KR101904087B1 - Fermented sorghum for preventing or treating inflammatory disease - Google Patents

Abstract

The present invention relates to fermented Sorghum bicolor which is obtained by conducting, using a Sorghum bicolor extract, incubation by Aspergillus oryzae as a starter and an enzymatic reaction by saccharification enzyme. The fermented Sorghum bicolor has functions of suppressing expression of vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), and cyclooxygenase-2 (COX-2) and functions of promoting expression of heme oxygenase 1 (HO-1), thereby showing effects of treating, preventing, or ameliorating various inflammatory diseases. Accordingly, the fermented Sorghum bicolor, according to the present invention, can be useful as an active ingredient in a composition for treating, preventing, or ameliorating inflammatory diseases such as inflammatory vascular disease, arthritis, inflammatory bowel disease, inflammatory skin disease, glomerulonephritis, retinitis, tonsillitis, sore throat, gastritis, pneumonia, hepatitis, and nephritis.

Description

[0001] Fermented sorghum for preventing or treating inflammatory diseases [0002]

The present invention relates to a fermented broth obtained by culturing a Sorghum bicolar extract with an enzymatic reaction by a saccharification enzyme and by culturing the seed culture by Aspergillus oryzae , wherein the fermented broth is selected from the group consisting of VCAM-1, ICAM-1, COX- 2 and the expression-enhancing effect of HO-1 at the same time, thereby exhibiting a remarkable effect in the treatment, prevention or amelioration of various inflammatory diseases.

An inflammatory reaction is a mechanism of restoring or regenerating a damaged tissue as one of biological tissue defense against physical, chemical stimulation or bacterial infection from the outside. Inflammatory cytokines (TNF-α, IL-6), inflammation-promoting enzymes (COX), and adhesion molecules (ICAM, VCAM). Enzymes such as prostaglandins (PGE2), nitric oxide (NO), and reactive oxygen species (ROS). The overexpression of the inflammatory mediator induces various inflammatory diseases such as inflammatory vascular disease, arthritis, inflammatory bowel disease, inflammatory skin disease, glomerulonephritis, retinitis, tonsillitis, sore throat, gastritis, pneumonia, hepatitis and nephritis

In addition, HO (heme oxygenase) is a protein that induces oxidation of heme protein. Among them, HO-1 is known to be induced by stimulation of immune cytokines and oxidative stress-related factors. HO-1 maintains the homeostasis of the cells and plays a role in reducing oxidative stress and inflammatory responses in tissues. Thus, the expression of HO-1 has been shown to inhibit the production of cytokines or chemokines induced by inflammatory mediators.

Therefore, a substance that promotes the expression of HO-1 while inhibiting the expression of an inflammatory mediator to treat, prevent, or ameliorate an inflammatory disease may exhibit a remarkable effect in the treatment, prevention and improvement of inflammatory diseases.

On the other hand, sorghum ( Sorghum bicolor ) is a perennial herbaceous herbaceous plant, which is one of the important grains following rice, barley, wheat and corn. According to recent research results, it has been reported that quercetin compounds, caffeoyl glycolic acid methyl ester, and 1-O-caffeoylglycerol isolated from the extracts of water are effective for the treatment and prevention of inflammatory diseases (see Patent Documents 1 and 2)

Korean Patent No. 10-1467395 Korean Patent No. 10-1648400

The present inventors have continued research on maximizing the antiinflammatory activity of aquatic extract as an environmentally friendly natural substance, and have succeeded in obtaining a fermented product through a fermentation process using a saccharifying enzyme and an outbreak of bacteria, It was confirmed that the anti-inflammatory efficacy was remarkably improved in comparison with the water extract, thereby completing the present invention.

Accordingly, it is an object of the present invention to provide a pharmaceutical composition for the treatment and prevention of inflammatory diseases in which a fermented product obtained through a specific fermentation process is contained as an active ingredient.

It is another object of the present invention to provide a food composition effective for preventing or improving inflammatory diseases in which a fermented product obtained through a specific fermentation process is contained as an active ingredient.

It is another object of the present invention to provide a method for producing a fermented product having an anti-inflammatory activity by performing a series of fermentation processes using a saccharifying enzyme and an outbreak of bacteria from a water extract.

In order to solve the above-mentioned problems, the present invention provides a method for the treatment and prevention of inflammatory diseases in which a fermented product obtained by fermenting Sorghum bicolar extract with an enzymatic reaction with a saccharifying enzyme and cultivating seeds by Aspergillus oryzae as an active ingredient There is provided a pharmaceutical composition for oral administration.

The present invention also relates to a food composition useful for preventing or ameliorating an inflammatory disease in which a fermented product obtained by culturing a Sorghum bicolar extract by a saccharifying enzyme and seed culture by Aspergillus oryzae as an active ingredient to provide.

In addition, the present invention provides a method for producing an enzyme, which comprises the step of subjecting Sorghum bicolar extract to an enzymatic reaction with a saccharifying enzyme; And culturing the enzyme reaction product by Aspergillus oryzae ; Wherein the activity of inhibiting the expression of an inflammatory mediator is inhibited.

The fermented broth provided by the present invention has an excellent activity of promoting the expression of HO-1 (heme oxygenase-1) while inhibiting the expression of VCAM-1, ICAM-1 and COX-2, The efficacy is remarkably excellent.

Therefore, the fermented broth provided by the present invention can be used for the treatment, prevention or prevention of various inflammatory diseases such as inflammatory vascular diseases, arthritis, inflammatory bowel disease, inflammatory skin disease, glomerulonephritis, retinitis, tonsillitis, sore throat, gastritis, pneumonia, hepatitis, And is useful as an active ingredient of medicines or health foods intended for improvement.

FIG. 1 shows the result of preparing the ethanol-extract inhibitory effect against the ethanol concentration.
FIG. 2 shows the results of confirming the anti-inflammatory effects (VCAM-1, ICAM-1 and COX-2 expression levels) and antioxidant efficacy (HO-1 expression level) of the fermented broth.
FIG. 3 shows the results of comparing ICAM-1, VCAM-1, COX-2, and HO-1 expression levels of extracts of SBE and FSE.
Figure 4 shows the results of comparing the expression levels of VCAM-1 and COX-2 in 23 active compounds isolated from sorghum.
FIG. 5 shows the results obtained by comparing the content of procyanidin C1 and procyanidin B1 contained in the hydrolyzed extract and the fermented broth, respectively.

Hereinafter, the present invention will be described in detail so that those skilled in the art can easily carry out the present invention.

The present invention relates to a method for inhibiting the expression of VCAM-1, ICAM-1, and COX-2 of the fermented broth containing the fermented broth obtained by carrying out a specific enzyme reaction and seed culture, And to a pharmaceutical composition or a health food composition having an effect of treating, preventing or ameliorating various inflammatory diseases on the basis of the activity of promoting the expression of HO-1 (heme oxygenase-1).

The fermented broth containing the active ingredient in the pharmaceutical composition or health food composition of the present invention is a natural product obtained from a natural raw material and has an advantage that the side effects are extremely low.

The method for preparing a fermented product comprising the active ingredient in the pharmaceutical composition or the health food composition of the present invention comprises the steps of: a) enzymatic reaction of sorghum bicolar extract with a saccharifying enzyme; And b) seeding the enzyme reaction product with an Aspergillus sp. Strain; .

The process for producing the fermented fermented product according to the present invention will be described in more detail below.

The hydrolyzed extract used in the production method of the present invention is prepared by extracting millet with a conventional extraction method, and there is no particular limitation on the above extraction method. However, since the content of the active ingredient differs depending on the extraction solvent, it is preferable to use an alcohol or alcohol aqueous solution having 1 to 4 carbon atoms as the extraction solvent in terms of the improvement in the affinity. As a result of screening for the expression level of the inflammatory substance in the extract obtained using ethanol or ethanol aqueous solution as an extraction solvent, it was found that when ethanol of 40 to 100% by weight concentration was used as an extraction solvent, Activity was maximized, and when 50% by weight ethanol was used as the extraction solvent, it was confirmed that the anti-inflammatory activity was the highest (see FIG. 1). Thus, in the example of the present invention, fermentation was carried out using a water extract extracted with 50% ethanol as a raw material.

The fermentation of the present invention consists of an enzymatic reaction by a saccharifying enzyme and a seed culture by an Aspergillus sp. Strain.

In the enzymatic reaction, one or two or more selected from the group consisting of cellulase, amylase and sucrase may be used. Among them, cellulase and amylase are used together as a saccharifying enzyme. As the saccharifying enzyme, it is more preferable to mix the cellulase and the amylase in a ratio of 1: 0.5 to 1.5 by weight. The enzymatic reaction can be carried out at a temperature of 40 to 60 ° C for about 3 to 12 hours by dissolving the water extract in water and adding a saccharifying enzyme. The amount of water used may be in the range of 1: 3 to 10 times the weight of the dry powder of the extract. In order to completely dissolve the extract in water, it may be heated to a temperature of 80 to 90 캜, if necessary. When the enzyme reaction is completed, the enzyme reaction solution may be further sterilized at 100 to 150 ° C. for 30 minutes to 3 hours.

Aspergillus sp. Strain was used for the seed culture, and seed culture using Aspergillus oryzae was performed in the example of the present invention. The seed culture may be carried out by a conventional method. Specifically, the seed culture may be carried out at a temperature of 30 to 40 DEG C and 50 to 200 rpm for 2 to 3 days. When the seed culture is completed, the seed culture may be further sterilized at a temperature of 80 to 100 ° C. for about 1 to 3 hours.

When the enzyme reaction and seed culture are completed, the culture is concentrated under reduced pressure, further sterilized at 80 to 100 ° C for about 1 to 3 hours, and spray dried to obtain a powdery fermented product.

In the present invention, procyanidin B1 and procyanidin C1, which have been found to be most effective in inhibiting the expression of inflammatory mediators among the active compounds isolated from sorghum in order to prepare for the treatment, prevention and amelioration of inflammatory diseases, And the content of the indicator substance was quantitatively analyzed to prepare for its efficacy. The fermented broth obtained through the fermentation process as described above contains 15 to 30 mg of procyanidin B1 and 1 to 5 mg of procyanidin C1 as indicator components based on 1 g of the fermented broth. In comparison with the extracts of the fermentation broth, the fermented broth contained about 5 to 6 times more of the content of procyanidin C1 and about 20 to 30 times more of the content of procyanidin B1.

In addition, the fermented broth was remarkably improved in its activity of inhibiting the expression of VCAM-1, ICAM-1, and COX-2 and promoting the expression of HO-1 (heme oxygenase-1) Efficacy. Therefore, the fermented broth can be used for the treatment, prevention and improvement of various inflammatory diseases, specifically inflammatory vascular diseases, arthritis, inflammatory bowel disease, inflammatory skin diseases, glomerulonephritis, retinitis, tonsillitis, sore throat, gastritis, pneumonia, hepatitis and nephritis And may be used as an active ingredient in a desired pharmaceutical composition or health food composition.

The pharmaceutical composition according to the present invention comprises a fermented product as an active ingredient, and the fermented product may be contained in a liquid or powder form. The fermented broth may be contained in an amount of 0.1 to 50% by weight based on the total weight of the pharmaceutical composition, but the present invention is not limited thereto.

When the pharmaceutical composition is used clinically, it may be formulated together with carriers customary in the pharmaceutical field to prepare pharmaceutical preparations customary in the pharmaceutical field, such as tablets, capsules, powders, granules, pills, liquids and suspensions Preparations for oral administration; Injectable preparations in the form of injectable solutions or suspensions, or ready-to-use injectable dry powders which can be used as distilled water for injection for injection; Or ointments, and the like. The pharmaceutical preparations comminuted with conventional carriers may be administered orally or parenterally, for example intravenously, subcutaneously, intraperitoneally or topically. Accordingly, the pharmaceutical composition of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of medicaments.

Examples of carriers, excipients and diluents that can be contained in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methylcellulose, hydroxymethylcellulose, microcrystalline cellulose, silicified microcrystalline cellulose, povidone, crospovidone, croscarmellose sodium, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, Colloidal silicon dioxide, lactose, talc, magnesium stearate, colloidal stearyl magnesium, mineral oil, and the like.

 In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, troches, rosin, capsules and the like, which may contain at least one excipient such as lactose, saccharose, sorbitol , Mannitol, starch, amylopectin, cellulose, calcium carbonate, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, elixirs, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like are used in addition to water and liquid paraffin, May be included. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol gelatin and the like can be used. Parenteral administration may be by subcutaneous injection, intravenous injection, intramuscular injection or intra-thoracic injection.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the age, body weight, degree of disease, drug form, route of administration, and duration of the patient, but can be appropriately selected by those skilled in the art. However, for the desired effect, the pharmaceutical composition of the present invention may be administered at a dose of 0.01 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg per day. The administration can be administered several times a day, preferably once or six times, at a predetermined time interval according to the judgment of a doctor or a pharmacist.

In addition, the health food composition according to the present invention includes a fermented product as an active ingredient for the purpose of preventing or improving an inflammatory disease, and the fermented product may be contained in a liquid or powder form.

The active ingredient may be ingested as a food prepared from tablets, capsules, powders, granules, pills, liquid preparations, suspensions, etc., or may be added to general foods. Unlike conventional medicines, the health food uses food as a raw material, so there is no side effect that may occur when a medicine is taken for a long time. The content of the active ingredient can be suitably determined according to its use purpose (for prevention or improvement). Generally, the active ingredient in the health food composition may comprise from 0.1 to 90% by weight. However, in the case of long-term consumption intended for health or hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the food. Examples of the foods to which the active ingredient can be added include food products in the conventional sense including a drink, a drink, an ionic drink, a dairy product, a meat, a sausage, a bread, a noodle, a candy, a snack, a gum, . The shape of the food is not particularly limited, and it may be a solid, semi-solid, gel, liquid, or powder.

The beverage can be prepared using the health food composition of the present invention. There are no particular restrictions on the selection of other ingredients other than the active ingredient as a component contained in the beverage, and it may contain various flavors or natural carbohydrates as additional ingredients such as ordinary beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. As other flavoring agents, natural flavoring agents (tau martin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) have. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition, the health food composition of the present invention may contain various kinds of nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and aging agents (cheese, chocolate, etc.) Salts, alginic acid and salts thereof, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks and the like. It can also contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not so important, but is generally selected in the range of 0.1 to 20% by weight in the health food composition of the present invention.

The present invention as described above is explained in more detail based on the following examples, but the present invention is not limited thereto.

[Example]

Preparation Example: Preparation of fermented broth

Syringes were purchased from farmers in Gangwon Province. The millet was milled by selecting the edible part (millet), and the milled product was extracted with 50% ethanol aqueous solution and spray dried to obtain milled ethanol extract powder.

100 g of the alcoholic extract powder obtained above was mixed with 600 g of water and then heated at 100 DEG C for 1 hour. After cooling to 50 DEG C, 5 g of cellulase and 5 g of amylase were added, and the mixture was subjected to enzymatic treatment at a temperature of 50 DEG C for 6 hours, followed by sterilization at 121 DEG C for 1 hour.

Aspergillus oryzae culture was added to the enzyme-treated extract and incubated at 30 ° C for 2 days at 100 rpm. The seed culture was added at 4 wt% based on the weight of the enzyme-treated extract. After the seed culture was completed, the cells were sterilized at 95 DEG C for 1 hour.

The seed culture was concentrated under reduced pressure, again sterilized at 95 ° C for 2 hours, and spray dried to obtain a fermented watery powder.

Example 1. Inflammation-inhibiting effect of extract of Suwon

In the extracts obtained in the above example, the concentration of ethanol used as the extraction solvent was varied, and the extracts of various concentrations were extracted to measure the inhibitory effect against inflammation. The anti-inflammatory effect was judged as the amount of mRNA expression of VCAM-1 through real-time PCR.

Specifically, human aortic smooth muscle cells (HASMC) were cultured in a 100 mm dish at a concentration of 1 × 10 ⁶ cells and cultured for 24 hours. TNF-α (10 ng / ml) (50 < RTI ID = 0.0 > ul / mL) < / RTI > and incubated with shaking for 12 hours. Cultured HASMC cells were washed twice with PBS and collected in tubes using TRIzol ™ reagent. The supernatant was separated by centrifugation (12,000, rpm, 4 ° C, 5 minutes) and the supernatant was mixed with 70% ethanol to precipitate RNA, followed by washing twice with ethanol to separate total RNA.

Real-time PCR was performed using cDNA, SYBR Green, and VCAM-1 primers. For PCR, cDNA was diluted 1/100 with distilled water, and 0.5 μl each of 10 pM primer, 12.5 μl ROX plus, and 4 μl distilled water were used to make a total volume of 25 μl. In the real-time PCR cycle, the initial denaturation was performed at 95 ° C for 10 minutes, denaturation at 95 ° C for 30 seconds, annealing at 60 ° C for 1 minute, and elongation reaction at 95 ° C for 1 minute. The melting curve was started at 55 ° C, increased to 0.5 ° C at the end of 95 ° C, and reacted at 80 ° C to detect the desired fluorescence value. The fluorescence signal was quantified using the MAXPro program. In order to confirm that the same amount of RNA was used in each RT-PCR, RT-PCR for β-actin, a housekeeping gene, was carried out together.

The results of confirming the anti-inflammatory effect of the ethanol extract by the above-mentioned experimental method are shown in FIG. 1 shows that the water extract obtained by using ethanol at a concentration of 50% showed the highest anti-inflammatory activity.

Example 2. Inflammation-inhibiting effect of fermented broth

(SBE) obtained from the above preparation example, the fermented fermented product (E_FSE) obtained by enzymatically treating the fermented extract, and the fermented fermented product (E + S_FSE) Respectively. ICAM-1 and COX-2, which are expressed in cells induced by inflammation, were measured by western blot, and HO-1, an antioxidative factor, was measured.

Specifically, human aortic smooth muscle cells (HASMC) were cultured in a 100 mm dish at a concentration of 1 × 10 ⁶ cells and cultured for 24 hours. TNF-α (10 ng / ml) Samples (100 [mu] l / mL) were treated and incubated with shaking for 12 hours. The protein extracts were added to the cultured HASMC cells, left for 1 hour on ice, and then centrifuged at 4 ° C for 12 minutes at 12,000 rpm. Protein quantification was performed using a BCA ™ protein analysis kit. Each sample was electrophoresed on a 10% SDS polyacrylamide gel and transferred to a nitrocellulose membrane. The transferred membrane was blocked in primary antibody solution (5% milk / TBST) at room temperature for 1 hour, reacted at 4 ° C for 16 hours, and then washed more than 3 times with TBST buffer. The membranes were incubated in a secondary antibody solution (5% nonfat milk / TBST) at a ratio of 1: 4000, reacted at room temperature for 1 hour, and then washed three times with TBST buffer. The ECL solution was reacted and the degree of protein expression was measured and quantified using a Chemidoc ™ image analyzer.

The results of confirming the anti-inflammatory effect of the fermented broth through the above-described experimental method are shown in FIG. FIG. 2 shows the expression levels of ICAM-1, VCAM-1, and COX-2, which are involved in the adhesion of vascular endothelial cells to monocytes or T lymphocytes and play an important role in inflammation, and HO- And the amount of expression was measured. According to Fig. 2, the difference in the degree of protein expression can be expressed by the difference in the degree of protein expression between the fermented extract (SBE), the fermented fermented product obtained by enzymatically treating the fermented extract (E_FSE), and the fermented fermented product (E + S_FSE) It can be seen that it is visible. (E + S_FSE) significantly reduced the expression of inflammatory mediators of ICAM-1, VCAM-1 and COX-2 when compared to the enzyme-treated fermented product (E_FSE) , And the expression level of HO-1 is almost the same level. In addition, the amount of inflammatory mediators of ICAM-1, VCAM-1 and COX-2 significantly decreased when the fermented product (E + S_FSE) It was confirmed that the expression level of HO-1 was remarkably increased.

Example 3. Preparation of anti-inflammatory and antioxidant efficacies of extracts and fermented products

100, and 200 μl / ml of each of the fermented extract (SBE) obtained in the above preparation example and the fermented product (E + S_FSE) obtained by treating the fermented extract with enzyme + And antioxidant efficacy.

FIG. 3 shows the expression levels of ICAM-1, VCAM-1, COX-2 and HO-1 in the extracts of SBE and E + S_FSE. FIG. 3 shows that VCAM-1, ICAM-1 and COX-2, which are factors involved in vascular inflammation in the fermented broth (E + S_FSE) HO-1, which shows antioxidant activity, was found to increase. Therefore, it can be seen that the fermented product (E + S_FSE) has better antiinflammatory activity and antioxidant activity in comparison with the water extract (SBE).

Example 4. Anti-inflammatory efficacy versus active compound isolated from sorghum

The activity of inhibiting inflammation was measured by measuring the expression levels of VCAM-1 and COX-2, which are factors involved in inflammation, by Western blotting according to Example 2, on 23 kinds of activated compounds isolated from sorghum.

23 active compounds were purchased and prepared. In Table 1 and FIG. 4, the expression levels of VCAM-1 and COX-2 were measured for the 23 active compounds.

Active compound VCAM-1 expression level COX-2 expression level 1: Protocatechusan 1.17453 0.88513 2: Propionic acid 1.349079 0.971686 3: Ela Gassan 0.440139 0.038847 4: Glacial 1.516353 0.508068 5: pyrogallol 0.482257 1.407342 6: 2-Hydroxybenzoic acid 1.505397 0.274096 7: 3,4-Dimethoxybenzoic acid 2.275129 2.838006 8: 4-Hydroxybenzoic acid 1.909411 2.302352 9: 3,4-Hvdbenzoic acid 1.346538 2.219189 10: 2,4-Hydroxyphenylpropionic acid 1.507763 3.038882 11: (-) - catechin 1.325128 2.550169 12: Salicylic acid 1.636275 3.074106 13: quercetin 1.14317 2.417996 14: Narinenin 1.351056 1.724399 15: (-) - epigallocatechin gallate 1.593008 0.776272 16: (+) - catechin 1.150211 1.196072 17: procyanidin A2 1.209382 0.945476 18: Camphor Roll 1.011413 1.05518 19: (-) - epigallocatechin 0.559079 0.622784 20: ruthenolenidine chloride 0.356787 0.283534 21: Procyanidin C1 0.236472 0.624956 22: procyanidin B1 0.269764 0.850006 23: procyanidin B2 0.439303 0.899642

According to Table 1 and FIG. 4, procyanidin C1 (No. 21) and procyanidin B1 (No. 22) among the 23 kinds of compounds isolated from sorghum inhibited the expression of VCAM-1 and COX-2 factors involved in inflammation The best.

Example 5: Comparison of the content of procyanidin C1 and procyanidin B1 in the hydrolyzed extract and fermented broth

To quantify the content of procyanidin C1 and procyanidin B1 contained in each of the fermented extract (SBE) obtained in the above preparation example and the fermented product (E + S_FSE) obtained by enzyme-seeded treatment of the fermented extract, high performance liquid chromatography (HPLC ) Were analyzed. The results are shown in Table 2 and FIG.

division Content (mg / g) Procyanidin B1 Procyanidin C1 Sorghum extract 4.17 0.10 Fermented water 23.69 2.96

According to the results of the HPLC analysis of Table 2 and FIG. 5, it can be seen that the content of procyanidin C1 and procyanidin B1 is higher in the fermented product (E + S_FSE) compared to the extract of water extract (SBE). In preparing the fermented extract, the content of procyanidin C1 was about 5.7 times that of the fermented broth, and the content of procyanidin B1 was about 29.6 times greater.

Claims (17)

An enzyme reaction using a mixture of cellulase and amylase as a saccharifying enzyme in Sorghum bicolar extract; And
A pharmaceutical composition for treating or preventing a vascular inflammatory disease, wherein the fermented product obtained by culturing a seed culture by Aspergillus sp. Strain Aspergillus oryzae is contained as an active ingredient.
The method according to claim 1,
1 g of the fermented broth contains 15 to 30 mg of procyanidin B1 and 1 to 5 mg of procyanidin C1 as indicator components.
delete The method according to claim 1,
The fermented broth comprises a procyanidin B1, procyanidin C1 or an active ingredient thereof at the same time.
An enzyme reaction using a mixture of cellulase and amylase as a saccharifying enzyme in Sorghum bicolar extract; And
A food composition useful for preventing or ameliorating a vascular inflammatory disease containing an active ingredient as a fermented product obtained by culturing a seed culture by Aspergillus oryzae strain as Aspergillus sp .
6. The method of claim 5,
1 g of the fermented broth contains 15 to 30 mg of procyanidin B1 and 1 to 5 mg of procyanidin C1 as indicator components.
delete 6. The method of claim 5,
Wherein the fermented broth comprises procyanidin B1, procyanidin C1 or an active ingredient thereof at the same time.
6. The method of claim 5,
Wherein the food composition is selected from the group consisting of a drink, a drink, an ionic drink, a dairy product, a meat, a sausage, a bread, a noodle, a candy, a snack, a gum, a tea and a vitamin complex.
a) extracting Sorghum bicol with an aqueous solution containing an alcohol having 1 to 4 carbon atoms to produce a water extract;
b) an enzymatic reaction using a mixture of cellulase and amylase as a saccharifying enzyme in the extract; And
c) seed culture of the enzyme reaction product by an Aspergillus oryzae strain, Aspergillus sp . ;
/ RTI >
Wherein the aqueous solution has an alcohol concentration of 40 to 60 wt%
A method for producing a fermented broth having an activity of inhibiting the expression of an inflammatory mediator of blood vessels.
delete 11. The method of claim 10,
Wherein the saccharifying enzyme further comprises sucrose. ≪ RTI ID = 0.0 > 21. < / RTI >
11. The method of claim 10,
Wherein the saccharifying enzyme is a mixture of a cellulase and an amylase in a weight ratio of 1: 0.5 to 1.5.
11. The method of claim 10,
Wherein the enzymatic reaction is carried out in a water solvent at a temperature of 40 to 60 占 폚.
11. The method of claim 10,
Wherein the fermented broth has an effect of inhibiting the expression of VCAM-1, ICAM-1 and COX-2 and increasing the expression of HO-1.
11. The method of claim 10,
Wherein the fermented broth contains procyanidin B1, procyanidin C1 or the like as an indicator component at the same time.
11. The method of claim 10,
Wherein 1 g of the fermented broth contains 15 to 30 mg of procyanidin B1 and 1 to 5 mg of procyanidin C1 as indicator components.

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