KR101426907B1 - Composition for preventing or treating hepatic injury comprising davallialactone - Google Patents

Abstract

The present invention relates to a composition for prevention or treatment and improvement of liver injury comprising multivalent lactone as an active ingredient. The multivalent lactone according to the present invention is useful for the reduction of hepatic enzyme (ALT, AST) levels, inhibition of liver tissue necrosis, inhibition of the production of reactive oxygen species, inhibition of lipid peroxidation, and the expression of SOD Reduction of oxidative stress such as reduction of hepatocyte proliferation, reduction of factors related to inflammation of hepatocytes such as TNF-a, iNOS, and COX-2, and reduction of hepatocyte apoptosis, It can be very useful for treatment.

Description

TECHNICAL FIELD The present invention relates to a composition for preventing or treating hepatotoxicity,

The present invention relates to a composition for prevention or treatment and improvement of liver injury comprising multivalent lactone as an active ingredient.

The liver is the organ in which the metabolism of the body is most active in the human body organs. It is located between the digestive system and systemic circulation in the human body. It metabolizes external substances as well as protein synthesis and internal metabolism, It is a very important organ to perform the function. In particular, since substances such as drugs and toxins that enter the living body are first metabolized by the liver, the liver is more likely to be exposed to toxic substances than other organs, and the probability of damage is very high. However, the liver is an organ with excellent regeneration ability. In the case of acute liver injury, it is asymptomatic or only slightly elevated in hepatic enzyme levels, and most of it goes through self-healing process. However, if the injury is continuously chronicized by exposure to toxic substances, irreversible hepatic necrosis or the like results in hepatic dysfunction and cirrhosis, making it difficult to recover to normal liver. In addition to chemical and biological factors such as pollutants, toxic substances, various drugs and toxins, fungi, and viruses due to industrialization, the mental stress of modern people increases the damage of the liver, causing abnormalities of the human immune system, It is also a cause. Since liver disease has no symptoms at first, it is only until the damage is detected that the clinical symptoms are found. Therefore, it is the highest cause of death not only in Korea but also in the world, but there is no effective treatment method. Therefore, there is a desperate need to develop a drug capable of preventing or treating liver damage while maintaining the structure and function of liver tissue.

On the other hand, silybin (silybum marianum), sililybin, silydiamine and silycristine extracted from natural materials and applied in clinical practice are used as a liver function protecting agent. Although there are silymarin formulations consisting of isomers, there are only a few cases in which clinical trials are in actual use as a therapeutic agent, despite the numerous efforts to develop a therapeutic agent for liver damage from natural products. But the analysis of the exact components and the content is insufficient. Accordingly, the present inventors have studied the hepatoprotective activities of polyvalent lactones extracted from Phellinus linteus and Tile mushroom for the purpose of searching for a substance having the effect of preventing or treating liver damage and for applying it to foods and medicines. The strong antioxidative and antiinflammatory effects of polyvalent lactones have been known, but the prevention or therapeutic effect of liver injury has not yet been reported in domestic and foreign literature.

The inventors of the present invention conducted studies to search for a substance having a preventive or therapeutic effect on liver injury. In the liver injury mice treated with carbon tetrachloride, Reduction of oxidative stress such as increase of SOD expression, and inflammation of hepatocytes such as TNF-α, iNOS, and COX-2. The effect of decreasing the factor and the effect of decreasing the apoptosis of the hepatocyte are excellent. Thus, the present invention has been completed.

Accordingly, the present invention is intended to provide a composition for preventing or improving liver damage, which comprises multivalent lactone as an active ingredient.

The present invention provides a pharmaceutical composition for preventing or treating liver injury comprising pluralactone as an active ingredient.

The present invention also provides a food composition for prevention or improvement of liver injury comprising pluralactone as an active ingredient.

The multivalent lactone according to the present invention is useful for the reduction of hepatic enzyme (ALT, AST) levels, inhibition of liver tissue necrosis, inhibition of the production of reactive oxygen species, inhibition of lipid peroxidation, and the expression of SOD Reduction of oxidative stress such as decrease of hepatocyte proliferation, reduction of factors related to inflammation of hepatocytes such as TNF-α, iNOS, and COX-2, and reduction of hepatocyte apoptosis, It can be very useful for treatment.

FIG. 1 is a graph showing the effect of the multivalent lactone of the present invention on ALT (alanine aminotransferase) and AST (aspartate aminotransferase) levels in liver damaged mouse serum.
Fig. 2 is a graph showing the necrosis-inhibiting effect of the liver injury mouse liver tissue of the multiligaractone of the present invention.
FIG. 3 is a graph showing the effect of the present invention on the reduction of DHE (dihydroethidium) (3A, 3B), MDA (malonyldialdehyde) reduction (3C) and Cu / Zn SOD (superoxide dismutase) 3D).
4 is a graph showing the effect of the multivalent lactone of the present invention on CYP2E1 (full name) expression.
FIG. 5 is a graph showing the suppressive effect (5B) of serum TNF-α reduction (5A) and inflammatory markers (iNOS, COX-2) in hepatocyte-deficient mice of the multivalent lactone of the present invention.
6 is a graph showing the effect of the polyvalyllactone of the present invention on the expression of HO-1.
FIG. 7 is a graph showing the effect of reducing the expression of caspase 3 (caspase 3), which affects hepatocyte apoptosis and hepatocyte apoptosis, in hepatic injury mice of the multivalent lactone of the present invention.

The present invention provides a composition for preventing or treating liver damage comprising multivalent lactone represented by the following formula (1) as an active ingredient.

 [Chemical Formula 1]

The composition comprises a pharmaceutical composition or a food composition.

Hereinafter, the present invention will be described in detail.

The polyvalent lactone, which is an active ingredient of the composition of the present invention, can be obtained by purchasing a commercially available product, or by extracting and separating from fruiting bodies of mushrooms.

In the present invention, polyvalent lactone can be obtained by extraction and separation in the following manner.

First, the fruiting body of the mushroom is crushed, and then the fruiting body of the crushed mushroom is immersed in water, a C1-C4 alcohol, or a mixed solvent thereof for 1 to 3 days. At this time, the mushroom fruiting body is preferably, but not limited to fruiting body mushroom fruiting body or tile mushroom fruiting body. The C1 to C4 alcohols may be selected from methanol or ethanol, preferably methanol. Thereafter, the mushroom fruit body extract is filtered, and the filtrate is concentrated under reduced pressure. The concentrate is then fractionated with hexane and ethyl acetate. It is obtained by isolating polyvalent lactone from the mushroom fruity ethyl acetate fraction.

The multivalent lactone of the present invention excellently suppresses hepatocellular damage and functional integrity of hepatocytes in liver damaged mice induced by hepatic injury inducer, resulting in a decrease in serum hepatic enzyme (ALT, AST) levels, hepatic necrosis Inhibition of the production of reactive oxygen species, suppression of lipid peroxidation, reduction of oxidative stress such as increased SOD expression, reduction of factors related to inflammation of hepatocytes such as TNF-α, iNOS, and COX-2 and hepatocyte apoptosis It is excellent in reducing effect.

The hepatic injury inducing agent may be any one or more compounds selected from the group consisting of 3,5-diethoxycarbonyl-1,4-dihydrocholidine (DDC), carbon tetrachloride (CCl ₄ ) and thioacetamide.

The carbon tetrachloride is a toxic substance commonly used to cause liver damage. The toxic liver damage model made by administration of carbon tetrachloride is similar to a typical clinical case of liver injury. Liver injury caused by carbon tetrachloride administration begins with the first generation of highly reactive free radicals. The CYP2E1 (cytochrome p450 2E1) enzyme in hepatocytes degrades carbon tetrachloride to produce trichloromethyl radicals, CCl ₃ ^* . The resulting CCl ₃ ^* spontaneously reacts with oxygen to produce trichloromethyl peroxy radical, an active oxygen species, CCl ₃ OO ^* , and CCl ₃ OO ^* is rapidly neutralized by the liver glutathione peroxidase system. The second is initiated by TNF-α, an inflammatory cytokine produced by activated macrophages. TNF-a directly causes cytotoxicity, or is associated with inflammation of the liver and hepatocyte apoptosis. In addition, TNF-α activates other non-parenchymal cells involved in the inflammation of the liver. Strong inflammatory mediators in liver damage induced by carbon tetrachloride include iNOS and COX-2. These liver inflammatory responses play an important role in the progression of liver injury, so liver protective effects can be predicted by measuring the degree of inflammation reduction such as reduction of oxidative stress and inflammatory mediators.

In general, liver damage is known to be caused by oxidative stress and inflammation. As described above, the polyvalent lactone of the present invention is excellent in reducing oxidative stress and inflammation-related factors, and thus can be used as medicines and health foods useful for preventing or ameliorating liver damage, and for treatment.

The composition of the present invention, together with polyvalent lactone, may contain at least one known active ingredient having a preventive or ameliorative effect on liver damage and a therapeutic effect.

The composition of the present invention may further comprise at least one pharmaceutically acceptable carrier in addition to the above-described effective ingredients for administration. The pharmaceutically acceptable carrier may be a mixture of saline, sterilized water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components. If necessary, an antioxidant, , And other conventional additives such as a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Further, it can be suitably formulated according to each disease or ingredient, using appropriate methods in the art or by the method disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA.

The composition of the present invention may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically) depending on the intended method, and the dose may be appropriately determined depending on the patient's weight, age, , Diet, administration time, method of administration, excretion rate, and severity of the disease. The daily dose of the polyvalent lactone is about 1 to 500 mg / kg, preferably about 50 to 100 mg / kg, and is preferably administered once a day to several times a day.

The composition of the present invention can be used alone or in combination with methods using surgery, hormone therapy, drug therapy and biological response modifiers for prevention or treatment and improvement of liver injury.

In addition, the composition of the present invention can be added to a health food for the purpose of preventing or improving liver damage. When the polyvalent lactone of the present invention is used as a food additive, the polyvalent lactone can be used as it is or can be used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). Generally, in the production of food or beverage, the polyvalent lactone of the present invention is added in an amount of 10% by weight or less, preferably 5% by weight or less based on the raw material. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .

There is no particular limitation on the kind of the food. Examples of the food to which the above substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may include natural fruit juice, fruit juice beverage, and flesh for the production of vegetable beverages. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples.

Example  1. longevity situation mushroom From fruity body Multivalent lactone  detach

Longevity condition Mushroom fruiting body was crushed and immersed in methanol for 24 hours and extracted twice. The methanol extract of mushroom fruiting bodies was filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The extract was partitioned with hexane to remove the fatty acid component. To the separated water layer, ethylacetate was added by the same amount, and the mixture was extracted with water. The ethyl acetate layer was concentrated under reduced pressure. The concentrated ethyl acetate layer was fractionated by reverse phase column chromatography. The elution solvent was washed with 30% methanol and 40% methanol was used. The eluted fractions were concentrated under reduced pressure, dissolved in 70% methanol, and subjected to Sephadex LH-20 column chromatography to separate and purify the active substance. The active substance was identified as pluralactone, and the structure of this compound is as follows.

[Chemical Formula 1]

Example  2. Tile mushroom  From fruiting body Multivalent lactone  detach

In Example 1, a multi-row lactone was separated from a tile layer mushroom fruiting body in the same manner as in Example 1 except that fruiting body mushroom fruiting bodies were used instead of long-lived mushroom fruiting bodies.

Experimental Example  One. Multivalactone Liver damage  Mouse serum AST  And ALT Impact Analysis

In order to confirm the effect of the multivalent lactone of the present invention on serum AST and ALT of liver damaged mice, the following experiment was conducted.

1. Preparation of experimental animals

8-week-old FVB / n mice were purchased from Koatech (Pyeongtaek, Korea) and used. The mice were divided into three groups and divided into a control group (physiological saline solution group), a test group 1 (50 mg / kg group) and an experimental group 2 (100 mg / kg group). Experimental groups 1 and 2 were each subcutaneously injected with 200 μl of water for 3 days in a dose of 50 mg / kg and 100 mg / kg, respectively, in the physiological saline solution. In the control group, 200 μl of physiological saline alone was subcutaneously injected for 3 days . One hour after the last subcutaneous injection, a 10% carbon tetrachloride solution mixed with carbon tetrachloride in mineral oil was intraperitoneally injected at a dose of 1 mg / kg. When mice were sacrificed at 12 hours, 24 hours, and 48 hours after the injection of carbon tetrachloride, the blood and liver were extracted and used in the following experimental examples.

2. Measurement of serum AST and ALT

Serum aspartate aminotransferase (ALT) and alanine aminotransferase (ALT) were measured by the Reitman-Frankel method using a commercially available assay kit (Asan Pharm, Seoul, Korea).

The results are shown in Fig.

As shown in Figure 1, serum ALT and AST levels were significantly reduced in a dose-dependent manner in the experimental group treated with multiple galactolone compared to the control group at 12 and 24 hours after the injection of carbon tetrachloride. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are enzymes that act as markers for cell membrane damage and loss of functional integrity of hepatocytes. Thus, from the above results, it can be seen that the multivalent lactone of the present invention excellently suppresses liver cell membrane damage and hepatic cell functional loss in liver damaged mice.

Experimental Example  2. Multivalactone Liver damage  Mouse Liver tissue  Analysis of effects on necrosis

The following experiment was carried out in order to confirm the effect of the present multilayer lactone on hepatic necrosis of liver damaged mice.

Liver tissues were fixed with 10% neutral formalin, and then fixed to alcohol and paraffin by concentration. Histopathologic evaluation of thin sections cut with a thickness of 4 μm was performed by H & E (hematoxylin and eosin) staining. Necrotic lesions were randomly selected from five areas.

The results are shown in Fig.

As shown in FIG. 2, the size of the necrotic lesion was dose-dependently decreased in the experimental group administered with poly (lactic acid) at 12 hours after injection of carbon tetrachloride (FIG. 2A-2C). In the tissue histology (FIG. 2D-2F) at 24 hours after injection of carbon tetrachloride, the size of the necrotic lesion was dose-dependently decreased in the experimental group treated with the multiple liacolactone, which is shown in FIG. 2G showing the size of the necrotic lesion in% It was clear. Therefore, it can be seen that the polyvalent lactone of the present invention is excellent in the effect of inhibiting hepatic tissue necrosis of liver damaged mice.

Experimental Example  3. Multivalactone  Liver Oxidative  Stress and its antioxidative effects

In order to examine the effect of the polyvalent lactone of the present invention on the oxidative stress of the liver and its antioxidant effect, the following experiment was conducted.

1. Detection of reactive oxygen species (DHE test)

After injection of carbon tetrachloride into the abdominal cavity of mice, liver injury was induced by using DHE (dihydroethidium, Invitrogen, Carlsbad, CA, USA) for detection of reactive oxygen species (ROS) in liver tissues. Frozen liver tissue sections were treated with 3 μmol / L of DHE at 37 ° C for 30 minutes and then examined by fluorescence microscopy (Axioskop 2 plus, Carl Zeiss, Göttingen, Germany).

The results are shown in Figs. 3A and 3B.

As shown in Figs. 3A and 3B, it was found that the amount of reactive oxygen species detected in the experimental group (Fig. 3B) was significantly smaller than that of the control group (Fig. 3A). Therefore, it can be seen that the polyvalent lactone of the present invention is excellent in the effect of suppressing the generation of active oxygen species and thus has an excellent effect of reducing oxidative stress.

2. Measurement of lipid peroxidation (MDA level)

MDA (malonyldialdehyde) is a natural by-product of lipid peroxidation, which is an indicator of oxidative stress and cellular damage. MDA was reacted with thiobarbituric acid (TBA) at a ratio of 1: 2 to prepare a compound. The oxidative stress causing lipid peroxidation was analyzed by measuring the level of MDA in the serum using a TBARS (Thiobarbituric acid reactive substance) assay kit (Cell Biolabs, Inc., San Diego, Calif., USA). First, the sample was reacted with the TBA reagent at 95 ° C for 1 hour and then cooled at room temperature. After centrifugation, the supernatant was collected and mixed with butanol, and centrifuged again. The optical density (OD) value of the butanol layer was measured at a wavelength of 540 nm in a 96-well microplate.

The results are shown in Figure 3C.

As shown in Fig. 3C, serum levels of MDA in the serum of the experimental group administered with polyvalent lactone were significantly lower than those of the control group. Therefore, the polyvalent lactone of the present invention is excellent in the effect of inhibiting lipid peroxidation, and thus has an excellent effect of reducing oxidative stress.

3. Measurement of Cu / Zn SOD (Superoxide dismutase) (Western blot)

Liver injury The liver tissues of the mice were placed in a lysis buffer supplemented with phosphatase-1 inhibitor, reacted at 4 ° C for 30 minutes, and homogenized. The homogenized liver tissues were collected and quantified with BCA (bicinchoninic acid) solution. Mixed with two times of sample buffer (5 mM EDTA, 4% SDS (sodium dodecyl sulfate), 20% glycerol, 200 mM Tris buffer, pH 6.8, 0.06% bromophenol blue) , Followed by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) at 10% gel. The electrophoretic gel protein was transferred to the nitrocellulose membrane using a semi-dry dectrotransfer system (0.8 mA / cm ² ) and then reacted with 5% skim milk at room temperature for 1 hour to inhibit nonspecific antibody response. The primary antibody specifically binding to Cu / Zn SOD was diluted 1: 1000 in TBS-T, reacted with nitrocellulose membrane at room temperature for 2 hours, washed with TBS-T 3 times for 10 minutes, HRP (horseradish peroxidase) (Anti-rabbit igG conjugated HRP) was diluted 1: 3000 with TBS-T, reacted at room temperature for 1 hour, and ligated with Cu / Zn SOD using ECL kit (Amersham Co. England) Antigen-antibody complexes were visualized.

The results are shown in Figure 3D.

As shown in FIG. 3D, expression of Cu / Zn SOD in the liver tissue was significantly decreased in the experimental group treated with polylactic acid compared with that in the control group. Therefore, it can be confirmed that the polyvalent lactone of the present invention is excellent in the effect of increasing the expression of Cu / Zn SOD, which is an antioxidant enzyme, and thus has excellent antioxidative effect.

Experimental Example  4. Multivalactone CYP2E1 ( Cytochrome P450  2 E1 ) On the expression of

The following experiment was carried out to confirm the effect of the polyvalent lactone of the present invention on the expression of CYP2E1. The effect of the multivalent lactone of the present invention on the expression of CYP2E1 was analyzed in the same manner as in Experimental Example 3 except that the primary antibody and the secondary antibody were used as antibodies specifically binding to CYP2E1 (cytochrome p450 2E1) . The CYP2E1 is an important enzyme that metabolizes carbon tetrachloride, but in the process it produces toxic radicals and causes liver damage.

The results are shown in Fig.

As shown in Fig. 4, the expression level of CYP2E1 was found to be similar in both the control and experimental groups. Therefore, it can be seen that the multivalent lactone of the present invention did not affect the expression of CYP2E1, and thus did not affect the mechanism of hepatic tissue damage associated with CYP2E1.

Experimental Example  5. Multivalactone Liver damage  Mouse Inflammatory factor  Analysis of effects on expression

The following experiment was carried out to confirm the effect of the polyvalent lactone of the present invention on the expression of an inflammatory factor in liver damaged mice.

1. Measurement of serum TNF-α (tumor necrosis factor-α)

Blood samples from liver injured mice were centrifuged to separate serum and TNF-α levels were measured. Serum levels of TNF-α were measured using a mouse TNF-α ELISA kit (enzyme-linked immunosorbent assay kit, KOMABIOTECH Inc., Seoul, Korea) according to the manufacturer's instructions.

The results are shown in Figure 5A.

As shown in Fig. 5A, the level of TNF-a in the serum of the experimental group was markedly decreased in a dose-dependent manner in the multivalent lactone, as compared with the control group. Therefore, it can be seen that the polyvalent lactone of the present invention is excellent in reducing the chain action that induces inflammation by decreasing the level of TNF-a.

2. Measurement of iNOS (inducible nitric oxide synthase) and COX-2

In the same manner as in Experimental Example 3, except that the primary antibody and the secondary antibody were used as an antibody specifically binding to iNOS (inducible nitric oxide synthase), COX-2, the polyvalent lactone of the present invention was treated with iNOS And COX-2 on the expression of COX-2.

The results are shown in Figure 5B.

As shown in FIG. 5B, the expression of iNOS and COX-2 was inhibited in the tissues of the test group to which polyvallareactone was administered, as compared with the control group. Therefore, it can be seen that the multivalent lactone of the present invention inhibits the expression of iNOS and selectively inhibits the expression of COX-2, thereby having a liver protecting effect for alleviating inflammation of damaged liver tissues.

Experimental Example  6. Multivalent lactone HO -1 expression

In order to confirm the effect of the polyvaleroactone of the present invention on the expression of HO-1 (heme oxygenase-1), the following experiment was conducted.

In the same manner as in Experimental Example 3, except that the primary antibody and the secondary antibody were used as antibodies specifically binding to HO-1, the polyvalent lactone of the present invention inhibited the oxidative damage of HO-1 And the effect on expression was analyzed.

The results are shown in Fig.

As shown in FIG. 6, the expression of HO-1 was dose-dependently increased in dose of polyvallare. Therefore, the multivalent lactone of the present invention can predict the excellent effect of protecting the liver tissue from oxidative damage by increasing the expression of HO-1 enzyme.

Experimental Example  7. Multivalactone Liver tissue  The distribution of macrophages and hepatocytes To apoptosis  Impact analysis

In order to confirm the effect of the multivalactactone of the present invention on the distribution of macrophages in hepatic tissues and on the apoptosis of hepatocytes, the following experiment was performed using immunofluorescence.

First, frozen liver tissue was cut to a thickness of 10 μm, fixed in cold methanol for 5 minutes at room temperature, and then incubated in a universal protein block solution (Golden Bridge International, WA, USA) for 10 minutes at room temperature. To determine the extent of macrophage distribution, the cells were reacted with anti-F4 / 80 antibody conjugated with Alexa Fluor 488 (Invitrogen, Carlsbad, Calif., USA) for 1 hour at room temperature. To determine the degree of hepatocyte apoptosis, we incubated overnight at 4 ° C with an antibody specifically binding to cleaved caspase 3. Visualization of isolated Caspase 3 was performed by diluting a secondary antibody with Rhodamine Red at 1: 200 and staining for 1 hour. Each incubation step was washed three times for 5 minutes with PBS except for the protein block step, and the stained slide glass was preserved by dropping the mounting solution (Lab Vision Corporation, CA, USA). The staining results were examined with a fluorescence microscope (Axioskop 2 plus, Carl Zeiss, Göttingen, Germany).

The results are shown in Fig.

As shown in Fig. 7, the numbers of mouse macrophages and the isolated caspase 3 were significantly decreased in the experimental group (B, E) administered with polyvalla- tone compared to the control (A, D). This means that the polyvalent lactone of the present invention inhibits macrophage induction and caspase 3 activation in hepatic tissues and thus inhibits apoptosis of hepatocytes, thereby excellently preventing liver damage.

Examples of formulations for the composition of the present invention are illustrated below.

Formulation example  1. Preparation of pharmaceutical preparations

1. Manufacturing of powder

100 mg of multivalactactone

Lactose 200mg

The above components were mixed and packed in airtight bags to prepare powders.

2. Preparation of tablets

100 mg of multi-vala lactone

100 mg of corn starch

100 mg of milk

2 mg of magnesium stearate

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

3. Preparation of capsules

100 mg of multi-vala lactone

100 mg of corn starch

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

Formulation example  2. Manufacturing of food

Foods containing the polyvalent lactones of the present invention were prepared as follows.

1. Preparation of cooking seasoning

And 0.001 to 10% by weight of polyvalent lactone.

2. Manufacture of tomato ketchup and sauce

0.001 to 10% by weight of polyvalent lactone was added to tomato ketchup or sauce to prepare tomato ketchup or sauce for health promotion.

3. Manufacture of flour food

0.001-5.0 wt.% Of polyvalent lactone was added to wheat flour, and bread, cake, cookies, crackers and noodles were prepared using this mixture to prepare foods for health promotion.

4. Manufacture of soups and gravies

0.001 ~ 5.0 wt% of polyvalent lactone was added to the soup and juice to prepare health promotion meat product, noodle soup and juice.

5. Manufacture of ground beef

0.001 ~ 5.0 wt% of polyvalent lactone was added to ground beef to prepare ground beef for health promotion.

6. Manufacture of dairy products

0.001 to 10% by weight of polyvalent lactone was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

Formulation example  3. Manufacturing of beverage

1. Manufacture of carbonated beverages

The syrup is prepared by mixing 5 to 10% of sugar, 0.05 to 0.3% of citric acid, 0.005 to 0.02% of caramel and 0.1 to 1% of vitamin C and 79 to 94% of purified water to prepare syrup, ° C for 20 to 180 seconds, mixed with cooling water at a ratio of 1: 4, and then carbonated gas was injected at 0.5 to 0.82% to prepare a carbonated drink containing the multivalent lactone of the present invention.

2. Manufacture of health drinks

The raw materials such as the liquid fructose (0.5%), the oligosaccharide (2%), the sugar (2%), the salt (0.5%) and the water (75%) were uniformly blended with the multi- , And plastic bottles were packaged in small containers.

3. Manufacture of vegetable juice

Healthy vegetable juice was prepared by adding 5 g of polyvallactone to 1,000 ml of tomato or carrot juice.

4. Manufacture of fruit juice

Healthy fruit juice was prepared by adding 1 g of polyvalent lactone to 1,000 ml of apple or grape juice.

Claims (5)

A pharmaceutical composition for the prevention or treatment of toxic liver injury comprising, as an active ingredient, davallialactone represented by the following formula (1) extracted and isolated from fruiting body mushroom fruiting body.
[Chemical Formula 1]

The method of claim 1, wherein the toxic liver injury is selected from the group consisting of carbon tetrachloride, 3,5-diethoxycarbonyl-1,4-dihydrocholidine (DDC), and thioacetamide Or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
delete delete A food composition for preventing or ameliorating toxic liver injury comprising, as an active ingredient, davallialactone represented by the following formula (1) extracted and isolated from fruiting body mushroom fruiting body.
[Chemical Formula 1]

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