KR102005031B1 - The Ginseng-Berry extract including alcoholic liver injury prevention functional ingredients and method for manufacturing it - Google Patents

Abstract

The present invention relates to a method for producing an extract containing a functional ingredient for preventing alcoholic liver damage from ginseng fruit, and more particularly, to a method for preparing an extract containing ginseng fruit, The enzyme reaction is carried out using at least one hydrolytic enzyme selected from the group consisting of celluloses and the extraction is carried out by the alcohol extraction method at a temperature of 45 to 55 DEG C to increase the total content of ginsenosides, The present invention relates to a method for producing a high-quality ginseng fruit extract having hepatocyte protective effect.

Description

Technical Field [0001] The present invention relates to a ginseng fruit extract containing a functional ingredient for preventing alcoholic liver damage and a method for manufacturing the same,

It is an object of the present invention to provide a method for extracting an extract containing an alcoholic liver injury-preventing functional ingredient from ginseng fruit.

Korea's ginseng has a historical record of about 1,000 years ago, and now it is a unique traditional herbal medicine or health functional food, which is shaped by a global market size of about $ 3.5 billion and domestic venture companies can enter the global market.

Ginseng is a perennial semisynthetic oily plant belonging to the acanthaceae family. Its efficacy has been widely known in various fields of China and Korea and has been used as herbal medicine in many fields since the past. The ingredients of ginseng include carbohydrate (60 ~ 70%), (12 to 16 percent), saponin (3 to 6 percent), fat soluble ingredients (1-2 percent), ash (4-6 percent) and vitamins (0.005 percent). The physiological activity of ginseng has recently been systematically pharmacologically approached, and it has been recognized that the ginseng root effect, sexual function and improvement of reproductive dysfunction, anti-hypertension and anti-arteriosclerosis effect, hematopoietic function and anemia treatment effect, blood glucose metabolism and diabetes improvement effect, Improvement of dysfunction, hangovers effect, prevention of parasitic infection, and analgesic action. It is widely known that ginseng contains a large amount of ginsenoside, and it has recently been used as a food for health food by applying it to foods in recent years. Ginseng usually takes more than 4 years to grow, and in order to obtain the highest quality ginseng, it has to be cultivated for 6 years. It takes a long time to grow, and during the cultivation, yields are higher than the original cultivation for various reasons such as various diseases and natural disasters. There is a disadvantage that it takes a lot of time to obtain ginseng.

Ginseng begins to bloom with ginseng in its third year or more, and greenish-colored fruit is produced in May. In the middle of July, the green fruits of ginseng ripen while ripening, and become fruitless.

However, ginseng cultivated farms mainly use ginseng roots. Therefore, in order to promote root growth rather than reproductive growth, when ginseng fruit begins to be produced, ginseng fruit is picked up.

However, it is known that ginsenoside Re is much more effective in the ginseng root than the ginseng root, and has a function of inhibiting the central nervous system, promoting DNA and RNA, activating plasmin, promoting adrenocorticotropic hormone secretion, , And Rb ₂ is known to have central nerve inhibitory action, DNA and RNA synthesis promoting action, prasmin activating action, adrenocorticotropic hormone secretion promoting action, and antidiabetic action.

Ginsenoside refers to saponin contained in ginseng, and ginseng contains various kinds of ginsenosides. The ginsenosides of ginseng include Panaxadiol (PD), Panaxatriol (PT), and Oleanane. In addition to the above ginsenoside components, ginsenosides include carbohydrates such as starch as antidiaminoside materials, antioxidative aromatic compounds of polyacetylene, gomisin N-A to protect liver, and acidic peptides that act like insulin.

The main pharmacological actions of ginsenoside affect the central nervous system, the endocrine system, the immune system, the metabolic system, etc., and exert various effects on the body regulating function. In addition, ginsenoside has a high ability to decompose fat, promoting nutrient absorption and digestion, promoting metabolism through activation of intracellular enzymes, increasing energy, restoring fatigue, feeling helpless, improving appetite, It is known that there is an effect such as acceleration.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made to solve the above problems, and an object of the present invention is to provide a method for efficiently producing ginseng fruit extract. More specifically, by extracting an extract from ginseng fruit, And to provide a high-quality ginseng fruit extract and an extraction method thereof.

The present invention also provides a ginseng fruit extract having a liver cell protection effect against alcohol and a method for producing the same.

 In order to achieve the above object, the present invention provides a method for extracting ginseng fruit extract comprising an alcoholic liver injury-preventing functional ingredient, comprising the steps of: a) preparing a ginseng fruit frozen lump by lyophilization of ginseng fruit, and b) Adding an enzyme to the frozen lysate to perform an enzyme reaction; and (c) extracting the enzyme-reacted mixture using the alcohol.

Also, in the present invention, the ginseng fruit is crushed and the seedless flesh is freeze-dried at -25 to -35 캜 for 3 days to produce a frozen crushed product.

Also, in the present invention, the enzyme used in the enzyme reaction includes at least one enzyme selected from the group consisting of pectinase, cellulase, and hemi-cellulase among enzymes capable of hydrolyzing pectin and cellulose .

In addition, the method of the present invention is characterized by comprising the step of reacting an enzyme containing pectinase having a pectinase activity of 80000 / g or more.

In the present invention, the enzyme is characterized in that the enzyme is reacted at a temperature of 40 to 65 ° C at a pH of 3.0 to 5.0.

In the present invention, the step of extracting the alcohol includes the step of extracting the enzyme-reacted mixture while maintaining the temperature at 45 to 55 ° C.

The method for preparing ginseng fruit extract according to the present invention can improve the yield of total ginsenoside obtained from ginseng fruit by using the ginseng fruit as a raw material by freeze-drying, and further, after the hydrolytic enzyme reaction, , It is possible to enhance the yield of total ginsenoside and antioxidant ability and to enhance the effect of preventing the damage of liver cells. Therefore, it is possible to provide a product of higher quality than the product which does not process the enzyme reaction .

FIG. 1 shows the pilot production process of ginseng fruit extract, showing that the production yield of the pilot scale for the raw material is 3.5 to 4.8%, and the higher the amount of enzyme added, the higher the yield.
FIG. 2 shows changes in liver enzymes (ALT, AST) according to treatment with ginseng fruit in HepG2 cells induced by alcoholic liver toxicity.
FIG. 3 (A) is a graph showing MTT assay results for Hovenia dulcis, (B) MTT assay results for ginseng fruit-A, and (C) MTT assay results for ginseng fruit- As shown in the graph.
FIG. 4 is a graph showing blood concentrations of GOT / AST, GPT / ALT and LDH, which are indexes of liver toxicity, by inducing liver toxicity by ethanol administration.
FIG. 5 is a bar graph showing the values of blood GOT / AST and GPT / ALT, respectively.
FIG. 6 is a bar graph showing the blood levels of LDH produced by Hovenia dulcis extract, Ginseng fruit-A and Ginseng fruit-B against alcoholic liver disease.
FIG. 7 is a histogram showing liver and spleen weights measured by administration of ethanol, after administration of Hovenia dulcis extract, ginseng fruit-A and ginseng fruit-B, or pretreatment.
FIG. 8 is a bar graph showing the body weight gain rate by the administration of ethanol extract, the administration of Hovenia dulcis extract, the ginseng fruit-A and the ginseng fruit-B, or the pretreatment.
FIG. 9 is a bar graph showing the measurement of the expression levels of SOD (superoxide dismutase) and catalase, which are antioxidative activity-related enzymes.
FIG. 10 shows the expression of inflammation-related molecules COX-2, HO-1, iNOS and TGF-β1 in the group treated with Hovenia dulcis extract, ginseng fruit-A and ginseng fruit-B and western blot Western blot).
FIG. 11 is a bar graph showing changes in blood liver levels according to administration of Hovenia dulcis extract, ginseng fruit-A and ginseng fruit-B.
FIG. 12 is an anatomical observation of liver tissues of mice treated with Hovenia dulcis extract, Ginseng fruit-A and Ginseng fruit-B.
FIG. 13 is an anatomical observation of liver tissue treated with ethanol after pretreatment of Hovenia dulcis extract, Ginseng fruit-A and Ginseng fruit-B.

Hereinafter, preferred embodiments of the present invention will be described in detail. It should be understood, however, that the invention can be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. It should be noted that the embodiments of the present invention described below are intended to sufficiently convey the spirit of the present invention to those skilled in the art.

According to an embodiment of the present invention, there is provided a method of preparing a ginseng fruit by freeze-drying a) ginseng fruit to prepare a frozen disintegration product of ginseng fruit, b) obtaining an enzyme reaction product by adding enzyme to the ginseng fruit freezing disintegration product, c) And extracting the ginseng fruit extract from the enzyme-reacted mixture. The ginseng fruit extract containing the alcoholic liver injury-preventing functional ingredient can be prepared.

In addition, the ginseng fruit may be pulverized to remove seeds, and the fruit pulp may be freeze-dried at -25 to -35 캜 for 3 days to produce a frozen crumbly product.

In addition, the enzyme comprises at least one enzyme selected from the group consisting of pectinase, cellulase, and hemi-cellulase among enzymes capable of hydrolyzing pectin and cellulose. Ginseng fruit extract can be prepared.

In addition, the step of obtaining the enzyme-reacted mixture may include the step of reacting the enzyme containing the pectinase in an amount of 80000 ug / g or more to prepare the ginseng fruit extract containing the alcoholic liver injury-preventing functional ingredient.

The step of obtaining the enzyme-reacted mixture comprises allowing the enzyme to react with the enzyme at a temperature of 40 to 65 ° C at a pH of 3.0 to 5.0, thereby preparing a ginseng fruit extract containing an alcoholic liver injury-preventing functional ingredient .

The extracting step may include extracting the ginseng fruit extract at a temperature of 45 to 55 ° C to prepare an extract of ginseng fruit containing an alcoholic liver injury-preventing functional ingredient.

Examples of the present invention may include ginseng fruit extract prepared by the above-described method.

In addition, examples of the present invention may include a liver damage-preventing functional health supplement including an extract prepared by the above-described method.

1. Ginseng fruit freeze drying and shredding

In order to increase the content of total ginsenosides contained in the ginseng fruit in the present invention, the ginseng fruit collected from ginseng is preferably washed with purified water and lyophilized at a temperature of -25 to -35 ° C . In the present invention, freeze-drying of the ginseng fruit is preferably carried out for 3 days so that the moisture content of the ginseng fruit is 4.55%. The inventors of the present invention prepared freeze-dried fruits of ginseng fruit by increasing the content of total ginsenosides by using the characteristics that the freeze-dried products have good crushability and at the same time minimize the change of substances such as nutrients of the raw materials.

2. Screening condition of extract of ginseng fruit frozen lysate

In order to increase the content of total ginsenosides and to obtain ginseng fruit extract having excellent antioxidative ability, the extraction conditions using the alcohol were set by the following method. 100% (v / v) of 95% alcohol was added to 1 kg of ginseng fruit frozen crumbs, and the mixture was extracted with a refluxing shaking water bath at 50 ℃ for 3 and 5 hours, filtered through nonwoven fabric, and then the filtrate was used as an analytical sample. The antioxidant activity of the ginseng fruit extracted with the alcohol was investigated. The results are shown in Table 1 below.

Type Processing time (hr) The DPPH radical scavenging activity IC ₅₀ (ppm) Control 0 0 449.3 Control group 3 3 321.12 Control group 5 5 342.91 Ethanol (EtOH) 3 355.98 5 328.16

As shown in Table 1, the DPPH radical scavenging IC ₅₀ of the ginseng fruit of the ginseng fruit was 449.3 ppm. The temperature change was maintained at 50 ° C. and the change with time was observed. As a result, the IC ₅₀ value was lowest at 321.12 ppm, and then increased slightly to 342.91 ppm at 5 hours. In the extraction of alcohol, the IC ₅₀ value tended to decrease as the extraction time was longer, unlike the control group. In case of extracting ginseng fruit frozen lumps, it can be predicted that increasing the extraction time increases the content of antioxidant components.

3. Selection of Ginseng Fruit Hydrolysates

In the present invention, in order to efficiently extract the functional ingredient for preventing alcoholic liver damage of ginseng fruit, enzyme extraction conditions were selected by the following method. After adding 100% (v / v) of purified water to 1kg of ginseng fruit frozen lysate, 0.05% and 0.2% of enzyme were added to each other and hydrolyzed in a 50 ℃ reflux shaking water bath for 3 and 5 hours respectively. The lysates were lyophilized and used as analytical samples. The antioxidant properties of pectin and cellulose hydrolytic enzymes were examined and the results are shown in Table 2 below.

Enzyme type Enzyme concentration (%) Enzyme treatment time (hr) The DPPH radical scavenging activity IC ₅₀ (ppm) Control 0 0 0 449.3 Control group 3 0 3 321.12 Control group 5 0 5 342.91 Enzyme A 0.05 3 314.3 5 381.5 0.2 3 329.3 5 377.6 Enzyme B 0.05 3 359.3 5 345.4 0.2 3 386.2 5 428.7

Table 2 above shows the IC ₅₀ values for DPPH radicals depending on the enzyme type, treatment concentration and time. The enzyme A in Table 2 is Sumizyme SPC, which is a combination of pectinase, cellulase, and hemicellulase among enzymes capable of hydrolyzing pectin and cellulose, and in particular, those containing more than 80000 u / g pectinase Feature. In this case, 1u means the amount of enzyme capable of producing 1μmol of galacturonic acid per minute by decomposing polygalacturonic acid under the conditions of pH 4.0 and 25 ° C by the sigma method. For enzyme A, the enzyme reaction is carried out at a temperature of 40 to 65 ° C and at a pH of 3.0 to 5.0.

The enzyme B in Table 2 is a mixture of pectinase and cellulase, which are enzymes capable of hydrolyzing pectin and cellulose, as aplantacease TCL. Particularly, when the pectinase is 190 u / g, the cellulase is 1495 U / g or more . At this time, 1 U means the amount of enzyme capable of producing 1 μmol of glucose per minute by decomposing the substrate carboxymethylcellulose. For enzyme B, the enzyme reaction is treated at a temperature of 40 to 60 ° C and a pH of 2.0 to 5.5.

In the case of enzyme A, the IC ₅₀ value tended to be low when the enzyme treatment was performed for a short period of time. Enzyme B also showed a lower IC ₅₀ value in the treatment period of 3 hours, but it was higher than that of the control group. It is predicted that the addition of enzyme A to the ginseng fruit through alcohol extraction will increase the contents of antioxidant components.

4. Changes of ginsenoside components by enzymatic hydrolysis and extraction of ginseng fruit juice

In order to efficiently extract total ginsenosides of the ginseng fruit, the content of ginsenosides was measured by extracting the ginseng fruit using an enzyme that can be used as a food and a diet, and the results are shown in Table 3 below.

Sample Compound Rg1 Re Rf F5 F3 Rb1 Rg2 Rh1 Rc Rb2 Rb3 F1 Rd F2 Total content Control 0 5.18 0.92 0.10 0.27 0.56 0.22 0.12 0.01 0.34 0.37 0.04 0.08 0.36 0.26 8.84 Control group 3 5.39 0.99 0.08 0.27 0.56 0.27 0.13 0.02 0.45 0.49 0.05 0.09 0.49 0.29 9.58 Control group 5 5.07 0.92 0.08 0.25 0.53 0.25 0.13 0.02 0.40 0.46 0.05 0.09 0.44 0.26 8.94 ethanol
3hr 6.74 0.96 0.09 0.27 0.56 0.26 0.21 ND 0.40 0.46 0.05 0.09 0.44 0.27 10.82 ethanol
5hr 6.60 0.95 0.10 0.28 0.56 0.28 0.17 0.01 0.44 0.51 0.06 0.09 0.49 0.28 10.84 Enzyme A 0.05%, 3 hr 5.87 1.09 0.09 0.25 0.52 0.19 0.12 0.02 0.43 0.47 0.06 0.10 0.52 0.31 10.03 Enzyme A 0.2%, 3 hr 6.32 1.16 0.09 0.19 0.60 0.19 0.14 0.02 1.26 1.58 0.18 0.18 2.03 0.43 14.39 Enzyme A 0.05%, 5hr 5.32 0.95 0.09 0.26 0.57 0.21 0.13 0.01 0.40 0.47 0.06 0.09 0.49 0.29 9.33 Enzyme A 0.2%, 3 hr 4.93 1.00 0.07 0.23 0.53 0.17 0.12 0.01 0.37 0.45 0.05 0.09 0.50 0.30 8.83 Enzyme B 0.05%, 3 hr 5.91 1.06 0.09 0.26 0.58 0.31 0.14 0.02 0.68 0.78 0.09 0.12 0.84 0.35 11.23 Enzyme B 0.2%, 3 hr 5.73 1.17 0.09 0.24 0.54 0.16 0.14 0.02 0.45 0.49 0.05 0.10 0.57 0.36 10.11 Enzyme B 0.05%, 5 hr 5.69 1.02 0.10 0.25 0.55 0.37 0.14 0.02 0.76 0.90 0.10 0.12 0.96 0.34 11.31 Enzyme B 0.2%, 3 hr 5.42 0.97 0.09 0.25 0.54 0.18 0.13 0.01 0.37 0.44 0.03 0.14 0.45 0.27 9.28

Table 3 shows changes in ginsenoside components by hydrolysis of the enzyme of ginseng fruit and extraction of the alcohol. To Table 3 by looking showed overall content of Rg ₁ highest _{Re, Rd, Rb 2, F3} , Rc, F2, F5, Rb 1, Rg 2, F 1, Rf, Rb 3, Rh 1 order Respectively. The total ginsenoside content was the highest at 9.58% in the control group for 3 hours and 10.82 ~ 10.84% for 3 hours and 5 hours in the alcohol extract group. Enzyme A, 0.2%, showed the highest level of 14.39% in the 3 hour period and the highest level was 11.31% in the enzyme B, 5 hour period. As a result of Example 4, the IC ₅₀ value of DPPH radical was lowest in enzyme A and the highest content of ginsenoside was detected. Thus, the enzyme A was used to hydrolyze ginseng fruit It is predicted that the method of alcohol extraction will increase the content of total ginsenosides.

5. Extraction of minimum test volume of hydrolysis of ginseng fruit

In the present invention, the extraction procedure of the minimum test production scale was conducted as shown in FIG. 1, in order to prepare the ginseng fruit extract having the effect of protecting the alcoholic liver damage by different concentrations of the enzyme A selected in the previous study. In this process, enzymes deactivation, enrichment and sterilization processes, which are necessary for quality control, were added to production and products, and spray drying was performed to reduce production costs.

Fig. 1 shows the production process of the minimum test production scale of ginseng fruit extract.

As shown in FIG. 1, it can be seen that the production yield of the minimum test production scale for this raw material is 3.5 to 4.8%, and the higher the enzyme addition amount is, the higher the yield is.

6. Measuring the protective effect of alcoholic liver damage on ginseng fruit

In order to examine the efficacy of the ginseng fruit for the protection against alcoholic liver damage, toxicity test using alcohol-induced HepG2 cells derived from human liver carcinoma among the cell lines used for the experiment related with alcoholic liver toxicity (ALT, AST) by the treatment of ginseng fruit, as shown in FIG. 2.

FIG. 2 shows changes in liver enzymes (ALT, AST) according to treatment with ginseng fruit in HepG2 cells induced by alcoholic liver toxicity.

As shown in FIG. 2, in vitro toxicity can be confirmed by measuring the release amount of transaminase in the culture medium of the cells. When hepatocytes such as HepG2 are treated with ethanol for 24 hours, ALT (Alanine transaminase) and Aspartate transaminase (AST). In this experiment, the levels of ALT and AST were increased in the ethanol treatment, and the difference in degree of ethanol treatment between the ginseng and the ethanol extract was found to be different according to the extraction condition of the sample. However, .

<Example 1> Cytotoxicity test on fruit extract of ginseng

To examine the cytotoxicity of ginseng fruit extract (hereinafter, "ginseng fruit-A") prepared by using enzyme A and ginseng fruit extract (hereinafter referred to as "ginseng fruit-B") prepared by using enzyme B, RAW 264.7 cells Hovenia japonica, Ginseng fruit-A and Ginseng-B samples were diluted to various concentrations and cultured for 48 hours. Then, cytotoxicity was measured by MTT assay.

FIG. 3 (A) is a graph showing MTT assay results for Hovenia dulcis, (B) MTT assay results for ginseng fruit-A, and (C) MTT assay results for ginseng fruit- As shown in the graph. In each graph, the abscissa is the dose (占 퐂 / ml) and the ordinate is the cell growth rate (%).

The cytotoxicity of Hovenia dulcis, Ginseng Fruit-A and Ginseng fruit-B samples was examined by the graph shown in FIG. 3, and it was confirmed that the cell growth rate was decreased when 500 μg / ml or more of all three samples were administered . As a result, it can be confirmed that the concentration of 250 g / ml of Hovenia dulcis, Ginseng fruit-A and Ginseng fruit-B samples is safe.

It is difficult to determine the dose of the sample to be used in the in vivo experiment based on the cytotoxicity experiment data described in the above-described in vitro experiment, but it is estimated that the dosage up to 2-3 mg / mouse is safe, The sample was administered at 2 mg / mouse.

<Example 2> Hepatic toxicity induction experiment by ethanol administration

Five-week-old male Balb / c mice were orally administered 25% ethanol at a dose of 5 g / kg once a day for a total of 7 days to induce alcoholic liver disease.

FIG. 4 is a graph showing blood concentrations of GOT / AST, GPT / ALT and LDH, which are indexes of liver toxicity, by inducing liver toxicity by ethanol administration. FIG. 4A is a graph showing the blood concentration of GOT / AST (U / I), FIG. 4B is a graph indicating the blood concentration of GPT / ALT (U / I) U / I). &Lt; / RTI &gt;

4, the blood concentration of GOT / AST, GPT / ALT, and LDH in an experimental group (EtOH only) administered with 25% ethanol compared to a control group (NONE) in which 25% ethanol was not administered at 5 g / It was confirmed that all of them increased.

Accordingly, it can be confirmed that administration of 25% ethanol damages liver cells and toxicity is expressed.

<Example 3> Test for inhibiting liver toxicity of fruit extract of ginseng

GOT / AST and GPT / ALT in the serum of mice given 10 times orally once a day at a dose of 0.5 mg / mouse and 1 mg / mouse from 3 days before the administration of ethanol, ginseng fruit-A and ginseng fruit- And the activity of inhibiting liver disease of the sample was confirmed by quantitative analysis. At this time, blood serum GPT / ALT, GOT / AST and LDH were quantified using the serum samples 1 day after completion of the ethanol administration.

FIG. 5 is a bar graph showing the values of blood GOT / AST and GPT / ALT, respectively. In FIG. 5 (A), the abscissa represents the groups in which ethanol was not administered (NONE), ethanol alone (EtOH only), ethanol and hovenoe extract 0.5 mg, ethanol and hinoki extract A group treated with ethanol and ginseng fruit-A 0.5 mg, a group administered with ethanol and ginseng fruit-A 1 mg, a group administered with ethanol and ginseng fruit-B 0.5 mg, And 1 mg of ginseng fruit-B, and the vertical axis represents the blood GOT / AST (U / I) value. Statistical significance was assessed by Student's two-tailed t-test on groups treated with ethanol only, * means p <0.05.

5 (B), the abscissa represents the groups in which ethanol was not administered (NONE), ethanol alone (EtOH only), ethanol and hovenoe extract 0.5 mg, ethanol, A group treated with ethanol and ginseng fruit-A 0.5 mg, a group administered with ethanol and ginseng fruit-A 1 mg, a group administered with ethanol and ginseng fruit-B 0.5 mg, , 1 mg of ethanol and ginseng fruit-B, and the vertical axis represents the GPT / ALT (U / I) level in the blood. Statistical significance was assessed by Student's two-tailed t-test on groups treated with ethanol only, * means p <0.05.

5, it was confirmed that the values of GOT / AST and GPT / ALT in the blood raised by the administration of ethanol were decreased in the experimental group administered with ginseng fruit-A and ginseng fruit-B. In particular, it was confirmed from FIG. 5 that the ginseng fruit-A exhibits an inhibitory effect equal to or higher than that of the comparative group, Hovenia dulcis.

In addition, using the fact that hepatocellular damage occurs and the blood concentration of LDH enzyme in liver cells increases, the extract of Hovenia dulcis extract, Ginseng fruit-A and ginseng The effect of inhibiting the liver damage of fruit-B was observed.

FIG. 6 is a bar graph showing the blood levels of LDH produced by Hovenia dulcis extract, Ginseng fruit-A and Ginseng fruit-B against alcoholic liver disease. In the graph of FIG. 6, the abscissa axis indicates the groups in which ethanol was not administered (NONE), ethanol alone (EtOH only), ethanol and 0.5 mg of Hovenia dulcis extract, ethanol and Hovenia dulcis extract Ethanol, ginseng fruit-A 0.5 mg, ethanol and ginseng-A 0.5 mg, ethanol and ginseng-A 0.5 mg, ethanol and ginseng-A 0.5 mg, And 1 mg of fruit-B together, and the vertical axis represents blood LDH (U / I). The statistical significance was examined by Student's two-tailed t-test for the ethanol-only group, * means p <0.05, ** means p <0.01 do.

As a result of observing the inhibitory effect on liver damage of Hovenia dulcis, Ginseng Fruit-A and Ginseng Fruit-B to the increase of blood LDH concentration, the ginseng fruit-A and the ginseng fruit- It is possible to confirm that the effect of suppressing the increase of LDH in blood is better than that of the extract, and thus it can be presumed that the occurrence of alcoholic liver disease can also inhibit liver cell damage.

On the other hand, in order to measure the level of toxicity to organs by administration of Hovenia dulcis extract, ginseng fruit-A and ginseng fruit-B or ethanol treatment after pre-treatment, the extracts of Hovenia dulcis, Ginseng fruit A and Ginseng- And the weight of liver and spleen of the ethanol-treated group after the pretreatment of the sample was measured.

FIG. 7 is a histogram showing liver and spleen weights measured by administration of ethanol, after administration of Hovenia dulcis extract, ginseng fruit-A and ginseng fruit-B, or pretreatment.

In FIG. 7 (A), the abscissa indicates the group administered with no ethanol (NONE), ethanol alone (EtOH only), 0.5 mg of Hovenia dulcis extract, and 1 mg of Hovenia dulcis extract B group, the ginseng fruit-B 0.5 mg group, and the ginseng fruit-B 1 mg group, and the vertical axis represents the group of the liver liver weight (g).

In FIG. 7 (B), the abscissa represents the groups in which ethanol was not administered (NONE), ethanol alone (EtOH only), 0.5 mg of ethanol and Hovenia dulcis extract, ethanol, A group treated with ethanol and ginseng fruit-A 0.5 mg, a group administered with ethanol and ginseng fruit-A 1 mg, a group administered with ethanol and ginseng fruit-B 0.5 mg, And 1 mg of ginseng fruit-B, and the vertical axis represents the weight (g) of the liver.

In FIG. 7 (C), the abscissa indicates the group from which ethanol was not administered (NONE), ethanol alone (EtOH only), 0.5 mg of Hovenia dulcis extract, and 1 mg of Hovenia dulcis extract (1 mg), ginseng fruit-B (0.5 mg), and ginseng fruit-B (1 mg), and the vertical axis represents the spleen weight (g) of spleen.

In FIG. 7 (D), the abscissa axis indicates the groups in which ethanol was not administered (NONE), ethanol alone (EtOH only), ethanol and 0.5 mg of Hovenia dulcis extract, ethanol, A group treated with ethanol and ginseng fruit-A 0.5 mg, a group administered with ethanol and ginseng fruit-A 1 mg, a group administered with ethanol and ginseng fruit-B 0.5 mg, And 1 mg of ginseng fruit-B together, and the vertical axis represents the weight (g) of the spleen.

7 (A) to 7 (D), the weight of the organs (liver, spleen) by the administration of Hovenia dulcis, Ginseng Fruit-A and Ginseng Fruit-B was reduced, and ethanol was administered after the pretreatment It can be confirmed that the weight loss of the organ (liver, spleen) is not significantly affected in one group.

Weight gain was also not significantly increased in the bar graph of FIG. 8, indicating that no significant weight gain occurred in the hovenose tree, ginseng fruit-A and ginseng fruit-B administration groups and ethanol treated group after pretreatment .

In the bar graph of FIG. 8, the abscissa axis indicates the group from which the ethanol was not administered (NONE), the ethanol alone group (EtOH only), the group administered 0.5 mg of Hovenia dulcis extract, and 1 mg of the Hovenia dulcis extract A group treated with ginseng fruit-A 0.5 mg, a group treated with ginseng fruit-A 1 mg, a group treated with ginseng fruit-B 0.5 mg, a group treated with ginseng fruit-B 1 mg, a group treated with ethanol and hinoki extract 0.5 , ethanol and ginseng fruit-1 mg, ethanol and ginseng fruit-A 0.5 mg, ethanol and ginseng-A 1 mg, ethanol and ginseng extract-1 mg, -B 0.5 mg together, ethanol and ginseng-B 1 mg together, and the vertical axis represents the rate of weight gain (% increase).

<Example 4> Antioxidant activity analysis in liver tissue

The expression levels of superoxide dismutase (SOD) and catalase (SOD), which are antioxidative enzymes, were determined and the relationship between the administration of Hovenia dulcis, Ginseng fruit A and Ginseng B and antioxidant activity was analyzed. At this time, SOD and catalase were analyzed by quantifying SOD and catalase in liver tissues by lysis of liver tissue on the first day after completion of ethanol administration.

FIG. 9 is a bar graph showing the measurement of the expression levels of SOD (superoxide dismutase) and catalase, which are antioxidative activity-related enzymes. In FIG. 9 (A), the abscissa represents the groups in which ethanol was not administered (NONE), ethanol alone (EtOH only), 0.5 mg of ethanol and Hovenia dulcis extract, ethanol, A group treated with ethanol and ginseng fruit-A 0.5 mg, a group administered with ethanol and ginseng fruit-A 1 mg, a group administered with ethanol and ginseng fruit-B 0.5 mg, And 1 mg of ginseng fruit-B, and the vertical axis represents the expression amount (S / ml) of superoxide dismutase (SOD).

In FIG. 9 (B), the abscissa indicates the groups in which ethanol was not administered (NONE), ethanol alone (EtOH only), ethanol and 0.5 mg of Hovenia dulcis extract, ethanol, A group treated with ethanol and ginseng fruit-A 0.5 mg, a group administered with ethanol and ginseng fruit-A 1 mg, a group administered with ethanol and ginseng fruit-B 0.5 mg, And 1 mg of ginseng fruit-B, and the vertical axis represents the expression amount (U / ml) of catalase. The statistical significance was examined by Student's two-tailed t-test for the ethanol-only group, * means p <0.05, ** means p <0.01 do.

9 (A) and 9 (B), there was no difference in the amount of SOD expressed by the administration of Hovenia dulcis, Ginseng Fruit-A and Ginseng Fruit-B, but the amount of catalase expression was slightly increased Can be confirmed. Accordingly, antioxidant activity can be expected to be increased by administration of Hovenia dulcis, Ginseng fruit-A and Ginseng fruit-B.

<Example 5> Analysis of inflammatory factors in liver tissue

In order to investigate the correlation between inflammation related factors such as Hovenia dulcis extract, ginseng fruit-A and ginseng fruit-B and COX-2, iNOS, HO-1 and TGF-β1 in liver cells, The expression levels of COX-2, iNOS, HO-1, and TGF-β1 were measured by Western blotting in the group administered with the ginseng extract-B and the group treated with ethanol after the pretreatment.

FIG. 10 shows the expression of inflammation-related molecules COX-2, HO-1, iNOS and TGF-β1 in the group treated with Hovenia dulcis extract, ginseng fruit-A and ginseng fruit-B and western blot Western blot).

10, no change was observed in the expression of COX-2, iNOS and TGF-? 1, but in the case of the HO-1 molecule having an inflammation-suppressing action, the ginseng fruit-A and the ginseng fruit- B pretreatment and then ethanol administration, the expression level of HO-1 molecule was increased.

Example 6: Tissue anatomical examination

In order to confirm the safety of Hovenia dulcis extract, Ginseng fruit A and Ginseng fruit B, changes in blood liver levels were measured by the administration of Hovenia dulcis extract, Ginseng fruit A and Ginseng fruit B.

In FIG. 11 (A), the abscissa indicates the group in which nothing was administered (NONE), ethanol alone (EtOH only), 0.5 mg of Hovenia dulcis extract, 1 mg of Hovenia dulcis extract , Ginseng fruit-A 0.5 mg, Ginseng fruit-A 1 mg, Ginseng fruit-B 0.5 mg and Ginseng fruit-B 1 mg, and the vertical axis represents GOT / AST (U / I).

In FIG. 11 (B), the horizontal axis represents the group in which nothing was administered (NONE), ethanol alone (EtOH only), 0.5 mg of Hovenia dulcis extract, 1 mg of Hovenia dulcis extract , Ginseng fruit-A 0.5 mg, ginseng fruit-A 1 mg group, ginseng fruit-B 0.5 mg group and ginseng fruit-B 1 mg group, and the vertical axis represents GPT / ALT (U / I).

Through the graphs shown in FIGS. 11A and 11B, the blood liver levels according to the administration of Hovenia dulcis, Ginseng Fruit-A and Ginseng Fruit-B did not change significantly compared with those in the non-administered group Therefore, it can be confirmed that there is no unusual toxicity.

12 is an anatomical observation of the liver tissue of the mouse. As a result of the anatomical observation according to FIG. 12, in the group administered with ethanol, slight cell damage was observed in the surrounding tissue of the blood vessels, but the extracts of Hovenia dulcis (0.5 mg and 1.0 mg), ginseng fruit-A (0.5 mg and 1 mg) It was confirmed that no remarkable cell damage was observed in the group administered with ginseng fruit-B (0.5 mg and 1 mg). In addition, no significant change was observed in the group treated with Hovenia dulcis extract, Ginseng fruit-A and Ginseng fruit-B alone.

FIG. 13 is an anatomical observation of liver tissue treated with ethanol after pretreatment of Hovenia dulcis extract, Ginseng fruit-A and Ginseng fruit-B. As a result of the observation according to FIG. 13, no remarkable liver damage was observed in the ethanol-treated group, but the liver was not damaged in the mice pretreated with Hovenia dulcis extract, Ginseng fruit-A and Ginseng fruit- I could confirm.

The present invention relates to a health supplement containing a ginseng fruit extract rich in the total content of ginsenoside produced by the above-mentioned method and having a high liver cell protection effect, wherein the health supplement is a liquid beverage formulation Can be provided.

As an example of the above-mentioned health supplement food, an extract of ginseng fruit may be pulverized using a method such as spray drying, and the powder may be provided as any one or more solid formulations selected from tablets, rings, powders, tablets and capsules have.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is clearly understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the present invention.

Claims (8)

a) preparing a ginseng fruit frozen lump by freeze-drying the ginseng fruit;
b) An enzyme consisting of cellulase, hemi-cellulase, and pectinase is added to the frozen crumb of the ginseng fruit at a concentration of 0.05 to 0.1% based on the total weight of the crushed product, and the mixture is heated at a temperature of 40 to 65 캜 at a pH of 3.0 to 5.0 Obtaining an enzyme-reacted mixture; And
and c) extracting the ginseng fruit extract from the enzyme-reacted mixture using the alcohol to obtain a ginseng fruit extract, and extracting the ginseng fruit extract as an active ingredient at a concentration of 200 to 300 μg / ml A method for preparing a ginseng fruit extract for the production of a composition for preventing alcoholic liver damage The method according to claim 1,
Wherein the ginseng fruit is crushed and the seedless flesh is freeze-dried at -25 to -35 캜 for 3 days to prepare a frozen crushed product. The method for producing the fruit extract of ginseng for the preparation of a composition for preventing alcoholic liver damage delete The method according to claim 1,
Wherein the step of obtaining the enzyme-reacted mixture comprises reacting an enzyme comprising 800,000 u / g or more of the pectinase. delete The method according to claim 1,
Wherein the extracting comprises:
And extracting the ginseng fruit extract at a temperature of 45 to 55 ° C. The method for producing a ginseng fruit extract for the production of a composition for preventing alcoholic liver damage. delete delete

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