Composition for improving liver function

Abstract

To provide a composition for improving liver functions by utilizing a plant-derived resorcinolic lipid.SOLUTION: A plant-derived resorcinolic lipid is used as an active ingredient of a composition for improving liver functions. The resorcinolic lipid is preferably an alkylresorcinol and is also preferably derived from a gramineae plant, also preferably from wheat and / or rye bran and / or whole grains.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a composition for improving liver function that utilizes a plant-derived component. [Background technology]

[0002] Resorcinol lipids are lipophilic substances having alkyl groups at the 2-, 4-, 5-, and / or 6-positions of the aromatic ring of resorcinol (1,3-dihydroxybenzene). Resorcinol lipids are naturally found in large amounts in the seeds of grasses, particularly in their outer skins (bran), ginkgo leaves, cashew nut shell oil, etc. In recent years, there have been reports of various functionalities of resorcinol lipids for humans, such as oral antibacterial properties, immunosuppression, antidiabetic properties, anti-inflammatory properties, intestinal flora regulation, anticholesterol properties, anti-obesity properties, sleep improvement properties, anti-aging properties, and circadian rhythm regulation (e.g., Patent Documents 1 to 10). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Patent No. 2613474 [Patent Document 2] JP 2013-091612 A [Patent Document 3] Patent No. 2014-139166 [Patent Document 4] Patent No. 2015-020993 [Patent Document 5] Patent No. 2015-218130 [Patent Document 6] Patent No. 2015-231986 [Patent Document 7] Patent No. 2016-132641 [Patent Document 8] Patent No. 2016-153387 [Patent Document 9] Patent No. 5926616 [Patent Document 10] Patent No. 5951448 Summary of the Invention [Problem to be solved by the invention]

[0004] However, there have been no reports to date of the effect of plant-derived resorcinol oils and fats in improving liver function.

[0005] An object of the present invention is to provide a composition for improving liver function, which utilizes resorcinol-containing oils and fats derived from plants and is useful, for example, for improving symptoms of non-alcoholic hepatitis. [Means for solving the problem]

[0006] In order to achieve the above object, in a first aspect, the present invention provides a composition for improving liver function, which contains a plant-derived resorcinol lipid as an active ingredient.

[0007] In the composition for improving liver function according to the present invention, the resorcinol lipid is preferably an alkylresorcinol.

[0008] In the composition, the resorcinol lipid is preferably derived from a grass family plant.

[0009] Also, in the composition, the resorcinol lipids are preferably derived from wheat and / or rye bran and / or whole grains.

[0010] In addition, in the composition, the composition for improving liver function preferably comprises a solvent extract of a plant containing resorcinol lipids.

[0011] In addition, in the composition, when the composition for improving liver function consists of a solvent extract of a plant containing resorcinol lipids, it is preferable that the solvent is ethanol and / or hexane.

[0012] In addition, regarding the composition, the composition for improving liver function is preferably one used for improving the symptoms of hepatitis.

[0013] In addition, regarding the composition, the composition for improving liver function is preferably one used for improving symptoms of non-alcoholic hepatitis.

[0014] In addition, regarding the composition, the composition for improving liver function is preferably one used for suppressing neutral fat accumulation in the liver caused by non-alcoholic hepatitis.

[0015] In addition, with regard to the composition, the composition for improving liver function is preferably provided as a food or drink, a food additive, a pharmaceutical, a supplement, or an animal feed.

[0016] To achieve the above object, in a second aspect, the present invention provides use of a plant-derived resorcinol oil or fat for preparing a composition for improving liver function.

[0017] In the use of resorcinolic oils and fats of vegetable origin according to the invention, the resorcinolic lipids are preferably alkylresorcinols.

[0018] In addition, in this use, the resorcinol lipid is preferably derived from a grass plant.

[0019] Also, in said use, it is preferred that said resorcinol lipids are derived from wheat and / or rye bran and / or whole grains.

[0020] In addition, in the use, the composition for improving liver function preferably comprises a solvent extract of a plant containing resorcinol lipids.

[0021] In addition, in the case where the composition for improving liver function is composed of a solvent extract of a plant containing resorcinol lipids, it is preferable that the solvent is ethanol and / or hexane.

[0022] In addition, in terms of its use, the composition for improving liver function is preferably used for improving the symptoms of hepatitis.

[0023] In addition, in terms of its use, the composition for improving liver function is preferably used for improving symptoms of non-alcoholic hepatitis.

[0024] In addition, in terms of its use, the composition for improving liver function is preferably used for suppressing neutral fat accumulation in the liver caused by non-alcoholic hepatitis.

[0025] In addition, in terms of its use, the composition for improving liver function is preferably provided as a food or drink, a food additive, a pharmaceutical, a supplement, or an animal feed. Effect of the Invention

[0026] According to the present invention, a composition for improving liver function, which is useful for improving symptoms of, for example, non-alcoholic hepatitis, can be provided by utilizing a plant-derived resorcinol oil or fat. [Brief description of the drawings]

[0027] [Figure 1] 1 is a graph showing the results of investigating the change in weight gain in each group over the test period in Test Example 2. [Diagram 2] 1 is a graph showing the results of investigating changes in feed intake in each group during the test period in Test Example 2. [Diagram 3]This is a diagram showing the results of investigating the transition of AST values ​​in each group during the test period in Test Example 3. The results are shown as the average value and its standard deviation for each group (n=5-6). In addition, the statistically significant difference from the control group (Group C) was determined by Dunnett's multiple comparison test, and in the figure, "*" indicates a significant difference at a risk level of p<0.05. [Figure 4] This is a diagram showing the results of investigating the transition of ALT values ​​in each group during the test period in Test Example 3. The results are shown as the average value and its standard deviation for each group (n=5-6). In addition, the statistically significant difference from the control group (Group C) was determined by Dunnett's multiple comparison test, and in the figure, "*" indicates a significant difference at a risk level of p<0.05. [Diagram 5] 1 is a chart showing the results of examining the amount of neutral fat in liver tissue for each group in Test Example 4. The results are shown as the average value and its standard deviation for each group (n=5 to 6). [Figure 6] 1 is a graph showing the results of examining liver tissues of each group by HE staining in Test Example 5. [Figure 7] 1 is a chart showing the results of examining the expression level of the PPARγ gene in liver tissue of each group in Test Example 6. The results are shown as the average value and its standard deviation for each group (n=5 to 6). [Figure 8A] 1 is a diagram showing the results of examining the expression level of the PGC-1α gene in liver tissue of each group in Test Example 7. The results are shown as the average value and its standard deviation for each group (n=5-6). In addition, the statistically significant difference from the control group (Group C) was determined by Dunnett's multiple comparison test, and in the figure, "*" indicates a significant difference at a risk level of p<0.05. [Figure 8B] 1 is a diagram showing the results of examining the expression level of the PPARα gene in liver tissue of each group in Test Example 7. The results are shown as the average value and its standard deviation for each group (n=5-6). In addition, the statistically significant difference from the control group (Group C) was determined by Dunnett's multiple comparison test, and "*" in the figure indicates a significant difference at a risk level of p<0.05. [Figure 8C]1 is a graph showing the results of examining the expression level of the adiponectin gene in liver tissue of each group in Test Example 7. The results are shown as the average value and its standard deviation for each group (n=5-6). In addition, the statistically significant difference from the control group (Group C) was determined by Dunnett's multiple comparison test, and in the graph, "*" indicates a significant difference at a risk level of p<0.05. [Figure 9] This is a graph showing the results of evaluating the number of mitochondria in the liver tissue of each group based on the ratio of mitochondrial DNA content to genomic DNA content in Test Example 8. The results are shown as the average value and its standard deviation for each group (n=5-6). In addition, the statistically significant difference from the control group (group C) was determined by Dunnett's multiple comparison test, and "*" in the figure indicates a significant difference at a risk level of p<0.05. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0028] In the present invention, a plant-derived resorcinol lipid is used as an active ingredient of a composition for improving liver function.

[0029] As described above, resorcinol lipids are lipophilic substances having alkyl groups at the 2nd, 4th, 5th, and / or 6th positions of the aromatic ring of resorcinol (1,3-dihydroxybenzene). In nature, they are often found in plants belonging to the Anacardiaceae, Ginkgoaceae, Proteaceae, Ardisiaceae, Primulaceae, Myristicae, Iridaceae, Araceae, Fabaceae, Poaceae, etc., and in mugwort of the Asteraceae family. Among these plants, Poaceae plants such as wheat and rye contain relatively abundant resorcinol lipids in their edible parts (seeds), and therefore can be suitably used as a raw material for the resorcinol lipids used in the present invention. For example, the bran and whole grains of the edible parts (seeds) of wheat, rye, etc. contain about 0.015 to 0.3 mass % of an alkylresorcinol compound represented by the following formula (I). Fermented products such as miso, which is made from wheat or rye, can also be exemplified as preferred raw materials for the resorcinol lipids used in the present invention.

[0030] The resorcinol lipid used in the present invention typically includes an alkylresorcinol compound represented by the following formula (I).

[0031] [ka]

[0032] In formula (I), R1 represents a saturated or unsaturated linear or branched alkyl group. The number of carbon atoms of the alkyl group represented by R1 is not limited, but is preferably 1 to 27, more preferably 3 to 27, and further preferably 5 to 27.

[0033] In formula (I), R1 is preferably a saturated or unsaturated linear alkyl group, more preferably a saturated linear alkyl group. Examples of the saturated linear alkyl group represented by R1 include methyl, n-propyl, n-pentyl, n-heptyl, n-nonyl, n-undecyl, n-tridecyl, n-pentadecyl, n-heptadecyl, n-nonadecyl, n-henicosyl, and n-tricosyl.

[0034] In formula (I), the position and number of unsaturated bonds in the unsaturated linear alkyl group represented by R1 are not particularly limited. Examples of the unsaturated linear alkyl group include the above-mentioned saturated linear alkyl group having an unsaturated bond at any position on the carbon chain.

[0035] In formula (I), the position or number of branches of the saturated or unsaturated branched alkyl group represented by R1, or the position or number of unsaturated bonds, is not particularly limited.

[0036] Preferred examples of the compound of formula (I) above include the following.

[0037] 5-Pentylresorcinol [Olivetol, or 1,3-dihydroxy-5-n-pentylbenzene (C5:0)] 5-heptylresorcinol [or 1,3-dihydroxy-5-n-heptylbenzene (C7:0)] 5-Nonylresorcinol [or 1,3-dihydroxy-5-n-nonylbenzene (C9:0)] 5-Undecylresorcinol [or 1,3-dihydroxy-5-n-undecylbenzene (C11:0)] 5-Tridecylresorcinol [or 1,3-dihydroxy-5-n-tridecylbenzene (C13:0)] 5-Pentadecylresorcinol [or 1,3-dihydroxy-5-n-pentadecylbenzene (C15:0)] 5-heptadecylresorcinol [or 1,3-dihydroxy-5-n-heptadecylbenzene (C17:0)] 5-Nonadecylresorcinol [or 1,3-dihydroxy-5-n-nonadecylbenzene (C19:0)] 5-Henicosylresorcinol [or 1,3-dihydroxy-5-n-henicosylbenzene (C21:0)] 5-tricosylresorcinol [or 1,3-dihydroxy-5-n-tricosylbenzene (C23:0)] 5-Pentacosylresorcinol [or 1,3-dihydroxy-5-n-pentacosylbenzene (C25:0)] 5-heptacosylresorcinol [or 1,3-dihydroxy-5-n-heptacosylbenzene (C27:0)]

[0038] The plant-derived resorcinol lipid used in the present invention may be, for example, a commercially available product, or may be extracted from a plant by a conventional method. For example, the commercially available product of the compound of formula (I) above can be purchased from ReseaChem GmbH, SIGMA-ALDRICH, etc.

[0039] Examples of the extraction method from plants include a method in which the raw material is immersed in an extraction solvent at room temperature or heated under normal or increased pressure while stirring as necessary, a reflux extraction method, etc. Before being added to the extraction solvent, the raw material may be cut, crushed, squeezed, dried, or a combination thereof as necessary.

[0040] Examples of the extraction solvent include alcohols that are liquid at room temperature, such as lower alcohols such as methanol, ethanol, n-propanol, isopropanol, and n-butanol, or polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, and glycerin; ethers such as diethyl ether and propyl ether; esters such as butyl acetate and ethyl acetate; ketones such as acetone and ethyl methyl ketone; hexane; and organic solvents such as chloroform. These solvents may be used alone or in combination of two or more. Among the above solvents, from the viewpoints of operability and environmental impact, it is preferable to use alcohols that are liquid at room temperature, such as lower alcohols having 1 to 4 carbon atoms, and it is more preferable to use ethanol from the viewpoint of safety due to residual solvents.

[0041] The extraction solvent may further include a water-containing organic solvent in which an aqueous component is contained in the organic solvent. From the viewpoint of maintaining high extraction efficiency, the content of the aqueous component in the water-containing organic solvent is usually 50% by volume or less, preferably 30% by volume or less, more preferably 20% by volume or less. The water-containing organic solvent may preferably be a water-containing alcohol in which an aqueous component is contained in the alcohol that is liquid at room temperature as described above, more preferably a water-containing ethanol.

[0042] The conditions such as temperature and time for extraction can be appropriately set depending on the type of extraction solvent used and the extraction conditions, but the extraction temperature is preferably 2 to 100°C and the extraction time is preferably about 30 minutes to 60 hours. The amount of the extraction solvent may be preferably about 200 to 2000 parts by mass per 100 parts by mass of the raw material. After the extraction process, the mixture containing the extract and the residue may be subjected to filtration or centrifugation, etc., as necessary, to remove the solid components that are the residue. Furthermore, the removed solid components may be subjected to an extraction operation using the extraction solvent again, and this operation may be repeated several times. The obtained extract may be subjected to liquid chromatography or the like to prepare an extract containing further purified resorcinol lipids. When purified in this way, the degree of purification may be, for example, typically in the range of 1% by mass or more, more typically in the range of 5% by mass or more, and even more typically in the range of 10% by mass or more, as the content of the resorcinol fats and oils derived from the above-mentioned plants in any form. For example, the content may typically be in the range of 100% by mass or less, more typically in the range of 90% by mass or less, and even more typically in the range of 80% by mass or less. The obtained extract may be used in the present invention as it is, or after further treatment such as concentration, freeze-drying, hot air drying, pulverization, powderization, classification, dilution, mixing with water, etc., as necessary.

[0043] As shown in the examples below, plant-derived resorcinol lipids have the effect of improving liver function. The present invention provides a composition for improving liver function based on the attributes of this resorcinol lipid. The "liver function" includes, for example, improvement of symptoms of hepatitis such as non-alcoholic hepatitis, alcoholic hepatitis, and viral hepatitis. In addition to hepatitis, the composition for improving liver function according to the present invention includes improvement of symptoms of liver diseases such as non-alcoholic liver disease, fatty liver, and cirrhosis. However, the application of the composition for improving liver function according to the present invention is not limited to these symptoms, and can be used for the purpose of improving liver function in general. The improvement of symptoms can be evaluated, for example, by measuring the aspartate aminotransferase (AST) value or alanine aminotransferase (ALT) value in plasma, which are general indicators of liver function.

[0044] The dosage of the composition for improving liver function according to the present invention may be appropriately changed depending on the species, symptoms, age, sex, etc. of the individual to which it is applied. The dosage for humans may usually be 0.01 to 10 g of the above-mentioned plant-derived resorcinol oil per adult per day. The daily dosage may be administered once or in several divided doses. It is more preferable to administer the composition orally. The subject of application is not limited to humans, and the composition may also be applied to animals such as dogs and cats.

[0045] The form of the composition for improving liver function according to the present invention is not particularly limited as long as it is a form in which the resorcinol-containing oil derived from the above plant can be ingested. For example, the form may be solid, semi-solid or liquid, or may be various forms such as tablets, chewable tablets, powders, capsules, granules, drinks, gels, syrups, liquid foods for tube-enteral nutrition, etc. In addition, the composition may be in the form of, for example, health foods, functional foods, foods for specified health uses, foods for sick people, food additives therefor, medicines, supplements, animal feeds for livestock, racehorses, ornamental animals, pets, etc. That is, by containing the resorcinol-containing oil derived from the above plant as a part of each form, it can be made into a form that is easy to ingest. On the other hand, the composition for improving liver function according to the present invention may be substantially composed of the resorcinol-containing oil derived from the above plant. Thus, for example, in any form, the content of the plant-derived resorcinol fat or oil may be, for example, typically in the range of 0.1% by mass or more, more typically in the range of 0.5% by mass or more, and even more typically in the range of 1.0% by mass or more, and may be, for example, typically in the range of 100% by mass or less, more typically in the range of 90% by mass or less, and even more typically in the range of 80% by mass or less.

[0046] Typical pharmaceutical forms include, for example, oral preparations such as tablets, capsules, granules, powders, syrups, dry syrups, liquids, and suspensions, inhalants, transdermal preparations, enteral preparations such as suppositories, and parenteral preparations such as drops and injections. The liquid preparations such as the above liquids and suspensions may be dissolved or suspended in water or other suitable medium immediately before administration, and the above tablets and granules may be surface-coated by a known method. The above injections may be solutions or suspensions of the above plant-derived resorcinol oils in sterile distilled water or sterile saline, optionally containing a solubilizing agent.

[0047] The above pharmaceutical forms may contain various pharma- ceutical acceptable carriers, such as excipients, stabilizers, and other additives, as necessary, and may further contain other medicinal ingredients, such as various vitamins, minerals, and herbal medicines.

[0048] Typical forms of food and beverages include tea beverages such as green tea, oolong tea, and black tea, coffee beverages, soft drinks, jelly drinks, sports drinks, milk beverages, carbonated drinks, fruit juice drinks, lactic acid bacteria beverages, fermented milk beverages, powdered beverages, cocoa beverages, alcoholic beverages, mineral water, and other beverages; spreads such as butter, jam, furikake, and margarine; mayonnaise, shortening, cream, dressings, bread, cooked rice, noodles, pasta, miso soup, tofu, milk, yogurt, soups or sauces, and sweets (e.g., biscuits, cookies, chocolate, candy, cake, ice cream, chewing gum, and tablets).

[0049] In the form of the above-mentioned food and beverage products, they may contain, as necessary, other food and beverage ingredients acceptable for food, various nutrients, various vitamins, minerals, amino acids, various oils and fats, various additives (for example, taste components, sweeteners, acidulants such as organic acids, surfactants, pH adjusters, stabilizers, antioxidants, colorants, flavors), etc., or they may further contain other medicinal ingredients such as various vitamins, minerals, herbal medicines, etc.

[0050] Typical food additives may have a composition and form that allows them to be incorporated into foods and beverages, as exemplified above.

[0051] Typical animal feeds can have compositions and forms that are almost similar to those of foods and drinks, as exemplified above, except that they are for animals.

[0052] Typical forms of the supplement include oral compositions among the above-mentioned forms, such as tablets, chewable tablets, powders, capsules, granules, drinks, gels, syrups, etc. EXAMPLES

[0053] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples.

[0054] <Preparation Example 1> ~Preparation of alkylresorcinol~ Rye bran (approximately 9.4 kg) was dried under reduced pressure (80°C, 16 hours), and 99% ethanol was added in an amount five times that of the dried raw material, followed by stirring and extraction under heating (60°C, 1 hour). After that, solid-liquid separation was performed to obtain filtrate A, and the same amount of 99% ethanol was added to the residue, which was then rinsed and further separated into solid and liquid to obtain filtrate B. The obtained filtrate A and filtrate B were combined and concentrated under reduced pressure to obtain 516 g of rye bran ethanol extract.

[0055] 463 g of rye bran ethanol extract was mixed with 5.8 L of purified water and 5.8 L of 99% ethanol, stirred for 1 hour on a 60°C water bath, and then 696 g of α-cyclodextrin was added and stirred for another 1 hour. This was spray-dried to obtain 649 g of spray-dried product, and further particle size adjustment (10 mesh pass) was performed to obtain 639 g of dried rye bran ethanol extract product.

[0056] The above dried product was dissolved in 2 L of hexane and purified by silica normal phase chromatography. The column used was Yamazen Corporation's Hi-Flash Column Silica 5 L Catalog No. W007 (column inner diameter x column length: 60 x 180 mm, packing amount: 250 g). The chromatography conditions were a flow rate of 70 mL / min, detection wavelength of 280 nm, and mobile phase (hexane / ethyl acetate: 90 / 10 (v / v) for 9 minutes → 80 / 20 (v / v) for 15 minutes → 60 / 40 (v / v) step gradient for 16 minutes), and the eluate was collected between 30 and 40 minutes of elution. The collected product was dried under reduced pressure to obtain 50 g of dried product.

[0057] <Test Example 1> -Analysis by high performance liquid chromatography- The amount of alkylresorcinol in the dried product obtained in Preparation Example 1 was quantified by high performance liquid chromatography. Hereinafter, alkylresorcinol may be referred to as "ARs" or "AR".

[0058] Specifically, the analytical column used was Inertsil ODS4 (GL Science), and the analytical conditions were a flow rate of 1 mL / min, detection wavelength of 275 nm, column temperature of 40°C, and mobile phase (gradient of 89 v / v% methanol → (5 min) → 92 v / v% methanol → (25 min) → 100% methanol). Commercially available Olivetol (AR with a linear alkyl chain of 5 carbon atoms: Sigma-Aldrich) was used as the alkylresorcinol standard. The elution positions of each AR with a different alkyl chain length from Olivetol were specified in advance, and the measured peak areas were converted by multiplying the molecular weight of each AR divided by the molecular weight of Olivetol as a coefficient.

[0059] As a result, it was confirmed that the dried product obtained in Preparation Example 1 contained 6.8 g of ARs per 50 g.

[0060] <Test Example 2> ~Animal test using hepatitis model mice~ Non-alcoholic steatohepatitis model mice (male C57BL / 6NASH mice, manufactured by Charles River Japan) were brought in at 6 weeks of age, and after a 2-day acclimation period, were housed 6 mice per cage. The mice were housed in a temperature- and humidity-controlled room maintained on a 12-hour light-dark cycle (light period: 8:00 a.m. - 8:00 p.m.), and after the acclimation period, they were allowed to freely consume the food appropriate for each test group shown below.

[0061] Control group (Group C): Control feed not containing the test substance Resveratrol intake group (R group): Feed containing 0.4% resveratrol (Tokyo Chemical Industry Co., Ltd.) by mass High ARs intake group (AH group): Feed containing 0.9% ARs by mass Low ARs intake group (AL group): Feed containing 0.3% ARs by mass

[0062] During the test period, body weight was measured once a day. Blood was collected once a week from the tail vein of the mouse to obtain plasma. The plasma was stored at -80°C until use in the measurement. On the final day of the test, body weight was measured, blood was collected under anesthesia with diethyl ether, and the liver was immediately removed after sacrifice, weighed, frozen on dry ice, and divided into halves, each placed in a 1.5 mL tube, and stored at -80°C.

[0063] FIG. 1 shows the change in weight gain over the test period for each group.

[0064] As shown in FIG. 1, the control group (group C) showed almost no weight gain during the test period. The mice used in this test example were 6 weeks old and in the growth stage, so they would normally gain weight, but because they were hepatitis model mice, it was thought that they had developed metabolic disorders of nutrition, which led to the suppression of weight gain. In contrast, weight gain was confirmed in the ARs low intake group (group AL) and the ARs high intake group (group AH). This was thought to be because hepatitis was suppressed by ARs intake, and as a result, the metabolic disorders of nutrition were improved. On the other hand, the group that ingested resveratrol (group R), which is known to have the effect of suppressing fat accumulation in organs such as the liver, was unable to improve the suppression of weight gain in the hepatitis model mice. This was thought to be because the action and effect of ARs was not simply the result of a general improvement in fat metabolism, but was a specific action and effect on the symptoms of hepatitis.

[0065] On the other hand, Figure 2 shows the change in feed intake for each group over the test period. As shown in Figure 2, there was no significant difference in feed intake among the groups.

[0066] <Test Example 3> - Measurement of AST and ALT values ​​in plasma - Plasma aspartate aminotransferase (AST) and alanine aminotransferase (ALT), which are common indicators of liver function, were measured using Transaminase CII-Test Wako (Fujifilm Wako Pure Chemical Industries, Ltd.) according to the manufacturer's instructions.

[0067] Figure 3 shows the changes in AST levels over the study period in each group.

[0068] As shown in Figure 3, in the control group (group C), AST values ​​increased from week 0 to week 4, confirming the progression of hepatitis. In contrast, in both the low ARs intake group (group AL) and the high ARs intake group (group AH), the increase in AST values ​​was suppressed. The degree of suppression was more pronounced in the high intake group than in the low intake group. Meanwhile, in the resveratrol intake group (group R), the values ​​remained at roughly the same level as the control group (group C).

[0069] Figure 4 shows the changes in ALT levels over the study period in each group.

[0070] As shown in Figure 4, in the control group (group C), ALT levels increased from week 0 to week 4, confirming the progression of hepatitis. In contrast, in both the low ARs intake group (group AL) and the high ARs intake group (group AH), the increase in ALT levels was suppressed. The degree of suppression was more pronounced in the high intake group (group AH) than in the low intake group (group AL). Meanwhile, in the resveratrol intake group (group R), levels remained at roughly the same level as the control group (group C).

[0071] As shown in the results of Figures 3 and 4, it was revealed that ARs have an intake-dependent effect on hepatitis. This effect is a specific effect on the symptoms of hepatitis that is not exerted by resveratrol, which is generally known to suppress fat accumulation in organs such as the liver.

[0072] <Test Example 4> - Measurement of neutral fat content in liver tissue - 50 mg of frozen liver tissue was placed in a tube, and 1 mL of a mixture containing chloroform and methanol in a liquid volume ratio of 2:1 was added. The tissue was ground into a paste using a homogenizer, and the entire mixture was stirred at room temperature for 15-20 minutes using a shaker. The homogenate was centrifuged at 15,000 rpm for 5 minutes, and the supernatant was collected. 0.5 mL of physiological saline was added to the collected supernatant and mixed for a few seconds. The mixture was centrifuged at 2,000 rpm for 1 minute, and the upper phase was removed. This operation was repeated twice. The lower chloroform phase was dried under reduced pressure, and the dried matter was suspended in ethanol in 100 μL portions, and 45 μL portions were added to ethanol and mixed. This was used as a sample to measure the amount of neutral fat.

[0073] The measurements were performed using Triglyceride E-Test Wako (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) according to the manufacturer's instructions. The results are shown in Figure 5.

[0074] As shown in Figure 5, the control group (group C) had a triglyceride level of 130 mg / g, a clear indication of fatty liver. In contrast, in the ARs low intake group (group AL) and the ARs high intake group (group AH), the levels were both halved to 60 mg / g or less. These values ​​were within the range of normal triglyceride levels. On the other hand, in the group that consumed resveratrol (group R), which is known to have the effect of suppressing fat accumulation in organs such as the liver, the triglyceride levels in the liver tissue of the hepatitis model mice could not be corrected. Therefore, the effect of alkylresorcinol intake was not simply a result of general improvement in fat metabolism, but was considered to be a specific effect on the symptoms of hepatitis.

[0075] <Test Example 5> ~Pathological evaluation of liver tissue~ The frozen liver tissue was embedded in paraffin and sliced. The sections were stained with hematoxylin and eosin (HE) by standard methods. The results of HE staining of the liver tissue from each group are shown in Figure 6.

[0076] As shown in Figure 6, empty follicles derived from lipid droplets were confirmed in the liver tissue of the control group (group C), and both the size and number of empty follicles were large, showing a clear image of fatty liver. In contrast, the number and size of empty follicles were suppressed in both the ARs low intake group (group AL) and the ARs high intake group (group AH). On the other hand, almost no effect was observed in the resveratrol intake group (group R). Therefore, as in the results of Test Example 4, the effect of correcting the amount of neutral fat in liver tissue by taking alkylresorcinols was not simply an effect resulting from general improvement of fat metabolism, but was considered to be a specific effect on the symptoms of hepatitis.

[0077] <Test Example 6> - Measurement of PPARγ gene expression level in liver tissue - Peroxisome Proliferators-Activated Receptor γ (PPARγ) is a protein belonging to the nuclear receptor superfamily and is known to play an important role in lipid metabolism. Pioglitazone (used as a diabetes drug), a ligand of PPARγ, is also effective as a treatment for non-alcoholic hepatitis. For this reason, enhancement of PPARγ function was considered to be a useful target for the treatment of non-alcoholic hepatitis. In this test example, the expression level of the PPARγ gene in liver tissue was examined. Specifically, it was measured by quantitative PCR as follows.

[0078] Total RNA was prepared from liver tissue using the NucleoSpin® RNA II Kit (Takara Bio Inc.) according to the manufacturer's instructions. cDNA synthesis from total RNA was performed using the Protoscript® II First strand cDNA Synthesis Kit (New England BioLabs) according to the manufacturer's instructions.

[0079] Quantitative PCR using the prepared cDNA was performed using the MyGo Mini Real Time PCR device (Funakoshi) and Luna universal LPCR Mix (NEW ENGLAND BioLabs) according to the manufacturer's instructions. The following primer sequences were used:

[0080] (for PPARγ) Forward: 5'-TGTCGGTTTCAGAAGTGCCTTG-3' (SEQ ID NO: 1) Reverse: 5'-TTCAGCTGGTCGATATCACTGGAG-3' (SEQ ID NO:2) (for β-actin) Forward: 5'-TGACAGGATGCAGAAGGAGA-3' (SEQ ID NO: 3) Reverse: 5'-GCTGGAAGGTGGACAGTGAG-3' (SEQ ID NO: 4)

[0081] The PCR reaction was carried out under the following conditions. (95℃ 30 sec → 95℃ 5 sec → 60℃ 30 sec) × 40 cycles → 95℃ 60 sec

[0082] The expression level of peroxisome proliferator-activated receptor gamma (PPARγ) gene in liver tissue was compared between the control group (group C) as 100%. The results are shown in FIG. 7.

[0083] As shown in Figure 7, the expression level of PPARγ gene in liver tissue of the high ARs intake group (AH) was increased by about 3.5 times compared to the control group (C group). This suggests that the liver function improving effect of alkylresorcinol is related to the action of enhancing the expression of PPARγ gene.

[0084] <Test Example 7> ~Measurement of expression levels of various genes in liver tissue~ The gene expression levels in liver tissue were examined for various genes, including Peroxisome Proliferators-Activated Receptor γ Coactivator-1α (PGC-1α), Peroxisome Proliferators-Activated Receptor α (PPARα), and Adiponectin, which are known to play important roles in lipid metabolism. Measurements were performed by quantitative PCR in the same manner as in Test Example 6. The primers used for this purpose were the following sequences:

[0085] (for PGC-1α) Forward: 5'-AAGTGTGGAACTCTCTGGAACTG-3' (SEQ ID NO: 5) Reverse: 5'-GGGTTATCTTGGTTGGCTTTATG-3' (SEQ ID NO: 6) (for PPARα) Forward: 5'-AAGTGCCTGTCTGTCGGGATG-3' (SEQ ID NO: 7) Reverse: 5'-CCAGAGATTTGAGGTCTGCAGTTTC-3' (SEQ ID NO: 8) (for adiponectin) Forward: 5'-GTCAGTGGATCTGACGACACCAA-3' (SEQ ID NO: 9) Reverse: 5'-ATGCCTGCCATCCAACCTG-3' (SEQ ID NO: 10) (for β-actin) Forward: 5'-TGACAGGATGCAGAAGGAGA-3' (SEQ ID NO: 3) Reverse: 5'-GCTGGAAGGTGGACAGTGAG-3' (SEQ ID NO: 4)

[0086] The PCR reaction was carried out under the following conditions. (95℃ 30 sec → 95℃ 5 sec → 60℃ 30 sec) × 40 cycles → 95℃ 60 sec

[0087] The gene expression levels in liver tissues were compared for each group relative to the control group (group C) which was set at 100%. The results are shown in Figures 8A to 8C.

[0088] As shown in Figures 8A-8C, the expression levels of the genes PGC-1α, PPARα, and adiponectin in liver tissue were significantly increased in the ARs intake group. In particular, the gene expression levels of PGC-1α and adiponectin were even higher than in the positive control resveratrol intake group (R group) (Figures 8A, C). This suggests that the liver function improvement effect of alkylresorcinols is related to the action of enhancing the expression of these genes.

[0089] <Test Example 8> ~Evaluation of intracellular mitochondrial number in liver tissue~ PGC-1α, whose gene expression was confirmed to be increased in the alkylresorcinol intake group in Test Example 7, is known to result in an increase in the number of mitochondria, promotion of oxidative phosphorylation, activation of the TCA cycle, promotion of lipid burning through β-oxidation, and promotion of gluconeogenesis and ketone body synthesis through enhanced gene expression.

[0090] In this test example, the number of mitochondria in liver tissue was examined. Specifically, the number of mitochondria in liver tissue was evaluated by comparing the ratio of the amount of mitochondrial DNA to the amount of genomic DNA measured by quantitative PCR performed as follows.

[0091] DNA extraction from liver tissue was performed using the NucleoSpin (registered trademark) Tissue Kit (Takara Bio Inc.) according to the manufacturer's instructions. The number of copies of each DNA was quantified using the Mouse Feeder Cell Quantification Kit (Takara Bio Inc.) according to the manufacturer's instructions. The following primer sequences were used:

[0092] (for mitochondrial DNA) Forward: 5'-AACCCCGCTCTACCTCACC-3' (SEQ ID NO: 11) Reverse: 5'-GTAGCCCATTTCTTCCCATTT-3' (SEQ ID NO: 12) (for genomic DNA) Forward: 5'-CGCGGTTCTATTTTGTTGGT-3' (SEQ ID NO: 13) Reverse: 5'-AGTCGGCATCGTTTATGGTC-3' (SEQ ID NO: 14)

[0093] The PCR reaction was carried out under the following conditions. (95℃ 30 sec → 95℃ 5 sec → 60℃ 30 sec) × 40 cycles → 95℃ 60 sec

[0094] The ratio of the amount of mitochondrial DNA to the amount of genomic DNA measured was compared for each group relative to the control group (group C) which was set at 100%. The results are shown in FIG.

[0095] As shown in Figure 9, the resveratrol intake group (R group) showed no change in the number of intracellular mitochondria compared to the control group (C group). In contrast, the number of intracellular mitochondria increased in the ARs low intake group (AL group) and the ARs high intake group (AH group). This suggests that the liver function improvement effect of alkylresorcinol intake is related to the action effect of increasing the number of mitochondria in liver tissue cells.

Claims

1. A composition for improving ALT and / or AST values, comprising a resorcinol lipid derived from a grass family plant as an active ingredient.

2. The composition of claim 1 , wherein the resorcinol lipid is an alkylresorcinol.

3. 3. The composition according to claim 1 or 2, wherein the resorcinol lipids are derived from wheat and / or rye bran and / or whole grains.

4. A composition according to any one of claims 1 to 3, comprising a solvent extract of a grass family plant containing resorcinol lipids.

5. The composition of claim 4, wherein the solvent is ethanol and / or hexane.

6. The composition according to any one of claims 1 to 5, which is provided as a food or drink, a food additive, a medicine, a supplement, or an animal feed.

7. Use of a resorcinol lipid derived from a grass family plant for the preparation of a composition for improving ALT and / or AST levels.

8. 8. The use according to claim 7, wherein the resorcinol lipid is an alkylresorcinol.

9. 9. Use according to claim 7 or 8, wherein the resorcinol lipids are derived from wheat and / or rye bran and / or whole grains.

10. The use according to any one of claims 7 to 9, wherein the composition comprises a solvent extract of a grass family plant containing resorcinol lipids.

11. The use according to claim 10, wherein the solvent is ethanol and / or hexane.

12. The use according to any one of claims 7 to 11, wherein the composition is provided as a food or drink, a food additive, a medicine, a supplement, or an animal feed.

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