JP2003137802A - Liver function-protecting agent or improving agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a liver function-protecting or improving agent, a liver function protecting or improving food, beverage or feed, an additive for a food, beverage or feed having the liver function-protecting or improving activity, and a method for protecting or improving the liver function of an animal. SOLUTION: This liver function-protecting or improving agent is characterized by containing a plant body or plant extract of Artemisia apiacea Hance or Apium graveolens L. var. rapaceum Mill. DC. as an active ingredient, and the food, beverage or feed obtained by adding the plant body or plant extract of the Artemisia apiacea Hance or Apium graveolens L. var. rapaceum Mill. DC. is also provided.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a liver function protecting or improving agent, a food or drink for liver function protecting or improving, or a feed,
The present invention relates to an additive for food and drink or an additive for feed having a liver function protecting action or an improving action, and a method for protecting or improving an animal liver function.

[0002]

2. Description of the Related Art The liver has various important functions such as metabolism and storage of three major nutrients, carbohydrates, proteins and lipids, and decomposition and detoxification of substances unnecessary for the living body. It is an organ. These functions are acutely or chronically damaged by excessive intake of alcohol, viral infection, disturbed eating habits, stress, smoking, etc., but when the damage progresses, for example, acute hepatitis, chronic hepatitis, liver sclerosis, Diseases such as alcoholic fatty liver, hepatitis B virus, liver cancer and the like.

When hepatocytes are damaged by viruses, alcohols, etc., aspartate aminotransferase (also referred to as glutamic acid-oxaloacetate transaminase; hereinafter abbreviated as GOT) and alanine aminotransferase (glutamic acid-pyruvate transaminase) in the cells are damaged. In the following, an enzyme such as GPT) comes out in the blood, so that the numerical values showing the activity of these enzymes increase. Therefore, these GOT and GPT activities in blood are known as indicators of liver dysfunction.

Examples of drugs used for the prevention or treatment of liver dysfunction include antiviral agents such as acyclovir,
Immunosuppressant (Ayumi no Kiso 171 Vol. 14 No. 957-11
58, 1994, Ikuyakusho Publishing, Glutathione (Protein Nucleic Acid Enzyme, Vol. 33, No. 9, 1625-1631, 1988).
Kyoritsu Shuppan Co., Ltd.) is known. In addition, as foods and drinks that are effective in protecting, enhancing, and improving liver function,
Turmeric, thistle, thistle, sesame lignan, oyster extract, liver extract, etc. are known (FOOD Style 21,
Volume 12, No. 12, 1998, Food Chemistry Newspaper).

However, it is not known that the plant or the plant extract of Pleurotus cornucopia or celery arc has a liver function protecting action or an improving action.

[0006]

It has been earnestly desired to develop a drug capable of effectively treating liver diseases and a healthy food or drink or animal feed capable of preventing or treating liver dysfunction by daily intake. The object of the present invention is a liver function protector or improver, a food or drink for liver function protection or improvement, a feed, a food or drink additive having a liver function protecting or improving effect, and a liver function of an animal. It is to provide a protection method or an improvement method.

[0007]

The present invention includes the following (1).
Regarding (6). (1) Saiko ( Artemisia apiacea Hance) or celery arc ( Apiumgraveolens L. var. Rapaceum Mil)
L. DC.) plant or an extract of the plant as an active ingredient.

(2) The liver function-protecting agent or improving agent according to (1), which is orally administered.

(3) Artemisia apiacea Ha
nce) or celery arc ( Apiumgraveolens L. var. r.
apaceum Mill. DC.) plant or a food or drink or feed for protecting or improving liver function, which is obtained by adding the plant extract.

(4) Pepper ( Artemisia apiacea Ha
nce) or celery arc ( Apiumgraveolens L. var. r.
apaceum Mill. DC.) plant or an extract of the plant, which is a food / beverage additive or feed additive having a liver function protecting or improving action.

(5) A method for protecting liver function of an animal, which comprises feeding the animal the protector or improver for liver function according to (1) or (2) above or the feed according to (3) above. Or liver function improvement method.

(6) The method according to (5) above, wherein the animal is livestock, poultry, or farmed fish.

[0013]

BEST MODE FOR CARRYING OUT THE INVENTION The liver function in the present invention means all functions of the liver and is not particularly limited. Specific liver functions include, for example, blood storage (regulation of circulating amount), treatment of hemoglobin (treatment and discharge of hemoglobin), bile production, enterohepatic circulation of bile pigment, plasma protein (acute phase protein, albumin, blood coagulation). Functions in blood and circulation such as the synthesis of factors, steroid binding proteins, other hormone binding proteins, etc., nutrients and vitamins (glucose and other sugars, amino acids, lipid-fatty acids, cholesterol, lipoproteins, fat-soluble vitamins, water-soluble Metabolic functions of nutrients such as metabolism of vitamins, various substances (toxins, steroids such as estrogen and androsterone,
Detoxification or decomposition function such as inactivation of other hormones, etc.
And immune function, etc.
March 31, 2), "New Clinical Nutrition" Revised 3rd Edition (May 20, 2000)]. All of these functions are impaired by excessive alcohol intake.

The plants used in the present invention are gypsum ( Ar
temisia apiacea Hance) or celery arc ( Apium g)
raveolens L. var. rapaceum Mill. DC.), and these also include plants bred from the plant. As the plant body, for example, wild plants, plants obtained by cultivation or plants obtained by culture such as tissue culture, for example,
Examples thereof include leaves, flowers, branches, stems, fruits, roots, seeds, cultured cells or organs, and callus, and various processed products obtained by directly or physically or chemically treating these. To be

Examples of physical / chemical treatment methods include drying treatments such as sun-drying and air-drying, freeze-drying treatments, pulverization treatments, and the like.
Examples include freeze-dried products and crushed products. Examples of the biological treatment method include a fermentation method, and examples of the biological treatment product include a fermentation treatment product.

Examples of the plant extract include extracts obtained from the above-mentioned plant by various extraction methods. Examples of the extraction method include various solvent extractions and supercritical fluid extractions. The extract is sedimentation separation, cake filtration, clarification filtration, centrifugal filtration, centrifugal sedimentation, compression separation, various solid-liquid separation methods such as filter press, various concentration methods, various drying methods, formulation methods such as granulation or powderization, You may process by various purification methods etc.

Examples of the purification method include solvent fractionation method, column chromatography method, recrystallization method and the like.
In particular, a column chromatography method using various carriers such as Diaion HP-20 (manufactured by Mitsubishi Chemical Corporation) and Sephadex LH-20 (manufactured by Pharmacia) is preferably used.

Examples of the concentration and drying methods include freeze drying, natural drying, hot air drying, ventilation drying, blast drying, spray drying, reduced pressure drying, sun drying, vacuum drying, fluidized bed drying, foam layer drying, drum dryer and the like. Drying methods such as coating film drying, ultrasonic drying, electromagnetic wave drying and the like can be mentioned, and spray drying and freeze drying are preferably used.

During the extraction and treatment of the extract, for example, an antioxidant and a preservative can be added. As the solvent used for solvent extraction, any solvent can be used as long as it can extract a substance exhibiting a liver function protecting or improving action of the present invention, for example, tap water, distilled water, water such as deionized water, an inorganic salt aqueous solution, Aqueous media such as buffers, monohydric alcohols such as methanol, ethanol, propanol and butanol, polyhydric alcohols such as propylene glycol and glycerol, hexane, toluene, petroleum ether, benzene, ethyl acetate, chloroform, dichloromethane, 1,1, Two
-Organic solvents such as trichloroethene, dimethylsulfoxide, and acetone are listed, and an aqueous medium and alcohol are preferably used.

Examples of the buffer solution include a phosphate buffer solution and a citrate buffer solution. Examples of the inorganic salt of the inorganic salt aqueous solution include sodium chloride, potassium chloride, calcium chloride and the like. The alcohol is preferably a monohydric alcohol, and the monohydric alcohol is preferably ethanol.

These solvents may be used alone or in combination of two or more. The mixed solvent is preferably hydrous alcohol, more preferably hydrous monohydric alcohol, and particularly preferably hydrous ethanol. The water content is preferably 70% by volume or less, more preferably 40% by volume or less.
As the solvent, carbon dioxide converted into a supercritical fluid can also be used.

The extraction is carried out using, for example, 0.1 part by weight to 10000 parts by weight, preferably 1 part by weight to 100 parts by weight of the solvent per 1 part by weight of the plant. The extraction temperature is not particularly limited, but is preferably 0 ° C to 100 ° C, for example, 20 ° C to 90 ° C.
Is more preferable. The extraction time is not particularly limited, but is preferably 1 minute to 1 week, more preferably 30 minutes to 1 day, for example.

As a preferable extraction method, there is a method of extracting the residue obtained by extracting the aforementioned plant, the physically / chemically treated product of the plant or the biologically treated product with water with alcohol or hydrous alcohol. As the extraction temperature when extracting with water and alcohol or hydrous alcohol,
For example, 0 ° C to 100 ° C is preferable, and 20 ° C to 90 ° C is more preferable. The extraction time is not particularly limited, but for example, 1
Minutes to 1 week is preferable, and 30 minutes to 1 day is more preferable.

There are no particular restrictions on the equipment used for extraction, but examples include containers, stirrers, reflux condensers, Soxhlet extractors, homogenizers, shakers, ultrasonic generators, etc. devised for efficient extraction. To be

The liver function-protecting agent or improving agent of the present invention contains a plant or an extract of the plant prepared by the above-mentioned method, and if necessary, one or more pharmaceutically acceptable carriers. If necessary, it may further contain other active ingredients for protecting, enhancing, improving liver function or preventing or treating liver function disorder.

The liver function protecting or improving agent of the present invention is produced by any method well known in the technical field of pharmaceutics by mixing a plant or an extract of the plant with a carrier as the case requires. It The dosage form of the preparation is preferably the one that is most effective for protecting or improving liver function, and may be oral administration or parenteral administration such as intravenous, intraperitoneal or subcutaneous administration. . Of these, oral administration is preferred.

Dosage forms to be administered include tablets, powders, granules, pills, suspensions, emulsions, dips, capsules, syrups, injections, liquids, elixirs, extracts, tinctures, flow extracts. Agents and the like. Suitable for oral administration, for example, an extract, a tincture, a flow extract is a plant obtained by extracting from a plant of Gypsophila or celery arc with a solvent such as water, ethanol or a mixed solution of water and ethanol. The extract can be prepared as it is or concentrated.

Liquid preparations suitable for oral administration such as syrups include water, sugars such as sucrose, sorbitol and fructose, glycols such as polyethylene glycol and propylene glycol, oils such as sesame oil, olive oil and soybean oil. , P-hydroxybenzoic acid esters and other preservatives, paraoxybenzoic acid derivatives such as methyl paraoxybenzoate, preservatives such as sodium benzoate, and flavors such as strawberry flavor and peppermint. .

Further, tablets, powders, granules and the like suitable for oral administration include sugars such as lactose, sucrose, glucose, sucrose, mannitol and sorbitol, starches such as potato, wheat and corn, calcium carbonate, calcium sulfate. , Inorganic substances such as sodium hydrogen carbonate and sodium chloride, crystalline cellulose, licorice powder, excipients such as plant powder such as gentian powder, starch, agar, gelatin powder, crystalline cellulose, carmellose sodium, carmellose calcium, calcium carbonate, Disintegrating agents such as sodium hydrogen carbonate and sodium alginate, magnesium stearate, talc, hydrogenated vegetable oil, lubricants such as macrogol and silicone oil, polyvinyl alcohol, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose,
It can be produced using a binder such as carmellose, gelatin, starch paste, a surfactant such as fatty acid ester, a plasticizer such as glycerin and the like.

Injectables suitable for parenteral administration preferably consist of sterile aqueous preparations containing the active compound which is isotonic with the blood of the recipient. For example, in the case of an injection, a solution for injection is prepared by using a carrier such as a salt solution, a glucose solution or a mixture of salt water and a glucose solution. Also in these parenteral agents, the above-mentioned preservatives, preservatives, surfactants and the like can be used.

The dose and frequency of administration of the liver function-protecting or improving agent of the present invention vary depending on the administration form, age and weight of patient, nature or severity of symptoms to be treated, etc., but are not particularly limited. In the case of oral administration, the dry weight of the plant or the dry weight of the plant extract, the dry weight of the plant or the dry weight of the plant extract is 0.
01 mg to 50 g, preferably 0.05 mg to 10 g, is administered once to several times a day. In the case of parenteral administration such as intravenous administration, the dry weight of the plant or the dry weight of the plant is 0.001 mg to 50 g, preferably 0.01 mg to 10 g per adult once or several times a day. Administer.

When administered to animals, it varies depending on the dosage form, age, type of animal, nature of symptoms, severity, etc. and is not particularly limited, but as a dry weight of the plant or a dry weight of the plant extract. , 0.1m per 1kg body weight
g to 10 g, preferably 1 mg to 1 g is administered once to several times a day. In the case of parenteral administration such as intravenous administration, the dry weight of the plant or the dry weight of the plant extract is 0.01 mg to 10 g, preferably 1 mg to 1 g per 1 kg of body weight once or several times a day. Administer.

Regarding the dose and the number of doses,
It varies depending on the various conditions described above. As the food or drink or feed for liver function protection or improvement obtained by adding the plant or the plant extract of the present invention, the plant or the plant extract of the present invention can be used as a general food or drink or feed or their raw materials. Other than the addition of, those produced by general methods for producing foods and drinks or feeds can be mentioned.

Examples of general foods and drinks or feeds to which the plant or the plant extract is added, and raw materials thereof are:
There is no particular limitation, and the plant or the plant extract may be contained, or the plant or the plant extract may not be substantially contained. By adding a plant or the plant extract to a food or a drink or feed containing the plant or the plant extract, it is possible to increase the liver function protecting action or improving action of the food or feed. it can.

The food or drink or feed may be processed by a general processing method used for food or drink such as molding and granulation. Molding / granulation methods include fluidized bed granulation, stirring granulation, extrusion granulation, tumbling granulation, air flow granulation, compression molding granulation, crushing granulation, spray granulation, and spray granulation. Method, pan coating, fluidized bed coating, coating method such as dry coating, puff dry, excess steam method,
Examples include a foam matting method, a swelling method such as a microwave heating method, and an extrusion method such as an extrusion granulator and an extruder.

The amount of the plant or the plant extract of the present invention to be added to the food or drink or the feed is particularly limited as long as the food or the feed can exhibit a liver function protecting action or an improving action. However, the dry weight of the plant or the dry weight of the plant is, for example, 0.001 to 1
It is added so as to contain 100% by weight, preferably 0.01 to 100% by weight, and more preferably 0.1 to 100% by weight.

Specific foods and drinks to which the plant or the plant extract is added include, for example, juices, soft drinks and soups to which the plant or the plant extract is added.
Eggs such as teas, lactic acid bacteria beverages, fermented milk, frozen desserts, butter, cheese, yogurt, processed milk, skimmed milk powder, dairy products such as ham, sausage, hamburger, smelted fish products, soup rolls, egg tofu, etc. Examples include products, cookies, jelly, snacks, chewing gum and other confectionery, breads, noodles, pickles, smoked products, dried foods, boiled vegetables, seasonings and the like.

The forms of food and drink include powdered foods,
Sheet food, bottled food, canned food, retort food,
Examples include capsule foods, tablet foods, liquid foods, drinks, and the like. The food or drink of the present invention can be used as a healthy food or drink or a functional food or drink for protecting or improving liver function.

When a food or drink containing the plant of the present invention or the plant extract as an active ingredient and having a liver function protecting action or an improving action is ingested, the intake amount is not particularly limited, but per adult , The dry weight of the plant or the dry weight of the plant is from 0.01 mg per day to
The amount is 50 g, preferably 0.05 mg to 10 g. This intake is continued for 1 day to 1 year, preferably 2 weeks to 3 months. However, this intake amount is merely a guideline, and can be adjusted appropriately within a suitable range according to the degree of symptoms, age, weight, etc. of the ingestor.

Specific feeds to which the plant or the plant extract is added include mammals, birds, reptiles, amphibians,
Any feed may be used for animals such as fish, for example, pet feed such as dogs, cats, rats, livestock feed such as cattle and pigs, poultry feed such as chickens and turkeys,
Examples include feed for farmed fish such as Thailand and yellowtail. The feed of the present invention can be produced by appropriately mixing a plant or a plant extract with a feed material. Examples of the feed raw material include cereals, rice bran, vegetable oil meal, animal feed raw material, other feed raw materials, and refined products.

The grains include, for example, milo, wheat, barley, oats, rye, brown rice, buckwheat, fluff, acne, knees,
Examples include corn and soybeans. For rice bran,
Examples thereof include rice bran, defatted rice bran, bran, powder, wheat, germ, wheat bran, corn bran, and corn germ. Examples of vegetable oil residue include soybean oil residue, soybean oil residue, linseed oil residue, cottonseed oil residue, peanut oil residue, safflower oil residue, palm oil residue, palm oil residue, sesame oil residue, sunflower oil residue, rapeseed oil residue. , Kapok oil residue, mustard oil residue, etc.

Examples of animal feed raw materials include fish meal (North Sea meal, imported meal, whole meal, coastal meal, etc.), fish solvel, meat meal, meat-and-bone meal, blood meal, broken hair, bone meal, by-product for livestock treatment, and feather meal. , Silkworm, skim milk powder, casein, dried whey and the like.

Other feed ingredients include plant foliage (alfalfa, hay cube, alfalfa leaf meal, black acacia powder, etc.), corn processing industry by-products (corn gluten, meal, corn gluten feed, corn stapler, etc.), starch processed products. (Starch, etc.), sugar, fermentation products (yeast, beer dregs, malt root, alcohol dregs, soy sauce dregs, etc.), agricultural production by-products (citrus processed dregs, tofu dregs, coffee dregs, cocoa dregs, etc.), etc. (cassava, Broad beans, Guam,
Seaweed, krill, spirulina, chlorella, minerals, etc.) and the like. As a purified product, protein (casein,
Albumin, etc.), amino acids, sugars (starch, cellulose, sucrose, glucose, etc.), minerals, vitamins and the like.

When an animal is ingested a feed containing the plant or the plant extract according to the present invention as an active ingredient and having a liver function protecting action or an improving action, the amount of ingestion is not particularly limited. The dry weight of the plant body or the dry weight of the plant body per 1 kg is 0.1 mg to 10 g, preferably 1 mg to 1 g per day. This intake for 1 day to 1 year, preferably 2 weeks
Continue to take for 3 months. However, this intake amount is a guide only, and can be appropriately adjusted within a suitable range according to the type, age, weight, etc. of the ingested animal.

A food or beverage additive or feed additive having a liver function protecting action or improving action, which comprises a plant or an extract of the plant as an active ingredient,
An additive containing a plant or a plant extract prepared by the above-mentioned method as an active ingredient and generally used for foods and drinks or feeds as needed, for example, Food Additive Labeling Handbook (Japan Food Additives Association, Heisei (Published January 6, 9)
, Sweeteners, colorants, preservatives, thickening stabilizers, antioxidants, color formers, bleaches, fungicides, gum bases, bitters, enzymes or enzyme sources, brighteners, acidulants, seasonings Additives such as ingredients, emulsifiers, toughening agents, manufacturing agents, flavors, spice extracts and the like may be added. Moreover, you may add the carrier illustrated in the above-mentioned liver function protection or improving agent.

The concentration of the plant or plant extract in the food or drink additive or feed additive is not particularly limited, but is preferably 1 to 100% by weight, and 10 to 100% by weight.
% Is more preferable, and 20 to 100% is particularly preferable. Examples of the additives include the following additives. Examples of the sweetener include aspartame, licorice, stevia, xylose, swingle and the like.

Examples of coloring agents include carotenoids, turmeric pigments, flavonoids, caramel pigments, shikon pigments,
Examples include spirulina pigment, chlorophyll, purple yam pigment, purple yam pigment, perilla pigment, and blueberry pigment. Examples of the preservatives include sodium sulfite, benzoic acids, udo extract, echinacea extract, sage extract, sorbic acid, propionic acids and the like.

Examples of the thickening stabilizer include gums such as gum arabic and xanthan gum, alginic acids, chitin, chitosan, kidachi aloe extract, guar gum, hydroxypropyl cellulose, sodium caseinate, corn starch, carboxymethyl celluloses, gelatin, agar,
Examples thereof include dextrin, methyl cellulose, polyvinyl alcohol, microfibrous cellulose, microcrystalline cellulose, seaweed cellulose, sodium polyacrylate, sodium polyphosphate, carrageenan, yeast cell wall, konjac potato extract, nata de coco, mannan and the like.

Examples of the antioxidant include vitamin C
, Sodium ethylenediaminetetraacetate, calcium ethylenediaminetetraacetate, erythorbic acid, oryzanol, catechin, quercetin, clove extract, enzyme-treated rutin, apple extract, sesame oil extract, dibutylhydroxytoluene, fennel extract, horseradish extract, Seri extract, tea extract, tempeh extract, dokudami extract, tocotrienol, tocopherols, rapeseed oil extract, green coffee bean extract, sunflower seeds, ferulic acid, butylhydroxyanisole, blueberry leaf extract, propolis extract , Hego ginkgo extract, hesperetin, pepper extract, spinach extract, gallic acid, bayberry extract, eucalyptus extract, rosemary extract and the like.

Examples of the color former include sodium nitrite and the like. Examples of the bleaching agent include sodium sulfite and the like. Examples of the fungicide include orthophenylphenol and the like. As the gum base, for example, methyl acetylricinoleate, urshiro,
Ester gum, elemi resin, auri cucumber wax, ozokerite, opopanax resin, kauri gum, carnauba wax, guaiac resin, guttakachu, guttahankan, guttaperka, glycerin fatty acid selenium, gaylou, copaoba balsam, copal resin, rubber, rice bran wax, sugar cane wax, shelkowell Jeruton, sucrose fatty acid ester, solver, sorbitan fatty acid ester, talc, calcium carbonate, dammar resin, chicle,
Cirte, tsuno, low molecular weight rubber, paraffin wax, far balsam, propylene glycol fatty acid ester,
Examples include powder pulp, powdered chaff, jojoba wax, polyisobutylene, polybutene, microcrystal wax, mastic, massaland chocolate, beeswax, calcium phosphate and the like.

As the bittering agent, for example, isoalpha-bitter acid, caffeine, agaricus extract, quina extract, yellowfin extract, gentian extract, enzyme-treated naringin,
Jamaica Cassia extract, deobromine, naringin, big oyster extract, wormwood extract, Hikokoshi extract, Himematsutake extract, bolapet, methylthioadenosine, litchi extract, olive tea, red sea bream extract,
Examples include hop extract and mugwort extract.

Examples of the enzyme or enzyme source include amylase, trypsin, rennet, lactic acid bacteria and the like. Examples of the brightening agent include Urushirou and Mokurou. Examples of the acidulant include adipic acid, itaconic acid, citric acid, succinic acid, sodium acetate, tartaric acid, carbon dioxide, lactic acid, phytic acid, fumaric acid, malic acid, phosphoric acid and the like.

Examples of the seasoning include asparagine, aspartic acids, glutamic acid, glutamine, alanine, isoleucine, glycine, serine, amino acids such as cystine, tyrosine, leucine, proline, sodium inosinate, sodium uridylate, sodium guanylate, cytidyl. Nucleic acid such as sodium acidate, ribonucleotide calcium, ribonucleotide sodium, citric acid,
Organic acids such as succinic acid, potassium chloride, low salt sodium salt solution in salt water, crude seawater potassium chloride, whey salt, tripotassium phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, diphosphate Examples include sodium hydrogen, trisodium phosphate, chlorella extract and the like.

Examples of the emulsifier include sucrose fatty acid ester and the like. Examples of the strengthening agent include zinc salts, vitamin Cs, various amino acids, 5-adenylic acid, iron chloride, hesperidin, various calcined calcium, various uncalcined calcium, dibenzoylthiamine, calcium hydroxide, calcium carbonate, thiamine hydrochloride, Dunaliella. Carotene, tocopherol, nicotinic acid, carrot carotene, palm oil carotene, calcium pantothenate, vitamin A, hydroxyproline, calcium dihydrogen pyrophosphate, ferrous pyrophosphate, ferric pyrophosphate, ferritin, heme iron, menaquinone, folic acid , Riboflavin and the like.

Examples of the manufacturing agent include processing aids such as acetone and ion exchange resins, fig leaf extract, ash straw extract, kaolin, glycerin fatty acid ester, mulberry extract, bone ash, perilla extract, ginger extract, various kinds. Examples include tannin, paffia extract, grape seed extract, ethanol and the like. Examples of the fragrance include vanilla essence and the like.

Examples of the spice extract include capsicum extract and the like. In addition, these various additives can also be used by adding them to the above-mentioned liver function protecting agent or improving agent, and food and drink or feed having a liver function protecting action or improving action. The present invention will be described in detail below based on examples. However, the following examples do not limit the present invention.

[0057]

Example 1 Production of freeze-dried powder of acetone extract of ginseng Dried ginseng ( Artemisia apiacea Hance) powder (manufactured by Tochimoto Tenkaido Co., Ltd.) 1 kg was added with 10 liters of acetone.
The extract was extracted twice (at room temperature, stirred for 1 hour), and the obtained extract was concentrated and freeze-dried to obtain 54.6 g of freeze-dried powder of an acetone extract of Ganoderma lucidum.

Example 2 Production of lyophilized powder of acetone extract of celery arc Commercial celery arc ( Apium graveolens L. var. Rapac)
eum Mill. DC.) 1 kg of powder obtained by dry pulverization was extracted twice with 10 liters of acetone (stirring at room temperature for 1 hour), and the resulting extract was concentrated and freeze-dried to give celery arc. Lyophilized powder of acetone extract 45
3.5 g was obtained.

Example 3 Preparation of Acetone Extract Fraction of Salmonella 5 g of the freeze-dried powder prepared in Example 1 was added to 500 m of water.
Suspended with l. The suspension was passed through a column packed with a hydrophobic adsorption resin (Diaion HP-20, void volume 100 ml, manufactured by Mitsubishi Chemical Corporation). Then 250m
l 33% by volume methanol was passed through the column, and 250 ml
Of 33% by volume of methanol, 250 ml of 66% by volume of methanol, and 250 ml of 6% by volume of methanol.
6% by volume of methanol was passed through the column, and 250 ml of 10
Pass through 0% methanol and add 250 ml of 100%
Methanol was passed through the tower. Then, 250 ml of 100% acetone was passed through the column, and the eluted solution was concentrated and dried to obtain 0.084 g of a concentrated dry solid.

Example 4 Preparation of Fraction of Celery Arc Acetone Extract 5 g of freeze-dried powder prepared in Example 2 was added to 500 m of water.
Suspended with l. The suspension was passed through a column packed with a hydrophobic adsorption resin (Diaion HP-20, void volume 100 ml, manufactured by Mitsubishi Chemical Corporation). Then 250m
l 33% by volume of methanol is passed through the column, and 250 ml is added.
Of 33% by volume of methanol, 250 ml of 66% by volume of methanol, and 250 ml of 6% by volume of methanol.
6% by volume of methanol was passed through the column, and 250 ml of 10
Pass through 0% methanol and add 250 ml of 100%
After passing through methanol, 250 ml of 100% acetone was passed through. Then, 250 ml of 100% acetone was passed through the column, and the eluted solution was concentrated and dried to obtain 0.0076 g of a concentrated dry solid.

Example 5 Diet containing eucommia powder A feed was produced by mixing the following components.

[0062]     CE-2 (manufactured by CLEA Japan, Inc.) 99% by weight     Powder produced in Example 1 1% by weight

Example 6 Feed containing celery arc powder A feed was prepared by mixing the following components. CE-2 (manufactured by CLEA Japan, Inc.) 99% by weight Powder produced in Example 2 1% by weight

Comparative Example 1 A feed was produced by mixing the following components.

[0065]     CE-2 (manufactured by CLEA Japan, Inc.) 99% by weight     Pa index (Matsuya Chemical Co., Ltd.) 1% by weight

Example 7 Inhibition of Acetaminophen (AAP) -Induced Primary Culture Hepatocyte Injury by Fractions of Acetone Extract of Salmon and Celery Arc Collagenase perfusion [Seglen, PO, method in cell bio Logistics (Methods in Cell Biol
ogy), 13 , 29 (1976)]. That is, a male SD weighing about 130 g
Rats were abdominated under anesthesia and preperfusion solution kept at 37 ° C. [Hanks' Balanced Salt Solution (Gibco): 9.5 g, HEPES (Nacalai Tesque): 2.38 g, EGTA (Manufactured by Sigma): 0.19
g, NaHCO ₃ (manufactured by Kishida Chemical Co., Ltd.): 0.35 g
It was dissolved in L water and adjusted to pH 7.2], and 400 ml of the liver was perfused through the portal vein at a flow rate of 30 ml / min. Next, the collagenase solution kept at 37 ° C. [Hanks' liquid Nissui (manufactured by Nissui Pharmaceutical Co., Ltd.): 9.8 g, HEPES (manufactured by Nacalai Tesque, Inc.): 2.38 g, NaHCO ₃ (manufactured by Kishida Chemical Co., Ltd.): 0.35 g, CaCl ₂ (manufactured by Kishida Chemical Co., Ltd.): 0.56 g, trypsin inhibitor (manufactured by Sigma): 0.02 g, collagenase (manufactured by Sigma): 0.
5 g dissolved in 1 L of water and adjusted to pH 7.5]
Was perfused with 200 ml.

After the liver was digested, the liver was collected in a Petri dish, 20 ml of S-MEM medium (manufactured by Gibco) was added, and the sliced into small pieces with a scalpel. Then, pipetting with a 10 ml tip thick Komagome pipette to disperse the hepatocytes, and then filtering the hepatocytes with a gauze and a cell strainer (manufactured by Ikemoto Rika Kogyo Co., Ltd.),
A hepatocyte dispersion was obtained. The hepatocytes obtained here are non-parenchymal cells other than hepatocytes, such as endothelial cells, Kupffer cells,
Since it contains Ito cells, stellate cells, etc., only hepatocytes were purified by centrifugation. Specifically, the obtained hepatocyte dispersion was centrifuged at 50 × G for 1 minute under cooling, and the hepatocytes precipitated on the bottom were collected. This operation was repeated 3 times to separate and collect hepatocytes with high purity.

The separated and collected hepatocytes were suspended in the following basic medium to a concentration of 1.2 × 10 ⁶ cells / ml, seeded in 1.4 ml on a 6-well plate coated with Matrigel, and cultured as follows. Primary culture was performed under the conditions. The medium is MB752 / 1 (Waymouth's MB752 / 1) medium of Way Mouse (manufactured by Gibco), 10% by volume of bovine serum (FBS), and 50 U.
/ Ml penicillin, 50 μg / ml streptomycin, 10 ^-8 mol / l insulin and 10 ^-6 mo
A medium containing 1 / l of dexamethasone (hereinafter sometimes referred to as basal medium) was used.

The culture was carried out in a CO ₂ incubator at 37 ° C., 5% by volume CO ₂ , 95% by volume air. The basal medium was removed 4 hours after the completion of seeding, washed with PBS, and 700 μl of a new basal medium was dispensed for further culturing. The concentrated dry solid obtained in Example 3 and the concentrated dry solid obtained in Example 4 were dissolved in dimethyl sulfoxide (hereinafter abbreviated as DMSO) to a concentration of 10 mg / ml, and this was dissolved. Dilute 50 times with the above basic medium to obtain a fraction concentration of 200
A test solution of μg / ml was prepared.

Twenty-four hours after cell seeding, 14 wells of test solution were placed in each well.
0 μl each was added (the final concentration of the fraction was 20 μg / m
It becomes l. ), And 1 hour later, 560 μl of 50 mmol / l acetaminophen aqueous solution (liver injury inducer) was added (final concentration of acetaminophen was 20 mmo).
It becomes 1 / l. ), This was made into the test area. In addition, 24 hours after cell seeding, 2% by weight DMSO-containing medium 140 was added to each well.
μl each, and 1 hour later, 560 μl of 50 mmol / l acetaminophen aqueous solution (liver injury inducer) was added (final concentration of acetaminophen was 20 mm).
It becomes ol / l. ), This was made into the 1st control section.

Twenty-four hours after seeding the cells, 140 μl of a 2 wt% DMSO-containing medium was added to each well, and 1 hour after that, 560 μl of distilled water was added to each well, which was used as a second control group. 48 hours after addition of acetaminophen aqueous solution or distilled water, MTT [3- (4,5-Dimethyl-2-thia
zolyl) -2,5-diphenyl-2H-tetrazolium bromide] assay, and the number of cells in each section was measured by absorbance. Specifically, the basal medium in each well was removed, and MTT (10 mg
/ ML) -dissolved PBS (phosphate buffer solution-containing physiological saline) in an amount of 10% by volume, Waymouth's MB75
1.4 ml of 2/1 medium was dispensed. After incubation for 1 hour in a CO ₂ incubator at 37 ° C, 4.2 m
After adding 1 l of DMSO and stirring vigorously, the absorbance at 570 nm was measured with a microplate reader (BioRad, model 3550). The evaluation was performed in duplicate. The hepatocellular injury suppression rate is expressed by the following formula (I)
Calculated by

[0072]

[Formula 1] The results are shown in Table 1.

[0073]

[Table 1]

As shown in Table 1, the concentrated dry solids obtained in Example 3 and the concentrated dry solids obtained in Example 4 were 201.4% and 41%, respectively, of hepatocellular damage due to acetaminophen. Suppressed by 8%.

Example 8 Inhibition of Galactosamine-Induced Primary Culture Hepatocyte Injury by Fractions of Acetone Extract of Cellulose, Celery Arc, and the concentrated dry solid obtained in Example 3 and the concentrated dry solid obtained in Example 4 Dissolve each in DMSO to 10 mg / ml,
Dilute this with basic medium 50 times to obtain a fraction concentration of 200μ
Fraction dilutions of g / ml were made.

Cells were seeded and primary cultured in the same manner as in Example 7. Two hours after seeding, the medium was removed, and after washing with PBS, 700 μl of a new basic medium was dispensed, and 140 μl of the fraction dilution liquid was added (the final concentration of the fraction was 20 μl.
g / ml. ). 2 hours later, dissolved in the medium 5
0 mmol / l galactosamine (liver injury inducer) 56
Add 0 μl (final concentration of galactosamine is 20 mm
It becomes ol / l. ), This was made into the test area.

24 hours after cell seeding, 140 μl of 2 wt% DMSO-containing medium was added to each well, and 1 hour after that, 560 μl of 50 mmol / l galactosamine (liver injury inducer) was added (final concentration of galactosamine was 20
It becomes mmol / l. ), This was made into the 1st control section. After 24 hours of cell seeding, 2% by weight DM was added to each well.
140 μl of SO-containing medium was added, and 1 hour later, 560 μl of distilled water was added, and this was used as the second control group.

48 hours after the addition of galactosamine or distilled water, the number of cells in each section was measured by absorbance by the MTT assay. Specifically, except the basal medium in each hole,
PBS containing 10 mg / ml MTT dissolved in 10% by volume
1.4 ml of Waymouth's MB752 / 1 medium contained was dispensed. After incubation for 1 hour in a CO ₂ incubator at 37 ° C, 4.2 ml of DMSO
Was added, and the mixture was vigorously stirred and then, in a microplate reader (BioRad, model 3550), 57
The absorbance at 0 nm was measured respectively. The evaluation was performed in duplicate. The hepatocyte damage inhibition rate was calculated by the above-mentioned formula (I).

The results are shown in Table 2.

[0080]

[Table 2]

As shown in Table 2, the concentrated dry solids obtained in Example 3 and the concentrated dry solids obtained in Example 4 were 36.5% and 1%, respectively, of hepatocellular damage due to galactosamine.
It was suppressed by 9.2%.

Example 9 Suppression of Galactosamine-Induced Rat Liver Injury by Ginseng Wistar (Wistar) Purchased from Japan SLC
A white rat (male, 150 ± 20 g) was acclimated for 3 days or longer under constant conditions (temperature: 24 ± 2 ° C., humidity: 60 ± 5%, light / dark cycle: 12 hours), and then produced in Example 5. The feed and the feed produced in Comparative Example 1 were each fed for a total of 4 days. On the third day, a galactosamine aqueous solution dissolved in physiological saline at a concentration of 40 mg / ml was intraperitoneally administered at a rate of 400 mg per kg body weight. Twenty-two hours after the administration of galactosamine, laparotomy was performed under anesthesia with Nembutal, and blood was collected.

Using the blood thus obtained, the activity of GPT in blood was measured by the following method as an index of liver function. After coagulating the collected blood, centrifuge,
Serum was obtained. The GPT in the obtained serum was measured using Transaminase CII-Test Wako (manufactured by Wako Pure Chemical Industries, Ltd.). The GPT activity was obtained by calculating the relative value (%) of the value obtained by ingesting the feed produced in Example 5 with the value obtained by ingesting the feed of Comparative Example 1 as 100. Values are shown as mean ± standard error, and statistically significant difference test was conducted by T-test.

The results are shown in Table 3.

[0085]

[Table 3]

As shown in Table 3, when the feed produced in Example 5 was ingested, the serum content, which is an index of liver dysfunction, was higher than that in the diet produced in Comparative Example 1. The GPT activity of was suppressed to a low value of 46.4%, indicating that liver dysfunction was suppressed. During the four-day feeding period, no difference was observed in weight gain and no abnormality in appearance or behavior was observed regardless of which diet was given.

Example 10 Preparation of eucommia acetone extract compounding agent A liver function protecting agent or liver function improving agent having the following composition was prepared.
It was prepared by mixing the components.

[0088]   49 g of eucommia acetone extract produced in Example 1   Paraindex # 3 49g   Ferric pyrophosphate (iron source) 0.1 g   Foscal EFC (calcium source; manufactured by Nikko Fine Products) 1 g   Vitamin mix (Merck) 1g

Example 11 Production of celery arc acetone extract compounding agent A liver function protecting agent or liver function improving agent having the following composition was prepared.
It was prepared by mixing the components.

[0090]   49 g of celery arc acetone extract produced in Example 2   Paraindex # 3 49g   Ferric pyrophosphate (iron source) 0.1 g   Foscal EFC (calcium source; manufactured by Nikko Fine Products) 1 g   Vitamin mix (Merck) 1g

Example 12 20 g of liver function-protecting agent or improving agent prepared in Example 10
Was dispersed in 180 ml of water to prepare a beverage for protecting or improving liver function.

Example 13 20 g of liver function-protecting agent or improving agent prepared in Example 11
Was dispersed in 180 ml of water to prepare a beverage for protecting or improving liver function.

Example 14 Production of cake containing eucommia acetone extract A cookie (30 pieces) was produced by the following formulation.

[0094]   Soft flour 100g   74 g starch   Water 14g   30 g of eucommia acetone extract produced in Production Example 10   2 tsp baking powder   2 tsp salt   1 egg   80g butter   2 tbsp milk

Example 15 Preparation of Celery Arc Acetone Extract Blend Cake A cookie (30 pieces) was produced according to the following formulation.

[0096]   Soft flour 100g   74 g starch   Water 14g   30 g of celery arc acetone extract produced in Production Example 11   2 tsp baking powder   2 tsp salt   1 egg   80g butter   2 tbsp milk

[0097]

INDUSTRIAL APPLICABILITY According to the present invention, a liver function protector or improver, a food or drink for liver function protection or improvement, a feed or food additive having a liver function protecting or improving effect, and an animal, and an animal It is possible to provide a method for protecting or improving liver function of the.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A23L 1/30 A23L 1/30 B A61P 1/16 A61P 1/16 (72) Inventor Yoko Kamiya Tsukuba, Ibaraki Prefecture 2nd Miyukigaoka City, Kyowa Fermentation Industry Co., Ltd. Tsukuba Research Institute F-term (reference) CA14 MA52 NA14 ZA75 ZC61

Claims (6)

[Claims] 1. A ginger ( Artemisia apiacea Hance)
Or celery arc ( Apium graveolens L. var. Rapac
eum Mill. DC.) plant or an extract of the plant as an active ingredient. 2. The liver function protecting agent or improving agent according to claim 1, which is administered orally. 3. A ginger ( Artemisia apiacea Hance)
Or celery arc ( Apium graveolens L. var. Rapac
Eum Mill. DC.) plant or food / beverage or feed for protecting or improving liver function, which is obtained by adding the plant extract. 4. A ginger ( Artemisia apiacea Hance)
Or celery arc ( Apium graveolens L. var. Rapac
eum Mill. DC.) plant or an extract of the plant, which is a food / beverage additive or feed additive having a liver function protecting or improving action. 5. A method for protecting liver function of an animal or a method for improving liver function, which comprises feeding the animal with the protector or improver for liver function according to claim 1 or 2, or the feed according to claim 3. . 6. The method according to claim 5, wherein the animal is livestock, poultry, or farmed fish.