JP2009263257A - Agent for preventing and/or improving fatty liver - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a highly safe agent for preventing and/or improving fatty liver, which has a long history of safe use in food and exerts inhibitory action against excessive lipid accumulation in the liver. <P>SOLUTION: The agent for preventing and/or improving fatty liver comprises olive or an extract thereof as an active ingredient. A liver fat synthesis inhibitor is also provided. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to an agent for preventing and / or improving fatty liver.

  Fatty liver is a disease that develops due to overeating, lack of exercise, excessive intake of alcohol, viral infection, and the like, and is a state in which excess lipid has accumulated in the liver. In Japan, the rate of fatty liver detection among health check-ups from 1989 to 1999 has more than doubled from 16% to 34% for men and from 7.6% to 19% for women. It has been reported that a high discovery rate has continued since 2000 (Non-patent Document 1). In other words, it can be said that many people in today's society are prone to developing fatty liver.

  In most cases, fatty liver has no subjective symptoms and the importance of onset is difficult to recognize. However, fatty liver is not only involved in the onset and exacerbation of lifestyle-related diseases, but also causes liver damage such as hepatitis and cirrhosis (Non-patent Document 2). Is considered very important in maintaining health.

  In the prevention and improvement of fatty liver, it is first thought to balance daily intake and consumption of energy by thorough daily nutrition management and appropriate exercise. However, in a busy modern society, it is considered that there are many difficulties such as maintaining time and motivation to continuously carry out complicated nutrition management and regular exercise. Therefore, there is a need for a method that can be expected to have a sufficient effect and can be practiced easily and safely.

As a means for solving this problem, a method of orally ingesting a component having an effect of preventing or improving fatty liver can be considered. As described above, since fatty liver is a state in which excess lipid has accumulated in the liver, it is important to prevent fatty liver synthesis by inhibiting lipid synthesis in the liver or by inhibiting lipid accumulation by enhancing lipid metabolism. It is effective for improvement. Actually, an effect of improving fatty liver by using a fibrate drug that is a ligand of PPARα and promotes a lipid burning reaction has been disclosed (Patent Document 1).
However, it can be said that there are problems in terms of convenience and safety because many drugs cannot be used easily and there are side effects such as impaired liver function. Therefore, development of safe food materials with few side effects is expected.

On the other hand, olive is a plant belonging to the genus Oleaceae, and is widely cultivated in the Mediterranean region. The fruits are used extensively all over the world for olive oil production and edible, and the food experience is very rich. It is known that olive fruits contain polyphenol components such as oleuropein and hydroxytyrosol (Non-patent Documents 3 and 4). It is reported that the polyphenol component contained in the olive extract and fruit has an anti-arteriosclerosis effect (Non-patent document 5), a hypertension inhibitory effect (Non-patent document 6), a bone mass decrease inhibitory effect (Non-patent document 7), and the like Has been. In addition, it has been reported that the olive extract has an antioxidant action, a whitening action, an anti-aging action on the skin, an antitumor action and the like (Patent Document 2). Furthermore, the catecholamine-inducing action of oleuropein and its derivatives (Patent Document 3) has been reported.
However, it has never been known that olives, olive extracts, and the polyphenol components contained in them have the effect of preventing and improving fatty liver by suppressing lipid accumulation and fat synthesis in the liver. .
JP 2002-220345 A JP 2002-186453 A JP 2005-179353 A Japanese Journal of Gastroenterology, vol.104, pp492-500 Intern Med, Vol. 46, No. 2: 101-103, J Agric Food Chem, vol.34, pp823-826, 1986 J Agric Food Chem, vol.52, pp479-484, 2004 Atherosclerosis, vol.188, No.1, pp35-42, 2006 Int J Food Sci Nutr, vol.56, No.8, 613-620, 2005 Clin Nutr, vol.25, No.5, 859-868, 2006

  The present invention relates to providing a preventive and / or ameliorating agent for fatty liver, which has abundant dietary experience, is highly safe, and has an action of suppressing excessive lipid accumulation in the liver.

  In view of the above-mentioned problems, the present inventors searched for active ingredients from natural products with abundant dietary experience and high safety. It was found to be effective for the prevention and improvement of fatty liver.

That is, the present invention provides a preventive and / or ameliorating agent for fatty liver comprising olive or an extract thereof as an active ingredient.
Moreover, this invention provides the liver fat synthesis inhibitor which uses an olive or its extract as an active ingredient.
Moreover, this invention provides the food / beverage products for the prevention and / or improvement of the fatty liver which contain olive or its extract 0.0001-10 mass% in conversion of a dry matter.
Moreover, this invention provides the food-drinks for liver fat synthesis | combination suppression which contain 0.0001-10 mass% of olives or its extract in dry matter conversion.

  The agent for preventing and / or improving fatty liver and the liver fat synthesis inhibitor (hereinafter also referred to as “fatty liver improving agent”) of the present invention effectively uses olives or extracts thereof that have abundant dietary experience and high safety. It is used as a component, and is useful as a pharmaceutical or a food or drink for suppressing lipid accumulation and fatty synthesis in the liver or preventing and / or improving fatty liver.

  In the present invention, olive means an olive of the genus Oleaceae (Olea europaea). The olive varieties include Manzanillo, La Tanche, Gentile di Chieti, Casaliba, Picudo, Tajaska, Colonneki, Pendrino, Nevadillo Blanco, Mission, Picual, Ohi Blanca, Arbequina, Cornica Bra, Gordal, Frantoio, Moloiolo, Retino, Colatina, Ascorana terrena, etc. are known, but any variety can be used in the present invention.

  Examples of the use site of olives include fruits, pericarps, seeds, seed coats, leaves, stems, buds and the like, but it is preferable to mainly use fruits. These can be used as they are or after dry pulverization.

  As the olive extract, in addition to the extract, fruit juice obtained by extracting the above-mentioned use site, preferably fruit, as it is or after being dried and cut into a suitable size or pulverized, A more active fraction (component) obtained by separation and purification is included. For example, it is possible to use a powder extract obtained by squeezing olive fruit, separating the obtained fruit juice into an oil layer (olive oil) and an aqueous layer by centrifugation, and spray-drying the obtained aqueous layer. it can. Furthermore, olive polyphenol can be concentrated and purified from the obtained powder extract with an organic solvent or the like. Alternatively, a product obtained by treating olive oil with an extraction solvent and separating and purifying components contained in olive oil may be used.

  The extraction from the use site is performed by using a distillation method such as steam distillation in addition to solvent extraction performed by impregnation with a solvent at room temperature or in a heated state or using an extraction device such as a Soxhlet extractor. A method, a supercritical extraction method in which carbon dioxide gas is brought into a supercritical state, a pressing method for obtaining an extract by pressing, or the like can be used. The fruit juice or the like can be used after being subjected to treatment such as spray drying.

As the extraction solvent used for solvent extraction, either a polar solvent or a nonpolar solvent can be used, and these can also be mixed and used. For example, water; alcohols such as methanol, ethanol, propanol and butanol; polyhydric alcohols such as ethylene glycol, propylene glycol and butylene glycol; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; tetrahydrofuran Linear and cyclic ethers such as diethyl ether; polyethers such as polyethylene glycol; halogenated hydrocarbons such as dichloromethane, chloroform and carbon tetrachloride; hydrocarbons such as hexane, cyclohexane and petroleum ether; benzene and toluene Aromatic hydrocarbons such as; pyridines; supercritical carbon dioxide; oils, waxes, other oils, and the like. These can be used alone or in combination of two or more, and can be repeated by changing the solvent. In . Of these, water, ethanol, propylene glycol, butylene glycol and the like are preferably used, and a water / ethanol mixed solution is more preferably used.
For the extraction from the above-mentioned use site, for example, 1 to 50 parts by mass of a solvent is used with respect to 1 part by mass of olive, and it is preferably immersed or refluxed at 3 to 100 ° C. for several hours to several weeks.

  Examples of the means for separating and purifying the extract include activated carbon treatment, liquid-liquid distribution, column chromatography, liquid chromatography, gel filtration, and precision distillation.

  The olive extract of the present invention may be used as it is, or may be used as a diluted solution diluted with an appropriate solvent, or as a concentrated extract or dry powder, or in a paste form It may be prepared as follows.

Olive includes olive polyphenols such as hydroxytyrosol, oleuropein and their glycosides, and other phenolic compounds such as tyrosol and its glycosides.
The olive extract of the present invention preferably contains one or more of these compounds, and more than 20% by mass of olive polyphenol with less impurities, preferably from the viewpoint of effectiveness, color tone, taste, etc. when blended with food. The content is preferably 30% by mass or more, more preferably 50% by mass or more, and still more preferably 50 to 100% by mass.
In addition, in this invention, olive polyphenol content is calculated | required by measuring OD280nm (J Chromatogr B Biomed Appl. 24; 665 (2): 383-9. (1995)), The content is made into catechin equivalent amount. It is calculated.
Examples of olive extracts rich in tyrosol, hydroxytyrosol, oleuropein or their glycosides include olive polyphenol (Olivex PHO6) from Gardonnenk (France).

  As shown in the examples described later, the olive of the present invention or its extract was confirmed to have an action of inhibiting lipid accumulation in the liver and an action of inhibiting fat synthesis in a feeding test of an olive extract-containing food using mice. Therefore, olive or an extract thereof can be used for preventing and improving fatty liver, and can further be used for producing an agent for preventing and / or improving fatty liver. In addition, olive or an extract thereof can be used to suppress fat synthesis in the liver, and can further be used to produce a liver fat synthesis inhibitor. The said fatty liver improving agent etc. can be used as a pharmaceutical for humans or animals, a quasi-drug, and food-drinks for preventing and improving fatty liver, and suppressing fat synthesis in the liver. In addition, the preventive and / or ameliorating agent for fatty liver and the inhibitor for liver fat synthesis are based on the concept of inhibiting excessive lipid accumulation in the liver, inhibiting fat synthesis, or preventing and improving fatty liver. It can also be used as a quasi-drug or the like displaying the effect.

  Examples of the dosage form when the fatty liver improving agent of the present invention is used as a pharmaceutical product or quasi-drug include oral administration using tablets, capsules, granules, powders, syrups, etc., or injections, suppositories, and inhalants. And parenteral administration by transdermal absorption agents, external preparations and the like. In order to prepare such various dosage forms, olives or extracts thereof can be used alone or other pharmaceutically acceptable excipients, binders, extenders, disintegrants, surface active agents. Agents, lubricants, dispersants, buffers, preservatives, flavoring agents, fragrances, coating agents, carriers, diluents and the like can be used in appropriate combinations. Of these dosage forms, the preferred form is oral administration, and when an oral liquid preparation is prepared, it can be produced by a conventional method by adding a flavoring agent, a buffering agent, a stabilizer and the like.

In the case of using the fatty liver improving agent or the like of the present invention as a food or drink, breads, cakes, noodles, confectionery, jelly, frozen foods, ice creams, dairy products, beverages, soups, edible oils In addition to various foods such as the above, the same forms (tablets, capsules, syrups, etc.) as the above-mentioned oral administration preparations are mentioned. Examples of the beverage include fruit juice beverages, carbonated beverages, tea-based beverages, near water, sports beverages, milk beverages, alcoholic beverages, and soft drinks. Moreover, a drink can be used as the container-packed drink with which the container was filled.
Furthermore, cooking oils and seasonings, mayonnaise, dressings, processed oils and fats such as margarine, pasta sauces and the like can be mentioned.
To prepare various forms of food, olives or their extracts alone or other food ingredients, solvents, softeners, oils, emulsifiers, preservatives, fragrances, stabilizers, colorants, antioxidants It can be used as a fatty liver preventive and / or improved food, liver fat synthesis-suppressed food, pet food and the like by appropriately combining agents, humectants, thickeners and the like.

  The compounding amount of olive or its extract with respect to these varies depending on the form of use, but in the form of food or drink, olive or its extract (in terms of dry matter, hereinafter the same) is usually 0.0001 to 10% by mass. Further, it is preferably 0.001 to 5% by mass, particularly preferably 0.002 to 2% by mass. For example, in the case of beverages, olive or extract thereof is 0.001 to 0.5% by mass, further 0.005 to 0.25% by mass, and particularly 0.01 to 0.1% by mass in the beverage. preferable. In the case of food tablets and / or capsules such as tablets, those containing 0.1 to 95% by weight, further 1 to 90% by weight, particularly 5 to 50% by weight of olive or an extract thereof are preferable. In the case of general foods such as breads and cakes, 0.001 to 95% by weight of olive or an extract thereof, further 0.001 to 10% by weight, particularly 0.01 to 5% by weight, and more preferably 0.8. What contains 02-3 mass% is preferable. In the case of edible oils such as cooking oil and processed oils and fats, the olive extract is contained in an amount of 0.01 to 2% by mass, further 0.02 to 1.0% by mass, particularly 0.05 to 0.5% by mass. What is doing is preferable. The olive or the extract thereof may be in a dissolved state or a dispersed state, and the presence state is not limited.

  In the case of pharmaceuticals other than the above, for example, oral solid preparations such as tablets, granules, capsules, and oral liquid preparations such as oral liquids and syrups, olives or extracts thereof (in terms of dry matter) are usually 0. 0.01 to 95% by mass, more preferably 5 to 90% by mass, and particularly preferably 10 to 50% by mass.

  The dose (effective intake) of the fatty liver improving agent and the like of the present invention is preferably 1 to 5000 mg / 60 kg body weight per day as olive or extract thereof (in terms of dry matter), and more preferably 5 to 3000 mg / day. 60 kg body weight, particularly 10 to 2000 mg / 60 kg body weight is preferable, and 50 to 1000 mg / 60 kg body weight is most preferable.

  The typical test examples and examples of the present invention are shown below.

Test Example Inhibition of lipid accumulation in liver by ingestion of olive extract Evaluation of the action of olive extract for inhibiting lipid accumulation in the liver was performed as follows. In addition, the olive fruit polyphenol PHO6 (Olivex PHO6) of Gardonnenk (France) was used for the olive extract.
The 21-week-old SAM-P1 male mice were divided into a control diet group and a test diet group so that there was no difference in body weight (8 mice in each group).
Breeding was carried out for 13 weeks using a control diet and a test diet prepared with the formulation shown in Table 1. There was no difference in the amount of food consumed during the breeding period between the groups. On the last day of the breeding test, the body weight was measured and then dissection was performed, the liver was collected, and the weight was measured. Thereafter, liver lipids were extracted from a portion of the collected liver according to the method of Folch et al., And the liver lipid content was measured (Folch, J., J. Bio. Chem., 23, 497-509 (1957)). Moreover, total RNA was extracted from the liver using an RNA extraction reagent, and expression analysis of fat synthesis-related genes was performed by a real-time PCR method. Here, Acetyl-CoA carbylase alpha (Acaca), Stearoyl-CoA desaturase 1 (Scd1) involved in fatty acid synthesis, and Peroxisome proliferator activated receptor (G) that is induced in fatty liver. The expression level of acid binding protein 4 (Fabp4) was analyzed. The primers used for the reaction are shown in Table 2. The expression level of each gene was corrected and compared based on the expression level of ACID RIBOSOMAL PHOSPHOPROTEIN P0 (Arbp), which is a kind of housekeeping gene.

  Compared with the control diet group, the test diet group had a smaller liver weight (Fig. 1), and when the lipid content in the liver was measured, the test diet group showed a significantly lower liver lipid content (Fig. 2). From these results, it was found that the olive extract has an action of suppressing lipid accumulation in the liver.

  Moreover, when the expression analysis of the fat synthesis type gene in the liver was conducted, it was confirmed that the expression of the fatty acid synthesis type gene was significantly low in the test meal group (FIG. 3). In addition, the expression of Pparg, a transcription factor that regulates liver lipid metabolism, and the expression of Fabp4, which is a target gene, is significantly low, and the expression of a series of genes involved in the liver fat synthesis system is tested. It was revealed that it was significantly suppressed in the diet group.

  From these results, it was found that the olive extract can suppress the fat synthesis system in the liver and suppress excessive fat accumulation in the liver. That is, it was confirmed that the olive extract is useful as an agent for preventing / ameliorating fatty liver.

Examples Preventive and / or ameliorating agents (1) to (11) for fatty liver were produced.
(1) Soft drinks Soft drinks were produced with the formulations shown in Table 3.

(2) Soft drinks Soft drinks were produced with the formulation shown in Table 4.

(3) Capsule The composition (300 mg) shown in Table 5 was enclosed in a capsule to produce a capsule.

(4) Tablets The composition shown in Table 6 (1 tablet = 250 mg) was tableted to produce tablets.

(5) Granule The composition shown in Table 7 (1 bag = 500 mg) was mixed to produce a granule.

(6) Food The composition shown in Table 8 (1 tablet = 1000 mg) was tableted to produce a chewable tablet food.

(7) Sports drink The sports drink of the present invention was prepared with the formulation and conditions shown in Table 9.

(8) Edible oil Cooking oil was prepared with the composition shown in Table 10.

(9) Edible oil Cooking oil was prepared with the composition shown in Table 11.

(10) Edible oil Cooking oil was prepared with the composition shown in Table 12.

(11) Edible oil Cooking oil was prepared with the composition shown in Table 13.

It is a graph which shows the liver weight after completion | finish of a breeding test. It is the graph which showed the lipid content contained in a liver. It is the graph which showed the expression level of the fat synthesis type | system | group gene in a liver.

Claims (8)

  A preventive and / or ameliorating agent for fatty liver comprising olive or an extract thereof as an active ingredient.   A liver fat synthesis inhibitor comprising olive or an extract thereof as an active ingredient.   The preventive and / or ameliorating agent for fatty liver and the liver fat synthesis inhibitor according to claim 1 or 2, wherein the olive extract contains 20% by mass or more of olive polyphenol.   The agent for preventing and / or improving fatty liver according to any one of claims 1 to 3, wherein the olive extract contains one or more selected from hydroxytyrosol, tyrosol, oleuropein and glycosides thereof, and Liver fat synthesis inhibitor.   Use of olive or an extract thereof for prevention of fatty liver and / or production of an improving agent.   Use of olive or an extract thereof for producing a liver fat synthesis inhibitor.   A food and drink for preventing and / or improving fatty liver, containing 0.0001 to 10% by mass of olive or an extract thereof in terms of dry matter.   A food or drink for suppressing liver fat synthesis containing 0.0001 to 10% by mass of olive or an extract thereof in terms of dry matter.

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