.93 6.16 ± 1.64 3.23 ± 0.12 10.86 ± 1.98 Methanol extract Normal 4.22 ± 0.24 5.27 ± 0.29 2.05 ± 0.02 8.87 ± 1.12 Positive control 9.65 ± 3.66 9.06 ± 1.68 5.67 ± 0.64 14.54 ± 3.66 50 8.44 ± 1.43 8.96 ± 2.44 5.56 ± 0.87 13.55 ± 3.12 8.15 ± 2.32 8.94 ± 2.79 5.63 ± 0.53 13.67 ± 2.98 100 7.51 ± 1.85 7.76 ± 1.61 4.87 ± 0.95 13.68 ± 7.43 7.77 ± 1.63 7.56 ± 3.66 4.95 ± 1.31 13.98 ± 6.43 200 7.33 ± 2.91 6.78 ± 3.21 4.33 ± 1.13 12.45 ± 3.43 7.46 ± 2.85 6.69 ± 3.78 3.96 ± 0.65 11.32 ± 1.22 70% spirit extract Normal 2.37 ± 0.05 4.92 ± 0.81 1.98 ± 0.04 7.75 ± 1.67 Positive control 9.43 ± 3.51 13.54 ± 1.89 6.43 ± 0.95 13.96 ± 2.54 50 8.53 ± 1.74 11.34 ± 1.89 5.98 ± 1.71 13.31 ± 1.56 8.32 ± 1.97 10.56 ± 1.59 5.56 ± 1.30 13.05 ± 1.12 8.15 ± 4.33 10.67 ± 3.54 5.32 ± 0.82 12.98 ± 3.34 100 7.86 ± 2.65 9.32 ± 1.43 5.17 ± 0.76 12.44 ± 1.49 7.96 ± 2.07 9.04 ± 1.60 5.09 ± 0.45 12.31 ± 1.11 7.43 ± 1.53 8.43 ± 0.92 4.95 ± 0.59 12.08 ± 5.63 200 6.87 ± 1.72 8.32 ± 1.08 4.43 ± 0.61 11.67 ± 3.44 6.54 ± 1.32 8.14 ± 1.5 4.12 ± 0.32 11.21 ± 2.32 5.43 ± 0.13 7.67 ± 0.85 3.85 ± 0.14 10.54 ± 1.85

Antioxidant Activity and Lipid Peroxidation in Liver Tissues Dose (mg / kg) Catalase (IU / mg protein) SOD (IU / mg protein) TBARS (IU / mg Protein) PCOOH (IU / mg protein) Water extract Normal 6.34 ± 0.54 19.7 ± 1.25 2.35 ± 0.78 17.54 ± 2.36 Positive control 10.31 ± 1.91 23.43 ± 3.1 5.43 ± 1.21 35.23 ± 5.65 50 1.15 ± 2.34 22.79 ± 2.56 5.21 ± 1.12 34.21 ± 6.43 10.04 ± 1.41 25.57 ± 3.86 4.90 ± 1.05 32.33 ± 5.82 9.87 ± 1.05 23.52 ± 2.12 5.13 ± 0.98 33.21 ± 6.05 100 9.99 ± 1.14 22.78 ± 1.49 4.67 ± 0.92 33.13 ± 5.65 9.77 ± 0.93 22.55 ± 1.21 4.15 ± 0.84 30.05 ± 6.32 9.74 ± 1.07 22.23 ± 2.78 4.13 ± 1.06 29.96 ± 5.43 200 8.96 ± 0.74 21.76 ± 2.84 3.93 ± 0.93 25.63 ± 3.98 8.26 ± 0.68 20.55 ± 2.53 3.23 ± 0.73 23.83 ± 5.32 8.13 ± 0.76 20.39 ± 1.55 3.15 ± 0.85 22.42 ± 4.56 Methanol extract Normal 5.19 ± 0.59 16.44 ± 2.85 3.23 ± 0.56 14.54 ± 1.38 Positive control 13.38 ± 2.97 20.22 ± 3.76 7.84 ± 1.25 27.80 ± 2.31 50 12.75 ± 1.66 20.45 ± 2.57 7.65 ± 1.14 26.64 ± 3.79 12.53 ± 2.34 20.13 ± 3.54 7.43 ± 0.97 26.32 ± 2.65 100 11.13 ± 1.21 19.77 ± 2.56 7.16 ± 1.05 24.55 ± 3.19 11.05 ± 1.17 19.55 ± 5.68 7.12 ± 1.36 21.23 ± 5.69 200 10.65 ± 2.43 17.51 ± 3.59 6.94 ± 0.95 20.23 ± 4.32 10.23 ± 1.56 18.65 ± 2.45 6.13 ± 0.76 18.69 ± 3.55 70% spirit extract Normal 8.43 ± 1.56 15.32 ± 1.44 3.64 ± 0.57 16.31 ± 2.37 Positive control 16.74 ± 3.86 21.43 ± 2.67 8.64 ± 1.44 36.30 ± 6.29 50 15.66 ± 2.93 20.43 ± 3.41 7.16 ± 0.65 34.21 ± 5.47 14.66 ± 3.14 20.25 ± 2.32 7.13 ± 0.73 33.10 ± 6.54 14.76 ± 2.79 20.12 ± 2.16 7.06 ± 0.85 25.54 ± 4.89 100 13.58 ± 2.34 19.76 ± 3.54 6.83 ± 0.61 31.36 ± 3.43 12.97 ± 1.3 19.26 ± 2.32 6.54 ± 0.95 28.23 ± 5.68 12.66 ± 1.79 19.17 ± 1.22 6.13 ± 0.45 26.30 ± 4.89 200 11.43 ± 1.55 18.13 ± 1.78 5.94 ± 0.85 23.48 ± 3.63 10.67 ± 1.15 17.65 ± 2.85 5.43 ± 0.61 21.59 ± 2.54 1.35 ± 2.31 17.13 ± 1.45 4.86 ± 0.66 21.36 ± 3.71

6-2-1. In case of water extract administration group

Antioxidant enzymes in serum significantly increased ( p <0.01) levels of catalase and SOD in the positive control group compared to the normal control group. In particular, the catalase so in all treated groups significantly compared to the control group (p <0.05 or p <0.01) in the eoteuna show a tendency to increase, stage control, copy control, and Posay 200 mg / kg administered group significantly compared to the positive control group (p < 0.05) showed a tendency to decrease. In SOD, all 50 or 100 mg / kg groups except for 100 mg / kg sieve showed a tendency to increase significantly ( p <0.05 or p <0.01) compared to the normal control group. The 200 mg / kg administration group of the pill and sieve showed a significant decrease ( p <0.05) compared to the positive control group (see Table 8).

In the liver tissues, antioxidant enzyme levels were similar to those of serum, and catalase increased significantly ( p <0.05 or p <0.01) in all treatment groups except for 200 mg / kg fossa group. In the case of the administration of 200 mg / kg of abdominal and sieves, there was a tendency to decrease significantly ( p <0.05) compared to the positive control group. In the SOD, the positive control group and the 50 or 100 mg / kg group except the 200 mg / kg group showed a tendency to increase significantly ( p <0.05 or p <0.01), and the 200 mg / Significant decreases ( p <0.05) were observed with the kg administration compared to the positive control (see Table 9).

6-2-2. Methanol extract administration group

Antioxidant enzymes in serum significantly increased ( p <0.01) levels of catalase and SOD in the positive control group compared to the normal control group. In particular, catalase showed a tendency to increase significantly ( p <0.05) in all the administration groups compared to the normal control group, but the increase in the Methanol extract administration group compared with the positive control group was suppressed concentration-dependently. So in all groups except for the 200 mg / kg treated group even in the SOD significance compared to the control group (p <0.05) in the eoteuna show a tendency to increase, stage control and recovery control 200 mg / kg administered group significantly compared to the positive control group (p < 0.05) showed a tendency to decrease (see Table 8).

In the liver tissues, antioxidant enzyme levels were similar to those of serum. Catalase showed a significant increase ( p <0.01) in all treatment groups, including the positive control group, but was not affected by the methanol extract of the epithelium. The concentration tended to be increased in a concentration-dependent manner. In SOD, the positive control group and the 50 or 100 mg / kg group except the 200 mg / kg group were significantly increased ( p <0.05). The results showed that the mono- and multi-dose 200 mg / kg administration resulted in a significant decrease ( p <0.05) compared to the positive control group (see Table 9).

6-2-3. Alcohol 70% extract administration group

Antioxidant enzymes in serum significantly increased ( p <0.01) levels of catalase and SOD in the positive control group compared to the normal control group. Catalase showed a tendency to increase significantly ( p <0.05 or p <0.01) in all treatment groups compared with the normal control group, but in the mono-, multi-drug and sieve 200 mg / kg group, it was significant ( p <0.01). Tends to decrease). So in all treated group even in the SOD significance compared to the control group (p <0.05 or p <0.01 ) eoteuna show a tendency to increase, 100 or significantly higher than the positive control in both 200 mg / kg group (p <0.05 or p < 0.01) showed a decreasing result (see Table 8).

In the liver tissues, antioxidant enzyme levels in catalase were significantly increased ( p <0.05 or p <0.01) in all treatment groups except the control group and the 200 mg / kg administration group. And sieve 100 mg / kg administration group (above p <0.05) and 200 mg / kg administration group showed a significant ( p <0.01) decrease compared to the positive control group. SOD levels showed a significant increase ( p <0.05 or p <0.01) in the positive control group and all the control groups except the control group and the 200 mg / kg injection group, and the single agent, the double agent, and the foam agent. The administration of 200 mg / kg showed a tendency to decrease significantly ( p <0.05 or p <0.01) compared to the positive control group (see Table 9).

Experimental Example 6. Determination of Lipid Peroxidation Level in Liver Fraction after Administration of Extract of the Present Invention

1.5 ml of 20% acetic acid, 0.225 ml of 8.1% SDS, 0.075 ml of distilled water, 0.075 ml, 1.2% TBA solution in a test tube according to the method of Suematsu et al. (Suematsu T et al, Clin. Chim. Acta. 79 , pp267-770, 1977) . 1 ml and liver homogenates, cytosol, or microsomes were added respectively. The reaction solution was heated at 100 ° C./30 minutes and centrifuged (3,000 rpm, 10 min) to measure the absorbance at 532 nm for the supernatant. The calibration curve was converted using MDA (malondialdehyde).

In addition, NADPH 0.1mM and ADP-Fe ²⁺ (ADP 0.5mM, Fe ²⁺ 0.02mM) were added to compare the lipid peroxidation degree of the microsomal membrane and the microsomes were added to the test tube at 37 ° C. After reacting for 0, 5, 10, 30 and 60 minutes, the MDA content was measured in the same manner as above.

Thiobarbituric acid-reactive substances (TBARS) content of liver tissue was measured using a method such as Buege et al. (Buege JA et al, Methods Enzymol . 52, pp302-310, 1978). Ie homogenize 1 g of liver in 4 ml of 50 nM Tris-HCl buffer (pH 7.4), and then add 1 g of thiobarbituric acid solution (15 g of TCA in 1 ml of homogenized 20% homogenate). (trichloroacetic acid) 50 ml of distilled water was added to the reagent to dissolve it, followed by addition of 0.375 g of TCA reagent, 25 ml of 1 N HCl, and 2 ml of ethanol solution containing 40 mg of BHT. Volume to 100 ml) 2 ml was added and mixed by vortex for 30 seconds, followed by agitation for 15 minutes in 100 ° C. water, followed by cooling at room temperature. After cooling, the mixture was centrifuged at 3000 g for 10 minutes, and the supernatant was measured for absorbance at 535 nm. The standard solution was used by dissolving tetraethoxypropane in methanol.

Phosphatidylcholine hydroperoxide (PCOOH) content of liver tissue was determined by Blight and Dyer's method (Bligh EG et al, Can J Biochem Physiol . 37, pp911-917, 1959), followed by Miyazawa T et. al, J Lipid Res . 33, pp1051-1058, 1992). 1 g of liver tissue was homogenized in 4 ml of 50 mM Tris-HCl buffer (pH 7.4). Add 3 ml of chloroform: methanol (2: 1, v / v) to 1 ml of homogenized 20% homogenate, mix with vortex for 30 seconds, and add 2 ml of chloroform: distilled water (1: 1, v / v) Mix well with vortex. The lower layer was separated by centrifugation at 3000 g for 10 minutes. 1.5 ml of chloroform was added to the remaining supernatant, mixed with Vortex for 30 seconds, and centrifuged at 3000 g for 10 minutes to separate the lower layer, and then combined with the lower layer obtained by the first centrifugation. The solvent was removed with nitrogen gas, and then dissolved in chloroform: methanol (1: 1, v / v) again, 20 μl of which was taken up by chemiluminescence-HPLC (Gilson, France.column: Finepak SIL NH 2-). 5, 4.61DX250mm, Jasko Corporation, Japan.Column temperature: 40 ° C detector: CLD-110, Tohoku Electric Company, Japan).

6-1. In case of water extract administration group

In the serum control group, TBARS and PCOOH levels were significantly increased ( p <0.05) in the positive control group compared to the normal control group. In particular, the tendency of the case of TBARS so in all treated groups significantly compared to the control group eoteuna indicate (p <0.05) increased tendency to, decreased stage control and the Posay 200 mg / kg administered group significantly (p <0.05) compared to the positive control Indicated. PCOOH also showed a tendency to increase significantly ( p <0.05) in all 50 mg / kg groups, but overall lower in the 100 or 200 mg / kg group, especially in the 200 mg / kg group. Showed a significant decrease ( p <0.05) compared to the positive control (see Table 8).

In the liver tissues, TBARS increased significantly ( p <0.05) in both the positive control group and the 50 mg / kg group, and in the 100 mg / kg group of the mono-drug group compared to the normal control group. The 200 mg / kg administration group of the pill and sieve showed a tendency to decrease significantly ( p <0.05) compared to the positive control group. PCOOH showed a tendency to increase significantly ( p <0.05 or p <0.01) compared to the normal control group in all groups except the 200 mg / kg administration group of emollients and sieves, and 100 mg / kg group of sieves (above p <0.05). ) And 200 mg / kg administration group showed a significant decrease ( p <0.01) compared to the positive control group (see Table 9).

6-2. Methanol extract administration group

In the serum control group, TBARS and PCOOH levels were significantly increased ( p <0.01) in the control group. TBARS showed a tendency to increase significantly ( p <0.05 or p <0.01) in all treatment groups except the 200 mg / kg dosing group, and only the 200 mg / kg dosing group was significantly higher than the positive control group. Decrease ( p <0.05). It allows for both In group in PCOOH significance compared to the control group (p <0.05 or p <0.01 ) increased tendency shown were, only control and recovery control significantly higher than the positive control at 200 mg / kg group (p <0.05) for the It showed a decreasing result (see Table 8).

Lipid peroxides in liver tissues were significantly increased ( p <0.01) in TBARS and PCOOH levels compared to normal controls. TBARS showed a tendency to increase significantly ( p <0.05 or p <0.01) in all the treatment groups except the 200 mg / kg dosing agent, and also in the PCOOH group compared to the normal control group. p <0.05 or p <0.01) increased. However, PCOOH showed a significant decrease ( p <0.05 or p <0.01) in monoclonal 200 mg / kg and monoclonal 100 or 200 mg / kg administration groups (see Table 9).

6-3. In case of 70% extract administration group

In the serum control group, TBARS and PCOOH levels were significantly increased ( p <0.01) in the control group. In particular, PCOOH showed a tendency to increase significantly ( p <0.05 or p <0.01) in all the treatment groups except the 200 mg / kg sieve group, and in the 200 mg / kg group, It showed a tendency to decrease significantly ( p <0.05) compared to the positive control group. In TBARS, all groups except monoclonal 100 mg / kg and foam 200 mg / kg showed a tendency to increase significantly ( p <0.05 or p <0.01) compared to the normal control group. The results were significantly ( p <0.05) decreased compared to the positive control (see Table 8).

On the other hand, lipid peroxides in liver tissues showed a tendency to increase significantly ( p <0.05 or p <0.01) in the positive control group and the 50 or 100 mg / kg administration group, but not in the monoclonal, oral, The 200 mg / kg sieve group showed a significant decrease ( p <0.05) compared to the positive control group. In PCOOH, the positive control group and the 50 or 100 mg / kg administration group showed a tendency to increase significantly ( p <0.01) compared to the normal control group. It showed a tendency to decrease significantly ( p <0.05 or p <0.01) compared to the control (see Table 9).

Experimental Example 7. Toxicity Test

In order to investigate the presence of toxicity by the oral administration to the rats of the sieve extract of Example 1 to the male and female rats of SD system at a dose of 1 g / kg, 2 g / kg and 5 g / kg 3 Five oral single doses were administered and 14 deaths, general symptoms, weight change and autopsy findings were observed. The result of this test is as follows.

(1) There were no deaths observed during the entire test period in all administration groups of the composition of the present invention during the test period, and there was no change in general symptoms specifically observed compared to the control group.

(2) Body weight change showed significant weight loss on day 3 after administration in 5 g / kg dose group, but was not different from control group in all other administration groups.

As a result, no clinical symptoms were observed in all test substance-treated groups compared to the death and control groups, and there were almost no specific symptoms in the anatomical findings at weight change and autopsy. In addition, since the composition did not show a clear toxicity at the dose of 5 g / kg or less, LD ₅₀ (lethal dose) in rats by oral administration is determined to be more than 5 g / kg.

Extract of the present invention can be administered in the following formulations, the formulation examples below are merely to illustrate the invention, whereby the content of the invention is not limited.

Formulation Example 1 Preparation of Powder

20 mg of extract of Example 4

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

10 mg of extract of Example 4

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

10 mg of extract of Example 4

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

10 mg of extract of Example 4

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO _4, 12H ₂ O 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

20 mg of extract of Example 4

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method of preparing a liquid solution, each component is added to the purified water to dissolve it, the lemon flavor is added appropriately, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by the addition of purified water, and then filled in a brown bottle. The solution is prepared by sterilization.

Formulation Example 6 Preparation of Healthy Food

1000 mg of extract of Example 4

Ferulic acid compound 1000 mg

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B ₁ 0.13 mg

Vitamin B ₂ 0.15 mg

Vitamin B ₆ 0.5 mg

0.2 μg of vitamin B ₁₂

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

Folate 50 ㎍

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

The vitamin and mineral mixtures are prepared by mixing the relatively suitable ingredients for health foods in a preferred embodiment, but the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to a general health food manufacturing method, and then granulated. It can be prepared and used in the manufacture of health food compositions according to conventional methods.

Formulation Example 7 Preparation of Healthy Drink

1000 mg of extract of Example 4

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

After mixing the above components according to the conventional healthy beverage manufacturing method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 ℃ container, sealed sterilization and refrigerated Used to prepare the healthy beverage composition of the invention.

 Although the composition ratio is a composition suitable for a preferred beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, use purpose.

As mentioned above, the composition of the present invention shows excellent hepatic function protection in carbon tetrachloride-induced liver failure, significantly lowers GOT and GPT levels in liver cirrhosis, and is stable to human body while reducing antioxidant activity and lipid peroxides. Therefore, it can be usefully used for medicines and health foods for protecting liver cells and preventing or treating liver damage.

Claims (7)

40 ~ 60% by weight of baekbaekpi, 10 ~ 20% by weight, brown root 10 ~ 20%, mold 3 ~ 8%, safflower 3 ~ 8%, shiho 3 ~ 8% and windproof 3 ~ 8% by weight A pharmaceutical composition for the prevention and treatment of hepatotoxicity, cirrhosis, fatty liver or acute hepatitis, comprising an extract as an active ingredient and comprising a pharmaceutically acceptable carrier, excipient or diluent. delete delete The pharmaceutical composition of claim 1, wherein the mixed extract is soluble in water, a lower alcohol having 1 to 4 carbon atoms, or a polar solvent selected from a mixed solvent thereof. 3 ~ 10 times of 10% NaCl aqueous solution was added to mixed pulverized powder containing baekbaekpi, bokyeong, brown root, mold opening, safflower, sea tiger and windproof for 30 ~ 52 hours, then dried, and then dried 3 ~ 10 times 15 A method of extracting the powder obtained by adding the aqueous solution of% ethanol for 48 to 96 hours and then drying to extract 3 to 10 times water, methanol or 70% alcohol. 40 ~ 60% by weight of baekbaekpi, 10 ~ 20% by weight, brown root 10 ~ 20%, mold 3 ~ 8%, safflower 3 ~ 8%, shiho 3 ~ 8% and windproof 3 ~ 8% by weight A dietary supplement for the prevention and improvement of hepatotoxicity, cirrhosis, fatty liver or acute hepatitis, which contains an extract as an essential active ingredient and which contains a food supplement which is food acceptable. delete