KR20200079212A - Composition for antioxidant and anti inflammation containing fermented product of Houttuynia cordata and scoria - Google Patents

Abstract

The present invention relates to an antioxidant and anti-inflammatory composition comprising a fermented product of Houttuynia cordata and scoria, and more particularly, to an antioxidant or anti-inflammatory food, cosmetic ingredient, and pharmaceutical composition comprising the fermented product. The fermented product of Houttuynia cordata and scoria according to the present invention has excellent radical scavenging activity and ability to inhibit production of nitric oxide generated due to an inflammatory reaction, and thus can be variously used in the fields of food, cosmetic ingredients and pharmaceuticals for antioxidant or anti-inflammatory activity.

Description

Composition for antioxidant and anti inflammation containing fermented product of Houttuynia cordata and scoria}

The present invention relates to a composition for anti-oxidation and anti-inflammatory, including fermented fish and volcanic pine, and more particularly, to a food, cosmetic, and pharmaceutical composition for anti-oxidation or anti-inflammatory, comprising the fermented product.

Recently, interest in materials that incorporate fermentation technology from functional natural materials is increasing. Fermented materials have the advantage of increasing the absorption rate of bioactive substances in the body by converting the physiologically active glycoside components into non-glycosides, while most of the nutrient components except the energy source are preserved (Bae et al., 2004).

8월경에 꽃이 피며, 특유의 독특한 냄새(비린내)를 함유한 생리활성 물질들이 있다. 어성초의 생리활성 성분은 플라보노이드 유도체인 퀘르세틴, 퀘르시트린, 레이노트린, 히페린, 루틴 등과 정유 성분으로 데카노일 아세트알데하이드, 메틸 노닐케톤, 라우린알데하이드, 미르센이 함유되어 있으며(Hong and Kim, 2004), 효능에 관한 연구로는 항균(Song et al., 2003), 항산화(Lee et al., 1993), 항백혈병 및 고지혈증 억제효과(Chung et al., 1999), 신경세포 보호작용(Jeong et al., 2010) 등이 보고되고 있다. Houttuynia cordata is a wild herb of perennial herbs belonging to the Saururaceae family, and is a wild herb native to Korea, China, and Japan (Jang et al., 1999). There are some differences depending on the region, but it usually buds around April, 6

It blooms around August, and there are physiologically active substances that contain a unique characteristic odor (smelly). The bioactive components of Eoseongcho are essential oils of flavonoid derivatives such as quercetin, quercitrin, raynotrin, hyperin, rutin, etc., and contain decanoyl acetaldehyde, methyl nonylketone, laurinaldehyde, and myrcene (Hong and Kim, 2004). ), Efficacy studies include antibacterial (Song et al., 2003), antioxidant (Lee et al., 1993), anti-leukemia and hyperlipidemia inhibitory effects (Chung et al., 1999), neuronal cell protective action (Jeong et. al., 2010).

Jeju volcanic cluster refers to a volcanic soil with a high porosity consisting of a calcined body calcined at high heat among porous materials, volcanic sand, and other volcanic ash mixed with various substances ejected when a volcanic eruption in a part of Jeju Island explodes. It is the only weakly alkaline resource similar to human blood, and is called'Scoria' in the Jeju-do dialect. The cluster is produced as a parasitic volcano (Oreum) and distributed throughout Jeju. The colors are largely divided into reddish brown, yellow brown, black and dark gray colors, and most commonly reddish brown clusters are most distributed. In Jeju Island, which has been a high temperature and humid environment since ancient times, it was used as a roof, inside and outside walls of houses using volcanic stone clusters, and used as heat preservation, insulation, soundproofing, and desiccants. It has been widely used, and has recently been developed as a soil improver, fertilizer additive, feed additive, natural dye, and cosmetic raw material. In particular, there are efficacy such as antibacterial, anti-inflammatory, and moisturizing effects, but systematic research on this is lacking.

Accordingly, the present inventors have completed the present invention by confirming that the fermentation product of Eoseongcho and volcanic pine pine has a remarkable antioxidant and anti-inflammatory effect as a result of studying a method for enhancing the efficacy of natural products.

Accordingly, an object of the present invention is to provide a composition for anti-oxidation or anti-inflammatory, comprising a fermented product of Houttuynia cordata and volcanic pine.

In order to achieve the above object, the present invention provides a food composition for anti-inflammatory comprising a seongseongcho and volcanic pine fermentation.

In addition, the present invention provides a cosmetic composition for anti-inflammatory, comprising a seongseongcho and volcanic pine fermentation.

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of inflammatory diseases, including Eoseongcho and volcanic pine fermentation products.

In addition, the present invention provides a pharmaceutical composition for antioxidant, comprising a fermented fish and volcanic pine.

In addition, the present invention provides a food composition for antioxidant, including seongseongcho and volcanic pine fermentation.

In addition, the present invention provides a cosmetic composition for antioxidant, including seongseongcho and volcanic pine fermentation.

Eoseongcho and volcanic pine fermentation products according to the present invention have excellent radical scavenging activity and inhibitory ability to produce nitric oxide due to an inflammatory reaction, and thus can be used in various fields in food, cosmetic, and pharmaceutical fields for antioxidant or anti-inflammatory.

1 is a view showing the results of measuring the DPPH radical scavenging activity of Eoseongcho and volcanic pine fermentation according to the present invention.
2 is a view showing the results of measuring the ABTS radical scavenging activity of Eoseongcho and volcanic pine fermentation according to the present invention.
3 is a view showing the results of evaluation of the cytotoxicity of Eoseongcho and volcanic pine fermentation products according to the present invention.
4 is a view showing the results of analyzing the inhibition of nitric oxide production ability of Eoseongcho and volcanic pine fermentation according to the present invention.

Hereinafter, the present invention will be described in detail.

According to an aspect of the present invention, there is provided a composition for anti-oxidation or anti-inflammatory, comprising a fermented vinegar ( Houttuynia cordata ) and volcanic pine.

In an embodiment of the present invention, it is preferable that Eoseongcho included in the fermentation product is Eoseongcho extract.

In the present invention, the'extract' is extracted using an appropriate solvent, and includes, for example, a crude extract, a polar solvent soluble extract, or a non-polar solvent soluble extract. In addition, the extract is a concept including all of the extracts, fractions, their diluents, concentrates or dried products obtained at each stage of extraction, fractionation or purification.

Eoseongcho extract, which is an active ingredient of the composition of the present invention, can be obtained by a conventional method or can be used by purchasing a commercially available one.

The Eoseongcho extract may use various parts of Eoseongcho, which is a raw material plant, and preferably uses leaves, stems, or roots of Eoseongcho.

The Eoseongcho extract may be obtained by extraction, separation and fractionation from nature using extraction, separation and fractionation methods known in the art.

Water or an organic solvent may be used as a suitable solvent for obtaining the Eoseongcho extract, and any pharmaceutically acceptable organic solvent may be used. For example, the solvent may be one or more selected from the group consisting of water, anhydrous or hydrous alcohol having 1 to 4 carbon atoms, ethyl acetate, acetone, glycerin, ethylene glycol, propylene glycol and butylene glycol. Preferably, methanol or ethanol may be used as the solvent, and more preferably, ethanol may be used.

The extraction temperature is preferably 50 to 100 ℃. In addition, various methods such as a hot water extraction method, a cold immersion extraction method, a reflux cooling extraction method, a solvent extraction method, a steam distillation method, an ultrasonic extraction method, an elution method, and a compression method may be used, but are not limited thereto, preferably a hot water extraction method or reflux It is to use the cooling extraction method.

The Eoseongcho extract can be obtained by liquefying, heating and filtering at room temperature by a conventional method known in the art, or additionally evaporating, spray-drying or freeze-drying the solvent. In addition, the Eoseongcho extract may be prepared in powder form by additional processes such as distillation under reduced pressure and freeze drying or spray drying. In addition, the extract or fraction is further purified by using various chromatography such as silica gel column chromatography, thin layer chromatography, high performance liquid chromatography, etc. You can also get

Therefore, the Eoseongcho extract used in the present invention is a concept that includes all of the extracts, fractions and refinements obtained from each step of extraction, fractionation or purification, their diluents, concentrates or dried products.

In an embodiment of the present invention, the fermentation product is fermented after mixing Eoseongcho extract and volcanic pine; Or it is a mixture of fermented Eoseongcho extract and volcanic pine, respectively, but may be, but is not limited thereto.

In a preferred embodiment of the present invention, it is preferable that the fermented fish and volcanic pine fermentation product fermented a mixture containing 0.1 to 10% by weight of Eoseongcho extract and 0.01 to 5% by weight of volcanic pine, and more preferably, the mixture is glucose 0.01 to 10% by weight, 0.01 to 1% by weight of yeast extract, 0.005 to 1% by weight of soy may further include.

In the present invention, "fermentation" refers to a phenomenon in which alcohol, organic acid, carbon dioxide, etc. are generated by breaking the carbon bond of a carbohydrate when a microorganism uses carbohydrate as an energy source. Examples of the microorganism include Lactobacillus Lactobacillus rhamnosus ), Lactobacillus paracasei , Lactobacillus arizonensis , Saccharomyces cerevisiae , Saccharomyces ellipsoides , Bacillus subtilis sublistilis Bacillus And Bacillus licheniformis and Leuconostoc mesenteroides .

In an embodiment of the present invention, the microorganism for fermentation was Bacillus subtilis , but is not limited thereto.

In an embodiment of the present invention, water or an organic solvent may be used as a suitable solvent for diluting the fermented fish and volcanic fern, and any pharmaceutically acceptable organic solvent may be used. For example, as the diluting solvent, one or more selected from the group consisting of water, anhydrous or hydrous alcohol having 1 to 4 carbon atoms, ethyl acetate, acetone, glycerin, ethylene glycol, propylene glycol and butylene glycol may be used. .

In an embodiment of the present invention, the fermented fish and volcanic pine fermentation product is preferably diluted 2 to 300 times with a diluting solvent, more preferably 5 to 200 times diluted, more preferably 10 to 100 times It may be diluted. If the dilution factor is more than 300 times, the efficacy of the extract is insignificant, which is undesirable.

According to an embodiment of the present invention, the fermented fish and volcanic pine fermentation products have a high total polyphenol and total flavonoid content, and excellent DPPH and ABTS radical scavenging activity. In addition, the fermented fish and volcanic pine fermentation products significantly inhibit the production of nitric oxide in cells inducing inflammation. Therefore, the fermented fish and volcanic fern of the present invention can be usefully used as a composition for antioxidant or anti-inflammatory.

In an embodiment of the present invention, the composition of the present invention may be a food or cosmetic composition.

When the composition of the present invention is used as a food composition, the food composition of the present invention refers to a food having a bioregulatory function, such as prevention and improvement of disease, biodefense, immunity, recovery after illness, and suppression of aging. When it should be harmless to the human body.

There are no particular restrictions on the type of food. Examples of foods to which the above substances can be added are meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, dairy products including gums, ice cream, various soups, beverages, teas, drinks, Alcoholic beverages and vitamin complexes, and all of the health foods in the ordinary sense.

In an embodiment of the present invention, the food composition of the present invention may be a food additive. The food additives may be added as the fermented fish and volcanic pear fermentation as it is or may be used in combination with other foods or food ingredients, and may be appropriately used according to conventional methods. The mixing amount of the active ingredient can be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment). In general, when preparing food or beverage, the fermented fish and volcanic stalk of the present invention are added in an amount of 15% by weight or less, preferably 10% by weight or less, based on the raw materials. However, in the case of long-term intake for the purpose of health and hygiene or for health control, it may be below the above range, and since there is no problem in terms of safety, the active ingredient may also be used in an amount above the above range.

In an embodiment of the present invention, the food composition of the present invention may be a health drink composition. The health drink composition may include various flavoring agents or natural carbohydrates, etc., as additional ingredients, such as a normal drink, in addition to the fermented fish and volcanic pine. The natural carbohydrates described above may include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharin and aspartame. The proportion of the natural carbohydrate is generally about 0.01 to 10 g per 100 ml of the composition of the present invention, preferably about 0.01 to 0.1 g.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, And carbonated agents used in carbonated beverages. In addition, the composition of the present invention may include flesh for the preparation of natural fruit juice, fruit juice beverage and vegetable beverage. These ingredients can be used independently or in combination. The proportion of these additives is not critical, but is generally selected from 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

When the composition of the present invention is used as a cosmetic composition, the cosmetic composition of the present invention is a conventional auxiliary and carrier such as antioxidants, stabilizers, solubilizers, vitamins, pigments, fragrances, etc., which are commonly used in cosmetic compositions in addition to the above-mentioned active ingredients. May be further included. For example, the cosmetic composition may further include auxiliary components such as glycerin, butylene glycol, polyoxyethylene cured castor oil, tocopheryl acetate, citric acid, panthenol, squalane, sodium citrate, allantoin, and the like. .

Since the cosmetic composition of the present invention is basically applied to the skin, it can be prepared in any formulation that is conventionally prepared with reference to cosmetic compositions in the art. For example, it can be formulated as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray, etc. It is not limited to this. More specifically, it can be prepared in the form of a flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, mask pack, spray or powder.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, trakant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. may be included as carrier components. have.

When the formulation of the present invention is a powder or spray, as a carrier component, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be included. In particular, in the case of a spray, additionally chlorofluorohydrocarbon, propane /Propellant such as butane and dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a carrier component may include a solvent, a solubilizing agent, an emulsifying agent, etc., specifically water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene Glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan, and the like.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol, and propylene glycol as carrier components; Suspension agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester; Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, trakant, and the like.

When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide as a carrier component Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, ethoxylated glycerol fatty acid esters, and the like.

The fermented fish and volcanic pine fermentation products are preferably contained in an amount of 0.01 to 95% by weight based on the total weight of the cosmetic composition, and more preferably 1 to 80% by weight. If the content is less than 0.01% by weight, the effect may be insignificant, and when it exceeds 95% by weight, the rate of increase in the effect compared to the amount of use may be low and uneconomical.

In an embodiment of the present invention, the composition of the present invention may be a pharmaceutical composition for anti-inflammatory. The pharmaceutical composition may be a pharmaceutical composition for the prevention or treatment of inflammatory diseases.

The inflammatory disease is a generic term for diseases in which inflammation is a primary lesion, but is not limited thereto, allergic diseases including allergic asthma, allergic rhinitis, allergic mucositis, urticaria and anaphylax, Myopathy, arthritis, atopic dermatitis, psoriasis, asthma, multiple sclerosis, sepsis, multiple chondritis, scleroderma, eczema, gout, periodontal disease, including systemic sclerosis, dermatomyositis and inclusion body myositis Diseases, Behcet's syndrome, edema, vasculitis, Kawasaki disease, diabetic retinitis, autoimmune pancreatitis, vasculitis, glomerulonephritis, acute and chronic bronchitis, influenza infection, and the like.

The pharmaceutical composition of the present invention can be used by formulating in various forms according to a conventional method. For example, it can be formulated into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, etc., and can be used in the form of external preparations, suppositories, and sterile injectable solutions.

The pharmaceutical composition of the present invention may be prepared by including at least one pharmaceutically acceptable carrier in addition to the active ingredient for administration. The pharmaceutically acceptable carrier included in the pharmaceutical composition of the present invention is commonly used in formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, but is not limited thereto It does not work. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components.

The dosage of the pharmaceutical composition of the present invention may vary depending on the method of formulating the pharmaceutical composition, the method of administration, the administration time and/or the route of administration, and the type and degree of reaction to be achieved by administration of the pharmaceutical composition. , Factors, including the type of subject, age, weight, general health condition, severity or severity of the disease, gender, diet, excretion, and other components of the drug used simultaneously or simultaneously with the subject, and It can be varied according to similar factors well known in the pharmaceutical field, and those skilled in the art can easily determine and prescribe an effective dosage for the desired treatment.

The dosage of the pharmaceutical composition of the present invention may be, for example, 1 mg/kg to 1,000 mg/kg per day, but the dosage does not limit the scope of the present invention in any way.

The route of administration and the mode of administration of the pharmaceutical composition of the present invention may be independent of each other, and are not particularly limited in the manner, and any route of administration and mode of administration as long as the pharmaceutical composition can reach the desired site. You can follow.

The pharmaceutical composition may be administered by oral administration or parenteral administration. The parenteral administration method includes, for example, intravenous administration, intraperitoneal administration, intramuscular administration, transdermal administration, or subcutaneous administration, and a method of applying, spraying, or inhaling the pharmaceutical composition to a diseased site may also be used. However, it is not limited thereto.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.

Example 1 Eoseongcho and volcanic cluster fermentation product (FHC-S)

Eoseongcho was purchased from the Gyeongdong market and coarsely crushed to 20 mesh or less, and extracted and filtered for 72 hours with 100% 70% ethanol (v/v) for 100 g of the sample (Whatman No2, Maidstone, England). The filtered ethanol extract was concentrated under reduced pressure (N-1000S-WD, Eyela Co., Tokyo, Japan) and then freeze-dried (FDU-1100, Eyela Co., Tokyo, Japan) to prepare Eoseongcho ethanol extract.

Volcanic clusters were purchased from Songi Industry Co., Ltd.

In addition, a Bacillus subtilis strain was used as the fermentation strain. The whole culture of the fermentation strain was cultured for 24 hours in a shake incubator having a temperature of 37°C using a nutrient medium (Difco, Detroit, MI, USA).

To prepare the fermentation product of Eoseongcho and volcanic pine, 5% by weight of Eoseongcho ethanol extract, 0.5% by weight of volcanic pine, 0.6% by weight of glucose, 0.3% by weight of yeast extract, and 0.1% by weight of soytone were mixed. The mixture was inoculated with Bacillus subtilis, a fermentation strain, and cultured with shaking at a temperature of 37°C for 3 days. The fermented product was filtered through a 0.2 μm filter to obtain a fermented fish and volcanic fern.

Comparative Example 1. Eoseongcho Extract (HC)

10% of 70% ethanol (v/v) was added to 100 g of Eoseongcho sample and extracted for 72 hours, followed by filtration (Whatman No2, Maidstone, England).

Comparative Example 2. Volcanic cluster extract (S)

After extracting for 72 hours by adding 10 times 70% ethanol (v/v) to 100 g of the volcanic cluster sample, it was filtered (Whatman No2, Maidstone, England).

Comparative Example 3. Eoseongcho and Volcanic Cluster Extract (HC-S)

After 10 hours 70% ethanol (v/v) was added to 100 g of the composite sample mixed with the eoseongcho and volcanic cluster in a ratio of 9:1, the mixture was extracted for 72 hours, and then filtered (Whatman No2, Maidstone, England).

Comparative Example 4. Eoseongcho Fermentation Extract (FHC)

Eoseongcho was purchased from the Gyeongdong market and coarsely crushed to 20 mesh or less, and extracted and filtered for 72 hours with 100% 70% ethanol (v/v) for 100 g of the sample (Whatman No2, Maidstone, England). The filtered ethanol extract was concentrated under reduced pressure (N-1000S-WD, Eyela Co., Tokyo, Japan) and then freeze-dried (FDU-1100, Eyela Co., Tokyo, Japan) to prepare Eoseongcho ethanol extract.

The fermentation strain was Bacillus subtilis strain. The whole culture of the fermentation strain was cultured for 24 hours in a shake incubator having a temperature of 37°C using a nutrient medium (Difco, Detroit, MI, USA).

To prepare the fermented product of Eoseongcho, 5% by weight of ethanolic extract of Eoseongcho, 0.6% by weight of glucose, 0.3% by weight of yeast extract, and 0.1% by weight of Soytone were mixed. The mixture was inoculated with Bacillus subtilis, a fermentation strain, and cultured with shaking at a temperature of 37°C for 3 days. The fermentation product was filtered through a 0.2 μm filter to obtain a fermented eoseongcho.

In the experimental examples to be described later, Eoseongcho and volcanic pine fermentation products of Example 1 and the extracts of Comparative Examples 1 to 4 were diluted (5X to 200X) with distilled water and used in the experiment.

Experimental Example 1. Measurement of total polyphenol and total flavonoid content

Method for measuring total polyphenol content

The content of total polyphenolic compounds of Eoseongcho and volcanic pine fermentation was analyzed. The analysis was performed by applying the Folin-Denis method (Folin and Denis, 1912). Each sample was mixed by adding 2 mL of a 2-fold diluted folin reagent (Sigma Co., St. Louis, MO, USA) to 2 mL of the solution diluted by concentration. After the mixture was left for 3 minutes, 2 mL of 10% Na ₂ CO ₃ (Sigma Co.) was added and reacted for 1 hour. Using UV/visible spectrophotometer (UVIKON 922, Kontran Co., Milan, Italy), the absorbance was measured at 700 nm of the reactant, and the total polyphenol content was obtained based on the standard curve.

The standard curve is prepared by measuring the absorbance of gallic acid (Sigma Co.) at 700 nm.

Method for measuring total flavonoid content

The total flavonoid content of Eoseongcho and volcanic pine fermentation was measured by applying the method of Nieva Moreno et al. (Nieva et al., 2000). 0.1 mL of each sample and 0.9 mL of 80% ethanol were mixed. To the mixture was added 0.1 mL of 10% aluminum nitrate (Sigma Co.), 1 M potassium acetate (Sigma Co.), and 4.3 mL of 80% ethanol, and left at room temperature for 40 minutes. Thereafter, the absorbance of the reactant was measured at 415 nm, and the total polyphenol content was obtained based on the standard curve. The standard curve was prepared by measuring the absorbance of quercetin (Sigma Co.) at 415 nm.

Table 1 shows the total polyphenol and total flavonoid content measurement results.

sample Total polyphenols ¹⁾
(μg/mL) Total flavonoids ²⁾
(μg/mL) Eoseongcho and Volcanic Cluster Fermentation 6.02±0.17 ³⁾ 10.85±0.45 Eoseongcho fermentation 3.84±0.32 8.55±0.17 ¹⁾ Milligrams of total polyphenol content/g of samples based on gallic acid as standard.
²⁾ Milligrams of total flavonoid content/g of samples based on quercetin as standard.
³⁾ Each value is mean±SD (n≥3).

As shown in Table 1, it was confirmed that Eoseongcho and volcanic pine fermentation products have higher total polyphenol and total flavonoid contents than Eoseongcho fermentation products.

Experimental Example 2. DPPH radical scavenging activity measurement

Free radical scavenging activity of Eoseongcho and volcanic pine fermentation was confirmed by measuring the reducing power for DPPH (1,1-diphenyl-2-picryl hydrazyl, Sigma Co.). Specifically, 800 μL of a diluted solution in which each sample was diluted in 99% methanol and 200 μL of a 0.15 mM DPPH solution dissolved in methanol were mixed and left at room temperature for 30 minutes. The absorbance was then measured at 517 nm. DPPH radical scavenging activity was calculated according to the following equation (1).

[Equation 1]

The results of measuring DPPH radical scavenging activity are shown in FIG. 1.

As shown in FIG. 1, it was confirmed that Eoseongcho and volcanic pine fermentation product (FHC-S) had significantly higher DPPH scavenging activity than Eoseongcho fermentation product (FHC) of Comparative Example 4.

Experimental Example 3. ABTS radical scavenging activity measurement

The antioxidant power measurement using the ABTS radical was performed by the ABTS ⁺ cationic decolorization analysis method (Re et al., 1999). 7 mM ABTS (Sigma Co.) and 2.45 mM potassium persulfate (Sigma Co.) were mixed to a final concentration and left in a cow at room temperature for 24 hours to form ABTS ⁺ . Then, it was diluted with PBS (pH7.4) so that the absorbance value at 732 nm was 0.70 (±0.02). After adding 10 μL of sample to 990 μL of the diluted solution, it was allowed to stand for exactly 1 minute and absorbance was measured. The results of measuring the ABTS radical scavenging activity are shown in FIG. 2.

As shown in Figure 2, Eoseongcho and volcanic cluster fermentation product (FHC-S) was confirmed that the radical scavenging ability is significantly higher than the extracts of Comparative Examples 1 to 4.

Experimental Example 4. Cytotoxicity evaluation

Prior to the analysis of nitric oxide production inhibitory ability, it was investigated whether treatment of LPS with Eoseongcho and volcanic pine fermentation affects cell survival.

4-1. Cell line culture

The microglial cell line BV-2 used for the evaluation of cytotoxicity was used for pre-sale at Harvard University. BV-2 cells were RPMI1640 with 10% fetal bovine serum (FBS) (Gibco, BRL, USA), 100 μg/mL penicillin (Gibco, BRL, USA) and 100 μg/mL streptomycin (Gibco, BRL, USA). The medium (Gibco, BRL, USA) was cultured once every 2-3 days in a 37° C. incubator with 5% CO ₂ .

4-2. Cytotoxicity assessment by measuring cell viability

In order to confirm the effect of Eoseongcho and volcanic pine fermentation on cell survival in LPS-stimulated BV-2 cells, the survival of the cells was determined by MTT(3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl. -tetrazolium bromide). Cells (1X10 ⁵ cell/mL) were dispensed in a 96-well plate in 180 μL increments and cultured in a CO ₂ incubator for at least 12 hours, and then treated according to each condition to incubate for 24 hours. After incubation, the culture solution was removed, and 200 μL of a medium containing 0.5 mg/mL MTT was added, followed by incubation for 4 hours to reduce MTT. Thereafter, the culture solution was removed and 100 μL of DMSO (dimethylsulfoxide) was added to dissolve the generated formazan crystals, and absorbance was measured at 540 nm using an ELISA reader. Cell viability was expressed as a percentage (%) compared to the control. The cytotoxicity evaluation results are shown in FIG. 3.

As shown in FIG. 3, when stimulation with LPS and treatment with Eoseongcho and volcanic pine fermentation products (FHC-S) were performed, it was confirmed that the cell viability was 95% or more. The above results indicate that the fermented fish and volcanic ferns are not cytotoxic.

Experimental Example 5. Analysis of nitric oxide production inhibitory ability in glial cells activated with LPS

The anti-inflammatory efficacy of Eoseongcho and volcanic pine mushrooms was analyzed. Specifically, by stimulating the glial cells by treating the inflammation-inducing factor LPS (100 ng / ml) was prepared. The stimulated glial cells were dispensed in a 24 well plate at a concentration of 5 x 10 ⁵ cells/mL. After adding LPS and samples to various cells, the cells were reacted in an incubator for 24 hours. After reacting with 50 μL of grease reagent (1% sulfanilamide/0.1% N-(1-naphthyl)- ethylenediamine dihydrochloride/2.5% H ₃ PO ₄ ) to the reacted cells, Sunrise Basic 96- with a wavelength of 540 nm Absorbance was measured using a well basic plate (Sunrise basic 96-well microplate spectrophotometer, TECAN AUSTRIA). The results of the nitric oxide production inhibitory activity analysis are shown in FIG. 4.

As shown in Figure 4, Eoseongcho and volcanic cluster fermentation product (FHC-S) treatment group had a higher inhibitory ability to produce nitric oxide than other comparative groups, especially in the 100X dilution treatment group, the concentration of nitrogen oxide was different from other comparative groups It was confirmed that it exhibited a superior inhibitory ability to produce nitric oxide by confirming a reduction of 50% or more compared to the control group (LPS treated group).

Hereinafter, the present invention will be described in more detail through formulation examples. The formulation examples are only for illustrating the present invention, and the scope of the present invention is not to be construed as being limited by the formulation examples.

Formulation Example 1. Preparation of pharmaceutical composition for anti-inflammatory or antioxidant

1-1. Preparation of powder

Eoseongcho and Volcanic Pine Fermentation 20 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight fabric to prepare a powder.

1-2. Preparation of tablets

Eoseongcho and Volcanic Pine Fermentation 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above ingredients, tablets are prepared by tableting according to a conventional tablet manufacturing method.

1-3. Preparation of capsules

Eoseongcho and Volcanic Pine Fermentation 10 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into a gelatin capsule to prepare a capsule.

1-4. Preparation of injection

Eoseongcho and Volcanic Pine Fermentation 10 mg

Mannitol 180 mg

Injectable sterile distilled water 2974 mg

Na ₂ HPO ₄ 2H ₂ O 26 mg

It is prepared with the above-mentioned ingredient content per ampoule (2 ml) according to the preparation method of a conventional injection.

1-5. Preparation of liquid

Eoseongcho and Volcanic Pine Fermentation 20 mg

Isomerized sugar 10 g

5 g of mannitol

Purified water

Each component is added to the purified water according to the manufacturing method of a conventional liquid, dissolved, and the lemon flavor is added in an appropriate amount.The above components are mixed, and then purified water is added to adjust the total to 100 ml. To prepare a liquid formulation.

Formulation Example 2. Preparation of antioxidant cosmetic composition

2-1. Preparation of flexible cosmetic lotion (skin lotion)

Eoseongcho and volcanic cluster fermentation products 0.5%

Beta-1,3-glucan 1.0%

Butylene glycol 2.0%

Propylene glycol 2.0%

Carboxyvinyl polymer 0.1%

PG-12 nonylphenyl ether 0.2%

Polysorbate 80 0.4%

Ethanol 10.0%

Triethanolamine 0.1%

Preservatives, pigments, flavors

Purified water to 100%

2-2. Preparation of nutrient makeup (milk lotion)

Eoseongcho and volcanic cluster fermentation products 0.5%

Beta-1,3-glucan 1.0%

Wax 4.0%

Polysorbate 60 1.5%

Sorbitan sesquioleate 1.5%

Liquid paraffin 0.5%

Caprylic/Capric Triglyceride 5.0%

Glycerin 3.0%

Butylene glycol 3.0%

Propylene glycol 3.0%

Carboxyvinyl polymer 0.1%

Triethanolamine 0.2%

Preservatives, pigments, flavors

Purified water to 100%

2-3. Preparation of nutrition cream

Eoseongcho and volcanic cluster fermentation products 1.0%

Beta-1,3-glucan 5.0%

Mill blade 10.0%

Polysorbate 60 1.5%

Easy Easy 60 Castor Oil 2.0%

Solbitan sesquioleate 0.5%

Liquid paraffin 10.0%

Squalane 5.0%

Caprylic/Capric Triglyceride 5.0%

Glycerin 5.0%

Butylene glycol 3.0%

Propylene glycol 3.0%

Triethanolamine 0.2%

Preservatives, pigments, flavors

Purified water to 100%

As described above, since a specific part of the present invention has been described in detail, it is obvious to those skilled in the art that this specific technique is only a preferred embodiment, and the scope of the present invention is not limited thereby. something to do. Therefore, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (11)

Food composition for anti-inflammatory, including Eoseongcho ( Houttuynia cordata ) and fermented volcanic pine.
According to claim 1,
The seongseongcho is water, anhydrous or hydrous alcohol having 1 to 4 carbon atoms, ethyl acetate, acetone, glycerin, ethylene glycol, propylene glycol, and butylene glycol extracted from one or more selected from the group consisting of, the composition.
According to claim 2,
The fermentation product is a fermented mixture containing 0.1 to 10% by weight of Eoseongcho extract, 0.01 to 5% by weight of volcanic pine, 0.01 to 10% by weight of yeast, 0.01 to 1% by weight of yeast extract, and 0.005 to 1% by weight of Soytone Phosphorus, composition.
According to claim 1,
The fermentation product is diluted 2 to 300 times with a diluting solvent.
According to claim 4,
The dilution solvent is at least one member selected from the group consisting of water, anhydrous or hydrous alcohol having 1 to 4 carbon atoms, ethyl acetate, acetone, glycerin, ethylene glycol, propylene glycol and butylene glycol.
According to claim 1,
The fermented product is Lactobacillus ramno suspension (Lactobacillus rhamnosus), Lactobacillus para Kasei (Lactobacillus paracasei), Lactobacillus ahrijo norbornene sheath (Saccharomyces cerevisiae), beer yeast (Saccharomyces ellipsoides) in MRS celebrity busy with (Lactobacillus arizonensis), Saccharomyces, Bacillus subtilis ( Bacillus subtilis ), Saccharomyces Bacillus licheniformis ( Bacillus licheniformis ) and Leuconostok mesenteroid ( Leuconostoc mesenteroides ) fermented with one or more strains selected from the group consisting of.
A cosmetic composition for anti-inflammatory comprising fermented fish and volcanic pine.
Pharmaceutical composition for the prevention or treatment of inflammatory diseases, including Eoseongcho and volcanic pine fermentation products.
The method of claim 8,
The inflammatory diseases are allergic diseases including allergic asthma, allergic rhinitis, allergic mucositis, urticaria and anaphylax, systemic sclerosis, dermatomyositis and inclusion body myositis Myopathy, arthritis, atopic dermatitis, psoriasis, asthma, multiple sclerosis, sepsis, multiple chondritis, scleroderma, eczema, gout, periodontal disease, Behcet syndrome, edema, vasculitis, Kawasaki disease, diabetic retinitis, autoimmunity A composition of at least 1 selected from the group consisting of pancreatitis, vasculitis, glomerulonephritis, acute bronchitis, chronic bronchitis and influenza infection.
Antioxidant food composition comprising a fermented fish and volcanic pine.
Antioxidant cosmetic composition comprising Eoseongcho and volcanic pine.

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