KR101769592B1

KR101769592B1 - Novel use of Limonium tetragonum

Info

Publication number: KR101769592B1
Application number: KR1020150056167A
Authority: KR
Inventors: 정은주
Original assignee: 경남과학기술대학교 산학협력단
Priority date: 2015-04-21
Filing date: 2015-04-21
Publication date: 2017-08-18
Also published as: KR20160125243A

Abstract

The present invention provides a new use of the magnifying glass. The present invention has the advantage of effectively treating, ameliorating and / or preventing at least one selected from hyperlipidemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation or alcoholic liver disease.

Description

Novel use of Limonium tetragonum

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a goniometer, and more particularly to a new use of goniometer.

Limonium tetragonum is an herbaceous plant of Plumbaginaceae, which is known to have an effect on Alzheimer's (Korea Patent No. 10-0965304).

On the other hand, the problems of hyperlipemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation or alcoholic liver disease are serious to modern people,

Korean Patent No. 10-0965304, (June 22, 2010), Claims

A problem to be solved by the present invention is to provide a new use of the sun goggle mirror.

The object of the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

The present inventors have surprisingly found that Limonium tetragonum has effects on hyperlipidemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation and / or alcoholic liver disease.

The present invention provides medicinal and / or food uses for the treatment, improvement and / or prevention of one or more selected from hypercholesterolemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation or alcoholic liver disease.

The at least one selected from the above-mentioned hyperlipemia, the hepatitis, the liver cirrhosis, the fatty liver, the liver cancer, the liver damage, the inflammation or the alcoholic liver disease is caused by dietary habit, for example, by alcohol consumption. Treatment refers to the amelioration or alleviation of symptoms associated with one or more selected from hyperlipidemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation or alcoholic liver disease, and prevention includes inhibition of developing into a disease at a pre- . The alcohol may be ethanol. The liver damage may be due to oxidative stress.

The alcoholic liver disease may be one or more selected from alcoholic fatty liver disease, alcoholic hepatitis, alcoholic chronic hepatitis, alcoholic liver cirrhosis, alcoholic liver cancer or alcoholic liver injury.

The present invention provides a pharmaceutical composition for treating or preventing at least one selected from hyperlipidemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation, or alcoholic liver disease, which comprises a gallbladder as an active ingredient.

The bacterium { Limonium tetragonum (Thunb) AA Bullock} is a biennial herb that originated in Korea, distributed in Korea and Japan, and grows on the beach. Such spikelets may have been harvested from nature or cultivated.

The goniocarpus may be an outpost or a part thereof, for example, a ground part.

The starch diameters may be at least one selected from the group consisting of germplasm diameters, dried germplasms, or gabapyrum solvent extracts.

The solvent may be polar or polar solvent.

The solvent may be at least one selected from water, alcohol or a mixed solvent thereof.

The alcohol may be a lower alcohol and may be at least one selected from among alcohols having 1 to 6 carbon atoms. In one embodiment, the alcohol may be methanol.

The extract may be a fraction extract obtained by extracting a crude extract obtained by extracting at least one selected from the raw egg yolk or the dried egg yolk with at least one selected from the group consisting of water, alcohol and a mixed solvent thereof.

The fraction extraction may be performed by fractionating the crude extract in the order of chloroform, ethyl acetate, and n-butanol, followed by obtaining an ethyl acetate fraction.

In addition, the extract of Prunus thermophilus may be obtained by extracting at least one selected from the raw egg yolk or the dried egg yolk with one of water, alcohol or a mixed solvent thereof to obtain a crude extract; Contacting the crude extract with water to obtain a contact; And extracting the contact with fractions of ethyl acetate.

The fraction extraction may be performed by fractionating the contact product in the order of chloroform, ethyl acetate, and n-butanol, followed by obtaining an ethyl acetate fraction. The fraction can be carried out by conventional fractionation methods known in the art.

The treatment or prophylaxis is effective for inhibiting lipid accumulation in liver tissue, decreasing the activity of aspartic acid aminotransferase (AST) in liver tissue, decreasing the activity of alanine aminotransferase (ALT) in liver tissue, By inhibition of cholesterol accumulation in blood, inhibition of triglyceride accumulation in blood, promotion of alcohol metabolism, increase of aldehyde dehydrogenase activity, increase of antioxidant enzyme activity in liver tissue, or inhibition of increase of lipid peroxidation in liver tissue .

The composition of the present invention may contain the active ingredient in an amount of 0.1 to 95% by weight based on the total weight of the composition.

The present invention also provides a food composition for improving or preventing at least one selected from hyperlipidemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation, or alcoholic liver disease, which comprises Rhizobium as an active ingredient.

Unless otherwise indicated, the food composition is equally applied to the pharmaceutical composition of the present invention, unless otherwise stated. This " improvement " is included in the " treatment " and means that the condition or symptom is improved.

The food composition may be variously contained in foods containing beverages, and may be in the form of beverages, gums, tea, health functional foods, etc. The health functional foods may be formulated into tablets, capsules and the like. The term "health functional food" as used herein refers to foods prepared (including processing) by using raw materials or ingredients having useful functions in accordance with the Korean Health Functional Foods Act No. 12669, and "functional" Refers to the structure and function of the human body to obtain nutritionally useful effects such as controlling nutrients and physiological functions. The food composition may comprise conventional food additives, and the food additives may be selected from natural compounds such as ketones, chemical compounds such as glycine, sodium citrate, nicotinic acid, cinnamic acid, etc., Additives, L-sodium glutamate preparations, noodle-added alkalies, preservative preparations, tar coloring preparations and the like. In addition, the food composition includes food additives and the like.

The present invention also provides one or more therapeutic or prophylactic agents selected from hyperlipidemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation, or alcoholic liver disease, comprising administering to a mammal, including a human, And for the manufacture of one or more therapeutic or prophylactic agents selected from hypercholesterolemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation or alcoholic liver disease. The germplasm to be administered may be an effective amount of the germinal barrier.

Unless otherwise stated, the terms of the pharmaceutical compositions, food compositions, methods, or uses of the invention are applied to the same extent as each other unless they are contradictory.

The starch capsule or the composition can be administered orally or parenterally to a mammal including a human, and the active ingredient can be formulated together with a pharmaceutically acceptable carrier to be formulated and administered. When formulating, fillers, extenders, binders, wetting agents, disintegrants, surfactants, diluents or excipients usually used may be used. Solid form preparations 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 and And / or gelatin. In addition, lubricants such as magnesium and talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. Various excipients such as wetting agents, sweetening agents, fragrances and / or preservatives, and the like are used in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include injectable solutions, suspensions, emulsions, lyophilisers, nasal cleansers, and suppositories. Injectable solutions, suspensions and emulsions may be prepared by mixing water, non-aqueous or suspensions, and active ingredients. Examples of non-aqueous and suspensions include vegetable oils such as propylene glycol, polyethylene glycol and olive oil, ethyl oleate , And the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol and / or gelatin can be used. For parenteral administration, it can be administered by subcutaneous injection, intravenous injection, or intramuscular injection.

The germplasm included in the composition of the present invention or used in the use or the method can be used in a dose of 1 to 1000 mg / kg, preferably 10 to 500 mg / kg on an adult basis. The administration can be administered once or several times a day. However, the scope of the present invention is not limited by the dose and the number of administrations.

The preparation of the starch can be carried out by adding a pharmaceutically or physiologically acceptable carrier, excipient or diluent such as Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA Reference can be made to the literature.

Accordingly, the composition of the present invention may further comprise a pharmaceutically acceptable additive or a pharmaceutically acceptable additive, and may be composed of the active ingredient and the pharmaceutically acceptable excipient or the pharmaceutically acceptable excipient.

The present invention can effectively treat, ameliorate and / or prevent one or more selected from hyperlipidemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation or alcoholic liver disease.

1 is a photograph showing a histopathological examination result of Example 1;
2 is a graph showing the blood alcohol concentration measurement result of Example 2. Fig.
3 is a graph showing the results of ADH measurement in Example 2. Fig.
4 is a graph showing the results of ALDH measurement in Example 2. Fig.

BRIEF DESCRIPTION OF THE DRAWINGS The advantages and features of the present invention and the manner of achieving them will be more apparent from the following detailed description taken in conjunction with the accompanying drawings. The present invention may, however, be embodied in many different forms and should not be construed as being limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. But is only provided to fully inform the owner of the scope of the invention, and the present invention is only defined by the scope of the claims.

Throughout the specification, "and / or" include each and every combination of one or more of the components mentioned.

The terminology used herein is for the purpose of illustrating embodiments and is not intended to be limiting of the present invention. In the present specification, the singular form includes plural forms unless otherwise specified in the specification. &Quot; comprises, " and / or "comprising" when used in this specification do not exclude the presence or addition of one or more other components, and / or steps, .

In the present specification, the term 'spawning column diameter' means a seedling spawning section or a part of a spawning column which has not undergone a separate drying process after collection, and the 'spawning column diameter' means that the spawning column diameter is dried.

In the following examples, an animal model in which liver disease and hyperlipemia were induced through administration of a high-fat diet containing high-concentration alcohol was used to evaluate the effects of alcoholic fatty acids, such as astringent fatty liver, hepatitis, liver cirrhosis, liver cancer, liver damage, inflammation and / And liver disease.

The reagents and the like used in the examples were the highest available from the market, and those purchased from Sigma Co. were used.

Example 1 Identification of Improvement, Treatment and / or Prevention of Hyperlipidemia, Hepatitis, Liver Cirrhosis, Liver Disease, Liver Cancer, Liver Damage, Inflammation or Alcoholic Liver Disease

1-1. Preparation

Wild horseshoe plague outpost from 2013.7. Collected from the shore of Shinan-gun, Korea and deposited with a voucher specimen (GNP-70) in Gyeongnam University of Science and Technology. The ground portion of the same diameter as that of the deposited sample was dried with a freeze dryer to obtain a dried product (2 kg), followed by crushing. The same amount of methanol as that of the dry sample was added to the mixture to conduct ultrasonic extraction for 3 hours at a low temperature of 25 ° C. And further extracted twice by the same method as the above method to obtain a secondary extract and a tertiary extract. The primary, secondary and tertiary extracts were mixed, filtered, and concentrated under reduced pressure to obtain 569.9 g of a concentrate. After the concentrate is suspended in distilled water, the same amount of chloroform is added, and the mixture is left in a funnel for 30 to 60 minutes. When the layer is completely separated, only the chloroform layer is obtained as the first fraction. The second fraction and the third fraction were obtained by two additional fractions in the same manner, and the first, second and third fractions were all mixed to obtain a chloroform fraction. The chloroform fraction was applied, and the ethyl acetate fraction and the normal-butanol fraction were sequentially obtained in the same manner as in the above chloroform fractionation in the remaining distilled water suspension. Among them, the ethyl acetate fraction was concentrated and then lyophilized to prepare a powdery extract of Rhododendron syrup.

1-2. Preparation of control substance etc.

Control substance: Silymarin (LEGALON CAP. 140, Bukwang Pharm)

Diseases Inductive substances: Ethyl alcohol

Excipient: 0.5% carboxymethylcellulose (CMC)

1-3. pharmacy

1-3-1. Preparation of test substances, control substances and excipients

A certain amount of test substance and control substance are weighed, placed in a 0.5% excipient and stirred with a stirrer for at least 10 minutes. For the control material, remove the capsules and weigh only the contents.

A certain amount of excipient is weighed and stirred with sterilized distilled water for at least 10 minutes with a stirrer to prepare a 0.5% excipient.

1-3-2. Preparation frequency and storage

The test substance is prepared once a week, and the preparation test substance is stored in the refrigerator.

1-4. Preparation of experimental animals

Species: Mouse (specific pathogen member (SPF))

System / Subsystem: C57BL / 6J Jms Slc

Number of animals received: 48 (48 males)

Number of animals administered: 48 (48 males)

Weekly range: Approximately 7 weeks of age. Approximately 8 weeks old animals are used at the start of the treatment.

Source: Central laboratory animals

Purification: Every animal has a five-day purification period to adapt to the laboratory environment.

Group separation: Select the animal that shows the proper weight without showing any clinical symptoms such as illness or injury, and use it for the test. Based on the most recently measured body weight, randomly place in the control and administration groups using the Path / Tox System.

Breeding: Breeding is carried out in a polycarbonate breeding box with no more than 6 animals during the refinement period and the administration period.

Animal room environment: Temperature (23 ± 3 degrees Celsius), Humidity (30 - 70%), Lighting 12 hour light / 12 hour dark cycle (lights off at 08:00 ~ 20:00), Illumination (150 - 300 Lux) - 20 times / hour)

Feed and water: During the purifying period, feed the animal feed (Lab Diet (R) # 5002 Lab Diet (R) # 5053 PMI Nutrition International, USA) with gamma irradiation. After the refinement period, special diets containing alcohol and control diets (Lieber-DeCarli ethanol diet # 710260, Lieber-DeCarli control diet # 710027, Dyets, USA) are prepared daily according to the experimental design. Water freely feeds the tap water that has passed through the microfilter and ultraviolet water sterilization device.

1-5. Experimental group

The experimental group was constructed as shown in the following table.

group Marital number Inducing substance Test substance Normal control group 12 Lieber-DeCarli
Control diet 0.5% CMC
10 mg / kg Positive control group 12 Lieber-DeCarli
Ethanol diet Silymarin
100 mg / kg Ethanol-treated group 12 Lieber-DeCarli
Ethanol diet 0.5% CMC
10 mg / kg Lepidopteran group 12 Lieber-DeCarli
Ethanol diet Example 1-1. extract
100 mg / kg

1-6. administration

1-6-1. Alcohol feed adaptation period

After group separation, the alcohol concentration in the feed is gradually increased for seven days to adapt the animal to the feed of the highest alcohol concentration. After separation of the group, Lieber-DeCarli ethanol diet containing ethyl alcohol as an inducing substance according to the following table is fed into a feeding tube and freshly fed daily to induce alcoholic fatty liver. The feed preparation of the normal control group is mixed with a blender at a ratio of 221.78 g of Lieber-DeCarli control diet and 1 L of cold water.

Work feed
(g) Alcohol
(mL) Cold water
(total volume; mL) density
(%) Day 1 106 8 800 One Day 2 106 16 800 2 Day 3 106 24 800 3 Day 4 106 28 800 3.5 Day 5 106 28 800 3.5 Day 6 106 32 800 4 Day 7 106 36 800 4.5 After 8th day 106 40 800 5

1-6-2. Induction of Alcoholic Liver Damage by Binge Drinking

Biberna et al., (2013) refer to the NIAAA model for single binge ethanol feeding during feeding with Leiber-DeCarli feed (NIA Protoc. Mar 2013; 8 (3): 627-637). Seven days before the autopsy, 40% EtOH 10 ml / kg is orally administered to each of the positive control, ethanol-treated group, and astrocytoma-treated group.

1-6-3. Method of administration

The feed of the test substance is started at the same time as the feed of the highest alcohol concentration is fed.

The test substance was administered for 8 weeks. The administration method was 0.5% CMC 10 ml / kg in the normal control group and the ethanol administration group based on the most recent body weight, silymarin 100 mg / kg in the positive control group, 100 mg / kg of extract is orally administered once a day. The administration is carried out at the same time in the morning each day, and the test substance is continuously stirred in the animal room before administration and during administration.

1-7. Blood collection and blood biochemical tests

1-7-1. Blood collection and testing methods

Plan Blood is collected before the autopsy on the test animals that are slaughtered. The animals fasted overnight on the day before the autopsy were inferred under CO ₂ anesthesia, and then 2 ml of blood was collected from the large vena cava and the whole blood was placed in a blood collection tube. The blood is gently mixed and left at room temperature for about 60 minutes. Separate the serum by centrifugation (3000 rpm, 10 minutes). Separated serum was transferred to a polypropylene tube and the blood biochemical parameters such as aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), blood urea nitrogen (BUN), creatinine ), Total cholesterol (TCHO), albumin / globulin ratio (A / G), total protein (TP), albumin (ALB), creatine kinase (CK), triglyceride (TG), total bilirubin (TBIL), phospholipid , Gamma Glutamyl transferase (GGT), Calcium (Ca), Inorganic phosphorus (IP), Chloride (Cl), Sodium (Na), Potassium (K)}. The above items were measured using a blood biochemical automatic analyzer (HITACHI Automatic analyzer 7180 Hitachi Co., Japan) according to the manufacturer's instructions.

For the statistical analysis of the data obtained, multiple comparison tests are performed. Levene equality analysis is performed on the test items. When all the significance is not recognized, it is tested by the ANOVA, which is a parametric method, with a significance level of 0.05. For the items with significance as a result of the test, Tukey post hoc test is used to determine whether there is a difference between the test groups. The significance level in the Levene test is Kruskal-Wallis, a nonparametric method. For the items with significant significance, rank order is given and Tukey post hoc test is performed. This analysis is performed using the SPSS 17.0 program.

Changes in blood glucose, serum total protein, blood albumin, blood AST, ALT in blood, ALP in blood, cholesterol in blood, triglycerides in blood may be associated with hyperlipidemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation or alcoholic liver disease Therefore, the present inventors have made it clear that the extract of Porphyromycetes is effective against hyperlipidemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation or alcoholic liver disease.

The results are shown in Table 3. Table 3 shows the blood test results.

GLU
(g / dL) TP
(g / dL) ALB
(g / dL) AST
(U / L) ALT
(U / L) ALP
(U / L) Chol
(mg / dL) TG
(mg / dL)
Normal control group

230.67
5.43
1.98
53.50
31.33
335.44
170.67
55.33
Ethanol-treated group

184.29 ^#
5.64
2.07
189.50 ^#
149.57 ^#
314.14
142.43 ^#
61.71
Lepidopteran group

159.25 ^*
5.61
2.06
141.25 ^*
101.13 ^*
310.50
122.14 ^*
49.38 ^*

# P <0.01 compared to normal control

* P <0.01 compared to the ethanol administration group

AST and ALT in blood are enzymes that are used as a general indicator of hepatic dysfunction because they are secreted from the liver into the blood when they are damaged due to liver alcohol and drugs and they can lower the activity of AST and ALT in blood It is known that certain substances can improve liver function, such as improving liver function. In addition, ALP separates various phosphate esters and promotes calcification in the skeleton, which is involved in phosphorus uptake in the intestine. It is elevated in bone diseases, liver and biliary diseases, pregnancy and malignant tumors (Korean J. Lab. Ani. Sci., 11, 141-145, 1995, J. J. Lab. Ani. Sci., 9, 71-82, 1993). In this example, the levels of AST and ALT in the serum were 189.50 and 149.57 U / L, respectively, which were significantly higher than those of the normal control group (#p <0.01). Compared with ethanol-treated group, AST and ALT levels of 141.25 and 101.13 U / L were significantly decreased (* p <0.01). ALP levels were slightly decreased in the ethanol treated group compared to the normal control group, but there was no significant difference. Albumin is a major protein produced in the liver that accounts for one quarter of the protein produced in the liver. Therefore, if the number of hepatocytes is not sufficient, the albumin is not sufficiently produced, the serum albumin level is lowered, and serious diseases such as liver disease, nephrotic syndrome and malnutrition can be suspected. No significant changes in total protein (TP) and albumin (ALB) levels were observed in the normal control group and ethanol treated group. The blood cholesterol level was slightly decreased in the ethanol - treated group compared with the normal control group, but it was judged to be similar when considering the weight gain of the normal control group administered with the high fat diet alone. Cholesterol levels were significantly (p <0.01) lower in ethanol-treated group than in ethanol-treated group. The triglyceride levels in the astringent group were lower than those in the ethanol - treated group and the normal control group, indicating that the astringent hypercapnia extract was effective in decreasing cholesterol and triglyceride in the blood.

From the above results, it can be seen that the induction of ethanol and the high fat diet increased the activity of AST and ALT, and the increase of cholesterol and triglyceride accumulation effectively induced liver diseases such as alcoholic hyperlipemia and fatty liver.

In the ethanol extract group, the activity of AST and ALT were significantly decreased and the levels of cholesterol and triglyceride in the blood were also significantly decreased compared to the ethanol group, even though ethanol was consumed in the same manner as the ethanol treatment group. From these results, it can be concluded that the coronary arteries can be used for the treatment of hyperlipidemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation or alcoholic liver diseases such as alcoholic hepatitis, alcoholic chronic hepatitis, alcoholic liver cirrhosis, Liver damage and the like.

From these results, it has been found that the gonadotropin-releasing hormone can be used for improving, treating and / or preventing hyperlipidemia, hepatitis, liver cirrhosis, liver cancer, liver damage, inflammation, fatty liver or alcoholic liver disease, Cirrhosis, liver cancer, liver damage, inflammation, fatty liver or alcoholic liver disease, etc. can be improved, treated and / or prevented.

1-8. Autopsy and pathologic examination

1-8-1. Autopsy and long-term weighing, fixation of tissue

Test animals are euthanized after blood sampling using CO ₂ aspiration, and the vena cava and abdominal aorta are cut to death. Before the autopsy, confirm the visual findings and check for complete blood loss. Observe the abnormalities of the abdominal cavity and thoracic cavity and remove the anterior lobe of the liver. The extracted liver is washed with distilled water (DW) and the remaining blood is removed. The weight is measured, and the portion to be used for the slide production is immediately fixed to the 10% neutral buffer formalin by trimming the fixation target position. The rest of the unfixed liver is placed in a 50 ml conical tube and stored in a deep freezer (-72 ° C).

1-8-2. Histopathological examination

Histological examination revealed pathologic findings such as proliferation of hepatocyte inflammatory cells, hepatocyte apoptosis, destruction of liver tissue structure, and fat accumulation by administration of ethanol and high fat diet. And to confirm the improvement effect.

Tissue specimens are fixed in 10% neutral buffered formalin and histological examination is performed after hematoxylin & eosin staining.

The results are shown in Fig. 1 is a photograph showing a histopathological examination result of Example 1;

As shown in FIG. 1, lipid droplet accumulation was markedly increased and inflammatory cells were increased in the ethanol-treated group compared with the normal control group. Therefore, it was confirmed that the induction substance (ethanol) And can be visually confirmed that it has been brought to a state where it can be or progressed. In addition, it can be seen that it was induced or progressed to hepatitis, liver cirrhosis, liver cancer, liver damage, inflammation, hyperlipidemia, alcoholic liver disease and the like. In the case of astaxanthin-treated group, it was shown that the fat globules were less than those in the ethanol-treated group, and the decrease of inflammatory cells and hepatocyte destruction were decreased. Histopathologic observations showed that hepatoprotective effects of hepatitis C, hepatitis, hepatocellular carcinoma, hepatocellular carcinoma, hepatocellular carcinoma, hepatocellular carcinoma, hepatocellular carcinoma Damage, inflammation, fatty liver or alcoholic liver disease.

As described above, it is possible to improve, treat and / or prevent hyperlipidemia, hepatitis, liver cirrhosis, liver cancer, liver damage, inflammation, fatty liver or alcoholic liver disease associated therewith by lipid accumulation inhibition action and anti- Treatment, and / or prevention of hyperlipidemia, hepatitis, liver cirrhosis, liver cancer, liver damage, inflammation, fatty liver or alcoholic liver disease caused by alcohol, and the like.

From the results of 1-7. ~ 1-8., It can be seen that ganoderma lucidum can inhibit lipid accumulation and alleviate inflammatory reaction in alcohol-induced hepatitis and fatty liver tissue.

From this, it can be said that astrocytes are useful for the improvement, treatment and / or prevention of hyperlipidemia, hepatitis, liver cirrhosis, liver cancer, liver damage, inflammation, fatty liver or alcoholic liver disease, more preferably hyperlipidemia, hepatitis, Liver injury, inflammation, fatty liver or alcoholic liver disease.

Therefore, the gonadal glands are useful for the improvement, treatment and / or prevention of hyperlipidemia, hepatitis, liver cirrhosis, liver cancer, liver damage, inflammation, fatty liver or alcoholic liver disease and the like, more preferably hyperlipidemia due to alcohol, hepatitis, Treatment, and / or prevention of liver damage, inflammation, fatty liver or alcoholic liver disease and the like.

Example 2 Confirmation of Improvement, Treatment and / or Prevention of Hyperlipidemia, Hepatitis, Liver Cirrhosis, Fatty Liver, Liver Cancer, Liver Damage, Inflammation or Alcoholic Liver Disease

2-1. Preparation

In the same manner as in Example 1-1., Extract of Rhodiola sativa extract was prepared.

2-2. Preparation of experimental animals

Experimental animals were used for the experiment after 7-week-old male Spargue-Dawley rats were adapted for 1 week. During the adaptation period, the environment of the animal room was controlled to maintain a light and dark cycle of 23 ± 1 ° C, humidity 50 ± 5%, and 12 hours. Feeds were fed with laboratory animals and water was given free of purified water.

2-3. Experimental animal treatment

The rats were divided into 3 groups of 8 rats adapted for 1 week and classified into the control group, the ethanol administration group, and the phytoplankton administration group. In the control group, 0.5% -MCMC (carboxymethylcellulose) solution was administered, and 200 mg / kg of Ganoderma lucidum extract prepared at 2-1. Was administered orally in 0.5% -CMC. One hour later, 10 mg / kg of 40% ethanol was orally administered to the astrocytoma group and the ethanol group. After 1, 2, and 4 hours after the ethanol administration, 1 mL of blood was collected from the tail vein of the control group, the ethanol-treated group, and the ethanol-treated group, and the liver was immediately extracted after the last blood collection.

2-4. Blood alcohol concentration measurement

The blood collected in 2-3. Was placed in a seperation tube, left at room temperature for 30 minutes, and then centrifuged at 3000 rpm for 10 minutes. After centrifugation, only the supernatant was taken and stored at minus 70 ° C. The stored supernatant was measured for alcohol concentration using an ethanol measurement kit (Bio-vision, Cat # K620-100) according to the manufacturer's instructions.

The results are shown in Fig.

2 is a graph showing the results of measurement of blood alcohol concentration in Example 2. The x-axis shows the elapsed time (time) and the y-axis shows the alcohol concentration (nM). The values in FIG. 2 represent the mean ± standard deviation, and * represents p <0.001 compared to the ethanol-treated group.

As shown in FIG. 2, the ethanol concentration increased after 4 hours of alcohol administration. The alcohol concentration in blood group was decreased rapidly compared to the group treated with ethanol.

It is an object of the present invention to provide a method for treating, ameliorating and / or ameliorating diseases and / or symptoms due to alcohol, such as hyperlipidemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation or alcoholic liver disease, Or prevention.

2-5. ALDH measurement in liver tissue

In order to investigate the mechanism by which the acceleration of blood alcoholysis is actually induced in vivo, the degree of ADH and ALDH activity involved in the metabolism of alcohol in the body was measured.

A certain amount of liver extracts from 2-3. Were added to 4 times as much 0.25M sucrose solution and a 20% (w / v) milling homogenate was prepared using a homogenizer. The homogenate was subjected to centrifugation at 600 × g for 10 minutes, and then the supernatant was centrifuged at 1,000 × g for 20 minutes and the supernatant was centrifuged at 15,000 × g for 1 hour. The activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in this fraction were measured using the ADH assay kit (Biovision Cat. # 787-100) and the ALDH assay kit (Biovision Cat # 731-100). The measurements followed the manufacturer's instructions.

The results are shown in Fig. 3 and Fig. FIG. 3 is a graph showing the results of ADH measurement in Example 2, and FIG. 4 is a graph showing the results of ALDH measurement in Example 2. FIG. The x-axis in Figs. 3 and 4 represents the group, the y-axis in Fig. 3 represents the ADH activity (NADH content per hour), and the y-axis in Fig. 4 represents the ALDH activity (NADH content per hour). Figures 3 and 4 show the mean ± standard deviation, # indicates p < 0.01 relative to the normal control, and * indicates p < 0.01 relative to the ethanol treated group.

As shown in FIG. 3, the ADH activity of the ethanol-treated group was higher than that of the normal control group, and the ADH activity of the control group was slightly higher than that of the normal control group but not significantly different from that of the ethanol treated group.

In the case of ALDH activity, as shown in FIG. 4, the ethanol-treated group showed an increasing tendency by ethanol administration, and the ethanol-treated group showed a significant increase in the ALDH activity group compared with the ethanol treated group.

These results suggest that the aldehyde - decomposing activity associated with alcohol metabolism in the gut is excellent.

The aldehyde-decomposing activity by the gonad size allows treatment, improvement and / or improvement of diseases and / or symptoms due to alcohol, such as hyperlipidemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation or alcoholic liver disease Prevention seems to be possible.

2-6. Antioxidant enzyme activity measurement in liver tissue

A certain amount of liver extracts from 2-3 were added to 4 times of 0.1 M phosphate buffer (pH 7.4), and the homogenate was subjected to homogenization using a homogenizer and then centrifuged at 3000 x g for 30 minutes to obtain supernatant. The activity of SOD (superoxide dismutase) was measured using the McCord and Fridovich method (J. Biol. Chem. 1969, 244, 6056-6063) and 200 μl of the supernatant was added to the reaction solution (0.2 mM hydroxylamine and hypoxanthine) were added in the same amount and reacted with 200ul of xanthine oxidase for 30 min at 37 ° C. After stopping the reaction, it was left at room temperature for 20 minutes and the absorbance was measured at 550 nm using a UV absorber. The activity units were determined by comparison with SOD standard value using human erythrocyte enzyme. The activity of GSH-px (glutathione peroxidase) was measured by cumene hydroperoxide (GSH) to glutathione disulfide (GSSG) method (Methods Enzymol. 1984, 105, 114-121). 200ul of the tissue supernatant was added to 880ul phosphate buffer (pH 7.2) containing 53ul of glutathione reductase, 133ul GSH, and 100ul of nicotinamide adenine dinucleotide phosphate (NADPH). The reaction solution was reacted at 37 ° C for 3 minutes and the degree of decrease in absorbance was measured at 340 nm for 2 minutes every 15 seconds. Catalase activity was measured by adding 200ul of the tissue supernatant to a microcuvette containing 150ul of 19mM hydrogen peroxide. The degree of depletion of hydrogen peroxide was measured at 240 nm using a UV absorber (see J. Bil. Chem. 1952, 195, 133-140).

The results are shown in Table 4. The values in Table 4 represent the mean ± standard deviation. As shown in the table, the activity of SOD, GSH-px and catalase was decreased in the ethanol-treated group compared to the normal control group. The activity of three antioxidant enzymes was increased in ethanol-treated group compared with ethanol-treated group. Especially, GSH-px and catalase recovered to normal control level.

group
Normal control group
Ethanol-treated group

Lepidopteran group
SOD
(mmol / mg protein)
102.79 ± 12.78
69.22 + - 8.89 ^#
78.34 + - 5.57 ^*
GSH-px
(nu / mg protein)
326.80 ± 25.57
259.61 + - 31.90 ^#
318.99 ± 22.28 ^*
Catalase
(U / g protein)
204.79 ± 19.89
165.77 ± 13.34 ^#
189.57 ± 17.20 ^*

# P <0.01 compared to normal control

* P <0.01 compared to the ethanol administration group

These results suggest that antioxidant enzyme activity in hepatic tissues caused by oxidative stress may cause hyperlipemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation or alcoholic liver disease, It is believed that the treatment, improvement and / or prevention of diseases and / or symptoms due to alcohol will be possible.

2-7. Measurement of lipid peroxidation in liver tissue

A predetermined amount of liver extracts from 2-3 were added to a 0.02M phosphate buffer (pH 7.4), and a homogenate of 10% (w / v) was prepared by using a homogenizer. Then, 0.02M phosphate buffer (pH 7.4) And diluted to a 5% (w / v) homogenate. The malondialdehyde (MDA) of the peroxidation product was measured by the thiobarbituric acid method in the post-mitochondrial supernatant obtained by centrifugation at 9000 xg for 30 minutes (Buege, JA; Aust, SD Methods Enzymol. 1978, 52, 302-310).

The results are shown in Table 5. The values in Table 5 represent the mean ± standard deviation. As shown in the table, the lipid peroxide concentration in the liver tissue was increased in the ethanol-treated group compared to the normal group, but decreased in the astaxanthin-treated group and was similar to that of the normal control group.

group
Normal control group
Ethanol-treated group

Lepidopteran group
MDA
(mmol / mg protein)
23.45 + 1.39
49.89 ± 3.45 ^#
35.44 + - 3.20 ^*

# P <0.01 compared to normal control

* P <0.01 compared to the ethanol administration group

From these results, it is possible to prevent, prevent, and / or prevent hyperlipidemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation or alcoholic liver disease by the action of inhibiting lipid peroxidation- Improvement is likely to be possible.

From the results of 2-4. ~ 2-7., It was found that the rice gonad might promote the alcohol metabolism, increase the activity of aldehyde dehydrogenase, increase the activity of antioxidant enzyme in liver tissues, inhibit the increase of lipid peroxidation by alcohol in liver tissue .

From this, it appears that it is possible to treat, ameliorate and / or prevent diseases and / or symptoms due to alcohol, such as hyperlipidemia, hepatitis, liver cirrhosis, fatty liver, liver cancer, liver damage, inflammation or alcoholic liver disease.

&Lt; Preparation Example 1 > Preparation of pharmaceutical composition

100 mg of cornstarch extract, 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate, prepared in the same manner as in Example 1-1, were filled in gelatin capsules to prepare capsules.

&Lt; Preparation Example 2 > Preparation of food composition

(0.5% by weight), oligosaccharide (2% by weight), sugar (2% by weight) and sodium chloride (0.5% by weight) prepared in the same manner as in Example 1-1 Were added, and the remaining amount was adjusted, and homogeneously blended and instant sterilized to prepare a health drink.

Claims

As the active ingredient,
The starch diameters are the starchy mildew solvent extract,
Wherein the extract is a fraction extract of a crude extract obtained by extracting at least one selected from a raw egg yolk or a dried egg yolk with at least one selected from the group consisting of water, alcohol, and a mixed solvent thereof, which is a fraction extract of ethyl acetate, for the treatment or prevention of hyperlipidemia Composition.

The pharmaceutical composition of claim 1, wherein said hyperlipidemia is due to alcohol ingestion.

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The pharmaceutical composition according to claim 1, wherein the alcohol is at least one selected from among alcohols having 1 to 6 carbon atoms.

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The pharmaceutical composition according to claim 1, wherein the treatment or prevention is by at least one selected from the group consisting of inhibition of accumulation of cholesterol in the blood or inhibition of accumulation of triglyceride in blood by the asthma clinic.

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As the active ingredient,
The starch diameters are the starchy mildew solvent extract,
Wherein the extract is a fractional extract obtained by fractionating a crude extract obtained by extracting at least one selected from a raw egg yolk or an egg yolk dried body with at least one selected from the group consisting of water, alcohol, and a mixed solvent thereof, and extracting the extract with ethyl acetate.