KR101418745B1 - A composition comprising extract of fermented ginseng using Lactobacillus plantarum CRNB-22 for treating or preventing atopic dermatitis - Google Patents

Abstract

The invention Lactobacillus plan tareom CRNB-22 (Lactobacillus plantarum CRNB-22, microorganism accession no.: KCTC11931BP). The present invention also relates to a composition for preventing or treating atopic dermatitis. The composition is excellent in inhibiting the activities of MCP-1 (monocyte chemoattractant protein-1), IL-8 (interleukin-8) and IL-6 (interleukin-6), and thus exhibits various symptoms of skin diseases such as atopic dermatitis Can be used as therapeutic agents for treatment or alleviation.

Description

TECHNICAL FIELD The present invention relates to a composition for preventing or treating atopic dermatitis comprising extracts of fermented ginseng using Lactobacillus plantarum cybenus-22.

The invention Lactobacillus plan tareom CRNB-22 (Lactobacillus plantarum CRNB-22, microorganism accession no.: KCTC11931BP). The present invention also relates to a composition for preventing or treating atopic dermatitis.

More particularly, the present invention relates to a method for inhibiting the activity of MCP-1 (monocyte chemoattractant protein-1), IL-8 (interleukin-8) and IL-6 (interleukin-6), Lactobacillus plantarum CRNB- 22 ( Lactobacillus plantarum CRNB-22, microorganism accession no.: KCTC11931BP). The present invention also relates to a composition for preventing or treating atopic dermatitis.

Although the pathogenesis of atopic dermatitis has not yet been fully elucidated, it has been known that atopic dermatitis is caused by a chronic inflammatory reaction caused by an irritant immune reaction (allergic reaction) to a common substance (allergen) in the environment. In addition, atopic dermatitis is a chronic inflammatory disease that occurs before the development of asthma and allergic diseases, and the symptoms vary depending on the concentration of chemokine, the quantitative change, and the degree of activity (Curr. Allergy Asthma Rep., 2005, 5, 284-290), and also refers to a phenomenon in which a Th2-type lymphocyte invades a lesional skin.

It is known that 0.5% to 1% of the world's population, especially 5% to 10% of children, suffer from atopic dermatitis. Environmental factors are known to be involved in the onset and deterioration. As an example, the incidence of atopic dermatitis is correlated with the amount of exposure to house dust mites, and it is known that the severity of atopic dermatitis also has a significant correlation. It is also known to reduce the severity of atopic dermatitis by reducing exposure to house dust mites in the environment. In addition, atopic dermatitis appears to be caused by a combination of dry skin, easily itchy skin, infection by bacteria, viruses, fungi, emotional factors, and environmental factors. In recent decades, the incidence of atopic dermatitis has been rising, and the number of patients in Korea is also increasing.

IL-8 (interleukin-8), one of the cytokines that cause atopic dermatitis, is an important inflammatory chemokine secreted from the airway epithelium. It is mainly expressed at the early stage of inflammation. IL-8 induces bronchial hyperresponsiveness. And bronchial asthma. As an example of this, treatment of THP-1 cells or EoL-1 cells with an extract of Phellinus linteus causing atopic dermatitis increases the amount of IL-8, a cytokine that causes atopic dermatitis.

The other cytokine that causes atopic dermatitis is MCP-1 (monocyte chemoattractant protein-1). MCP-1 binds to CCR2 (chemokine receptor) and MCP-1 gene- monocytes have been observed to be impaired in resistance to certain bacterial infections and have been reported to be similar to those observed in animals with the CCR2 gene removed. In addition, MCP-1 converts Th0 cells into cells that secrete Th2 cytokines in the test tube, and when intravenously injected with MCP-1, the production of IL-12 is decreased and the production of IL-4 is increased have.

On the other hand, therapeutic agents such as steroids, antihistamines, and antibiotics have been widely used as treatments for atopic dermatitis. Among these, steroids are the most widely used therapeutic agents for inflammatory diseases such as bronchial asthma, but the mechanism by which steroids respond in the body is still unknown. On the other hand, some patients with bronchial asthma or rheumatoid arthritis show steroid resistance that does not respond to steroids. In addition, atopic dermatitis is often chronicized rather than completely treated with a steroid-based drug, resulting in side effects such as denaturation and fibrosis of the tissue. On the other hand, when resistance to steroids increases, various cytokines such as TGF-beta, IL-4 and IL-13 related to fibrosis are secreted through various cells in the chronic inflammatory reaction. The secretion of IL-6, which activates the fibroblast, is increased, resulting in the proliferation of many fibroblasts and the over-production of extra cellular matrix, resulting in cell and tissue transformation and fibrosis .

Therefore, therapeutic agents for atopic dermatitis such as steroid agents are gradually restricted due to various side effects as described above, and a composition for treating a new concept of atopic dermatitis which is effective for atopic dermatitis but has no side effects is required have.

Panax ginseng ) is a perennial herbaceous plant belonging to the genus Ginseng ( Araliaceae ). It has been widely used as medicines or health foods in Korea and the Orient for thousands of years. In addition, ginseng can be used in various forms such as ginseng and red ginseng. Especially, it is known that red ginseng processed by steaming steam ginseng contains various ginsenosides useful for human body.

Saponin, which represents the major efficacy of ginseng, is composed of more than 30 kinds of ginsenosides. Ginsenosides are used to enhance immunity, antiinflammatory action, antiallergic action, anticancer effect, effect on erectile dysfunction, Action, anticholesterol action, antithrombotic action, adult disease and aging.

Saponin of ginseng is a neutral glycoside in which glucose, arabinose, xylose, and rhamnose are bonded to a triterpenoid-based dammarane skeleton. When extracting ginseng with water and alcohol, 20-O-? -D-glucopyranosyl-20 (S) -protonanaxadiol 20-O-? -D-glucopyranosyl-20 (S) -protopanaxadiol; Rb ₁ ), 20-O- [? -L-arabinopyranosyl (1? 6) -? - D-glucopyranosyl] - [α-L-arabinopyranosyl (1 → 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol Rb ₂ , 20- β-D-glucopyranosyl] -20 (S) -protopanaxadiol; Rc (S) -protopanaxadiol (20-O-α-L-arabinofuranosyl (20 (S) -protopanaxatriol; Rg ₁ ) and 20 (S) -protopanaxatriol (Rg ₁ ).

On the other hand, saponin is degraded and absorbed by intestinal microorganisms in the body (Planta. Med., 1996, 62 (5), 453 ~ 457). In the case of ginsenoside Rb ₁ , ginsenoside Rd, compound K (compound K) and protopanaxadiol. However, intestinal microorganisms that degrade ginseng saponin have different existence and existence degree depending on the constitution and eating habits of humans, and there is a difference in conversion to metabolites, so that there may be a difference in efficacy after taking ginseng. In other words, since intestinal microflora vary from person to person, the conversion and bioavailability of ginsenosides are different (J. Pharm. Pharmacol. 1998, 50 (10), 1155-1160).

Therefore, in order to eliminate the difference in the absorption rate of ginsenosides in ginseng due to differences in intestinal microorganisms, various kinds of microorganisms are used to ferment ginseng in advance, thereby increasing the absorption of ginsenoside into the body and providing a new type of useful ginseng fermentation There is an ongoing attempt to do so.

However, when the components and contents of ginsenosides in ginseng fermented using the above-mentioned microorganisms were confirmed, it was confirmed that the conversion of ginsenosides was not as high as expected. Therefore, microorganisms having a novel ginsenoside conversion activity There is a need for development.

On the other hand, the inventors of the present invention have found that a strain of Lactobacillus plantarum CRNB-22 (microorganism accession number: KCTC11931BP) is excellent in the ability to convert ginsenosides and thus has high functionality including a large amount of ginsenosides having a high intestinal absorption rate (Patent Application No. 10-2011-0043207). Accordingly, the inventors of the present invention have completed the present invention by confirming that the extract of fermented ginseng has an excellent effect of preventing or treating atopic dermatitis, while studying the activity of fermented ginseng using Lactobacillus plantarum CRNB-22.

Pivarcsi, A. et al., Chemokine networks in atopic dermatitis: traffic signals of disease., Curr. Allergy Asthma Rep., 2005, 5, 284-290. Hasegawa, H. et al., Main ginseng saponin metabolites formed by intestinal bacteria, Planta. Med., 1996, 62 (5), 453-457. von Moltke L.L. et al., Intestinal bacterial hydrolysis is required for the appearance of compound K in rat plasma after oral administration of ginsenoside Rb1 from Panax ginseng., J. Pharm. Pharmacol. 1998, 50 (10), 1155-1116. Lee J.S. , House dust mite, Dermatophagoides pteronissinus increases expression of MCP-1, IL-6 and IL-8 in human monocytic THP-1 cells. Cytokine, 2008, 42 (3), 365-371.

It is an object of the present invention to provide a pharmaceutical composition containing Lactobacillus plantarum CRNB-22 plantarum CRNB-22, microorganism accession no.: KCTC11931BP) as an effective ingredient for preventing or treating atopic dermatitis.

More specifically, it is an object of the present invention to provide a method of inhibiting the activity of MCP-1 (monocyte chemoattractant protein-1), IL-8 (interleukin-8) and IL- CRNB-22 ( Lactobacillus plantarum CRNB-22, microorganism accession no.: KCTC11931BP) as an effective ingredient for preventing or treating atopic dermatitis.

The present invention relates to a composition for preventing or treating atopic dermatitis containing an extract of fermented ginseng using Lactobacillus plantarum CRNB-22.

The extract may be an extract obtained by extracting fermented ginseng using Lactobacillus plantarum CRNB-22 with water, C1 to C4 lower alcohols, or a mixed solvent thereof.

Fermented ginseng using Lactobacillus plantarum CRNB-22 was prepared by inoculating 100 parts by weight of ginseng with 0.001 to 1.0 part by weight of Lactobacillus plantarum CRNB-22 strain And fermenting at 20 to 50 ° C for 5 to 9 days.

The present invention also relates to a health functional food for preventing or ameliorating atopic dermatitis containing the extract of fermented ginseng using Lactobacillus plantarum CRNB-22.

Hereinafter, the present invention will be described in more detail.

The extract of fermented ginseng using Lactobacillus plantarum CRNB-22 was prepared by adding 2 to 20 times (w / w) weight of the fermented ginseng to the fermented ginseng using Lactobacillus plantarum CRNB-22, 1 to 4 A lower alcohol or a mixed solvent thereof.

The extract of fermented ginseng using Lactobacillus plantarum CRNB-22 may be extracted with an organic solvent (alcohol, ether, acetone, etc.), an organic solvent (n-hexane, ethyl acetate, n-butanol) , A method by column chromatography, a known method used for separation and extraction of a plant component, etc., alone or in combination, can be further purified and, if necessary, further purification can be carried out according to a conventional method.

The chromatography used in the present invention includes silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, thin layer chromatography Thin layer chromatography (TLC), medium pressure liquid chromatography, and high performance liquid chromatography may be used.

Fermented ginseng using Lactobacillus plan tareom CRNB-22 of the present invention is inoculated to 100 parts by weight of ginseng Lactobacillus plan tareom CRNB-22 strain 0.001 ~ 1.0 parts by weight, 20 ~ 50 ℃, preferably at 25 ~ 45 ℃ 5 And fermentation may be performed by adding a carbon source such as rice bran, corn, starch, glucose, sucrose, fructose, lactose or the like, if necessary.

Ginseng is a perennial plant belonging to the genus Panax (Panax) for use in the production of the fermented ginseng, ginseng (Panax ginseng), hwagisam (Panax quinquefolia), jeonchilsam (thirty-seven, Panax notoginseng , Panax japonicus , Panaxa pseudoginseng ). Vietnamese Panax vietnamensis ), Panax elegatior , Panax wangianus ) and Panax bifunlatipidus ( Panax bipinratifidus , and Panax angustifolium . However, the present invention is not limited thereto. These ginsengs may be used alone or in combination of two or more.

Meanwhile, the ginseng preferably uses ginseng roots of 4 to 6 years old.

The present invention also provides a pharmaceutical composition for preventing or treating atopic dermatitis containing extract of fermented ginseng using Lactobacillus plantarum CRNB-22. The pharmaceutical compositions may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method. Examples of carriers, excipients and diluents that can be contained in the composition containing the extract of fermented ginseng using Lactobacillus plantarum CRNB-22 include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch , Hydroxypropyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, hydroxypropylmethyl cellulose, hydroxypropylmethyl cellulose, hydroxypropylmethyl cellulose, hydroxypropylmethyl cellulose, hydroxypropylmethyl cellulose, hydroxypropylmethyl cellulose, hydroxypropylmethyl cellulose, hydroxypropylmethylcellulose, . 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. The solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like. These solid preparations can be prepared by adding to the extract of fermented ginseng using Lactobacillus plantarum CRNB-22 at least one excipient such as starch , Calcium carbonate, sucrose or lactose, and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like 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. Examples of suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like.

The dose of the extract of fermented ginseng using the above-mentioned Lactobacillus plantarum CRNB-22 is determined by the age, sex, and weight of the subject to be treated, the severity of the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, Will vary. Dosage determinations based on these factors are within the level of ordinary skill in the art and generally the dosage ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferable dosage is 0.1 mg / kg / day to 500 mg / kg / day. The administration may be carried out once a day or several times. The dose is not intended to limit the scope of the invention in any way. The extract of fermented ginseng using Lactobacillus plantarum CRNB-22 of the present invention can be administered to mammals such as rats, livestock, and humans in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection. Also, since the extract of fermented ginseng using Lactobacillus plantarum CRNB-22 of the present invention is a composition derived from a natural product, it has little toxicity and side effects, and thus can be safely used for long-term use even for prophylactic purposes.

The present invention also provides a health functional food for preventing or ameliorating atopic dermatitis containing the extract of fermented ginseng using Lactobacillus plantarum CRNB-22. Food-acceptable food-aid additives may be added to the health functional food. The health functional food may be in the form of tablets, capsules, pills or liquids. The health functional foods include dairy products such as drinks, meat, sausage, bread, candy, snacks, noodles, It may include nutritional products, including beverages, alcoholic beverages and vitamin complexes, including ionic beverages.

In addition, the fermented ginseng extract using Lactobacillus plantarum CRNB-22 can be directly commercialized as a health functional food, and various additives can be mixed and commercialized to enhance flavor and flavor. Examples of additives include various saccharides, emulsifiers, sweeteners, acidulants, juices, and the like. Specific examples thereof include saccharides such as glucose, sucrose, fructose, and acacia, sugar alcohols such as sorbitol, xylitol, erysuricol, lactitol, and paratinide, fatty acid esters such as glycerin fatty acid esters, and emulsifiers such as lecithin In addition, various vitamins such as vitamin A, vitamin B, vitamin C and vitamin E, plant extract, cereal ingredient and vegetable ingredient can be used as additives.

The present invention relates to a composition for preventing or treating atopic dermatitis containing an extract of fermented ginseng using Lactobacillus plantarum CRNB-22 (Microorganism Accession Number: KCTC11931BP). The composition is excellent in inhibiting the activities of MCP-1 (monocyte chemoattractant protein-1), IL-8 (interleukin-8) and IL-6 (interleukin-6), and thus exhibits various symptoms of skin diseases such as atopic dermatitis Can be used as therapeutic agents for treatment or alleviation.

FIG. 1 shows the results of confirming the inhibitory effect of extracts of fermented ginseng on MCP-1 activity using Lactobacillus plantarum CRNB-22.
FIG. 2 shows the results of confirming the inhibitory effect of extract of fermented ginseng on IL-6 activity using Lactobacillus plantarum CRNB-22.
FIG. 3 shows the results of confirming the inhibitory effect of the extract of fermented ginseng on IL-8 activity using Lactobacillus plantarum CRNB-22.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the concept of the invention to those skilled in the art.

≪ Example 1: Isolation and identification of microorganism &

Example 1-1. Isolation of microorganisms

The microorganism of the present invention was isolated from Kimchi and Mung oil of Mongolia. Kimchi samples and horse oil were diluted in sterilized water and 1 ml of the diluted solution was divided into BCP agar plates (see composition of Table 1) and cultured at 30 ° C and 37 ° C for 3 days. After culturing, strains forming a yellow circle around a colony on a BCP agar plate were selected and cultured subcultured five times on BCP agar to be purely isolated.

In order to isolate strains having saponin-decomposing ability, the isolates were inoculated into Esculin-agar (see composition in Table 2) medium to observe the color change in the medium. The strain having saponin-decomposing ability produces β-glucosidase and esculetin produced by the reaction of β-glucose to esculin is produced by ferric ammonium citrate ferric ammonium citrate) to form a black circle around the colonies. Therefore, strains with black circles around the colonies were regarded as strains having? -Glucosidase activity, and as saponin-decomposing ability was determined, they were selected as isolates of saponin. Through these processes, six microorganisms were isolated from kimchi and ten microorganisms from Mongolian horse.

matter Content (g / ℓ) Yeast extract (Extract Yeast Dried) 2.5 Peptone 5.0 glucose 1.0 L-cysteine 0.1 Tween 80 1.0 Bromocresol Purple
(Bromcresol purple) 0.04 Agar powder 15.0

matter Content (g / ℓ) Casein extract
(Pancreatic digest of casein) 13.0 NaCl 5.0 Yeast extract 5.0 Esculin 1.0 Ferric ammonium citrate
(Ferric ammonium citrate) 0.5 Agar powder 15.0

Example  1-2. Identification of isolated microorganisms

Glucosidase activity and saponin decomposition ability among the strains selected in Example 1-1 were selected and microorganisms having the highest activity of? -Glucosidase among the strains selected as saponin-degrading microorganisms were selected for the 16S rDNA sequence Based on molecular systematic taxonomic analysis. Thereafter, the strain DNA was extracted and subjected to PCR using a universal primer (5'-GAGTTTGATCCTGGCTCAG-3 ', 5'-AGAAAGGAGGTGATCCAGCC-3') to amplify 16S rDNA (16S rRNA coding gene) . Amplification conditions were 96 ℃ for 2 min, denaturation at 96 ℃ for 10 sec, annealing at 50 ℃ for 5 sec, extension at 60 ℃ for 25 min, and final extension at 60 ℃ for 2 min. Amplified 16S rDNA was purified using a PCR product purification kit (Qiagen) and then determined using a genetic analyzer 377 (Perkin-Elmer).

As a result, the strain having the highest β-glucosidase activity and saponin degrading ability was designated as Lactobacillus plantarum CRNB-22 and deposited in the Korean Microorganism Resource Center (Accession No. KCTC11931BP) 10-2011-0043207).

< Example  2. Preparation of fermented ginseng>

0.1 g of the Lactobacillus plantarum CRNB-22 culture obtained in Example 1 was added to 100 g of 6-year old ginseng and fermented at 37 ° C for 7 days to prepare fermented ginseng.

< Example  3. Preparation of Fermented Ginseng Extract>

Example  3-1. Fermented ginseng water extract

The fermented ginseng of Example 2 was dried to a moisture content of 14%, 1 kg of the dried fermented ginseng was added to 10 kg of water, and the mixture was extracted at 90 ° C for 8 hours. Then, the filtrate from which the solid content was removed was lyophilized to prepare a fermented ginseng water extract .

Example  3-2. Fermented ginseng 70% ethanol aqueous solution extract

The fermented ginseng of Example 2 was dried to a moisture content of 14%, 1 kg of the fermented ginseng dried product was added to 4 kg of 70% ethanol aqueous solution, and the mixture was extracted at 50 ° C for 8 hours. Then, the filtrate was subjected to vacuum distillation and freeze- Extract of 70% ethanol aqueous solution of ginseng was prepared.

Example  3-3. 100% ethanol extract of fermented ginseng

The fermented ginseng of Example 2 was dried to a moisture content of 14%, 1 kg of the dried fermented ginseng was added to 4 kg of 100% ethanol, and the mixture was extracted at 50 ° C for 8 hours. Then, the filtrate with the solid content removed was distilled under reduced pressure, The extract was prepared.

< Comparative Example  1. Preparation of ginseng extract>

Comparative Example  1-1. Preparation of ginseng water extract

The extract was prepared in the same manner as in Example 3-1 except that 6-year-old white ginseng (moisture content of 14% or less) was used instead of the fermented ginseng dried product.

Comparative Example  1-2. Preparation of 70% Ethanol Extract of Ginseng

The extract was prepared in the same manner as in Example 3-2 except that 6-year-old white ginseng (moisture content of 14% or less) was used instead of the fermented ginseng dried product.

< Comparative Example  2. Preparation of red ginseng extract>

Comparative Example  2-1. Production of red ginseng water extract

The extract was prepared in the same manner as in Example 3-1, except that red ginseng (manufactured from 6-year-old ginseng) having a moisture content of 14% or less was used instead of the fermented ginseng dried ginseng.

Comparative Example  2-2. Preparation of 70% Ethanol Extract of Red Ginseng

The extract was prepared in the same manner as in Example 3-2 except that red ginseng (manufactured from 6-year-old root ginseng) having a moisture content of 14% or less was used instead of fermented ginseng dried ginseng.

< Experimental Example  1. Cell culture and treatment of fermented ginseng extract>

Human monocyte THP-1 cells (human acute monocytic leukemia cells) were cultured in RPMI 1640 medium, antibiotics (penicillin 10 ⁴ U / ml, streptomycin 10 mg / ml, amphotericin B 25 μg / fetal bovine serum) and incubated at 37 ° C and 5% CO ₂ for 3 days. Then, the THP-1 cells were divided into 24 well plates at a concentration of 2.0 x 10 &lt; ⁶ &gt; / ml using trypsin-EDTA to measure the activities of MCP-1, IL- in RPMI with FBS contain from 37 ℃, 5% CO ₂ condition were cultured for 16 hours. After culturing, the fermented ginseng extract (200 μg / ml) of Example 3, the ginseng extract of Comparative Example 1 (200 μg / ml) and the red ginseng extract of Comparative Example 2 (200 μg / ml), dexamethasone ), Tick extracts (purchased from Yonsei University Medical School, 1 ㎍ / ㎖) for 14 hours and then analyzed by enzyme-linked immunospecific assay (Cytokine. 2008, 42 (3), 365-371) 1, IL-6, and IL-8 were measured in the same manner as described above. In normal group, physiological saline, which is a water extract, was treated (the same result was obtained even when DMSO, which is a solvent of the ethanol treatment group, was treated).

As shown in FIGS. 1 to 3, when mite extracts known to induce atopy in THP-1 cells were treated, secretion of MCP-1, IL-6 and IL-8 was increased. Dexamethasone used as a positive control was found to lower the secretion of MCP-1 to almost normal level. In the cells treated with fermented ginseng extract using Lactobacillus plantarum CRNB-22 of the present invention, MCP-1, IL-6 and It was confirmed that the secretion of IL-8 was remarkably reduced, and thus the fermented ginseng extract showed excellent therapeutic effect of atopic dermatitis. On the other hand, the extracts of ginseng and red ginseng which had not undergone fermentation showed less inhibitory effect on the secretion of MCP-1, IL-6 and IL-8 than the extract of the present invention.

< Experimental Example  2. Toxicity test>

Experimental Example  2-1. Acute toxicity

This experiment was conducted to investigate the toxicity of the fermented ginseng extract using Lactobacillus plantarum CRNB-22 of the present invention to an animal body in an acute (within 24 hours) when an excessive amount of the fermented ginseng extract was consumed in a short period of time and to determine the mortality rate . Twenty ICR mice were injected into the control group and 30 mice, respectively. In the control group, nothing was administered. In the experimental group, 2.0 g / kg of the extract of fermented ginseng using Lactobacillus plantarum CRNB-22 prepared in Examples 3-1 to 3-3 per 10 animals (used in general animal experiments About 50 times the dose) was orally administered. After 24 hours of administration, the mortality rate was examined. As a result, the control group and the experimental group administered with the extract of fermented ginseng using Lactobacillus plantarum CRNB-22 prepared in Examples 3-1 to 3-3 at a concentration of 2.0 g / kg Survived.

Example  5-2. Experimental group  And control organ organs and tissue toxicity experiments

In order to investigate the effect of C57BL / 6J mice on the organs (tissues) of animals, the experimental group administered with the extract of fermented ginseng using Lactobacillus plantarum CRNB-22 prepared in Example 3 and the control group The blood was collected from the animals for 8 weeks and the blood levels of glutamate-pyruvate transferase (GPT) and blood urine nitrogen (BUN) were measured using Select E (Vital Scientific NV, Netherland). As a result, GPT, which is known to be related to hepatotoxicity, and BUN, which is known to be related to renal toxicity, showed no significant difference compared to the control group. In addition, liver and kidney were cut from each animal and histological observation was carried out with an optical microscope through a conventional tissue section production process. No abnormal abnormalities were observed.

< Examples  1. Pharmaceutical Formulation example >

Examples  1-1. Manufacture of tablets

20 g of fermented ginseng extract using Lactobacillus plantarum CRNB-22 prepared in Example 3-1 was mixed with 175.9 g of lactose, 180 g of potato starch and 32 g of colloidal silicic acid. To this mixture was added a 10% gelatin solution, which was pulverized and passed through a 14-mesh sieve. This was dried, and a mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate was made into tablets.

Examples  1-2. Injection preparation

2 g of fermented ginseng extract using Lactobacillus plantarum CRNB-22 prepared in Example 3-1 was dissolved in distilled water to make 100 ml. This solution was placed in a bottle and sterilized by heating at 20 DEG C for 30 minutes.

< Examples  2. Food Manufacturing example >

Examples  2-1. Manufacture of cooking seasonings

Healthy cooking sauce was prepared using 0.2 to 10% by weight of fermented ginseng extract using Lactobacillus plantarum CRNB-22 prepared in Example 3-1.

Examples  2-2. Manufacture of flour food products

The extract of fermented ginseng using Lactobacillus plantarum CRNB-22 prepared in Example 3-1 was added to flour at a concentration of 0.1-5.0 wt%, and the mixture was used to prepare bread, cake, cookies, crackers and noodles To produce health promotion foods.

Examples  2-3. soup  And juicy ( gravies )

The health-enhancing meat product, soup soup and juice were prepared by adding the fermented ginseng extract of Lactobacillus plantarum CRNB-22 prepared in Example 3-1 to the soup and the juice at 0.1 to 1.0 wt%.

Examples  2-4. dairy product( dairy products )

The extract of fermented ginseng using Lactobacillus plantarum CRNB-22 prepared in Example 3-1 was added to milk in an amount of 0.1-1.0 wt%, and various dairy products such as butter and ice cream were prepared using the milk.

< Examples  3. Drink Manufacturing example >

Examples  3-1. Vegetable juice  Produce

Healthy vegetable juice was prepared by adding 0.5 g of fermented ginseng extract to 1,000 ml of tomato or carrot juice using Lactobacillus plantarum CRNB-22 prepared in Example 3-1.

Examples  3-2. Fruit juice  Produce

Health enhancing fruit juice was prepared by adding 0.1 g of fermented ginseng extract to 1,000 ml of apple or grape juice using Lactobacillus plantarum CRNB-22 prepared in Example 3-1.

Claims (6)

Prevention or prevention of atopic dermatitis comprising extract of fermented ginseng with water, C1 to C4 lower alcohol, or a mixed solvent thereof, using Lactobacillus plantarum CRNB-22 (microorganism accession number: KCTC11931BP) &Lt; / RTI &gt; delete The method according to claim 1,
In the fermented ginseng using the above-mentioned Lactobacillus plantarum CRNB-22 ( Lactobacillus plantarum CRNB-22, microorganism accession number: KCTC11931BP), 0.001 to 1.0 part by weight of Lactobacillus plantarum CRNB-22 strain was inoculated into 100 parts by weight of ginseng , &Lt; / RTI &gt; for 5 to 9 days. &Lt; RTI ID = 0.0 &gt; 5. &lt; / RTI &gt; Prevention or prevention of atopic dermatitis comprising extract of fermented ginseng with water, C1 to C4 lower alcohol, or a mixed solvent thereof, using Lactobacillus plantarum CRNB-22 (microorganism accession number: KCTC11931BP) Health functional foods for improvement. delete 5. The method of claim 4,
In the fermented ginseng using the above-mentioned Lactobacillus plantarum CRNB-22 ( Lactobacillus plantarum CRNB-22, microorganism accession number: KCTC11931BP), 0.001 to 1.0 part by weight of Lactobacillus plantarum CRNB-22 strain was inoculated into 100 parts by weight of ginseng , &Lt; / RTI &gt; for 5 to 9 days. &Lt; RTI ID = 0.0 &gt;

Images