KR102136484B1 - Composition for preventing or treating liver injury and method for preparing the same - Google Patents

Abstract

The present invention relates to a composition for preventing or treating liver injury, and a method for preparing the same. The method for preparing a composition for preventing or treating liver injury includes the steps of: (a) dipping Korean thistle in n-hexane to remove grease and extracting the degreased Korean thistle with ethyl acetate to prepare an extract of Korean thistle; (b) adding water to pulverized Hovenia dulcis Thunb fruits, carrying out centrifugal separation, introducing ethyl acetate to the supernatant, followed by shaking, and separating and concentrating the ethyl acetate layer to prepare an extract of Hovenia dulcis Thunb. fruits; (c) extracting Lycium chinense Miller fruits with water at 65-85°C to prepare an extract of Lycium chinense Miller fruits; and (d) mixing 25-45 wt% of the extract of Korean thistle, 25-45 wt% of the extract of Hovenia dulcis Thunb. fruits and 25-45 wt% of the extract of Lycium chinense Miller fruits.

Description

Composition for preventing or treating liver injury and method for preparing the same}

The present invention relates to a composition for preventing or treating liver damage and a method for manufacturing the same, and more particularly, to a composition for preventing or treating liver damage, including a thistle extract, a terrestrial extract and a goji berry extract, and a method for manufacturing the same.

The liver is the largest organ in the human body, and plays a role in the synthesis of substances such as albumin and gallbladder, the detoxification of harmful drugs or toxic substances, hormone decomposition and metabolism, digestion and excretion, and storage of nutrients. Since the liver has excellent regenerative ability and is involved in various metabolism, it is called a'chemical plant in the body', and it is also called a'silent organ' because it does not develop pain in the sense of pain because it does not develop pain. If you feel pain in the liver, it is important to prevent liver damage because liver damage has progressed significantly and recovery is impossible.

There are many causes of liver damage, such as infection by viruses, toxic substances caused by drugs, alcohol intake, smoking, and abnormalities in the human immune system. Intermediate and reactive oxygen species that are highly reactive through repeated inflammatory reactions caused by them (reactive oxygen species, ROS) is produced. Reactive oxygen species refers to hydroxyl radicals, hydrogen peroxide (H ₂ O ₂ ), superoxide radicals, singlet oxygen, etc. as a by-product of metabolic processes in vivo. It is unstable and highly reactive, so it reacts easily with surrounding compounds, and causes oxidative damage such as damaging DNA and RNA in the cell or combining with unsaturated fatty acids in the cell membrane to induce lipid peroxidation to destroy the cell membrane. Lipid peroxidation reaction by free radical is a representative mechanism of oxidative damage of hepatocytes, and oxidative stress has been reported to cause various diseases such as diabetes, aging, liver fibrosis and mutation.

Therefore, in order to prevent and treat liver damage caused by oxidative stress, there is a need to develop a material that has an effect of improving liver damage from natural products, and efforts to discover functional food materials derived from natural products that have a preventive effect related to liver disease are also required. Is being made.

Republic of Korea Patent Registration No. 1865360 discloses a composition for liver treatment and maintenance of liver health, including Artemisia extract and Schisandra extract.

Korean Registered Patent No. 1817282 contains sprouted barley, edible barley mixed extract including raw barley, hawthorn fruit, etc., raw juice concentrate, browned concentrate, pear citron concentrate, unfermented embryo fermentation extract, sputum fruit concentrate, milk thistle extract powder, mixed mixture of biscuits and Disclosed is a method for preparing a mixed drink of sprouted barley with improved liver function and antioxidant activity, which is prepared by mixing honey.

Thistle is a perennial plant belonging to the family Asteraceae, and more than 15 species of narrow leaf thistles, spiny thistles, and white thorn thistles are native to Korea. As a foreign species, white pattern thistle is also called milk thistle. It has pharmacology such as detoxification, Bo Brain, interpolation, Idam, and anti-radiation. It is known to have activity and is also known to have a pharmacological effect on anti-inflammatory action and antioxidant action. In particular, Silymarin separated from milk thistle is polyphenolic flavolignan, which is silybin, isosilybin, silydianin, silycristin, and taxifolin. taxifolin) and promotes the regeneration of hepatocytes by activating the protein synthesis of hepatocytes, and is known to have an effect of preventing liver damage and enhancing liver function due to its ability to remove free radicals.

Earthia (Hovenia dulcis Thumb fructus) is a fruit of the hawthorn (Hovenia dulcis Thumb), which is effective in protecting liver functions such as jaundice, fatty liver, and cirrhosis of the liver due to alcohol resolution and liver detoxification. It is known to be excellent. As a component isolated from the hawthorn, ampelopsin as an antidote to hepatoxin from leaves and fruits, and hoduloside, jujuboside, saponin hovenoside, saponin, 3-hydroxydammarane as antisweet substances in leaves have been reported.

Lychee fructus is a fruit of the goji tree belonging to the branch family, betaine, rutin, zeaxanthin, physalien, beta-sitosterol, and choline ) Contains a large amount of functional ingredients. Goji berry is known to remove inflammation and activate functions when taken in chronic hepatitis, cirrhosis, etc., and is known to be effective in reducing vision due to ophthalmic diseases.

Based on thistle, sesame seed, and goji berry having the above-mentioned efficacy, the inventors of the present invention have developed a method capable of extracting pharmacologically active ingredients contained in thistle, sesame seed and goji berry to the maximum and damage the liver with these extracts A composition for prevention or treatment has been developed to reach the present invention.

Korea Registered Patent No. 1865360 (2018.05.31.) Republic of Korea Registered Patent No. 1817282 (Jan. 2018.04.)

An object of the present invention is to provide a composition for preventing or treating liver damage.

Another object of the present invention is to provide a method for preparing a composition for preventing or treating liver damage.

Another object of the present invention is to provide a method for preparing or treating liver damage.

The present invention to achieve the above object,

Provided is a composition for preventing or treating liver damage, including 25 to 45% by weight of thistle extract, 25 to 45% by weight of the extract of giza and 25 to 45% by weight of the wolfberry extract, based on the total weight of the composition.

In one embodiment of the present invention, the thistle extract may be prepared by immersing thistle in n-hexane to degrease and then extracting the degreased thistle with ethyl acetate or water.

In one embodiment of the present invention, the terrestrial extract may be prepared by adding water to the ground terminator and centrifuging to add ethyl acetate to the supernatant, shaking, separating and concentrating the ethyl acetate layer.

In one embodiment of the present invention, the wolfberry extract may be prepared by extracting wolfberry with water at 65 to 85°C.

In one embodiment of the present invention, the composition for preventing or treating liver damage may further include 200 to 400 parts by weight of pear juice with respect to 100 parts by weight of the composition.

In one embodiment of the present invention, the composition for preventing or treating liver damage may be prepared in a soft extract, pill, tablet or high dosage form.

The present invention to achieve the above other objects,

(a) preparing thistle extract by immersing thistle in n-hexane to degrease and then extracting the degreased thistle with ethyl acetate or water;

(b) adding water to the ground earth, centrifuging, adding ethyl acetate to the supernatant, shaking, separating the ethyl acetate layer, and concentrating to prepare a earth earth extract;

(c) preparing wolfberry extract by extracting wolfberry with water at 65 to 85°C; And

(d) 25 to 45% by weight of the thistle extract, 25 to 45% by weight of the terrestrial extract and 25 to 45% by weight of the wolfberry extract provides a method for preparing a composition for preventing or treating liver damage.

In one embodiment of the present invention, in the step (a), the degreased thistle was extracted with ethyl acetate or water for 6 to 10 hours, and the extracted extract was concentrated at a temperature of 50° C. or less to prepare a thistle extract. May be

In one embodiment of the present invention, it may be to further mix 200 to 400 parts by weight of pear juice with respect to 100 parts by weight of the composition in step (d).

The present invention in order to achieve another object described above,

Provided is a method for preventing or treating liver damage by using a hot bath method for preventing or treating liver damage.

The composition for preventing or treating liver damage of the present invention has a liver protective effect by having a glutathione (GSH) value significantly increased and a malondialdehyde (MDA) value being significantly reduced to have antioxidant activity.

According to the manufacturing method of the present invention, extraction of the pharmacologically active ingredient contained in thistle, sesame seeds and goji berries is maximized to increase the hepatoprotective activity efficacy of the composition for preventing or treating liver damage of the present invention.

The composition for preventing or treating liver damage of the present invention has fewer side effects to the human body by being made of a material derived from natural products.

The composition for preventing or treating liver damage of the present invention may be used in beverages, dietary supplements, or pharmaceutical compositions.

1 is a graph showing the effect of the composition for preventing or treating liver damage of the present invention on SOD (Superoxide Dismutases) levels.
2 is a graph showing the effect of the composition for preventing or treating liver damage of the present invention on Catalase levels.
3 is a graph showing the effect of the composition for preventing or treating liver damage of the present invention on glutathione (GSH) levels.
4 is a graph showing the effect of the composition for preventing or treating liver damage of the present invention on malondialdehyde (MDA) levels.
5 is a graph showing the effect of the composition for preventing or treating liver damage of the present invention on serum transaminase levels.
6 is a graph showing the effect of the composition for preventing or treating liver damage of the present invention on leukocytes.
7 is a graph showing the effect of the composition for preventing or treating liver damage of the present invention on red blood cells (Erythrocyte) and platelets (Thrombocyte).
8 is a photograph showing the effect of the composition for preventing or treating liver damage according to the present invention histologically on liver-damaged rats.

The composition for preventing or treating liver damage of the present invention is characterized in that it contains 25 to 45% by weight of thistle extract, 25 to 45% by weight of the extract of the earth and 25 to 45% by weight of the wolfberry extract, based on the total weight of the composition.

In the composition for preventing or treating liver damage of the present invention, if the thistle extract is less than 25% by weight, there is a problem in that the liver has a poor protective effect, and if it exceeds 45% by weight, there is a problem that the functionality is poor. In addition, if the extract of the earth is less than 25% by weight, there is a problem that the liver protective effect is insufficient, and if it exceeds 45% by weight, there is a problem that the functionality is poor. Furthermore, if the goji berry extract is less than 25% by weight, there is a problem of poor functionality, and if it exceeds 45% by weight, there is a problem that the liver protective effect is reduced.

The composition for preventing or treating liver damage of the present invention is more preferably composed of 30 to 40% by weight of the thistle extract, 30 to 40% by weight of the terrestrial extract, and 30 to 40% by weight of the wolfberry extract.

In the present invention, the thistle is preferably fruit and/or seed, and powder form is preferably used.

In one embodiment of the present invention, the thistle extract is preferably prepared by immersing thistle in n-hexane (normal hexane) to degrease (脫脂) and then extracting the degreased thistle with ethyl acetate or water. By going through the process of degreasing in the above, it is possible to facilitate the elution of the pharmacological component contained in thistle. When extracting degreased thistle with water, it is preferable to extract hot water.

In one embodiment of the present invention, the terrestrial extract is preferably prepared by adding water to the ground terminator and centrifuging to add ethyl acetate to the supernatant, shaking, separating and concentrating the ethyl acetate layer.

In one embodiment of the present invention, the wolfberry extract is preferably prepared by extracting wolfberry with water at 65 to 85°C.

In the above, when the wolfberry extract temperature is less than 65°C, there is a problem in that the extraction efficiency of the effective pharmacological component contained in the wolfberry is lowered, and when it exceeds 85°C, there is a non-economical problem without additional advantage. More preferred extraction temperature is 70 to 80°C.

In one embodiment of the present invention, the composition for preventing or treating liver damage may further include 200 to 400 parts by weight of pear juice with respect to 100 parts by weight of the composition.

Pear has about 80% moisture and contains 10-13% sugar (fructose and sucrose), organic acids such as malic acid, tartaric acid and citric acid, vitamins B and C, fiber and fat. The stomach is effective for bronchial diseases, so it is good for colds, relieves asthma, etc., and helps with bowel movement and diuretic action. In addition, it is known that pear removes phlegm and cough, relieves symptoms when the throat is resting or when the stomach is cold and sore, helps to cure boils, has a detoxifying effect, and is also effective in removing hangovers.

In the present invention, if the pear juice is less than 200 parts by weight, there is a problem that the functionality is poor and there is a problem that the dough mixing is not smooth during high production, and when the pear juice exceeds 400 parts by weight, the liver protective effect is reduced. There is this. A more preferable amount of pear juice is 250 to 350 parts by weight based on 100 parts by weight of the composition.

The composition for preventing or treating liver damage of the present invention may be prepared in a soft extract, pill, tablet, or high dosage form.

Next, a method for preparing a composition for preventing or treating liver damage of the present invention will be described. In describing the method for preparing a composition for preventing or treating liver damage of the present invention, the same application as the composition for preventing or treating liver damage will be omitted.

Method for preparing a composition for preventing or treating liver damage of the present invention,

(a) preparing thistle extract by immersing thistle in n-hexane to degrease and then extracting the degreased thistle with ethyl acetate or water;

(b) adding water to the ground earth, centrifuging, adding ethyl acetate to the supernatant, shaking, separating the ethyl acetate layer, and concentrating to prepare a earth earth extract;

(c) preparing wolfberry extract by extracting wolfberry with water at 65 to 85°C; And

(d) 25 to 45% by weight of the thistle extract, 25 to 45% by weight of the terrestrial extract and 25 to 45% by weight of the wolfberry extract.

To more specifically describe the method for preparing a composition for preventing or treating liver damage of the present invention, first, thistle extract, earthworm extract, and goji berry extract are respectively prepared.

The production process of thistle extract is as follows: After immersing thistle in n-hexane, defatted, this is extracted with ethyl acetate or water to prepare thistle extract (step (a)).

In one embodiment of the present invention, in the step (a), the degreased thistle was extracted with ethyl acetate or water for 6 to 10 hours, and the extracted ethyl acetate or water extraction solution was concentrated at a temperature of 50° C. or lower to obtain thistle. It is preferred to prepare the extract.

If the extraction time of thistle is less than 6 hours, the extraction efficiency of the active pharmacological component contained in thistle is decreased, and if it exceeds 10 hours, there is an uneconomical problem without additional advantage. The extraction time is more preferably 7 to 9 hours.

In addition, there is a problem in that the effective pharmacological component may be deteriorated when the extraction solution extracted from the above is concentrated to a temperature above 50°C. The more preferred concentration temperature is 30 to 45°C.

The process of manufacturing the earth extract is as follows. Water is added to the ground earth, centrifuged, ethyl acetate is added to the supernatant, shaken, and the ethyl acetate layer is separated and concentrated to prepare a earth earth extract (step (b)).

The process of manufacturing the wolfberry extract is as follows: wolfberry extract is extracted with water at 65 to 85°C to prepare a wolfberry extract (step (c)).

In the above, when the wolfberry extract temperature is less than 65°C, there is a problem in that the extraction efficiency of the effective pharmacological component contained in the wolfberry is lowered, and when it exceeds 85°C, there is a non-economical problem without additional advantage. More preferred extraction temperature is 70 to 80°C.

Subsequently, a mixture was prepared by mixing the above-prepared thistle extract, sesame seed extract, and goji berry extract, but 25 to 45% by weight of thistle extract, 25 to 45% by weight of the sesame seed extract, and 25 to 45% by weight of the wolfberry extract Mix% (step (d)). As such, 25 to 45% by weight of thistle extract, 25 to 45% by weight of the extract of the earth and 25 to 45% by weight of the extract of the wolfberry can be mixed to prepare a composition for preventing or treating liver damage of the present invention.

In one embodiment of the present invention, 200 to 400 parts by weight of pear juice may be further mixed with respect to 100 parts by weight of the composition for preventing or treating liver damage in step (d).

The composition for preventing or treating liver damage of the present invention may be prepared in a high dosage form using a hot water method.

According to the manufacturing method of the present invention, the extraction of pharmacologically active ingredients contained in thistle, earth and goji berries is maximized, so that the composition for preventing or treating liver damage produced by the method of the present invention has excellent liver protective activity effect.

Hereinafter, a method for preparing a composition for preventing or treating liver damage of the present invention will be described in more detail through the following examples.

<Example> Preparation of composition for preventing or treating liver damage

1-1. Thistle extract preparation

The white patterned thistle was dried and powdered. The powdered thistle powder 10g was immersed in n-hexane for 4 hours to degrease. The defatted thistle powder was extracted with reflux cooling for 8 hours using ethyl acetate as an extraction solvent using a soxhlet extractor. The extracted ethyl acetate extract was concentrated under reduced pressure for 1 hour with a rotary vacuum evaporator at 50° C. or less to prepare a powdered thistle extract.

1-2. Preparation of earth extract

The earthworm (fruit of hut tree) was dried and then ground. 100 ml of distilled water was added to 15 g of ground terminator, the sample was dissolved, and then centrifuged at 4,000 rpm for 10 minutes. The centrifuged supernatant was added to a separatory filter, and 100 ml of ethyl acetate was added and shaken. The ethyl acetate layer was separated and concentrated under reduced pressure for 1 hour to prepare a powdery earth extract.

1-3. Wolfberry extract preparation

The dried goji berries were powdered. 100 g of distilled water at 70 to 80° C. was added to 10 g of powdered goji berry powder, and reflux extraction was performed for 1 hour, followed by reflux extraction for another 1 hour. The extract was filtered, concentrated under reduced pressure for 1 hour, and then freeze-dried at -70°C to prepare a goji berry extract in powder form.

1-4. Preparation of a composition for preventing or treating liver damage

A composition for preventing or treating liver damage (hereinafter, referred to as a composition for alleviating liver damage) was prepared by mixing the thistle extract powder, earth extract powder, and goji berry extract powder prepared above in a ratio of 1:1:1 by weight.

<Example 2> Preparation of liver for preventing or treating liver damage

The liver damage mitigating composition prepared in Example 1 was kneaded by mixing the juice extract of 3 times by weight. The dough was placed in a pot. Onggi was placed in a water-filled water bath. The primary bath was performed while maintaining the temperature of 98 to 102°C with an automatic temperature control system for 3 days, and the water in the bath was maintained as an automatic water replenishment system so that the pottery could be locked by 80% or more. After the 1st bath, the 1st aging process was carried out while leaving at room temperature for 1 day. Again, for the first day, the second jungtang was performed under the same conditions as the first jungtang, and then, during the first day, the second aging process was performed under the same conditions as the first aging to prevent or treat liver damage.

After diluting 4 times the water based on the weight produced in the above, the residue was filtered through a filter to obtain 80% of the supernatant from the top of the filtrate. The supernatant was concentrated under reduced pressure until 1/4, and then lyophilized at -70°C to obtain a dried sample. The following experiment was conducted using the dry sample as a composition for alleviating liver damage.

<Experimental Example> Liver protective activity against D-galactosamine (GalN) induced liver damage

1-1. Experiment method

1) Animal

The experimental animals were 150-160 g male Sprague-Dawley-type rats purchased from Samtaco, Korea, and were bred in an experimental environment controlled at a temperature of 21±5℃ and a humidity of 60±10%. Three cages were placed in each cage, and water and feed (Samtaco, Korea) were allowed to be freely consumed by the experimental animals.

2) Preparation of drug samples

The liver damage relief composition prepared in Example was prepared by diluting with distilled water to a concentration of 100 mg/kg, 200 mg/kg, and 400 mg/kg.

As a positive control for comparing the efficacy of this sample, Urusa (Daewoong Pharmaceutical), which was used for the purpose of protecting liver function, was used and was prepared by diluting with distilled water to a concentration of 300 mg/kg.

3) Causes liver damage

For liver damage, the method of Jonker et al ( Jonker AM, Dijkhuis FW, Kroese FG, Hardonk MJ, Grond J. Immunopathology of acute galactosamine hepatitis in rats. Hepatology. 1990;11(4):622-7 ) was applied. D-galactosamine (galactosamine, GalN) (Sigma, USA) was intraperitoneally injected at a concentration of 400 mg/kg body weight of experimental animals to induce liver damage.

4) Group separation (n=6)

Normal group without any treatment (Normal), untreated control after inducing liver damage (Control), positive control (U300) with urusa at a concentration of 300 mg/kg after inducing liver damage, and 100 mg/kg concentration after inducing liver damage Experimental group 1 (M100) to which the liver damage-relieving composition was administered, and experimental group 2 (M200) to which the liver-damage-reducing composition was administered at a concentration of 200 mg/kg after inducing liver damage, and to a liver-relieving composition at a concentration of 400 mg/kg after inducing liver damage The group was separated into the administered experimental group 3 (M400).

5) Drug sample administration

The treatment drug was supplied once a day for 14 days from the third day of D-galactosamine induction until the end of the experiment.

6) Blood test and serum separation

WBC, RBC, HGB, and HCT were measured by injecting about 100 μl of blood obtained by blood collection into an EDTA-bottle and immediately injecting it into a blood analyzer (Multispecies Hematology Analyser) (950, Hemavet, USA). This was centrifuged for 20 minutes at 3,000 rpm in a high-speed centrifuge (VS 6000CFI, Vision, Korea) to separate serum for use in measurement.

7) Serum analysis

The separated serum was stored at 4°C until measurement, and biochemical measurements were GOT, GPT, albumin (Albumin, ALB), total bilirubin (TBIL), total protein (TP), glucose (Glucose, GLU) 6 items were measured using a biochemical analyzer (Clinical Chemistry Analyzer) (DRI-Chem 4000ie, Fujifilm Corporation, Japan).

8) Measurement of SOD (Superoxide Dismutases) activity

In order to measure the activity of SOD, a superoxide dismutases kit (BIOMATIK, USA) was used, and serum isolated from experimental animals was used as a sample. For preparation before measurement, 180 μl PBS (phosphate buffer saline) is diluted in 20 μl of the sample, and 990 μl PBS is diluted once more in 10 μl of the diluted sample. Add 100 µl of standard and sample to a 96 well plate and react at 37°C for 1 hour. Then, add 100 μl of Detection Reagent A reagent and store it at 37° C. for 1 hour. After washing 3 times, add 100 μl of Detection Reagent B reagent and store at 37° C. for 30 minutes. After washing 5 times, add 90 µl of Substrate Solution reagent and give color reaction at 37℃ for 20 minutes. Finally, 50 µl of Stop Solution was added to stop the color reaction and measured at 450 nm using a Microplate reader (EZ Reader 400, Biochrom, US).

9) Catalase activity measurement

In order to measure the activity of catalase, a catalase kit (BIOMATIK, USA) was used, and serum isolated from experimental animals was used as the sample. To prepare before measurement, dilute 180 μl PBS to 20 μl of the sample and use it. Add 100 µl of standard and sample to a 96-well plate, and react at 37°C for 1 hour. Then, add 100 μl of Detection Reagent A reagent and store at 37° C. for 1 hour. After washing 3 times, add 100 μl of Detection Reagent B reagent and store at 37° C. for 30 minutes. After washing 5 times, add 90 µl of Substrate Solution reagent and give color reaction at 37℃ for 20 minutes. Finally, 50 µl of the stop solution was added to stop the color reaction and measured at 450 nm using a microplate reader (EZ Reader 400, Biochrom, US).

10) Glutathione (GSH) measurement

In order to measure the activity of GSH, a glutathione kit (BIOMATIK, USA) was used, and serum isolated from experimental animals was used as a sample. Dilute PBS in the sample for preparation before measurement to find a dilution factor suitable for measurement. Add 50 µl of standard and sample to 96 well plate, add 50 µl of Detection Reagent A reagent on it, shake the plate well, and store at 37℃ for 1 hour. After washing 3 times, add 100 μl of Detection Reagent B reagent and store at 37° C. for 30 minutes. After washing 5 times, add 90 µl of Substrate Solution reagent and give color reaction at 37℃ for 20 minutes. Finally, 50 µl of the stop solution was added to stop the color reaction and measured at 450 nm using a microplate reader (EZ Reader 400, Biochrom, US).

11) Malondialdehyde (MDA) measurement

To measure the activity of MDA, a malondialdehyde kit (BIOMATIK, USA) was used, and serum isolated from experimental animals was used as the sample. Dilute PBS in the sample for preparation before measurement to find a dilution factor suitable for measurement. First, after washing the 96-well plate twice, 50 µl of the standard and sample are added, and 50 µl of an antibody reagent is added thereto, and then stored at 37°C for 45 minutes. After washing 3 times, 100 μl of SABC reagent is added and stored at 37° C. for 30 minutes. After washing 5 times, 90 µl of the TMB Substrate reagent is added and color development is performed at 37°C for 20 minutes. Finally, 50 µl of the stop solution was added to stop the color reaction and measured at 450 nm using a microplate reader (EZ Reader 400, Biochrom, US).

12) Histological observation

Liver tissue extracted from laboratory animals was fixed with bouin solution (Sigma, Germany) for 24 hours, followed by 2 in order of 70%, 80%, 90%, 100%, and ethanol/xylene (1:2) solution. After soaking and dehydrating for a period of time, the liver tissues were immersed in a solution of xylene/hard paraffin (2:1), xylene/hard paraffin (1:2), and hard paraffin in a 55°C incubator for 3 hours. Paraffin was embedded. The embedded liver tissue was sliced to a thickness of 5 μm using a microtome, and then subjected to HE (Hematoxylineosin-Eosin) staining process, and observed using an optical microscope (Nikon Eclipse 80i, Japan) at 400 magnification.

13) Statistics processing

The test results were expressed as the mean value and standard error (mean±S.E.). For statistical analysis between each experimental group, Mann-Whitney U test was performed among nonparametric methods using SPSS for Windows (version 10.05, SPSS). Statistical significance of the whole experiment was given significance in the confidence interval P<0.05.

1-2. Experiment result

1) Effect on antioxidant

① Change of SOD value

1 and Table 1 show the results showing the effect of the liver damage mitigating composition of the present invention on SOD levels. As a result of observing the effect of the sample administration on the SOD, the control group showed a significant decrease compared to the normal group, and the SOD values of all the experimental groups increased as compared to the control group.

② Change of Catalase

2 and Table 2 show the results showing the effect of the composition for alleviating liver damage of the present invention on catalytic levels. As a result of observing the effect of the sample administration on the catalase, the control group increased compared to the normal group, and the catalytic values of all the experimental groups decreased as compared to the control group.

③ GSH level change

3 and Table 3 show the results showing the effect of the liver damage mitigating composition of the present invention on GSH levels. As a result of observing the effect of sample administration on GSH, the control group showed a significant decrease compared to the normal group, and the GSH values of all experimental groups were significantly increased as compared to the control group. In addition, each experimental group increased the GSH level depending on the concentration of the liver damage-relieving composition.

④ MDA value change

4 and Table 4 show the results showing the effect of the liver damage mitigating composition of the present invention on the MDA level. As a result of observing the effect of sample administration on the MDA, the control group showed a significant increase compared to the normal group, and the MDA values of all the experimental groups were significantly decreased than the control group, like the positive control group.

2) Effect on serum transaminase levels

5 is a graph showing the effect of the composition for alleviating liver damage of the present invention on serum transaminase levels. As a result of observing the effect of sample administration on serum transaminase, the experimental group 3 (M400) showed a significant decrease compared to the control group in the ALT level, and the experimental group 2 (M200) and the experimental group 3 (M400) in the total bilirubin level. ) Showed a significant decrease compared to the control group.

3) Effect on leukocyte levels

6 is a graph showing the effect of the composition for alleviating liver damage of the present invention on leukocyte levels. As a result of observing the effect of the sample administration on the white blood cells, the control group increased in comparison with the normal group in white blood cell, neutrophil, and lymphocyte levels, and all experimental groups decreased as compared to the control group, as in the positive control group. Did.

4) Effect on Erythrocyte and Thrombocyte levels

7 is a graph showing the effect of the composition for alleviating liver damage of the present invention on Erythrocyte and Thrombocyte levels. In the platelet (PLT) level, the experimental group 3 (M400) showed a significant increase compared to the control group.

5) Histological observation

8 is a photograph showing the effect of the liver damage-relieving composition of the present invention on pathological changes (HE staining) in liver-damaged rats. As a result of observing the effect of the sample administration on histological changes, the control group showed a greater damage compared to the normal group, and the treatment groups improved the damage compared to the control group, especially the experimental groups 1 to 3 tended to improve in a concentration-dependent manner.

Although specific examples of the present invention have been presented above, the present invention is not limited to the above, and various modifications are possible within the technical scope of the present invention, and such modifications will belong to the claims of the present invention described below.

Claims (4)

Providing a composition for preventing or treating liver damage, including 25 to 45% by weight of thistle extract, 25 to 45% by weight of the extract of giza and 25 to 45% by weight of the wolfberry extract, based on the total weight of the composition,
The thistle extract is prepared by immersing thistle in n-hexane to degrease and then extracting the degreased thistle with ethyl acetate or water,
The extract of the earthworm is prepared by adding water to the ground earth, centrifuging, adding ethyl acetate to the supernatant, shaking, separating and concentrating the ethylacetate layer,
The wolfberry extract is prepared by extracting wolfberry with water at 65 to 85°C,
The composition for preventing or treating liver damage further comprises 200 to 400 parts by weight of pear juice with respect to 100 parts by weight of the composition.
delete (a) immersing thistle in n-hexane, degreasing, and then extracting the degreased thistle with ethyl acetate or water to prepare a thistle extract; Shaking the ethyl acetate into ethyl acetate, separating and concentrating the ethyl acetate layer to prepare a terrestrial extract, (c) extracting goji berry with water at 65 to 85°C to prepare a goji berry extract, and (d) the thistle Mixing 25 to 45% by weight of the extract, 25 to 45% by weight of the earth extract and 25 to 45% by weight of the wolfberry extract,
In the step (a), the degreased thistle was extracted with ethyl acetate or water for 6 to 10 hours, and the extracted extract was concentrated at a temperature of 50° C. or less to prepare a thistle extract,
A method for preparing a composition for preventing or treating liver damage, wherein 200 to 400 parts by weight of pear juice is further mixed with respect to 100 parts by weight of the composition in step (d). delete

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