WO2025159204A1 - Composition - Google Patents

Abstract

The present invention relates to a composition for improving the function of the liver, said composition containing a Japanese horseradish (wasabi) extract obtained using an organic solvent.

Description

composition

The present invention relates to a composition.

In recent years, various lifestyle-related diseases have become a problem due to changes in eating habits, unbalanced diets, stress, etc. Among these are problems such as a decline in liver function and the accumulation of fat in the liver.

For example, Non-Patent Document 1 discloses that sulforaphane, which is found in large amounts in broccoli sprouts, etc., has the function of lowering ALT, a liver function enzyme level in the blood. Non-Patent Document 2 discloses that curcumin and bisacron derived from autumn turmeric have the function of improving γ-GTP, AST, and ALT, which are liver function enzyme levels.

Kagome Co., Ltd., “Kagome Mail Order Health Direct Delivery Sulforaphane,” [online], accessed November 2, 2023, Internet <URL: https://shop.kagome.co.jp/lp2/sulforaphane/019r_hk/?adid=SPlsh0j&argument=Yv9hMnx2&dmai=a639acce7da98b&gad=1&gclid=EAIaIQobChMI2Ian-oaIgAMVmAN7Bx0O1Q9bEAAYASAAEgL3VvD_BwE> House Wellness Foods Co., Ltd., "Curbisa," [online], retrieved November 2, 2023, Internet <URL: https://curbisa.house-wf.co.jp/>

However, it was not known that this organic solvent-based wasabi extract had the effect of improving liver function and liver function enzyme levels, preventing fat accumulation in the liver, or reducing liver fat.

Therefore, one of the objects of the present invention is to provide a novel composition containing this wasabi extract extracted with an organic solvent, which exhibits effects related to liver function.

As a result of extensive research, the inventors discovered that a composition containing this wasabi extract extracted with an organic solvent could solve the above-mentioned problems, leading to the completion of the present invention.

That is, the present invention is as follows.
[1]
A composition for improving liver function, comprising the wasabi extract with an organic solvent.
[2]
A composition for improving liver function enzyme levels, comprising the wasabi extract obtained by organic solvent extraction.
[3]
The composition according to [2], wherein the liver function enzyme value is an AST or ALT value.
[4]
A composition for preventing fat accumulation in the liver and/or reducing liver fat, comprising the wasabi extract with an organic solvent.
[5]
The composition according to any one of [1] to [4], for oral administration.

The present invention also includes the following aspects.
[A1]
A method for improving liver function, comprising administering to a subject in need thereof a physiologically effective amount of the present wasabi extract in an organic solvent.
[A2]
A method for improving liver function enzyme levels, comprising administering to a subject in need thereof a physiologically effective amount of the present wasabi extract in an organic solvent.
[A3]
The method according to [A2], wherein the liver function enzyme value is an AST or ALT value.
[A4]
A method for preventing liver fat accumulation and/or reducing liver fat, comprising administering to a subject in need thereof a physiologically effective amount of the wasabi extract in an organic solvent.
[A5]
The method according to any one of [A1] to [A4], wherein the wasabi extract prepared with an organic solvent is orally administered.

[B1]
1. The wasabi extract with an organic solvent for use in a method for improving liver function.
[B2]
1. The wasabi extract with an organic solvent for use in a method for improving liver function enzyme levels.
[B3]
The wasabi extract obtained by the organic solvent according to [B2], wherein the liver function enzyme value is AST or ALT.
[B4]
10. The wasabi extract with an organic solvent for use in a method for preventing fat accumulation in the liver and/or reducing liver fat.
[B5]
The wasabi extract obtained by the organic solvent according to any one of [B1] to [B4], for oral administration.

[C1]
Use of the present wasabi extract with organic solvent to improve liver function.
[C2]
Use of the present wasabi extract with organic solvent to improve liver function enzyme levels.
[C3]
The use described in [C2], wherein the liver function enzyme value is an AST or ALT value.
[C4]
Use of the present wasabi extract with an organic solvent to prevent fat accumulation in the liver and/or reduce liver fat.
[C5]
The method according to any one of [C1] to [C4], wherein the wasabi extract prepared with an organic solvent is orally administered.

[D1]
Use of the present wasabi extract with an organic solvent in the manufacture of a composition for improving liver function.
[D2]
Use of the present wasabi extract with an organic solvent in the manufacture of a composition for improving liver function enzyme levels.
[D3]
The use described in [D2], wherein the liver function enzyme value is an AST or ALT value.
[D4]
Use of the present wasabi extract with an organic solvent in the manufacture of a composition for preventing fat accumulation in the liver and/or reducing liver fat.
[D5]
The use according to any one of [D1] to [D4], wherein the composition is taken orally.

[E1]
A food for improving liver function, containing the wasabi extract extracted with an organic solvent.
[E2]
A food product for improving liver function enzyme levels, containing this wasabi extract extracted with an organic solvent.
[E3]
The food described in [E2], wherein the liver function enzyme value is an AST or ALT value.
[E4]
A food for preventing fat accumulation in the liver and/or reducing liver fat, comprising the wasabi extract with an organic solvent.
[E5]
The food product according to any one of [E1] to [E4], which is intended for oral administration.

[F1]
A drug or quasi-drug for improving liver function, containing this wasabi extract extracted with an organic solvent.
[F2]
A drug or quasi-drug for improving liver function enzyme levels, containing this wasabi extract extracted with an organic solvent.
[F3]
The pharmaceutical or quasi-drug according to [F2], wherein the liver function enzyme value is an AST or ALT value.
[F4]
A pharmaceutical or quasi-drug for preventing fat accumulation in the liver and/or reducing liver fat, comprising the wasabi extract extracted with an organic solvent.
[F5]
A pharmaceutical or quasi-drug according to any one of [F1] to [F4], for oral administration.

In the above [1] to [5] and in each of the above [A1] to [F5] aspects, the wasabi extract may be an organic solvent extract from crushed wasabi powder, an organic solvent extract from the roots and/or rhizomes of wasabi, or an organic solvent extract from fine powder of the roots and/or rhizomes of wasabi.

According to the present invention, a composition for improving liver function containing this organic solvent-derived wasabi extract can be provided. It is also possible to provide a composition for improving liver function enzyme levels containing this organic solvent-derived wasabi extract. It is also possible to provide a composition for preventing fat accumulation in the liver and/or reducing liver fat containing this organic solvent-derived wasabi extract.

A schematic diagram of Test 1 is shown. The results of macroscopic observation of the liver, HE staining of hepatocytes, and Sirius red staining in Test 1 are shown below. The measurement results of liver TG, blood ALT, blood AST, liver steatosis score, and NAS in Test 1 are shown. The results of measurements in Test 2, including changes in average body weight, average food intake, average calorie intake, body weight at necropsy, liver weight, and relative liver weight, are shown. The results of measurement of liver TG, blood ALT, and blood AST in Test 2 are shown. The results of HE staining of hepatocytes, and the measurement results of hepatic steatosis score and NAS in Test 2 are shown. The results of measurement of inflammatory markers in Test 3 are shown below.

The following describes in detail the embodiments for implementing the present invention. Note that the present invention is not limited to the following embodiments, and can be implemented in various modifications within the scope of its gist.

(composition)
The composition of this embodiment contains the present wasabi extract with an organic solvent.
In this specification, the present wasabi refers to Wasabi japonica (Wasabi japonica or Eutrema wasabi).

In this embodiment, the wasabi extract obtained with an organic solvent refers to an extract obtained by extracting wasabi with an organic solvent. The part of wasabi used for extraction is not particularly limited, and for example, parts of rootlets, rhizomes, stems, leaves, flower stalks, flowers, etc., or combinations thereof can be used. From the perspective of more effectively and reliably achieving the effects of the present invention, it is preferable to extract wasabi containing roots and/or rhizomes with an organic solvent.
The organic solvent may be any solvent containing an organic solvent, and may be, for example, a mixed solvent with water.

The state of the real wasabi used for extraction is not particularly limited, but from the viewpoint of extraction efficiency, it is preferably in a pulverized state by methods such as freeze-pulverization, grating, or mashing. Real wasabi may be used immediately after harvest, or may be stored for a certain period of time after harvest. When real wasabi is stored for a certain period of time, the storage temperature is not particularly limited, and it may be stored at, for example, −78°C to 100°C.
The wasabi used for extraction may be crushed powder of the wasabi or crushed powder of the root and/or rhizome of the wasabi. When crushed powder of the wasabi root and/or rhizome is used, the crushed powder may include parts other than the root and/or rhizome.

The organic solvent used for the wasabi extraction is not particularly limited, and known organic solvents can be used. Examples of organic solvents include polar solvents and nonpolar solvents. Examples of polar solvents include, but are not limited to, methanol, ethanol, 1-butanol, 2-butanol, 1-propanol, 2-propanol, glycerin, edible oils and fats, acetone, ethyl methyl ketone, diethyl ether, ethyl acetate, methyl acetate, and dichloromethane. Examples of nonpolar solvents include, but are not limited to, cyclohexane, butane, and propane.
Among these, from the viewpoint of more effectively and reliably achieving the effects of the present invention, polar solvents may be used, such as ethanol, acetone, diethyl ether, or ethyl acetate. Furthermore, the organic solvent may be used alone or in combination of two or more kinds.
The temperature at which the wasabi is extracted is not particularly limited, but may be, for example, above the melting point and below the boiling point of the organic solvent used, and extraction can be performed under conditions such as room temperature, ice cooling, or heating.

After extracting wasabi with an organic solvent, the extract may be concentrated to obtain an organic solvent-based wasabi extract. The method for concentrating the extract is not particularly limited, but examples include evaporation, membrane concentration, and freeze concentration. Furthermore, before concentrating the extract, the extract may be dried with a drying agent such as anhydrous sodium sulfate. Furthermore, the extract may be filtered before concentrating.

After extracting wasabi with an organic solvent, the extract may be adsorbed or encapsulated in a carbohydrate such as dextrin to obtain an organic solvent-based wasabi extract. The carbohydrate such as dextrin is not particularly limited, but cyclodextrin is preferred, with α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin being more preferred. Alternatively, the extract may be concentrated and then adsorbed onto a carbohydrate, or the extract may be adsorbed onto a carbohydrate and then the carbohydrate to which the extract is adsorbed may be dried to obtain an organic solvent-based wasabi extract.

The components contained in the organic solvent-based wasabi extract are not particularly limited as long as they are derived from wasabi, but examples include proteins, lipids, carbohydrates, salts, etc. Ingredients contained in the organic solvent-based wasabi extract also include isothiocyanates derived from wasabi, and preferably include 6-methylsulfinylhexyl isothiocyanate (hereinafter also referred to as "hexarafan").

The content of each component contained in the present wasabi extract using an organic solvent is not particularly limited, but examples are as follows:
The organic solvent-derived wasabi extract preferably contains 2.5 to 3.5% by mass of protein, and more preferably 2.7 to 3.2% by mass. The organic solvent-derived wasabi extract preferably contains 0.3 to 1.5% by mass of lipids, and more preferably 0.5 to 1.2% by mass. The organic solvent-derived wasabi extract preferably contains 0.5% by mass or more of hexarafan, and more preferably 0.7% by mass or more. The organic solvent-derived wasabi extract preferably contains 85 to 96% by mass of carbohydrates, and more preferably 87 to 94% by mass. Note that when the organic solvent-derived wasabi extract is obtained by extracting wasabi with an organic solvent and then adsorbing or inclusion-forming the extract on a carbohydrate such as dextrin, the carbohydrate used for adsorption or inclusion is also included in the carbohydrate content.
The wasabi extract obtained with an organic solvent preferably contains 0.0015 to 0.0035% by mass, and more preferably 0.0020 to 0.0030% by mass, of sodium salts, and preferably contains 1.0 to 3.0% by mass, and more preferably 1.5 to 2.5% by mass, of sugars, based on 100% by mass of the wasabi extract obtained with an organic solvent.
In this specification, the content of sodium as a salt may be understood as the content of sodium salts as a sodium equivalent, and may also be understood as the content of sodium ions.
The content of each of the above components may be the content per 100% by mass of the present wasabi extract obtained by extracting the present wasabi with an organic solvent and then adsorbing or including the extract in a carbohydrate such as dextrin.
As an example of the present wasabi extract using an organic solvent, the following may be contained per 100% by mass of the present wasabi extract:
The protein content is preferably 2.5 to 3.5% by mass, more preferably 2.7 to 3.2% by mass,
The lipid content is preferably 0.3 to 1.5% by mass, more preferably 0.5 to 1.2% by mass,
The content of hexaraphane is preferably 0.5% by mass or more, more preferably 0.7% by mass or more,
The carbohydrate content is preferably 85 to 96% by mass, more preferably 87 to 94% by mass,
The sodium salt is preferably contained in an amount of 0.0015 to 0.0035% by mass, more preferably 0.0020 to 0.0030% by mass.
As an example, the content of each component may be any combination of a preferred amount and a more preferred amount.
Furthermore, as part of the carbohydrate content, sugars may be contained in an amount of 1.0 to 3.0% by mass, or 1.5 to 2.5% by mass, per 100% by mass of the wasabi extract obtained with an organic solvent.

The form of the wasabi extract obtained using an organic solvent is not particularly limited, but examples include liquid, solid, powder, etc., with powder being preferred.

(Improvement of liver function)
The composition of this embodiment has the effect of improving liver function. In this embodiment, improving liver function is not particularly limited, but examples thereof include improving liver function enzyme levels. Improving liver function enzyme levels means that liver function enzyme levels are reduced compared to before taking the composition of this embodiment. It may also mean reducing liver function enzyme levels that are higher in the healthy range. Examples of liver function enzyme levels include AST (GOT), ALT (GPT), γ-GTP, etc. The composition of this embodiment preferably improves AST and ALT levels.

(Preventing fat accumulation in the liver, reducing liver fat)
The composition of this embodiment has the effect of preventing fat accumulation in the liver and/or reducing liver fat. In this embodiment, preventing fat accumulation in the liver means, but is not limited to, for example, that the liver weight does not increase or the number of fat droplets does not increase after ingesting the composition of this embodiment compared to before ingesting the composition of this embodiment. Furthermore, reducing liver fat in this embodiment means, but is not limited to, for example, that the liver weight or the number of fat droplets decreases after ingesting the composition of this embodiment compared to before ingesting the composition of this embodiment. Furthermore, preventing fat accumulation in the liver and/or reducing liver fat means, but is not limited to, for example, that the triglyceride (TG) level, which is also an indicator of liver function, is improved. Generally, an increase in TG in the liver can cause fatty liver.

Furthermore, the effect of improving liver function and the effect of preventing fat accumulation in the liver and/or the effect of reducing liver fat may be related to each other. In other words, improving liver function may result in the effect of preventing fat accumulation in the liver and/or the effect of reducing liver fat, and preventing fat accumulation in the liver and/or the effect of reducing liver fat may result in the effect of improving liver function.

Furthermore, the effects of improving liver function, preventing fat accumulation in the liver, and reducing liver fat are more pronounced when this wasabi extract is administered using an organic solvent than when hexarafan is administered alone.

The composition of the present embodiment can improve, alleviate, treat, and/or prevent, for example, fatty liver, NAFLD (non-alcoholic fatty liver disease), etc. Generally, the etiology and progression mechanism of NAFLD are complex. However, for example, a pathway is known in which oxidative stress is caused by fatty liver or metabolic abnormalities in the liver, resulting in increased inflammatory signals (e.g., inflammatory markers, etc.) and increased levels of liver function enzymes in the blood, leading to the progression of NAFLD (e.g., Reference Material 1: Kenichi Ikeshima, Journal of the Japanese Society of Internal Medicine, Vol. 105 (2016), No. 1, pp. 15-24; Reference Material 2: Tomomi Ogiso, Journal of the Japanese Society of Internal Medicine, Vol. 105 (2016), No. 1, pp. 31-37). Here, the composition of the present embodiment can, for example, reduce liver fat, thereby preventing the production of oxidative stress and improving, alleviating, treating, and/or preventing NAFLD. Furthermore, the effect of improving, alleviating, treating and preventing fatty liver, NAFLD and the like may be exerted by the effect of improving liver function, the effect of preventing fat accumulation in the liver and/or the effect of reducing liver fat.
NAFLD may be a disease called metabolic dysfunction associated steatotic liver disease (MASLD).

The composition of this embodiment is not particularly limited, but is preferably taken orally or parenterally, and is more preferably taken orally. The composition of this embodiment is not particularly limited, but is preferably for human use.

(Food and Pharmaceuticals)
The composition of the present embodiment may be used as a food or food additive, and a pharmaceutical or quasi-drug.
In this embodiment, the dosage or intake of the composition is not particularly limited, and may be determined depending on, for example, the age and weight of the patient or user, symptoms, administration time, dosage form, administration method, drug combination, etc.
When ingested as a food product, the total amount of the composition is not particularly limited, but is preferably 10 to 1000 mg per adult per day, more preferably 20 to 800 mg, even more preferably 30 to 700 mg, and even more preferably 40 to 200 mg/day.
When orally administered as a pharmaceutical, the total amount of the composition is not particularly limited, but is, for example, preferably 10 to 1000 mg per adult per day, more preferably 20 to 800 mg, even more preferably 30 to 700 mg, and even more preferably 40 to 200 mg/day. It may be administered once a day or in divided doses (2 to 5 times a day).

The wasabi extract obtained with the organic solvent of this embodiment can be used as a food or food additive. Examples of the food include, but are not limited to, general foods, health foods, functional foods, nutritional supplements, health-promoting foods (e.g., foods for specified health uses, foods with functional claims, and foods with nutrient functions), and foods for special dietary uses (e.g., foods for infants, foods for pregnant women, and foods for sick people).
Furthermore, the wasabi extract using the organic solvent of this embodiment may be provided and sold as a food product labeled with uses related to improving liver function, improving liver function enzyme levels, improving AST or ALT levels, preventing fat accumulation in the liver, reducing liver fat, etc.
Such "display" acts include all acts for informing consumers of the above-mentioned uses, and any expression that can evoke or infer the above-mentioned uses falls under the category of "display" acts in this embodiment, regardless of the purpose of the display, the content of the display, the object or medium on which it is displayed, etc.

The labeling is not particularly limited, but is preferably one that claims to improve liver function, improve liver function enzyme levels, improve AST or ALT levels, prevent fat accumulation in the liver, reduce liver fat, etc. in healthy individuals, and more preferably, the labeling is for a food that has been approved or notified as necessary (a food for specified health uses, a food with functional claims, a food with nutrient function claims, or a food for special dietary uses, etc.). Foods that have been approved or notified for functional claims can be distinguished from general foods.

The form of the food or food additive is not particularly limited, but examples include orally ingestible forms such as solutions, suspensions, powders, and solid moldings. Such forms include, but are not limited to, supplements (powders, granules, soft capsules, hard capsules, tablets, chewable tablets, rapid-disintegrating tablets, syrups, liquids, etc.), beverages (carbonated drinks, lactic acid drinks, sports drinks, fruit juice drinks, vegetable drinks, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks, nutritional drinks, alcoholic drinks, etc.), sweets (gummy candies, jellies, gum, chocolate, cookies, candy, caramel, Japanese sweets, snacks, etc.), instant foods (instant noodles, retort pouch foods, canned foods, microwave foods, instant soups, miso soups, freeze-dried foods, etc.), oils, oily foods (mayonnaise, dressings, butter, cream, margarine, etc.), flour products (bread, pasta, noodles, cake mixes, breadcrumbs, etc.), seasonings (sauces, tomato-processed seasonings, flavor seasonings, cooking mixes, soups, etc.), and processed livestock products (livestock hams, sausages, etc.).

Foods may contain, but are not limited to, various nutrients, various vitamins (vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin C, vitamin D, vitamin E, vitamin K, etc.), various minerals (magnesium, zinc, iron, sodium, potassium, selenium, etc.), dietary fiber, stabilizers such as dispersants and emulsifiers, sweeteners, taste components (citric acid, malic acid, etc.), flavors, royal jelly, propolis, agaricus, etc.

The composition of this embodiment can be used as a pharmaceutical or quasi-drug. The dosage form of the pharmaceutical or quasi-drug is not particularly limited, but examples include powders, granules, pills, soft capsules, hard capsules, tablets, chewable tablets, rapid-disintegrating tablets, syrups, liquids, and suspensions. Furthermore, pharmaceuticals or quasi-drugs may be administered orally or parenterally, but oral administration is preferred.

In preparing the formulation, a drug or quasi-drug in the desired dosage form can be produced by a conventional method.
In preparing formulations, additives may be used in addition to the active ingredient. Examples of additives include, but are not limited to, animal and vegetable oils such as soybean oil, safflower oil, olive oil, germ oil, sunflower oil, beef tallow, and sardine oil; polyhydric alcohols such as polyethylene glycol, propylene glycol, glycerin, and sorbitol; surfactants such as sorbitan fatty acid esters, sucrose fatty acid esters, glycerin fatty acid esters, and polyglycerin fatty acid esters; excipients such as purified water, lactose, starch, crystalline cellulose, D-mannitol, lecithin, gum arabic, sorbitol solution, and sugar solution; sweeteners, colorants, pH adjusters, flavorings, binders, disintegrants, stabilizers, and lubricants. Liquid formulations may be dissolved or suspended in water or other suitable media at the time of administration. Tablets and granules may be coated by known methods.

The present embodiment will be explained in more detail below using examples and comparative examples, but the present embodiment is not limited to these examples.

<Test 1>
A study was conducted to determine the mouse model to use for the study.
[Test method]
Approximately 1-year-old mice (C57BL/6J, male, purchased from Jackson Laboratory Japan) were divided into an LFD group fed a normal diet and an HFD group fed a high-fat diet on day 0 of the study. Each group consisted of five mice. The normal diet contained soybean oil as fat, with approximately 12% of the calories coming from fat. On the other hand, the high-fat diet contained soybean oil and lard as fats, with approximately 60% of the calories coming from fat.
From day 0 to day 62 of the study, the rats were fed approximately 5 g of normal or high-fat food once a day and allowed to eat ad libitum. Fasting began in the evening of day 62, and autopsies were performed on day 63. The test method was based on Ishizuka, J Gastroenterol Hepatol. 2020.

[Test Results]
Macroscopic liver observation revealed that the HFD group had larger livers and more accumulated fat than the LFD group. Furthermore, hematoxylin-eosin (HE) staining of hepatocytes revealed that the HFD group had a greater number of white lipid droplets than the LFD group. Sirius red staining of hepatocytes revealed no staining in either the HFD or LFD groups, indicating no progression of fibrosis (Figure 2).
When liver triglycerides (TG) were measured, it was found that the HFD group had increased TG levels compared to the LFD group. Furthermore, when blood ALT and blood AST were measured, it was found that the HFD group had increased blood ALT and blood AST levels compared to the LFD group, and blood liver function enzyme levels were elevated. Furthermore, with reference to Hepatology. 2005 Jun; 41 (6): 1313-21, liver steatosis score and NAS (NAFLD Activity Score) were evaluated according to the table below, and it was found that the HFD group had a higher score compared to the LFD group (Figure 3).

In light of the above, it was determined that the model used in <Test 1> was appropriate as an animal model for evaluating liver function and liver function enzyme levels, and it was decided to use the same model as in <Test 1> in <Tests 2 and 3>.

<Test 2>
Tests were conducted to confirm the inhibitory effect of ingestion of this wasabi extract using organic solvents on hepatic fat deposition and on the suppression of increases in blood liver function enzyme levels.
[Test method]
Similar to Experiment 1, a 55-week-old mouse model was used. Each group was fed approximately 5 g of normal or high-fat diet once daily ad libitum from Day 0 to Day 64 or 65 of the experiment, according to the table below. The organic solvent-based wasabi extract used was Kinjirushi Wasabi Sulfinyl (registered trademark), sold by Kinjirushi Wasabi Co., Ltd. Kinjirushi Wasabi Sulfinyl (registered trademark), a wasabi extract containing 2.9 g of protein, 0.7 g of lipids, 93.8 g of carbohydrates, 25.0 mg of sodium, and 1.8 g of sugars per 100 g of Kinjirushi Wasabi Sulfinyl (registered trademark). Kinjirushi Wasabi Sulfinyl (registered trademark) also contains 0.8% or more hexaraphane and cyclic oligosaccharides as carbohydrates.

[Test Results]
The mean body weight transition, mean food intake, mean calorie intake, body weight at necropsy, liver weight, and relative liver weight were measured for each group (Figure 4). No differences were observed in mean food intake, calorie intake, or body weight at necropsy between the high-fat diet groups. However, liver weight was low in the LW and MW groups, and particularly low in the MW group.
Next, liver TG, blood ALT, and blood AST were measured (Figure 5). As a result, it was found that the high-fat diet increased liver TG and blood liver function enzyme levels. On the other hand, liver TG, blood ALT, and blood AST levels were low in the LW and MW groups, and especially in the MW group.
Next, the hepatic steatosis score and NAS (NAFLD Activity Score) were evaluated in the same manner as in <Test 1> ( Figure 6 ). The scores were calculated using the HE staining results of hepatocytes shown in Figure 6 . As a result, the LW and MW groups had lower hepatic steatosis score and NAS than the HFD group.

<Test 3>
A test was conducted to confirm the effect of ingestion of this wasabi extract in organic solvent on gene expression related to inflammation.
[Test Method]
For the mice used in <Test 2>, RNA was extracted from the livers collected at autopsy according to the ISOGEN II protocol, and real-time PCR was performed using the intercalator method to measure the gene expression levels in the liver of the inflammatory markers IL-6, Tnf, Ccl2, and Ccr2. TATA binding protein (Tbp) was used as an endogenous control, and the expression level ratio was calculated (Figure 7).
As a result, an increase in inflammatory markers was observed in the HFD group. On the other hand, the LW and MW groups showed a suppressed increase in inflammatory markers compared to the HFD group, indicating that oxidative stress was prevented and suppressed. Therefore, it was found that this wasabi extract using an organic solvent can suppress liver inflammation involved in the progression of NAFLD and is effective in the treatment and prevention of NAFLD.

Claims (5)

A composition for improving liver function, containing this wasabi extract extracted with an organic solvent. A composition for improving liver function enzyme levels, containing this wasabi extract extracted with an organic solvent. The composition of claim 2, wherein the liver function enzyme level is AST or ALT. A composition for preventing fat accumulation in the liver and/or reducing liver fat, comprising this wasabi extract extracted with an organic solvent. The composition described in any one of claims 1 to 4, for oral administration.