KR20030028234A - Extracts of buckwheat and/or buchwheat chaff with anti-inflammatory effects and composition comprising of the same - Google Patents

Abstract

PURPOSE: A composition containing an extract of buckwheat, buckwheat husk and a mixture thereof as an active ingredient is provided which has a suppression effect on inflammation generated by endotoxin and outer inflammation-inducing material. CONSTITUTION: The composition having antiinflammatory activity is prepared by extracting buckwheat, buckwheat husk and a mixture thereof in a solvent including water, ethanol, acetone or methanol and then subjecting to chromatography using 40 to 80% ethanol aqueous solution to obtain fractions having antiinflammatory activity.

Description

Buckwheat and / or buckwheat bark extract with anti-inflammatory activity, and anti-inflammatory composition comprising the same as an active ingredient

The present invention relates to an anti-inflammatory composition comprising an extract of a substance selected from the group consisting of buckwheat, buckwheat bark, and mixtures thereof and having an anti-inflammatory activity as an active ingredient.

Buckwheat, one of the traditional foods in Korea, has high nutritional biochemical value, rich in protein, essential amino acids, niacin, various vitamins, calcium, iron, and so on. It is used in food and recipes (Jewish, food compatibility, 1996). Traditional foods using buckwheat are known to have the effect of preventing and treating hypertension, diabetes mellitus, obesity, etc. as folk remedies (Jung Dong-hyo, et al., Food History, Advanced Culture History, 1998). On the other hand, studies on the physiological activity of conventional buckwheat extract, blood pressure control efficacy (hypnosis, Korean Journal of Nutrition and Nutrition, 20: 300-305, 1991), diabetes treatment efficacy (hypnosis, Korean Journal of Nutrition and Nutrition, 21: 478-483, 1992), antimutagenic efficacy (in seungseung, research report on antimutagenicity of buckwheat leaf ethanol extract, 32: 1141-1148, 1994), efficacy on kidney improvement (Yokozawa, Biosci. Biotechnol. Biochem. , 65: 396- 400. 2001), there are no other research reports on other biological activities. In particular, no studies have been conducted on whether buckwheat and buckwheat husk fractions that have been used in various foods have inhibitory effects on inflammation caused by bacterial endotoxins and external inflammatory agents.

Inflammation is a bacterium-derived abscess that actively infiltrate tissues with purulent fungi that white blood cells devour, resulting in the death of white blood cells, tissues damaged by bacteria, and cell debris, which accumulate in the tissues. The pathological state of the place, and the anti-inflammatory effect is to inhibit the growth of invading bacteria by antibacterial agents, or to activate the macrophage (macrophage) that phages the foreign matter accumulated in the abscesses to enhance the ability to digest and excrete them Abscess is the process of treatment. In general, an inflammatory response is a defense process of a living body that attempts to repair and repair the damaged area when an invasion that causes some organic change in a cell or tissue of the living body is applied. Therefore, such a series of reactions include local blood vessels, various tissue cells of body fluids, immune-involved cells, and the like.

To date, anti-inflammatory agents have been shown to inhibit the biosynthesis of prostaglandins activated by physicochemical and physiological stimuli from the cell membranes of damaged tissues or leukocytes inoculated by chemotactic factors in acute inflammation. (Moyazawa, K and Mikami, T., Japan J. Pharmacol., 38, 199, 1985). However, these drugs have a therapeutic effect on the symptoms of acute inflammation, but in chronic inflammation such as rheumatoid arthritis, only the primary inflammation is reported, and immunological therapeutic effects are not expected. Recently, researches on arachidonic acid have been actively conducted on hydroxyeicosatetraenoic acid (HETE), leukotriene B4, and leukotriene C4, which are substances produced by lipoxygenase. This has been shown to have some effect on chronic inflammation (Huskisson, ED et al., Inflammation, Mechanism and Treatment, MTP, Lancaster, England, p.55, 1980).

Therefore, it is necessary to study the substances that can be safely ingested without additional purification process, including natural plant extracts, can inhibit inflammation, and can be easily used in food.

It is an object of the present invention to provide an extract of a substance selected from the group consisting of buckwheat, buckwheat husk, and mixtures thereof having anti-inflammatory activity.

It is another object of the present invention to provide a solvent extract of a material selected from the group consisting of buckwheat, buckwheat husk, and mixtures thereof having anti-inflammatory activity.

It is another object of the present invention to provide a fraction extract obtained by performing adsorption chromatography on a solvent extract of a material selected from the group consisting of buckwheat, buckwheat bark, and mixtures thereof having anti-inflammatory activity.

Still another object of the present invention is an agent, a processed food, a functional food, a food additive, a functional beverage comprising an extract of a material selected from the group consisting of buckwheat, buckwheat bark, and mixtures thereof having the anti-inflammatory activity as an active ingredient. Or to provide a beverage additive.

1 is a measure of the effect of the buckwheat fraction extract according to the present invention inhibits rat mortality caused by lipopolysaccharide (LPS; lipopolysaccharide).

Figure 2 is a measure of the efficacy of buckwheat shell fraction extract according to the present invention inhibits rat mortality caused by lipopolysaccharide.

Figure 3 measures the extent to which the buckwheat fraction extract according to the present invention inhibits ear inflammation in rats induced by TPA (12- ο- tetradecanoylphorbol-13-acetate).

Figure 4 is a buckwheat shell fraction extract according to the present invention is to measure the efficacy of inhibiting ear inflammation of the rat induced by T.P.

Figure 5 is a comparison of the buckwheat fraction extract, buckwheat shell fraction extract, and epigallocatechin gallate (EGCG; epigallocatechingallate) according to the present invention to determine the efficacy of inhibiting ear inflammation in rats induced by T.P.

In order to achieve the above object, the present invention has an anti-inflammatory activity, buckwheat, buckwheat bark, and an extract of a substance selected from the group consisting of a mixture thereof, and pharmaceutical, processed food, functional food, food comprising the same as an active ingredient Provided are additives, functional beverages, or beverage additives.

Hereinafter, the present invention will be described in detail.

The inventors have confirmed that the extract of buckwheat and buckwheat husk is excellent in inhibiting inflammation caused by bacterial endotoxin and inflammation-causing substances in animal experiments, and completed the present invention based on this.

The present invention relates to extracts of substances selected from the group consisting of buckwheat, buckwheat husk, and mixtures thereof, having anti-inflammatory activity. In addition, the extract may be a solvent extract, preferably a fraction extract obtained by performing adsorption chromatography on a solvent extract of a material selected from the group consisting of buckwheat, buckwheat husk, and mixtures thereof.

In relation to the solvent extract, conventional solvent extraction methods known to those skilled in the art can be used to extract materials selected from the group consisting of buckwheat, buckwheat husk, and mixtures thereof. Preferably, the solvent is ethanol, acetone, or methanol, and in order to apply to food, the solvent is more preferably ethanol, but is not limited thereto. In addition, the solvent extract preferably contains 20 to 80% of a solvent such as ethanol, acetone, or methanol, more preferably an aqueous solution containing 50% of a solvent.

In relation to the fraction extract, the solvent extract may be loaded on an adsorption column and eluted with a solvent to obtain a fraction extract, and all of the usual adsorption chromatography known to those skilled in the art may be applied. For example, the adsorption column may use a styrene-based porous adsorption resin, a hydrophobic filler XAD-4 or HP-20. In one example of the present invention, after filling the solvent extract in the adsorption column, it can be prepared by washing with 10 to 30% ethanol solution, solvent fractionation with 40 to 80% ethanol. More preferably, the fraction extract of the present invention can be prepared by washing with 20% ethanol and solvent fractionation with 40% ethanol.

In another aspect, the present invention provides an anti-inflammatory composition comprising an extract of a material selected from the group consisting of buckwheat, buckwheat bark, and mixtures thereof having the anti-inflammatory activity as an active ingredient. The anti-inflammatory composition may be used as a drug, processed food, functional food, food additives, functional drinks, or beverage additives.

The extract of the material selected from the group consisting of buckwheat, buckwheat bark, and mixtures thereof is preferably included in an amount of 0.01 to 10% by weight based on the total composition. In addition, the composition containing the extract as an active ingredient is preferably administered orally.

The extract of the material selected from the group consisting of the buckwheat, buckwheat bark, and mixtures thereof may be prepared by a known method in the pharmaceutical field, to be used as a medicament for treating inflammation, or a pharmaceutically acceptable carrier It may be mixed with a carrier, a forming agent, a diluent, and the like to prepare a powder, granule, tablet, capsule, or injection formulation. Specifically, the drug may be used for inflammatory diseases and the like, and may be preferably used as an effective therapeutic agent for inflammation caused by bacterial endotoxins and external inflammation causing substances.

In addition, when used as processed foods, functional foods, food additives, functional foods, or food additives containing an extract of a substance selected from the group consisting of buckwheat, buckwheat bark, and mixtures thereof having the anti-inflammatory activity as an active ingredient, It may be prepared as a food as it is, or mixed with various components used in the conventional food. The foods to be prepared can be prepared in various food forms such as solid foods, creamy or jam-like semi-liquid foods, gel foods, and beverages, and are commonly used in the form of capsules, granules, tablets, drinks, and the like. Any substrate can be used to prepare soft drinks, juices, coffee, tea, liqueurs, milk, whey beverages, lactic acid bacteria beverages, candies, chewing gum, chocolate, gummi, yogurt, ice cream, puddings, water yolks and the like.

Noodles can be prepared as a processed food comprising an extract of a substance selected from the group consisting of buckwheat, buckwheat bark, and mixtures thereof having an anti-inflammatory activity of the present invention as an active ingredient, mixing flour and water, salt, and kneading In a conventional noodle prepared by adding the buckwheat fraction extract or buckwheat husk fraction extract of the present invention to 1 part by weight to 70 parts by weight based on the total weight of noodles.

Therefore, in order to confirm that the buckwheat and buckwheat bark extract according to the present invention has anti-inflammatory effects, the effect on bacterial endotoxin and inflammation induced by TP was investigated.

As can be seen in Figures 1 and 2, when oral administration of buckwheat fraction extract and buckwheat husk fraction extract to the rat, the mouse cooking rate caused by bacteria endotoxin lipopolysaccharide (LPS; lipopolysaccharide) The inhibitory effect was superior to the control group orally administered the same amount of phosphate solution.

3 through 4, the buckwheat fraction extract and the buckwheat husk fraction extract have inhibitory effects on ear inflammation in rats induced by dermal inflammation causing TPA (12- ο- tetradecanoylphorbol-13-acetate). As shown in FIG. 5, the ear inflammation inhibition rate was similar to that of epigallocatechin gallate of green tea, which is known as a conventional anti-inflammatory substance.

According to the present invention, an extract of a material selected from the group consisting of buckwheat, buckwheat husk, and mixtures thereof can effectively suppress inflammation caused by endotoxins and external inflammation causing substances secreted by bacteria. In addition, the extract can be used in food grade, so that it can be safely ingested without a separate purification process to remove the toxicity, so having an anti-inflammatory activity, an extract of a material selected from the group consisting of buckwheat, buckwheat bark, and mixtures thereof When added to foods can be obtained anti-inflammatory benefits. In particular, the economy can be improved by recycling the enormous amount of buckwheat husk that is being discarded.

Hereinafter, preferred examples are provided to help understanding of the present invention, but the following examples are merely to illustrate the present invention, and the scope of the present invention is not limited to the following examples.

EXAMPLE

Example 1 Preparation of Buckwheat Fraction Extract

50% ethanol (Duksan, Korea) aqueous solution was added about 6 times to 1 part of dried buckwheat and refluxed for 6 hours using a stirrer at room temperature. The supernatant obtained by centrifugation (Vision, Korea) for 30 minutes at 8,000 rpm, was collected, and the residue was repeatedly extracted once in the same manner. The extract thus obtained was filtered with filter paper (whatmann 41), and then freeze-dried to obtain a buckwheat solvent extract (hereinafter referred to as "buckwheat solvent extract").

The obtained buckwheat solvent extract was subjected to adsorption column chromatography to prepare a fraction extract of buckwheat solvent extract (hereinafter referred to as "buckwheat fraction extract"). Specifically, the buckwheat solvent extract is adsorbed onto XAD-4 (Sigma, USA) or HP-20 (Mitsbish, Japan), which is a hydrophobic filler, and 20% ethanol is poured to fractions containing nutrients such as proteins or sugars. Removed. 40% ethanol was poured again into the remaining fractions to obtain a buckwheat fraction extract.

Example 2 Preparation of Buckwheat Peel Fraction Extract

Except for using buckwheat husk as a starting material, the same method as in Example 1 was carried out to obtain a solvent extract of buckwheat husk (hereinafter referred to as "solvent extract of buckwheat husk"), and from the solvent extract of buckwheat husk Fraction extract of buckwheat husk (hereinafter, fraction extract of buckwheat husk) was prepared by adsorption column chromatography.

Example 3 Inhibitory Effect of Inflammation Caused by Bacterial Endotoxin

The inhibitory activity against inflammation caused by endotoxin of bacteria of the buckwheat fraction extract and the buckwheat husk fraction extract obtained in Examples 1 and 2 was measured. As a method for measuring this, it was carried out according to the following known method (Yang et al., J. Nutr., 128: 2334-2340, 1998).

First, 25-30 g of BALB / C mice were prepared in 25 groups per group and fasted for 12 hours before the experiment. Then, the buckwheat fraction extract and the buckwheat shell fraction extracts obtained in Examples 1 and 2 were treated at 1 g / kg. Orally administered to. On the other hand, only the same amount of phosphate solution was orally administered as a control. After 30 minutes of anesthesia with xylazine hydrochloride, 40 mg / kg lipopolysaccharide was administered intraperitoneally after 5 minutes. Survival was measured every 2 hours until 30 hours, and the degree of anti-inflammatory was measured according to the difference in survival numbers for the control group.

The buckwheat fraction extract and the buckwheat husk fraction extract obtained in Examples 1 and 2 have inhibitory effects on the mortality rate of rats caused by lipopolysaccharide, a bacterial endotoxin and an inflammatory substance, respectively. 2 is shown.

1 shows that when the buckwheat fraction extract of Example 1 is orally administered to rats, the buckwheat fraction extract inhibits rat mortality caused by lipopolysaccharide. Figure 2 shows that when the buckwheat husk fraction extract of Example 2 orally administered to the rat, the buckwheat husk fraction extract inhibits rat mortality caused by lipopolysaccharide.

The buckwheat fraction extract and the buckwheat husk fraction extract obtained in Examples 1 and 2 through FIGS. 1 and 2 were orally administered to rats at a concentration of 0.5 g / kg, respectively, and 52% and 30%, respectively, compared to the control at 30 hours. A 40% mortality reduction was observed, and at 12 hours, 44% and 44% mortality reduction effects, respectively.

Example 4 Inhibitory Effect of Inflammation Induced by TPA

The inhibitory activity against the inflammation caused by the buckwheat fraction extract and the buckwheat husk fraction extract obtained in Examples 1 and 2 by TPA (12- ο- tetradecanoylphorbol-13-acetate) was measured. As a method for measuring this, it was carried out according to the following known method (Romay C., et al., Inflamm. Res. 47: 334-338, 1998).

First, 25 to 30 g of BALB / C mice were prepared per group and fasted for 12 hours before the experiment, and then 0.01 g / kg of the buckwheat fraction extract and the buckwheat shell fraction extract obtained in Examples 1 and 2, Oral administration was performed at the respective concentrations of 0.1 g / kg, 0.5 g / kg and 1.0 g / kg. After 30 minutes of anesthesia with xylazine hydrochloride, after 5 minutes, TPA was applied to both ears at 25 μg per ear. After 5 hours the right and left ears were cut out with a 6 mm punch and weighed. Anti-inflammatory inhibition rate was used to reduce the weight increase of the buckwheat fraction extract, buckwheat husk fraction extract and TPI. As a control, an increase in the weight of rats in which only the same amount of TPE was added was used. All plots were averaged and statistical significance was tested by ANOVA method according to two-tailed Dunnett's t-test at significance level * P <0.05, ** P <0.01.

The buckwheat fraction extract and the buckwheat husk fraction extract obtained in Examples 1 and 2 are shown in Figure 3 and 4 the effect of inhibiting the inflammation caused by the inflammatory TIP.

Figure 3 shows that when the buckwheat fraction extract of Example 1 orally administered to the rat, the buckwheat fraction extract inhibits ear inflammation of the rat caused by TPI, a dermatitis-inducing substance. Figure 4 shows that when the buckwheat husk fraction extract of Example 2 orally administered to the rat, the buckwheat husk fraction extract inhibits ear inflammation in rats induced by Typhi.

The buckwheat fraction extract and the buckwheat husk fraction extract obtained in Examples 1 and 2 of the present invention through Figures 3 and 4, respectively, when the oral administration to rats at the concentration of 0.1 g / kg, respectively, 61% and 59% Oral administration at a concentration of 0.5 g / kg showed a similar inhibitory effect, showing an inhibition rate of 69% and 73%, respectively. In addition, this anti-inflammatory effect is found to increase in proportion to the concentration of buckwheat fraction extract and buckwheat shell fraction extract.

Comparative Example 1

0.3 g / kg of buckwheat fraction extract obtained in Example 1, 0.3 g / kg of buckwheat hull fraction extract obtained in Example 2, and epigallocatechingallate (EGCG) 0.1 of green tea, which is well known as an anti-inflammatory substance After oral administration at a concentration of g / kg, by using the same method as in Example 4 was measured the inhibitory effect on the inflammation caused by TI. The results are shown in FIG.

Example 5: Toxicity Test

Toxicity test was performed on the buckwheat fraction extract and the buckwheat husk fraction extract of the present invention as follows. Specifically, each of the fraction extracts was dissolved in dimethylsulfoxide (DMSO; dimethylsulfoxide), diluted with water, and then administered to the mice (10 mice per group), respectively, 1 g / kg, and observed for 7 days, but there was no death.

As described above, the solvent extract and fraction extract of the material selected from the group consisting of buckwheat, buckwheat bark, and mixtures thereof of the present invention have anti-inflammatory activity, in particular to endotoxins and external inflammation causing substances secreted by bacteria. Inflammation caused by this can be effectively suppressed. In addition, the extract can be used as a food grade, so that it can be safely ingested without a separate purification process to remove toxicity, when added to the food can be obtained anti-inflammatory effect. In particular, the economic efficiency can be improved by recycling the enormous amount of buckwheat husk that is discarded.

Claims (8)

Extract of a substance selected from buckwheat, buckwheat husk, and mixtures thereof having anti-inflammatory activity. The method of claim 1, Extract characterized in that the extract is a solvent extract. The method of claim 1, Extract is characterized in that the extract is a fraction extract obtained by performing the adsorption chromatography on a solvent extract. The method of claim 3, wherein After the solvent extract is filled in adsorption chromatography, washed with 10 to 30% ethanol solution, the extract is characterized in that the solvent fraction extracted with 40 to 80% ethanol aqueous solution. The method of claim 2 or 3, Extract, characterized in that the solvent is 20 to 80% ethanol, acetone, or methanol aqueous solution. An anti-inflammatory composition comprising an extract of a material selected from buckwheat, buckwheat husk, and mixtures thereof having an anti-inflammatory activity according to any one of claims 1 to 3 as an active ingredient. The method of claim 6, The anti-inflammatory composition is an anti-inflammatory composition is a drug, processed food, functional food, food additives, functional beverages, or beverage additives. The method of claim 7, wherein Anti-inflammatory composition wherein the processed food is noodles.