KR20190050667A - Composition for anti-oxidation or anti-inflammation comprising barley sprouts extract as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for anti-oxidation or anti-inflammation comprising a barley spout extract as an active ingredient, and to a production method of a high content saponarin-containing germanium barley extract, which comprises a step of adding a fermented alcohol having an alcohol frequency of 30(±3) %(v/v) to the barley sprout and circulating and extracting a mixture at room temperature for a total of 9(±0.5) hours. The barley spout extract contains the high content of saponarin by produced by the unique method of the present invention, and has remarkable antioxidant and anti-inflammatory effects.

Description

TECHNICAL FIELD The present invention relates to an antioxidant or anti-inflammatory composition comprising an extract of bud from barley as an active ingredient,

The present invention relates to a composition for antioxidant or antiinflammation comprising an extract of sprout barley as an active ingredient and a fermented alcohol aqueous solution of 30 (± 3)% (v / v) ) For a period of time equal to or longer than a predetermined period of time.

Enzymes such as mitochondria, peroxisome, xanthine oxidase (XOD), NADPH oxidase and cyclooxygenase (COX) present in the cells are reactive oxygen species (ROS) such as O ₂ ^- , H ₂ O ₂ and OH This ROS is the main cause of oxidative damage. In addition, reactive nitrogen species (RNS) such as NO, HNO ₂ , and ONOO ^- are produced by the immune response of macrophages, neutrophils, and other immune cells during inflammatory reactions, et al., Oxidative injury in the nervous system, Acta. Neurol. Scand., 98, pp 145-153, 1998; Brune B et al., Nitric oxide, oxidative stress, and apoptosis Kidney Int. -24, 2003).

Our body balances prooxidants and antioxidants to maintain homeostasis. If this balance is broken and tilted toward oxidation promotion, it will have harmful effects on cells, which is called oxidative stress. This oxidative stress is basically related to aging, and it causes DNA damage in the body, damage of cell membrane due to lipid peroxidation, oxidation of protein and lipid, and the like, and is caused by atherosclerosis, cancer, cataract, senile dementia, Parkinson's disease (Chung IM et al., Screening of Korean medicinal and food plants with antioxidant activity. Kor J. Med. Sci., 6, pp311-322, 1998).

Control of the inflammatory response is known to be extremely complex, a series of active mechanisms to reduce the enhancement and impairment of the in vivo recovery system, and is well known to be well regulated in all tissues. However, persistent inflammatory response by repeated tissue damage or regeneration leads to excessive ROS and RNS production in inflammation-related cells, resulting in permanent genetic modification (Azad N et al., Inflammation and lung cancer : roles of reactive oxygen / nitrogen species, J. Toxicol. Environ. Health., B. Crit. Rev., 11 (1), pp11-15, 2008). Thus, ROS and RNS are deeply involved in the inflammatory response that regulates the action of various cells in vivo.

Specifically, inflammation is one of the defensive reactions of biopsies against external stimuli. Clinically, symptoms such as fever, fever, swelling, pain, and dysfunction are observed, and cytokines, PGE2, lysosomal enzyme ), Free radicals, and the like. The activation of NF-κB, which is a transcription factor of the inflammatory response, is activated by the activation of some of the major inflammatory proteins including IL-1β, IL-6, IL-8 and TNF-α , And regulates the activation of COX-2 and iNOS, thus playing an important role throughout the inflammatory response (Chung HY et al., Dietary modulation of prostanoid synthesis in the aging process: role of cyclooxygenase-2. Ageing. Dev., 111 (2-3), pp. 97-106, 1999). Most of the factors involved in NF-κB activation, such as lipopolysaccharide (LPS), are known to cause ROS, and this NF-κB activation has been studied to be inhibited by chemically broad antioxidants. These studies show that NF-κB activation is mostly promoted by the oxidation reaction, indicating a close link between oxidation and inflammation.

In recent years, efforts have been made to develop antioxidants from natural sources as side effects of synthetic antioxidants have been revealed. In addition, there has been an increase in needs to use multifunctional natural materials for health promotion purposes .

Accordingly, the inventors of the present invention have conducted research to discover safe natural materials showing multifunctionality, and have found that the extract of the present invention has remarkable antioxidative and anti-inflammatory effects, .

Accordingly, an object of the present invention is to provide a food composition, a cosmetic composition and a pharmaceutical composition for antioxidant or antiinflammation, which comprises an extract of bud of barberry as an active ingredient.

It is still another object of the present invention to include a step of circulating and extracting a fermented juice aqueous solution of 30 (± 3)% (v / v) alcoholic alcohol at a room temperature for a total of 9 (± 0.5) And a saponin-rich shoot-barley extract prepared by the above-described method.

In order to achieve the above object, the present invention provides a food composition, a cosmetic composition and a pharmaceutical composition for antioxidant or antiinflammation, which comprises an extract of bud of barberry as an active ingredient.

In order to achieve still another object of the present invention, the present invention provides a fermented fermented juice aqueous solution of alcohol (30 (± 3)% (v / v)) added to bud of barley and circulatingly extracted at room temperature for a total of 9 The present invention also provides a method for producing a saponin-rich germanium barley extract and a saponin-rich germanium barley extract prepared by the method.

Hereinafter, the present invention will be described in detail.

The shoot barley is a barley ( Hordeum vulgare L.). In the present invention, the term " young plant " Normally, the bud barley can be obtained by dipping the barley in water for one day, then removing water and germinating under the condition of preventing loss of water, and then growing for 7 to 10 days. The sprout barley can be used after it is used without being rounded, or in a pretreatment process in consideration of a process speed and a process (manufacturing) efficiency that a person skilled in the art intends to take into consideration. As the pre-treatment process, for example, Washing, cutting, pulverizing, drying, and the like.

The sprout barley extract of the present invention comprises a step of adding a fermented fermented juice aqueous solution of 30 (± 3)% (v / v) of alcohol (ethanol) to bud of barley and circulatingly extracting the mixture at room temperature for a total of 9 (± 0.5) Which is manufactured by a manufacturing method. Due to the extraction method peculiar to the present invention, it is possible to obtain a barley extract having a high content of saponin. It has been difficult to extract more than 15 mg / g (per g of barley g) of saponin from the bud of barley through the conventional extraction method known in the prior art. However, through the extraction process unique to the present invention, (High content) saponin can be extracted. Preferably from 12 mg / g to 18 mg / g of saponin. Most preferably from 15 mg / g to 17.73 mg / g of saponin.

The specificity of such a method for producing an extract of the present invention is well illustrated in Example 1 of the specification of the present invention. The present inventors first compared the saponin extraction efficiency with various solvents. As a result, the degree of saponarin extraction in acetone or ethyl acetate, even in a polar solvent, was smaller than isopropyl alcohol, which is a nonpolar solvent. Therefore, it was confirmed that saponin had the extraction specificity for water or fermented alcohol. In addition, as shown in Table 2, in contrast to the conventional method in which the extraction temperature is raised or the polarity is increased in order to increase the extraction efficiency of the component materials, the alcohol (ethanol) The higher the concentration, the lower the saponin extraction efficiency, and the higher the temperature, the lower the efficiency of saponin extraction. From these results, it can be concluded that a high yield of saponin is difficult due to conventionally known extraction efficiency condition tendency, and that a specific process is required for obtaining saponin, and therefore, The extraction efficiency of saponin extract from the barley was significantly increased by adding an aqueous solution of 30 (± 3)% (v / v) fermented juice and circulating at room temperature for a total of 9 (± 0.5) hours.

Preferably, the fermentation alcohol may be added at a weight of 10 (+/- 3) times the weight of the raw material (bud barley). The term room temperature (rt) may preferably mean 20 (± 5) ° C. The circulation extraction may be repeated one to ten times, preferably two to three times, within the above-described time range.

After the extraction process, the process may further include a purification process for increasing the purity of the extract, such as filtration and centrifugation. The concentration may be further carried out by a concentration apparatus or a concentration method known in the art. The concentration may be, for example, precipitation concentration, evaporation concentration, concentration under reduced pressure, ultrafiltration, reverse osmosis , Centrifugation, and may be preferably concentrated under reduced pressure.

The extract of the present invention can be used in the form of a liquid by filtration and / or concentration, and solidified through a conventional drying process such as spray drying or freeze drying. In the drying step, dextrin or the like may be mixed and dried before spray drying or freeze drying.

The extract of the present invention has remarkable antioxidant and anti-inflammatory effects. This is well illustrated in the specification of the present invention. Accordingly, the present invention provides a food composition, a cosmetic composition and a pharmaceutical composition for antioxidant or anti-inflammation, which comprises the above-mentioned germanium barley extract as an active ingredient.

The food composition of the present invention includes all forms such as functional food, health function food, nutritional supplement, health food and food additives. Food compositions of this type may be prepared in a variety of forms according to conventional methods known in the art.

For example, as a health food, the above-mentioned germanium barley extract may be prepared in the form of tea, juice and drink and then taken for drinking, granulated, encapsulated and powdered. In addition, it can be prepared in the form of a composition by mixing together the known active ingredients known to have the effect of improving the wrinkles, anti-aging, skin elasticity and skin moisturizing effect of the present invention.

Functional foods also include beverages (including alcoholic beverages), fruits and their processed foods (e.g., canned fruits, bottled, jam, maalmalade, etc.), fish, meat and processed foods such as ham, Etc.), breads and noodles (eg udon, buckwheat noodles, ramen noodles, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, Protein, retort food, frozen food, various seasonings (eg, soybean paste, soy sauce, sauce, etc.).

Specifically, the present invention provides a health functional food comprising the above-described germanium barley extract and a pharmaceutically acceptable carrier or additive. The health functional food defined in the present invention has been newly established by the Act on Health Functional Foods, amended in 2008, so that the function and safety of the human body is sufficiently established, and the health functional food as defined in the Korean Food and Drug Administration Notice No. 2008-72 This means that the product is a health functional food contained in the regulation on food functional raw material qualification.

Such health functional foods can be formulated into conventional health functional foods formulations known in the art. The health supplements may be prepared, for example, as powders, granules, tablets, pills, capsules, suspensions, emulsions, syrups, infusions, solutions, exposures, gums, tea, jellies or drinks. As such a pharmaceutically acceptable carrier or additive, any carrier or additive known to be usable in the art for the preparation of the formulation to be prepared may be used.

The health functional food of the present invention can be used as a nutritional supplement, a vitamin, a mineral (electrolyte), a flavor such as a synthetic flavor and a natural flavor, a colorant and an enhancer (cheese, chocolate etc.), a pectic acid and its salt, , Organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like.

It may further contain various flavors or natural carbohydrates. Examples of the natural carbohydrate include conventional sugars such as monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugars such as xylitol, sorbitol, erythritol, Of sugar alcohols may be contained. In addition, as a flavoring agent, a natural flavoring agent (e.g., tau Martin, stevia extract, etc.) and a synthetic flavoring agent (e.g., saccharin, aspartame, etc.) may be contained.

In order to use the germanium barley extract of the present invention in the form of a food additive, it may be prepared in the form of powder or concentrate.

The preferred content of the germanium barley extract in the food composition of the present invention is 0.0001 to 90% by weight, preferably 0.01 to 50% by weight based on the total weight of the food composition.

The cosmetic composition of the present invention can be prepared by any of the formulations conventionally produced in the art. In addition to the sprout barley extract of the present invention, the cosmetic composition of the present invention can be used in topical application Or in the form of adjuvants which can be applied systemically.

In addition, the cosmetic composition of the present invention may further contain, in addition to the germanium barley extract, a lipid, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, Water, ionic or non-ionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics Such as, for example, cosmetic or dermatological ingredients, such as, for example, cosmetic or dermatological ingredients. And the above ingredients may be introduced in amounts commonly used in the dermatology field.

Formulations of suitable cosmetic compositions include, for example, solutions, gels, solid or paste anhydrous products, emulsions obtained by dispersing the oil phase in water, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes) In the form of a foliar dispersant, in the form of a cream, a skin, a lotion, a powder, an ointment, a spray or a conceal stick. It can also be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.

Examples of the cosmetic composition to which the cosmetic composition of the present invention can be added include but are not limited to skin lotion, skin softener, skin toner, convergent lotion, softening longevity, nutritional lotion, astringent, lotion, milk lotion, Cream, Massage Cream, Nourishing Cream, Moisture Cream, Hand Cream, Essence, Nutrition Essence, Pack, Soap, Shampoo, Cleansing Foam, Cleansing Lotion, Cleansing Cream, Body Lotion, Body Cleanser, Treatment, Serum, Milk, Press Powder, Loose powder, eye shadow, and the like.

The content of the germanium barley extract contained in the cosmetic composition of the present invention may be in the range of 0.0001 to 50% by weight, preferably 0.01 to 30% by weight based on the total weight of the cosmetic composition.

The pharmaceutical composition according to the present invention may further contain the above-mentioned bud-shaped extract of the present invention alone or in combination with one or more pharmaceutically acceptable carriers, excipients or diluents. The term " pharmaceutically acceptable " as used herein means a non-toxic composition which is physiologically acceptable and which, when administered to humans, does not inhibit the action of the active ingredient and does not normally cause an allergic reaction such as gastrointestinal disorder, dizziness, .

The pharmaceutically acceptable carrier may further include, for example, a carrier for oral administration or a carrier for parenteral administration. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. In addition, the carrier for parenteral administration may contain water, a suitable oil, a saline solution, an aqueous glucose and a glycol, and may further contain a stabilizer and a preservative. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, etc. in addition to the above components. Other pharmaceutically acceptable carriers and preparations can be found in Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, Pa., 1995).

The composition of the present invention can be administered to mammals including humans by any method. For example, it can be administered orally or parenterally. Parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual or rectal administration Lt; / RTI >

The pharmaceutical composition of the present invention can be formulated into oral preparations or parenteral administration preparations according to the administration route as described above. In the case of a preparation for oral administration, the composition of the present invention may be formulated into a powder, a granule, a tablet, a pill, a sugar, a tablet, a liquid, a gel, a syrup, a slurry, . For example, an oral preparation can be obtained by combining the active ingredient with a solid excipient, then milling it, adding suitable auxiliaries, and then processing the mixture into a granular mixture. Examples of suitable excipients include, but are not limited to, sugars including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol, and starches including corn starch, wheat starch, rice starch and potato starch, Cellulose such as methylcellulose, sodium carboxymethylcellulose and hydroxypropylmethyl-cellulose and the like, fillers such as gelatin, polyvinylpyrrolidone and the like. In addition, crosslinked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may optionally be added as a disintegrant. Further, the pharmaceutical composition of the present invention may further comprise an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent and an antiseptic agent.

 In the case of a preparation for parenteral administration, it can be formulated by a method known in the art in the form of injection, cream, lotion, external ointment, oil, moisturizer, gel, aerosol and nasal aspirate. These formulations are described in Remington's Pharmaceutical Science, 19th ed., Mack Publishing Company, Easton, Pa., 1995, which is a commonly known formulary for all pharmaceutical chemistries.

The total effective amount of the composition of the present invention may be administered to a patient in a single dose and may be administered by a fractionated treatment protocol administered over a prolonged period of time in multiple doses. In the pharmaceutical composition of the present invention, the content of the active ingredient may be varied depending on the degree of the disease. Preferably, the total preferred dose of the pharmaceutical composition of the present invention may be from about 0.01 μg to 1,000 mg, and most preferably from 0.1 μg to 500 mg, per kilogram of patient body weight per day. However, the dosage of the pharmaceutical composition may be determined depending on various factors such as the formulation method, administration route and frequency of treatment, as well as the patient's age, body weight, health condition, sex, severity of disease, diet and excretion rate, It will be possible to determine the appropriate effective dose of the composition of the present invention by those of ordinary skill in the art in view of this point. The pharmaceutical composition according to the present invention is not particularly limited to its formulation, administration route and administration method as long as the effect of the present invention is exhibited.

The germanium barley extract of the present invention is produced by a unique method of the present invention and contains not only high saponin content but also remarkable antioxidative and antiinflammatory effects.

Fig. 1 shows a method for producing the dried barley extract of the present invention.
FIG. 2 shows the results of MTT assay to examine the change of cell survival rate when oxidative stress was induced in t-BHP cells and treated with the extract of the present invention (### p <0.001, etc.). no treatment; *** p <0.001, vs. t-BHP treatment; Student's t-test).
FIG. 3 shows the results of confirming the effect of inhibiting apoptosis by Annexin V / PI staining and FACS technique when treating the cells with oxidative stress caused by t-BHP and treating the extract with the present invention (FIG. 3a shows FACS Data, Fig. 3b was quantified from the viewpoint of cell apoptosis, Fig. 3c was quantified from the viewpoint of cell viability, and the results were ### p <0.001, vs. no treatment; *** p <0.001, BHP treatment; Student's t-test).
FIG. 4 shows the results of oxidative stress induced by t-BHP on the cells and JC-1 staining when treated with the present invention barley extract (p <0.05, *** p <0.001, and so on). Fig. 4a shows FACS data, Fig. 4b shows quantitative data, -BHP treatment; Student's t-test)
Figure 5 shows the change of ARE activity in the treatment of the barley extract of the present invention (*** p &lt;0.001;Student's t-test).
FIG. 6 shows the results of quantitative changes of Nrf2 protein over time when treated with the extract of the present invention (FIG. 6A shows the quantitative change of Nrf2 protein in the cytoplasm and FIG. 6B shows the quantitative change of Nrf2 protein in the nucleus).
FIG. 7 shows the results of confirming the effect of the present invention on the expression of HO-1, a second phase antioxidant enzyme regulated by Nrf2 transcription factor, according to the present invention.
FIG. 8 shows the results of measuring changes in NF-kB activity when macrophages induced inflammation with LPS and treated with the present invention barley extract (### p <0.001, vs. no treatment; *** p < 0.001, vs. LPS treatment; Student's t-test).
FIG. 9 shows the results of confirming the change in the amount of NO produced when inflammation was induced in macrophages with LPS and treated with the extract of the present invention (### p <0.001, vs. no treatment; *** p &lt; 0.001, vs. LPS treatment; Student's t-test).
10 shows the results of examining changes in the expression levels of COX-2 and iNOS, which are typical inflammation-related proteins, when macrophage cells were infected with LPS and treated with the extract of the present invention (Fig. 10 (a) FIG. 10C shows quantitative data for COX-2, Values with different letters are significantly different from one another (ANOVA followed by Newman-Keuls multiple range test P <0.05)

Hereinafter, the present invention will be described in detail.

However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

< Example  1>

Saponin High content  Establishment of process for the production of germanium barley extract

<1-1> Preparation of raw barley

The seed barley used in this study was cultivated in Yeonggwang-gun, Jeollanam-do, Korea. The raw bud of spring buds grown at around 20cm using the original rice nori barbara (buckbone) is provided at Sookmyung in Yeonggwang-gun, Jeollanam-do.

<1-2> Identification of extraction solvent specificity

The dried seed buds were pulverized and then subjected to circulation extraction by adding various solvents as shown in Table 1 to the weight of the raw material as shown in Table 1. The specific temperature and time conditions for the preparation of each extract were as shown in Table 1 below. After the circulation extraction, the mixture was filtered using a 75 μm wound filter and subjected to centrifugation (7,800 rpm) to remove the germ extract.

In order to qualitatively and quantitatively analyze saponarin (or isovitexin-7-O-glucoside) compounds, the triterpenoid saponin family as an effective functional ingredient and indicator component of barley extract, USP 35 (p.1362-1364) 9, p. 903-905), and analyzed by LC-MS / MS according to the Codex Alimentarius Commission (CAC) regulation and AOAC method.

NO. Solvent Extract condition
(Temperature / Time) Content of Saponarin
(mg / 100g) Remarks One Water rt, 20 h 1,087 Episode 2
Average 2 Methanol rt, 20 h 643 3 Fermented Ethanol (70%) rt, 20 h 1,407 4 Isopropyl alcohol rt, 20 h 91 5 Acetone rt, 20 h 75 6 Ethylacetate rt, 20 h 74 7 n-Hexane rt, 20 h 68 8 CO2 + Fermented Ethanol
(33% + 77%, V / V%) rt, 3 h 419

In order to extract saponarin, the functional component of barley extract, extractive solvent specificities were investigated using various polar and non - polar solvents. As a result, as shown in Table 1, saponarin content was high when extracted with purified water or 70% fermented ethanol aqueous solution. Among these, saponarin content of 1,407 mg / 100g was the best when extracted at room temperature for 20 hours with 70% fermented alcohol. Interestingly, when extracted from a 70% fermented alcoholic beverage, the saponarin content was lower than expected (Table 1, entry 8). The degree of saponarin extraction in acetone or ethyl acetate, even in polar solvents, was smaller than isopropyl alcohol, a nonpolar solvent. Therefore, it was confirmed that saponin had the extraction specificity for water or fermented alcohol.

<1-3> Saponin High content  Condition setting

Based on the results of the solvent analysis of the above <1-2>, in order to establish an extraction condition with higher extraction efficiency of saponin, detailed extraction conditions as shown in Table 2 were established for water and fermented spirits, And the extract was prepared by circulation extraction method. After the circulation extraction, the mixture was filtered using a 75 μm wound filter and subjected to centrifugation (7,800 rpm) to remove the germ extract. The saponin analysis method as an indicator component is as described in <1-2> above.

NO. Solvent % of Solvent Extract condition
(Temperature / Time) Content of Saponarin, (mg / 100g) Note One Water 100% rt, 3 h 1,233 Episode 2
Average 2 rt, 20 h 1,087 3 50 ° C, 3 h 1,224 4 80 ° C, 3 h 1,097 5 Fermented Ethanol 10% rt, 3 h 1,357 6 30% rt, 3 h 1,469 7 rt, 9 h 1,773 8 rt, 9 h (temperature rise) 1,753 9 40% rt, 3 h 1,463 10 rt, 20 h 1,422 11 50 ° C, 3 h 1,377 12 80 ° C, 3 h 1,311 13 50% rt, 3 h 1,491 14 rt, 9 h 1,722 15 rt, 9 h 1,613 16 70% rt, 3 h 1,453 17 rt, 20 h 1,407 18 50 ° C, 3 h 1,300 19 80 ° C, 3 h 1,326 20 80 ° C, 9 h 1,468 21 95%
(The amount of the drug) rt, 20 h 339

As a result, as shown in Table 2, saponarin content was 1,773 mg / 100 g when extracted at room temperature for 9 hours using a fermented alcohol solution of 30% (v / v) of alcohol (ethanol) (Table 2, entry 7).

Particularly, as shown in Table 2, in contrast to the general case where the extraction temperature is increased or the polarity is increased in order to increase the extraction efficiency of the component material in general, when the reference concentration of the fermentation broth is high On the contrary, the extraction efficiency of saponin was lowered, and when the temperature was increased, many saponin extraction efficiency decreased. From these results, it is suggested that the high yield of saponin according to the conventionally known extraction efficiency condition tendency is difficult, and a specific condition process is required for obtaining saponin.

<1-4> Saponin High content  Establishment of extract manufacturing process

Based on the above results, the inventors of the present invention conducted a circulation extraction (2-3 times) for a total of 9 (± 0.5) hours at room temperature by adding an aqueous fermented alcohol solution of 30 (± 3)% (v / v) ) Were used to establish the extraction process. After the circulation extraction, the mixture was filtered using a 75 μm wound filter and subjected to centrifugation (7,800 rpm) to remove the germ extract. Each extract was then concentrated by vacuum evaporation (55-58 ° C) to remove residual solvent, which was then concentrated to 35 brix. Dextrin was blended and dissolved therein, sterilized at 95 ° C for 30 minutes, and spray dried (liquid temperature: 75 to 80 ° C, blowing temperature: 190 ° C, atomizer: 15, 100 rpm) to prepare a dried barley extract powder. The thus-produced saponin-rich germanium barley extract powder was referred to as 'BS' in this specification, and a specific process is referred to that shown in FIG.

< Example  2>

Saponin High content  Bud of barley extract (BS) Antioxidant ability  evaluation

&Lt; 2-1 > The t- On BHP  Of Oxidative Stress

HepG2 cells, a homo sapiens (human) origin, were cultured in MEM medium containing 10% fatal bovine serum (Capricom scientific, Ebsdorfergrund, Germany) and 1% Penicillin / Streptomycin (GenDEPOT, And incubated at 37 ° C in a 5% CO 2 incubator. The culture medium was changed every two days while the medium was changed.

HepG2 cells were treated with 300 μM t-BHP to induce oxidative stress. BS was treated with concentrations of 0.1, 0.2, 0.5 or 1 mg / ml, respectively. After 12 hours, the mitochondrial and cell membrane damage of the cells was measured by MTT assay. In addition, apoptosis of the cells was confirmed by fluorescence-activated cell sorting (FACS) using AnnexinV and PI, and JC-1 staining was performed to measure the mitochondrial membrane potential.

The MTT assay results are shown in FIG. As shown in FIG. 2, the cells were oxidatively stressed by the t-BHP treatment, and the survival rate of the cells was remarkably reduced. In the BS treatment group of the present invention, however, cell damage was inhibited in a concentration-dependent manner. Especially at a concentration of 0.5 mg / ml or more, it was confirmed that the cells were proliferated.

FACS results are shown in Figs. 3A to 3C. As shown in FIGS. 3A to 3C, apoptosis was significantly increased by t-BHP treatment, but apoptosis was inhibited by 1 mg / ml BS treatment.

JC-1 staining results are shown in Figs. 4A and 4B. When apoptosis occurs, these mitochondrial membrane potentials are destroyed. When t-BHP was treated, apooptosis proceeded and mitochondrial membrane potential was found to be problematic. At this time, JC-1 is present as a monomer in cytoplasm. In contrast, the group treated with the BS of the present invention showed an effect of restoring the mitochondrial membrane potential in a concentration-dependent manner.

&Lt; 2-2 > The ARE-reporter gene activity of BS, Nrf2  And Second phase  Evaluation of effects on the expression of antioxidant enzymes

HepG2 cells transfected with the ARE-luciferase gene were treated with BS for 12 hours at different concentrations. At this time, transformation of HepG2 cells using the ARE-luciferase gene was performed using a method known in the art.

 The ARE activity was measured by Luciferase assay system. The expression level of HO-1 protein as Nrf2 and the second phase antioxidant enzyme was confirmed by using the antibody against each of them, using conventional Western blotting. At this time, Nrf2 was performed on cytoplasm (cy) and nucleus (Nu).

The results of ARE activity measurement by Luciferase assay are shown in Fig. As can be seen from FIG. 5, the activity of inducing the ARE-reporter gene at a concentration of 0.5 mg / ml or more was remarkably observed in the BS, and it was confirmed that the ARE activity was increased more than 4 times at 1 mg / ml treatment.

As shown in FIGS. 6A and 6B, when the BS of the present invention was treated with time in HepG2 cells, the Nrf2 protein was confirmed in the cytoplasm and nucleus. As shown in FIG. 7, the representative antioxidant enzyme regulated by the Nrf2 transcription factor Heme oxygenase-1 (HO-1) was found to be increased depending on the BS treatment concentration of the present invention.

< Example  3>

Saponin High content  Inflammation of bud of barley extract (BS) Inhibition  evaluation

Murine macrophage cell line Raw264.7cell was used in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fatal bovine serum (Capricom scientific, Ebsdorfergrund, Germany) and 1% Penicillin / Streptomycin (GenDEPOT, TX, USA) And cultured in a 5% CO 2 incubator at 37 ° C. The medium was changed once every two days.

Raw264.7 cells were treated with LPS, which is an inflammation inducer, and treated with BS for 12 hours. The specific treatment concentration is the same as described in each of the following drawings. For each experimental group, NF-kB activity was evaluated by Luciferase assay system and the degree of NO production, COX-2 and iNOS expression level were evaluated.

As shown in FIG. 8, it was confirmed that the increased NFkB activity by treatment with LPS was decreased in a dose dependent manner in the BS treatment group of the present invention. As shown in FIG. 9, NO production after LPS stimulation was significantly decreased by the BS treatment of the present invention, and COX-2 and iNOS, which are typical inflammation related proteins, were found in LPS , But it was confirmed that BS treatment of the present invention significantly reduced the expression. As a result, it was confirmed that the BS of the present invention has excellent anti-inflammatory activity.

As described above, the present invention provides a composition for antioxidant or antiinflammation comprising an extract of sprout barley as an active ingredient, and a fermented spirits aqueous solution of alcohol (30 (3%) (v / v) And a step of circulating and extracting the extracts for a total of 9 (± 0.5) hours.

The barberry extract of the present invention is produced by a unique method of the present invention and contains not only high saponin content but also remarkable antioxidant and anti-inflammatory effect, so that it is highly industrially applicable in the field of medicines, cosmetics, foods and the like.

Claims (6)

A food composition for antioxidant or antiinflammation characterized by containing an extract of barley as an active ingredient.
The method according to claim 1, wherein the sprout barley extract comprises a step of circulating the sprout barley for a total of 9 (± 0.5) hours at room temperature by adding an aqueous fermented alcohol solution of 30 (± 3)% (v / v) &Lt; / RTI &gt; by weight.
A cosmetic composition for antioxidant and antiinflammation, which comprises an extract of barley barley as an active ingredient.
A pharmaceutical composition for antioxidant and antiinflammation, which comprises extract of barley as an active ingredient.
Characterized in that it comprises circulating and extracting a fermented liquor aqueous solution having a degree of alcohol of 30 (± 3)% (v / v) in the barley for a total of 9 (± 0.5) hours at room temperature. &Lt; / RTI &gt;
A saponin-rich germanium barley extract prepared by the method of claim 5.

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