Nitric oxide production inhibitor and food composition for inhibiting nitric oxide production

Abstract

To provide a nitrogen monoxide production inhibitor that is safe to a living body.SOLUTION: A nitrogen monoxide production inhibitor contains one or more water extract of spice selected from a group consisting of celery seed, star-anise, caraway, marjoram, long pepper, pepper, Kaffir lime leaf, allspice, anise seed, oregano, cumin and sage as an active ingredient.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a nitric oxide production inhibitor and a food composition for inhibiting nitric oxide production. [Background technology]

[0002] Nitric oxide (hereinafter, sometimes referred to as NO) is a physiologically active substance that exhibits a variety of functions in the body. NO is produced by macrophages and other cells, and when produced in large quantities in the body, it is harmful to the body, causing the destruction of autologous tissues, inflammation, and rheumatism and diabetes. Therefore, a search for NO production inhibitors that suppress the production of NO has been ongoing, and cherry blossom extract (Patent Document 1), isohumulone (Patent Document 2), sphingoglycolipids (Patent Document 3), and the like have been reported. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] JP 2012-229170 A [Patent Document 2] JP 2013-177369 A [Patent Document 3] JP 2016-108314 A Summary of the Invention [Problem to be solved by the invention]

[0004] However, since nitric oxide is involved in various physiological functions, it is required that it does not exert an excessive inhibitory effect and is safe for the living body. Therefore, an object of the present invention is to provide a nitric oxide production inhibitor which is safe to the living body. [Means for solving the problem]

[0005] The present inventors conducted extensive research into nitric oxide production inhibitors that are safe for the body and, as a result, surprisingly found that water extracts of celery seed, star anise, caraway, marjoram, long pepper, pepper, kaffir lime leaf, allspice, anise seed, oregano, cumin, or sage have the effect of inhibiting nitric oxide production. The present invention is based on these findings. Thus, the present invention provides [1] A nitric oxide production inhibitor comprising, as an active ingredient, one or more water extracts of spices selected from the group consisting of celery seed, star anise, caraway, marjoram, long pepper, pepper, kaffir lime leaf, allspice, anise seed, oregano, cumin, and sage; [2] The nitric oxide production inhibitor according to [1], which is for anti-inflammatory purposes. [3] A food composition for inhibiting nitric oxide production, comprising, as an active ingredient, one or more water extracts of spices selected from the group consisting of celery seed, star anise, caraway, marjoram, long pepper, pepper, kaffir lime leaf, allspice, anise seed, oregano, cumin, and sage; and [4] The food composition for inhibiting nitric oxide production according to [3], which is for anti-inflammatory purposes. Regarding. Effect of the Invention

[0006] According to the nitric oxide production inhibitor of the present invention, it is possible to suppress the production of nitric oxide, which is harmful to the living body, and further, to suppress inflammation caused by the production of nitric oxide. [Brief description of the drawings]

[0007] [Figure 1] 1 is a graph showing the inhibition of NO production and cell viability by water extract and hot water extract of celery seed. [Diagram 2] 1 is a graph showing the inhibition of NO production and cell viability by water extract and hot water extract of star anise. [Diagram 3] 1 is a graph showing inhibition of NO production and cell viability by water and hot water extracts of caraway. [Figure 4] 1 is a graph showing the inhibition of NO production and cell viability by water extract and hot water extract of marjoram. [Diagram 5] 1 is a graph showing the inhibition of NO production and cell viability by water extract and hot water extract of long pepper. [Figure 6] 1 is a graph showing inhibition of NO production and cell viability by water and hot water extracts of pepper. [Figure 7] 1 is a graph showing the inhibition of NO production and cell viability by a water extract and a hot water extract of kaffir lime leaf. [Figure 8] 1 is a graph showing inhibition of NO production and cell viability by water and hot water extracts of allspice. [Figure 9] 1 is a graph showing the inhibition of NO production and cell viability by water extract and hot water extract of aniseed. [Figure 10] 1 is a graph showing inhibition of NO production and cell viability by water and hot water extracts of oregano. [Figure 11] 1 is a graph showing the inhibition of NO production and cell viability by water extract and hot water extract of Cumin. [Figure 12] 1 is a graph showing the inhibition of NO production and cell viability by water extract and hot water extract of sage. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0008] [1] Nitric oxide production inhibitors The nitric oxide production inhibitor of the present invention contains, as an active ingredient, one or more water extracts of spices selected from the group consisting of celery seed, star anise, caraway, marjoram, long pepper, pepper, kaffir lime leaf, allspice, anise seed, oregano, cumin, and sage.

[0009] 《Water extract》 The spices used for extracting the active ingredient of the nitric oxide production inhibitor of the present invention may be used raw or dried. In the examples described later, the active ingredient is extracted from dried spices, but the active ingredient contained in the dried spices is considered to be contained in raw spices as well, and is considered to be extracted by a water extraction method or the like. In order to improve the extraction efficiency, the raw or dried spices may be processed into a crushed or powdered state before extraction. The extraction of the active ingredient can be performed using a conventional extraction method used for water extraction of ingredients derived from plants, etc. Specific examples include water extraction, hot water extraction, and steam distillation. Examples of the extraction liquid include water, physiological saline, and buffer solutions. Examples of the buffer solution include phosphate buffer, sodium phosphate buffer, sodium carbonate buffer, citrate buffer, citrate phosphate buffer, acetate buffer, and Tris buffer. The pH of the extraction liquid is not particularly limited and is, for example, 3 to 10, preferably 5 to 8, and more preferably 6.5 to 7.5.

[0010] The extraction temperature of the water extract is not particularly limited as long as the active ingredient is extracted into the extraction liquid (water, etc.), but is preferably, for example, 0°C to 150°C, more preferably 0°C to 100°C, and most preferably 5°C to 100°C. It is preferable to carry out the extraction with stirring or shaking so as to improve the extraction efficiency. The extraction time can be appropriately determined depending on the part to be used, such as the root, stem, leaf, flower, fruit, or seed. The extraction time can also be appropriately determined depending on the state of the spice, that is, whether it is raw or dried, or when it is processed into a crushed or powdered state. The extraction time can also be appropriately determined depending on the extraction conditions, such as the temperature of the extract, or the presence or absence of stirring or shaking. The extraction time is usually 1 minute to 72 hours, preferably 1 hour to 48 hours, and most preferably 12 hours to 36 hours.

[0011] The extract may be from a single spice or a combination of two or more of the following spices: celery seed, star anise, caraway, marjoram, long pepper, pepper, kaffir lime leaf, allspice, anise seed, oregano, cumin, and sage.

[0012] (Water extraction method) In the water extraction method, for example, 2 to 100 times the amount of distilled water is added to the raw or dried powder of the spice, and the mixture is stirred for 1 to 72 hours. The obtained extract is centrifuged to recover the supernatant. The supernatant is filtered by suction and used as the extract as is, or freeze-dried before use. The temperature of the water can be, for example, 0°C or higher and lower than 50°C.

[0013] (Hot water extraction method) In the hot water extraction method, for example, 2 to 100 times the amount of distilled water is added to the raw or dried powder of the spice, and the mixture is stirred for 1 to 72 hours. The obtained extract is centrifuged to recover the supernatant. The supernatant is filtered by suction and used as the extract as is, or freeze-dried before use. The temperature of the water can be, for example, 50°C or higher and 100°C or lower.

[0014] (Steam distillation method) The steam distillation method is a method in which steam is passed through a raw material packed in a column, and aroma components distilled with the steam are condensed together with the steam. As a distillation method, any of pressurized steam distillation, normal pressure steam distillation, and reduced pressure steam distillation can be used.

[0015] The extract may be liquid or solid and may be included in the nitric oxide production inhibitor of the present invention. As the liquid, a water extract or a hot water extract may be used as it is. As the solid, the extract from which water has been removed may be used. Specific examples of the solid include pellets and dried products. As the dried product, for example, a freeze-dried product may be used. Examples of the water removal treatment include natural drying treatment, heat drying treatment, and freeze-drying treatment.

[0016] The nitric oxide production inhibitor of the present invention may consist of the water extract of the spice or may contain the water extract of the spice. When the nitric oxide production inhibitor of the present invention contains the water extract of the spice, it may contain other additives. The dosage form of the nitric oxide production inhibitor of the present invention is not particularly limited, and examples thereof include oral preparations such as powders, fine granules, granules, tablets, capsules, suspensions, emulsions, syrups, extracts, or pills, or parenteral preparations.

[0017] The oral preparations can be manufactured in accordance with standard methods using excipients such as gelatin, sodium alginate, starch, corn starch, white sugar, lactose, glucose, mannitol, carboxymethylcellulose, dextrin, polyvinylpyrrolidone, crystalline cellulose, soybean lecithin, sucrose, fatty acid esters, talc, magnesium stearate, polyethylene glycol, magnesium silicate, anhydrous silicic acid, or synthetic aluminum silicate, binders, disintegrants, surfactants, lubricants, flow enhancers, diluents, preservatives, colorants, fragrances, flavorings, stabilizers, moisturizers, preservatives, or antioxidants. Examples of parenteral preparations include injections. In preparing injections, in addition to the active ingredient, for example, water-soluble solvents such as physiological saline or Ringer's solution, water-insoluble solvents such as vegetable oils or fatty acid esters, isotonicity agents such as glucose or sodium chloride, solubilizers, stabilizers, preservatives, suspending agents, or emulsifiers may be optionally used.

[0018] The nitric oxide production inhibitor of the present invention may contain, for example, 90% by weight or more, 50% by weight or more, 10% by weight or more, or 1% by weight or more of the water extract of spices as a dry matter. The nitric oxide production inhibitor may contain a water extract of a single spice, or may contain a water extract of two or more spices, celery seed, star anise, caraway, marjoram, long pepper, pepper, kaffir lime leaf, allspice, anise seed, oregano, cumin, and sage. It may also contain a water extract extracted from a combination of two or more spices.

[0019] (celery seed) Celery (Apium graveolens var. dulce) is an annual or biennial herb of the Apiaceae family native to Europe. The stems are about 30-75 cm long, and from June to September they produce small, spherical white flowers. They are classified as light-colored vegetables, with colors ranging from green to pale green to yellow-white. The stems and leaves are eaten raw, and are also used as a flavoring for meat dishes. Dried celery berries are called celery seeds, and are used as a spice. The extraction temperature for the water extract of celery seed used as an active ingredient of the nitric oxide production inhibitor of the present invention is not limited, but a relatively low temperature of, for example, about 0°C to 50°C is preferred.

[0020] (Star Anise) Star anise is a spice made from the dried fruit of the Illicium verum plant. Illicium verum is an evergreen tall tree of the Illicium genus native to southern China and northeastern Vietnam, and grows to a height of about 15m. It bears yellow-green, deep pink, or maroon flowers and is widely cultivated in southern China, southern India, and the Indochina Peninsula. In China, the fruit is harvested twice a year, from September to October and March to April. Star anise is also called star anise or star aniseed, and has a scent similar to that of the Apiaceae family. The extraction temperature for the water extract of star anise used as an active ingredient of the nitric oxide production inhibitor of the present invention is not limited, and for example, extraction is performed efficiently at about 0°C to 100°C, and the extract contains almost no substances harmful to cells.

[0021] (Callaway) Caraway, also known as Carum carvi, is a biennial herb of the Apiaceae family native to Western Asia. It grows to a height of 30-60 cm, and at the end of the second year, it produces flowers in an umbel and bears yellowish-brown crescent-shaped fruits about 3 mm long. These fruits are used as a spice. The extraction temperature of the water extract of caraway used as an active ingredient of the nitric oxide production inhibitor of the present invention is not limited, and for example, extraction is performed efficiently at about 0°C to 100°C, and the extract contains almost no substances harmful to cells.

[0022] (Marjoram) Marjoram (Origanum majorana) is a perennial plant of the mint family. It grows to about 30cm, with reddish branches and paired round white leaves. In early summer, it produces numerous white flowers at the tips of its branches. The leaves are used as a spice, and essential oil is extracted from them. As a spice, it is known as an herb for meat dishes, and is used in all kinds of meat dishes, such as sausages, pates, and roast chicken and pork. The extraction temperature of the water extract of marjoram used as an active ingredient of the nitric oxide production inhibitor of the present invention is not limited, but a relatively low temperature of, for example, about 0° C. to 50° C. is preferable. Even a small amount has a relatively strong nitric oxide production inhibitory effect.

[0023] (Long Pepper) Long pepper, also known as piper longum L., is a woody climbing plant of the Piperaceae family native to southern India and Southeast Asia. It is cultivated in India, Indonesia, and Malaysia. The fruit has a flavor similar to pepper and is used as a spice. The extraction temperature for the aqueous extract of long pepper used as an active ingredient in the nitric oxide production inhibitor of the present invention is not limited, but a relatively low temperature, for example, of about 0°C to 50°C, is preferred, as this allows the active ingredient to be extracted efficiently and the extract contains almost no substances harmful to cells.

[0024] (pepper) Pepper, whose Japanese name is kosho, is a spice made from the fruit of a climbing plant in the Piperaceae family. Depending on the drying method, there are black pepper, white pepper, etc. Black pepper is made by drying the green fruit, including the skin, for a long time before it is fully ripe, taken from the pepper plant, until it turns black. White pepper is made by harvesting the fully ripe red fruit, drying it, soaking it in water, and then peeling off the softened skin. Pepper (Piper nigrum) is a vine plant of the Piperaceae genus native to India. It is grown in India, Indonesia, Malaysia, Vietnam, Sri Lanka, Brazil, Cambodia, and other countries. The extraction temperature for the water extract of pepper used as an active ingredient in the nitric oxide production inhibitor of the present invention is not limited, but a relatively high temperature, for example, of about 50°C to 100°C, is preferable, as this allows the active ingredient to be extracted efficiently and the extract contains almost no substances harmful to cells.

[0025] (Kaffir lime leaf) Kaffir lime leaves are the leaves of the kaffir lime tree. Its Japanese name is Citrus hystrix, a citrus fruit native to Thailand and Malaysia. It has a refreshing citrus scent similar to lemon, and is often used in cuisines (soups, curries, chicken dishes, fish dishes, etc.) in Southeast Asian countries such as Thailand and Indonesia. The leaves have wings on either side of the petiole, but because there is little difference in size between the petiole and the blade, the entire leaf appears to be two-tiered. The extraction temperature for the water extract of kaffir lime leaf used as an active ingredient in the nitric oxide production inhibitor of the present invention is not limited, and for example, extraction is performed efficiently at about 0°C to 100°C, and the extract contains almost no substances harmful to cells.

[0026] (Allspice) Allspice (Pimenta dioica) is a plant of the Myrtaceae family native to Central and South America, whose fruits and leaves are used as a spice. The fruits are harvested while unripe and dried for use as a spice. The extraction temperature for the water extract of allspice used as an active ingredient of the nitric oxide production inhibitor of the present invention is not limited, but a relatively high temperature of, for example, about 50°C to 100°C is preferred.

[0027] (Aniseed) Anise seeds are the fruit of the anise plant. Anise (Pimpinella anisum) is an annual plant of the Apiaceae family. It is native to the eastern Mediterranean region, including the Anatolian Peninsula, Greece, and Egypt. During the flowering season, the flower stalk extends and grows to a height of about 50 cm. The fruit is a bicarp about 5 mm long, consisting of two joined carpels, and has a strong fragrance. The extraction temperature for the water extract of aniseed used as an active ingredient in the nitric oxide production inhibitor of the present invention is not limited, but a relatively high temperature, for example, of about 50°C to 100°C, is preferable, as this allows the active ingredient to be extracted efficiently and the extract contains almost no substances harmful to cells.

[0028] (oregano) Oregano (Origanum vulgare) is a perennial plant of the mint family native to the Mediterranean coastal region of Europe. It germinates from a seed and grows to about 30-60cm. The stem is square, creeps slightly along the ground, stands upright, branched and hairy. The leaves are used as a spice, are egg-shaped and about 1.5cm long, and come in both smooth and hairy varieties. From summer to autumn (July-October), numerous spike-shaped, pale pink with a purple tinge, flowers measuring 4-7mm in diameter, bloom at the tip of the stem. The extraction temperature of the water extract of oregano used as the active ingredient of the nitric oxide production inhibitor of the present invention is not limited, but the active ingredient can be efficiently extracted at a relatively high temperature, for example, about 50° C. to 100° C. On the other hand, by extracting at a relatively low temperature of 0° C. to 50° C., the extract contains almost no substances harmful to cells.

[0029] (cumin) Cumin (Cuminum cyminum) is an annual or biennial herb of the Apiaceae family native to the eastern Mediterranean coast. It grows to a height of 20-40 cm and is hairless throughout the plant. The petiole is short, about 1 cm long, and has a needle-shaped sheath. The leaves are long and needle-shaped, and are pinnately bilobed. The flowers are umbrella-shaped and 2-3 cm in diameter. The petals are pink or white. The petals are oblong, slightly chipped at the tips. The seeds are oblong, narrow at both ends, about 6 mm long and 1.5 mm wide, and covered with white bristles. The plant flowers in April and produces seeds in May. Cumin seeds are generally recognized as seeds, but botanically they are fruits. These cumin seeds are often used as a spice. The extraction temperature for the water extract of cumin used as an active ingredient of the nitric oxide production inhibitor of the present invention is not limited, but a relatively low temperature of, for example, about 0°C to 50°C is preferable.

[0030] (Sage) Sage (Salvia officinalis) is a perennial or evergreen shrub of the Lamiaceae family native to the Mediterranean. It grows to a height of about 50-70cm, and produces purple or white lip-shaped flowers in May-July. Its long, stalked leaves grow opposite each other and have fine curls on the surface. They are used as a spice. The extraction temperature for the water extract of sage used as an active ingredient of the nitric oxide production inhibitor of the present invention is not limited, but a relatively low temperature of, for example, about 0°C to 50°C is preferred.

[0031] The nitric oxide production inhibitor of the present invention can be used for anti-inflammatory purposes, such as cancer, autoimmune diseases, neurodegenerative diseases, and pruritic skin diseases. The dosage of the nitric oxide production inhibitor of the present invention can be appropriately determined depending on the type of inflammation, the patient's age, sex, body weight, the severity of symptoms, or the administration method, and can be, for example, 1 to 3000 mg / kg body weight / day, preferably 5 to 2000 mg / kg body weight / day, more preferably 10 to 1500 mg / kg body weight / day, even more preferably 15 to 1000 mg / kg body weight / day, even more preferably 20 to 800 mg / kg body weight / day, and most preferably 25 to 400 mg / kg body weight / day as a lyophilized product. Of course, the above administration method is an example, and other administration methods may be used. It is desirable that the administration method, dosage, administration period, administration interval, etc. of the nitric oxide production inhibitor to humans are determined by a controlled clinical trial.

[0032] The spice extract, which is the active ingredient in the present invention, can be administered to a subject [animal, preferably a mammal (especially a human)] in an effective amount, either alone or preferably together with a pharmaceutically or veterinarily acceptable usual carrier or diluent. It can also be given to animals other than humans in the form of food or drink as feed. Examples of animals other than humans include pets such as dogs, cats, rabbits, hamsters, guinea pigs, and squirrels; livestock such as cows and pigs; laboratory animals such as mice and rats; and animals kept in zoos, etc.

[0033] [2] Food composition for inhibiting nitric oxide production The food composition for inhibiting nitric oxide production of the present invention contains, as an active ingredient, one or more water extracts of spices selected from the group consisting of celery seed, star anise, caraway, marjoram, long pepper, pepper, kaffir lime leaf, allspice, anise seed, oregano, cumin, and sage. The food composition for inhibiting nitric oxide production of the present invention may contain, for example, the nitric oxide production inhibitor of the present invention. That is, in this specification, the food composition means one that contains the nitric oxide production inhibitor of the present invention and a food or beverage. The food composition of the present invention contains the nitric oxide production inhibitor of the present invention, and therefore can be used as a food composition for inhibiting nitric oxide production.

[0034] Specific examples of foods include fresh prepared foods such as salads; cooked foods such as steaks, pizzas, and hamburgers; stir-fried foods such as stir-fried vegetables; vegetables such as tomatoes, bell peppers, celery, bitter melon, carrots, potatoes, and asparagus, and prepared foods made from these vegetables; confectioneries such as cookies, bread, biscuits, dry bread, cakes, rice crackers, yokan, puddings, jellies, ice cream, chewing gum, crackers, chips, chocolate, and candies; noodles such as udon, pasta, and soba; fish paste products such as kamaboko, ham, and fish sausages; dairy products such as cheese, cream, and butter; condiments such as miso, soy sauce, dressing, ketchup, mayonnaise, soup base, noodle soup, curry powder, mirin, roux, and seasoning spices; soy foods such as tofu; konjac; and supplements. The food product is preferably a roux (eg for a curry or stew), a seasoning spice, or a supplement.

[0035] Examples of beverages include coffee beverages, cocoa beverages, vegetable juices obtained from the above-mentioned vegetables, fruit juice beverages such as grapefruit juice, orange juice, grape juice, and lemon juice, tea beverages such as green tea, black tea, green tea, and oolong tea, alcoholic beverages such as beer, wine (red wine, white wine, sparkling wine, etc.), sake, plum wine, sparkling wine, whiskey, brandy, shochu, rum, gin, and liqueurs, milk beverages, soy milk beverages, liquid diets, and sports beverages. The beverage is preferably spiced tea (e.g., herbal tea or chai).

[0036] The food or beverage includes feed for animals, such as primates, including humans, cows, pigs, sheep, goats, horses, dogs, cats, rabbits, rats, and mice.

[0037] If desired, these foods or beverages may contain food additives and food ingredients, such as antioxidants, flavorings, acidulants, colorants, emulsifiers, preservatives, seasonings, sweeteners, spices, pH adjusters, stabilizers, antioxidants, vegetable oils, animal oils, sugars and sugar alcohols, vitamins, organic acids, fruit juice extracts, vegetable extracts, grains, beans, vegetables, meats, seafood, etc., either alone or in combination of two or more. The amounts of these food ingredients and food additives to be added may be appropriately determined within the range that does not impair the object of the present invention.

[0038] These foods or beverages can be subjected to common sterilization treatments, such as heat-pressure sterilization using retort and autoclave, batch sterilization, plate sterilization, electric heating sterilization, microwave heating sterilization, and steam sterilization such as injection and infusion.

[0039] Foods and beverages include functional foods (beverages) and health foods (beverages). In this specification, "health foods (beverages)" refers to foods or beverages that have some effect on health or that can be expected to have an effect, and "functional foods (beverages)" refers to foods or beverages that, among the above-mentioned "health foods (beverages), have been designed and processed to fully express a bioregulatory function (i.e., an immunostimulatory function). Functional foods and health foods can be in the form of granules, solids, liquids, capsules, gels, or tablets.

[0040] The food composition for inhibiting nitric oxide production of the present invention can be used for anti-inflammatory purposes by taking advantage of its effect of inhibiting NO production. EXAMPLES

[0041] The present invention will be specifically described below with reference to examples, but the scope of the present invention is not limited thereto.

[0042] Example 1 In this example, the inhibitory effect of a water extract and a hot water extract of celery seed on nitric oxide production was examined. 20 g of dried celery fruit powder was added to 10 times the amount of distilled water (200 g) and mixed. The mixture was stirred at 4°C for 12 hours using a stirrer. The extract was centrifuged at 3200 rpm for 10 minutes. The supernatant was collected and suction filtered through a filter with a maximum pore size of 20 to 25 μm. The resulting supernatant was freeze-dried to obtain a dried water extract.

[0043] 20 g of dried celery fruit powder was added to 10 times the amount of distilled water (200 g) and mixed. The mixture was stirred at 95°C for 1 hour using a stirrer. The extract was centrifuged at 3200 rpm for 10 minutes. The supernatant was collected and suction filtered through a filter with a maximum pore size of 20 to 25 μm. The resulting supernatant was freeze-dried to obtain a dried hot water extract.

[0044] The inhibitory effect of the dried water extract and the dried hot water extract on NO production in macrophages was examined. Macrophage-like cell line (RAW264.7) subcultured in 10% FBS-DMEM medium was plated in a 96-well plate at 2 × 10 5 The cells were seeded at a cell density of 100 cells / well and cultured for 20 hours at 37°C in a 5% CO2 incubator to allow the cells to settle. Each sample was added at a concentration of 0.008%, 0.04%, or 0.2% by weight. In addition, the bacterial toxin Lipopolysaccharides (LPS) from E. coli O26:B6 was added at a concentration of 100 μg / mL to stimulate iNOS (inducible nitric oxide synthase) in RAW264.7. 5% CO 2 The cells were cultured in an incubator at 37°C for 24 hours. The amount of NO production was measured by collecting the culture medium, and measuring the amount of nitric oxide (NO - Nitrite (NO 2- ) was measured. After the end of the culture, the viability of the cells on the plate was measured using the MTT Cell Viability Assay Kit (Biotium). As shown in FIG. 1, both the water extract and the hot water extract dose-dependently inhibited the production of nitric oxide.

[0045] Example 2 In this example, the inhibitory effect of a water extract and a hot water extract of star anise on nitric oxide production was examined. The procedure of Example 1 was repeated, except that star anise fruit dried powder was used instead of celery fruit dried powder, to obtain dried water extract and hot water extract, and the inhibitory effect on NO production was examined. As shown in Figure 2, both the water extract and the hot water extract dose-dependently inhibited nitric oxide production.

[0046] Example 3 In this example, the inhibitory effect of a water extract and a hot water extract of caraway on nitric oxide production was examined. The procedure of Example 1 was repeated, except that the dried powder of caraway fruit was used instead of the dried powder of celery fruit, to obtain dried products of the water extract and the hot water extract, and the inhibitory effect on NO production was examined. As shown in Figure 3, both the water extract and the hot water extract dose-dependently inhibited the production of nitric oxide.

[0047] Example 4 In this example, the inhibitory effect of a water extract and a hot water extract of marjoram on nitric oxide production was examined. The procedure of Example 1 was repeated, except that dried marjoram leaf powder was used instead of dried celery fruit powder, to obtain dried water extract and dried hot water extract, and the inhibitory effect on NO production was examined. As shown in Figure 4, both the water extract and the hot water extract dose-dependently inhibited nitric oxide production.

[0048] Example 5 In this example, the inhibitory effect of the water extract and hot water extract of long pepper on nitric oxide production was examined. The procedure of Example 1 was repeated, except that the dried powder of long pepper fruit was used instead of the dried powder of celery fruit, to obtain dried products of the water extract and the hot water extract, and the inhibitory effect on NO production was examined. As shown in Figure 5, both the water extract and the hot water extract dose-dependently inhibited the production of nitric oxide.

[0049] Example 6 In this example, the inhibitory effect of a water extract and a hot water extract of black pepper on nitric oxide production was examined. The procedure of Example 1 was repeated, except that the dried powder of black pepper fruit was used instead of the dried powder of celery fruit, to obtain dried products of the water extract and the hot water extract, and the inhibitory effect on NO production was examined. As shown in Figure 6, both the water extract and the hot water extract dose-dependently inhibited the production of nitric oxide.

[0050] Example 7 In this example, the inhibitory effect of a water extract and a hot water extract of kaffir lime leaf on nitric oxide production was examined. The procedure of Example 1 was repeated, except that dried powder of kaffir lime leaves was used instead of dried powder of celery fruit, to obtain dried products of the water extract and the hot water extract, and the inhibitory effect on NO production was examined. As shown in Figure 7, both the water extract and the hot water extract dose-dependently inhibited the production of nitric oxide.

[0051] Example 8 In this example, the inhibitory effect of a water extract and a hot water extract of allspice on nitric oxide production was examined. The procedure of Example 1 was repeated, except that the dried powder of allspice fruit was used instead of the dried powder of celery fruit, to obtain dried products of the water extract and the hot water extract, and the inhibitory effect on NO production was examined. As shown in Figure 8, both the water extract and the hot water extract dose-dependently inhibited the production of nitric oxide.

[0052] Example 9 In this example, the inhibitory effect of a water extract and a hot water extract of aniseed on nitric oxide production was examined. The procedure of Example 1 was repeated, except that the dried powder of aniseed was used instead of the dried powder of celery fruit, to obtain dried products of the water extract and the hot water extract, and the inhibitory effect on NO production was examined. As shown in Figure 9, both the water extract and the hot water extract dose-dependently inhibited the production of nitric oxide.

[0053] Example 10 In this example, the inhibitory effect of a water extract and a hot water extract of oregano on nitric oxide production was examined. The procedure of Example 1 was repeated, except that the dried powder of oregano leaves was used instead of the dried powder of celery berries, to obtain dried products of the water extract and the hot water extract, and the inhibitory effect on NO production was examined. As shown in Figure 10, both the water extract and the hot water extract dose-dependently inhibited the production of nitric oxide.

[0054] Example 11 In this example, the inhibitory effect of a water extract and a hot water extract of cumin on nitric oxide production was examined. The procedure of Example 1 was repeated, except that cumin fruit dry powder was used instead of celery fruit dry powder, to obtain dried water extract and hot water extract, and the inhibitory effect on NO production was examined. As shown in Figure 11, both the water extract and the hot water extract dose-dependently inhibited nitric oxide production.

[0055] Example 12 In this example, the inhibitory effect of a water extract and a hot water extract of sage on nitric oxide production was examined. The procedure of Example 1 was repeated, except that dried powder of sage leaves was used instead of dried powder of celery berries, to obtain dried products of the water extract and the hot water extract, and the inhibitory effect on NO production was examined. As shown in Figure 12, both the water extract and the hot water extract dose-dependently inhibited the production of nitric oxide. [Industrial Applicability]

[0056] The nitric oxide production inhibitor and the food composition for inhibiting nitric oxide production of the present invention can suppress inflammation.

Claims

1. A nitric oxide production inhibitor comprising as an active ingredient one or more water extracts of spices selected from the group consisting of celery seed, star anise, marjoram, long pepper, pepper, allspice, anise seed, oregano, cumin, and sage.

2. The nitric oxide production inhibitor according to claim 1, which is for anti-inflammatory purposes.

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