KR20160096967A - Composition for preventing respiratorydisease, immune-enhancing, anti-obesity containing fermented Platycodon grandiflorum - Google Patents

Abstract

The present invention relates to a health tea composition for preventing a bronchial disease, enhancing immunity, and preventing obesity including a balloon flower root fermented material as an active ingredient. According to the present invention, a balloon flower root fermented material is confirmed to have an effective antimicrobial activity with respect to Klebsiella pneumoniae subsp. Pneumoniae, bronchial bacteria. In addition, growth of a human immune cell is promoted, and cytotoxicity is low, thereby reducing fat accumulation without affecting cell viability to prevent obesity. Therefore, the balloon flower root fermented material can be usefully used as bronchial disease preventing, immunity enhancing, and obesity preventing compositions. In addition, cytotoxicity does not have toxic and side effects with respect to a composition. Therefore, the composition can be safe to be used when being taken for a long period of time. In addition, the composition is stable in vivo.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a health tea composition for preventing bronchial disease, immune enhancement, and anti-obesity comprising an effective amount of a fermented product of Platycodon grandiflorum,

The present invention relates to a health care composition comprising bronchial asthma, immune enhancement, and anti-obesity-inhibiting activity, which comprises a fermented product of Doraji as an active ingredient.

Cough is a defensive period that occurs reflexively by airway mucous membrane stimulation. When dust or irritants come in from the outside with the same principle as cough, the bronchus of our body pushes out with saliva along with saliva, It is caused by the inflammation of the bronchial tubes and lungs, causing a purulent sputum.

Cough receptors such as the larynx, trachea, bronchial tubes, pharynx, and diaphragm are stimulated by physicochemical factors, and stimulation is transmitted to the cough center of the training site to cause cough reflex. These coughs have been used to control cough by inhibiting the center of relaxation of the soft tissues or by controlling stretch receptors at the periphery. Drugs that inhibit the growth of drugs include codeine, dextromethorphan, noscapine, ephedrine, and zipeprol, which are often called thrombotic responses , Myocardial infarction, respiratory depression, and gastrointestinal disorders. The drugs that regulate the peripheral stretch receptors that have been developed by complementing these drawbacks include levopropizine, benzonate, benzozamine, benzyl alchol, and the like. .

Or the parasympathetic nervous system is activated by physiochemical factors, the bronchial smooth muscle contracts, resulting in bronchospasm or bronchoconstriction leading to coughing. At this time, drugs such as clenbuterol, bambuterol, and formoterol have been used as β-2-adrenergic receptor agonists that relax smooth muscle.

Bronchial asthma is the most common allergic respiratory disease, and allergens that are inhaled by the airways secrete several kinds of interleukines from various immune cells in vivo. By these mast cells, Leukotrienes, prostaglandins, and histamines, which are primarily inflammatory reactions that cause acute bronchoconstriction, followed by secondary bronchoconstriction. Disodium cromoglycate, cromolyn sodium, nedcromil sodium, ketotifen, and the like have been used to treat asthma. In recent years, Development of medicines using inhibitors or metabolic inhibitors of inflammatory mediators focusing on various inflammatory reactions involved in bronchial relaxants and asthma is also under way.

Most of the expectorants have been used as medicinal medicines or experience prescriptions by using plants, and recently, cysteine derivatives such as carbocysteine and N-acetylcysteine have been used. However, since these drugs do not have completely satisfactory effects, and they are toxic and chemically unstable, there has been a demand for development of an antipyretic agent having low toxicity, no side effects, and excellent dandruff effect.

Obesity is one of the most prevalent diseases in which the pattern of disease is rapidly changing to a developed country type as the hygiene environment is improved due to the improvement of the living standard and the average life span is extended due to the westernization of the diet. Thus, adult diseases have emerged as the biggest medical problem today, and the obesity, which is a major cause of these adult diseases, is increasing rapidly. Obesity refers to the phenomenon that extra calories are accumulated in the body in the form of fat by consuming excessive calories compared to the calories consumed. Obesity is thought to be caused by various causes such as genetic influences, environmental influences due to westernized diet, and psychological effects due to stress. However, the precise cause and mechanism of obesity has not been established yet. However, since obesity can act not only as a problem of obesity itself, but also as a cause of diseases such as cardiovascular disease or diabetes (Manson et al., New England J. Med., 333, pp677-685, 1995; JAMA, 282, pp1523-1529, 1999). There is a growing interest in the treatment of obesity worldwide.

In addition, studies on the regulation of the human immune system against the natural extract have been actively conducted because the immune system abnormality breaks health and causes various diseases. Therefore, the development of immunomodulators for safe natural products that can minimize side effects can be a very important material for the development of food, cosmetics and pharmaceuticals. For example, mulberry bark and shiitake mushrooms are used as immunostimulants, Starfish and Ganoderma mushrooms as anti-cancer adjuvants, and licorice, licorice glycyrrhizin, sea cucumber, Research is underway on the differentiation materials. The development of immunomodulatory agents using these natural materials has attracted many people's interest and interest because they can safely treat cancer and chronic inflammatory diseases without side effects.

Thus, the present applicant has been able to prepare a bellflower health tea composition having a bronchial disease, immunity enhancement and anti-obesity prevention effect from natural extracts.

Korean Patent No. 10-1162710 Korean Patent No. 10-0495315

Accordingly, the present inventors have found that a bellflower fermented product has an antibacterial activity against bronchial disease bacteria, is effective for enhancing immune cell growth, has an anti-obesity-inhibiting effect, and includes a bronchial disease, The present invention has been completed by preparing a health tea composition having an anti-obesity effect.

Accordingly, an object of the present invention is to provide a composition for preventing or treating a bronchial disease, which comprises a bellflower as an active ingredient.

Another object of the present invention is to provide a composition for enhancing immunity comprising a bellflower fermented product as an active ingredient.

Another object of the present invention is to provide an anti-obesity composition comprising a bellflower fermented product as an active ingredient.

Another object of the present invention is to provide a method for producing a bellflower fermented product.

It is another object of the present invention to provide a method for manufacturing a bellflower fermented tea.

In order to accomplish the above object, the present invention provides a composition for preventing or treating bronchial diseases, which comprises a bellflower fermented product as an active ingredient.

In one embodiment of the present invention, the bronchial disease is selected from the group consisting of cold, flu, tonsillitis, acute bronchitis, chronic bronchitis, pneumonia, bronchial asthma, nasopharyngitis, otitis media, tonsil and adenoid disease Lt; / RTI >

The present invention provides a composition for enhancing immunity, comprising a borage broth as an active ingredient.

The present invention provides an anti-obesity composition comprising a bellflower fermented product as an active ingredient.

In one embodiment of the present invention, the bellflower fermented product is selected from the group consisting of Saccharomyces cerevisiae, Bacillus amyloliquefaciens, and Lactobacillus plantarum Wherein the composition is fermented using one or more selected microorganisms.

In one embodiment of the present invention, the bellflower fermentation tea is fermented using Saccharomyces cerevisiae, Bacillus amyloliquefaciens, and Lactobacillus plantarum ≪ / RTI >

Further, the present invention provides a method for preparing a dough, comprising the steps of: Inoculating the fermented broth to the cooked broth; And fermenting the inoculated platycodon. ≪ Desc / Clms Page number 3 >

The present invention relates to a process for preparing a starch, Inoculating the fermented broth to the cooked broth; Fermenting the inoculated platycodon; Naturally drying the fermented broth; And aging the naturally dried bellflower cornflower.

According to an embodiment of the present invention, the step of steamed brocade may be a heat treatment in steam at a temperature of 115 to 125 ° C for 15 to 25 minutes.

In one embodiment of the present invention, the step of inoculating the bacteria may be carried out after cooling the cooked broth to 35 to 40 ° C.

In one embodiment of the present invention, the fermentation step may be performed by fermenting the seedlings in the incubator at 33-37 ° C for 6-8 days.

In one embodiment of the present invention, the naturally drying step is naturally dried at room temperature for 1 to 3 days, and the aging step is performed at room temperature for 6 to 8 days.

The present invention relates to a health tea composition having bronchial disease, immunity enhancement and anti-obesity-preventing effect, which comprises a fermented product of Doraji as an active ingredient. The platelet-like fermented product according to the present invention is a bronze-inducing bacterium, Klebsiella pneumoniae subsp. pneumoniae , exhibited stimulation of growth of human immune cells, low cytotoxicity, and did not affect cell viability, decreased fat accumulation, and thus had anti-obesity effect, thus preventing bronchial diseases, immune enhancement and anti-obesity And can be usefully used as a composition. In addition, there is no cytotoxicity, toxicity to the drug, and side effects are avoided, so that they can be used safely even when taken for a long period of time and have a stable effect in the body.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a process diagram showing a process for producing a bellflower fermented product. FIG.
Fig. 2 shows the results of the bronchial-induced bacterium Klebsiella pneumoniae subsp. As a result of comparing antimicrobial activities of Pseudomonas aeruginosa with Pseudomonas aeruginosa against Pneumoniae, A shows antibacterial activity of Pseudomonas aeruginosa and B shows antimicrobial activity of Pseudomonas aeruginosa Fermented tea.
BF: Fermentation of Bacillus amyloliquefaciens
YF: Saccharomyces cerevisiae fermentation
LF: Lactobacillus plantarum fermentation
MF: fermentation with three strains
NF: No fermentation
FIG. 3 is a graph showing the cell growth using a human immune cell T cell in order to verify the effect of enhancing immune function on the borage fermented product and the fermented tea. FIG. 3 is a graph showing the immunoenhancing effect on the borage fermented product, It is a graph showing the immunity enhancement effect on tea.
BF: Fermentation of Bacillus amyloliquefaciens
YF: Saccharomyces cerevisiae fermentation
LF: Lactobacillus plantarum fermentation
MF: fermentation with three strains
NF: No fermentation
FIG. 4 is a graph showing the cell growth using the human immune cell B cells in order to verify the immune function enhancing effect on the platycodon fermented product and the fermented tea. In FIG. 4, A is a graph showing the immunoenhancing effect on the platycodon fermented product, It is a graph showing the immunity enhancement effect on tea.
BF: Fermentation of Bacillus amyloliquefaciens
YF: Saccharomyces cerevisiae fermentation
LF: Lactobacillus plantarum fermentation
MF: fermentation with three strains
NF: No fermentation
FIG. 5 is a graph showing the cell viability of 3T3-L1 cells for the measurement of cytotoxicity against platycodon fermented products and fermented tea. FIG. 5A is a graph showing the cytotoxicity of platelet fermented products, The results are shown in Fig.
BF: Fermentation of Bacillus amyloliquefaciens
YF: Saccharomyces cerevisiae fermentation
LF: Lactobacillus plantarum fermentation
MF: fermentation with three strains
NF: No fermentation
FIG. 6 is a graph showing the inhibitory effect of lipid accumulation on the fermented broodstock and fermented tea, wherein A is the lipid accumulation rate of the fermented broth and B is the graph of the lipid accumulation rate of the fermented broth.
BF: Fermentation of Bacillus amyloliquefaciens
YF: Saccharomyces cerevisiae fermentation
LF: Lactobacillus plantarum fermentation
MF: fermentation with three strains
NF: No fermentation
FIG. 7 is a photograph showing the effect of inhibiting adipocyte differentiation on morula fermented using Oil red O staining.
BF: Fermentation of Bacillus amyloliquefaciens
YF: Saccharomyces cerevisiae fermentation
LF: Lactobacillus plantarum fermentation
MF: fermentation with three strains
NF: No fermentation
FIG. 8 is a photograph showing the effect of inhibiting adipocyte differentiation on morula fermented tea using Oil red O staining.
BF: Fermentation of Bacillus amyloliquefaciens
YF: Saccharomyces cerevisiae fermentation
LF: Lactobacillus plantarum fermentation
MF: fermentation with three strains
NF: No fermentation
FIG. 9 shows the results of α-amylase inhibition activity of the platycodon fermented product and the fermented tea, wherein A is the α-amylase inhibitory activity of the platycodon fermented product, and B is the α-amylase inhibitory activity Graph.
BF: Fermentation of Bacillus amyloliquefaciens
YF: Saccharomyces cerevisiae fermentation
LF: Lactobacillus plantarum fermentation
MF: fermentation with three strains
NF: No fermentation
FIG. 10 shows the results of inhibition activity of α-glucosidase on the fermentation broth and fermented tea. FIG. 10 shows the results of inhibition activity of α-glucosidase on the fermentation broth, Graph.
BF: Fermentation of Bacillus amyloliquefaciens
YF: Saccharomyces cerevisiae fermentation
LF: Lactobacillus plantarum fermentation
MF: fermentation with three strains
NF: No fermentation
FIG. 11 shows the results of inhibition of procine pancreatic lipase activity in the fermentation broth of platycodon and fermented tea, A shows the inhibitory activity of procine pancreatic lipase in the fermented broth, and B shows the inhibitory activity of procine pancreatic lipase in the fermented broth Graph.
BF: Fermentation of Bacillus amyloliquefaciens
YF: Saccharomyces cerevisiae fermentation
LF: Lactobacillus plantarum fermentation
MF: fermentation with three strains
NF: No fermentation

The present invention provides a health tea composition having bronchial disease, immunity enhancement and anti-obesity-inhibiting effect, which comprises a fermented product of Dorotus as an active ingredient.

As a material for a composition for a health food having an effect of preventing bronchial disease, immunity enhancement and anti-obesity according to the present invention, the bellflower is also called Gakyung, Dorat, Gilbang, White Pig, Vinegar, and Mountain bollard. Roots are thick, stems grow straight, white juice comes out. The height is 40 ~ 100cm. Leaves are alternate phyllotaxis, long ovate or broad basal, serrate on edge, no petiole. The tip of the leaf is sharp and the base is wide. The front surface of the leaf is green and the back side is grayish blue. It has no hairs, length 47cm, width 1.54cm. The flower is white or purple in July ~ August. It is a bell-shaped bell with the tip end spreading. It is 45cm in diameter and the tip is divided into 5 pieces. The calyx is also divided into five, and the branch is bulky. There are 5 stamens, 1 pistil, 5 ovaries and 5 pistils. The fruit is ovate with ovate and ripened with calyx. Breeding is good with seeds. In the spring and autumn, take roots, eat raw or eat as herbs. The main component of the bellflower is saponin. The herbaceous ginseng root is peeled off or dried as it is, and it is used in diarrhea for the treatment of diarrhea. It is called for. Albiflorum for the white flower, double flower for the double flower, and double flower for the white flower. Flower language is eternal love. It is distributed in Korea, Japan, China and others.

Such bellflower has been used for a long time not only for edible use but also for medicinal use. It is said that the roots are dried in the sun and dried in the sun to rule the sore throat. The taste is spicy, spicy and slightly poisonous. It is said that the lungs of Mokko dominate the chest and poison the insects. In the Japanese anchovy plant, it is said that it excretes the agaric acid in the lungs of the lungs. It is said that it is effective in the current private prescription for cold, cough, cold disease, abdominal pain, swelling, diarrhea, postpartum illness, women's disease, insomnia, throat qataritis, bronchitis, dysmenorrhea, dysentery.

The inventors of the present invention have confirmed through experiments that the abovementioned platycodon fermented product has an effect of preventing bronchial disease, immune enhancement and anti-obesity. In particular, the present inventors have found the effect that the platelet-shaped fermented product can be easily prepared as tea.

More specifically, according to one embodiment of the present invention, three kinds of lactic acid bacteria, yeast, and bacillus which have been identified as excellent bacillus were selected and cultured in order to produce a belletized fermented product.

In order to prepare the bellflower, the selected bellflower was washed and dehydrated and then chopped. The mixture was steamed at 121 ° C for 20 minutes, cooled to 35-40 ° C, Each 1% of the fermenting bacteria was inoculated and mixed evenly. At this time, three kinds of fermenting bacteria were fermented individually or mixed with three kinds of bacteria, and they were placed in a container and fermented at 35 ° C for 7 days. After the fermented bellflower was naturally dried in a well-ventilated place, it was thoroughly dried and then wrapped in Korean paper, and the mixture was aged at room temperature for aging.

The inventors of the present invention conducted an experiment to verify the antimicrobial activity against the bronchial disease bacteria of the fermented broodstock and the fermented tea by using the fermented tea strains and the fermented tea strains as a sample, Respectively.

Klebsiella pneumoniae subsp. As a result of the test of antimicrobial activity of the prototype against pneumoniae , it was confirmed that the borage fermented tea had a relatively higher activity than the non-fermented tea cultivar (see Fig. 2).

The present inventors have to verify the immunity reinforcement effects for bellflower prototype through a different one embodiment, this effect is a result of verification using the T cell and B cell human immune cells, Bacillus amyl Lowry Quebec Pacific Enschede (Bacillus amyloliquefaciens) and three kinds of mixed fermentation tea showed higher growth promotion (see FIGS. 3 and 4).

Furthermore, the present inventors evaluated α-amylase inhibitory activity, α-glucose inhibitory activity, and pancreatic lipase inhibitory activity to test anti-obesity inhibitory effect of Doraji fermented tea using 3T3-L1 cell, Both of the fermented broth and fermented tea showed relatively high inhibitory activity in the fermentation strain of Bacillus strains (see FIG. 9). In the evaluation of the inhibitory activity of α-glucose, (See FIG. 10). In addition, the inhibitory activities of the three kinds of mixed fermentation tea were relatively high in the evaluation of inhibition activity of pancreatic lipase, (See FIG. 11).

In addition, the present inventors confirmed that cytotoxicity was low through 3T3-L1 cell cytotoxicity measurement (see FIG. 5). The oil red O staining resulted in a decrease in lipogenesis in Bacillus and yeast fermentation tea, (See Fig. 7).

Therefore, the inventors of the present invention have found that a health tea composition containing the fermented broth of the present invention as an active ingredient can be used as a food for preventing bronchial diseases, immune enhancement, and anti-obesity based on the experimental results obtained in the following examples there was.

Accordingly, the present invention provides a health tea composition having bronchial disease, immune enhancement, and anti-obesity-inhibiting effect, which comprises a fermented product of Dorotus as an active ingredient.

Specifically, the bronchial diseases to which the present invention of the present invention can be applied include, but are not limited to, cold, flu, tonsillitis, acute bronchitis, chronic bronchitis, pneumonia, bronchial asthma, nasopharyngitis, otitis media, And bronchial disease in which the bronchial mucosa is inflamed.

In the present invention, the term "immune enhancement" refers to a cell-growth promoting action of T cells and B cells, and may include immune cells other than the above-described immune cells.

In the present invention, the term "anti-obesity" refers to the ability of cells to inhibit α-amylase, α-glucose, pancreatic lipase and adipocyte differentiation, and the concentration of the composition for anti-obesity may be 10 to 1000 μg / ml.

In the present invention, 'obesity' means that the amount of fat in the body is excessively increased. Obesity is caused by an imbalance in the synthesis and degradation of fat in adipocytes. Obesity, which is caused by excess fat accumulation, increases the risk of developing chronic degenerative diseases and increases the risk of coronary artery disease, hypertension, stroke, hyperlipemia and diabetes .

In the present invention, " prevention " means all actions that inhibit obesity or delay progression by administration of the composition of the present invention.

&Quot; Improvement " in the present invention means all the actions of improving or ameliorating symptoms of obesity by administration of the composition of the present invention.

In the present invention, " treatment " means, unless otherwise indicated, reversing, alleviating, inhibiting or preventing the disease or disorder to which the term applies, or one or more symptoms of the disease or disorder And the term " treatment " as used herein refers to an act of treating when " treating " is defined as described above.

The term " administration " as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.

As used herein, the term fermentation refers to the fact that complicated organic matter is decomposed by the action of enzymes such as fungi, yeast, bacteria and the like to become simple other organic materials. Fermentation in the presence of enzymes is called aerobic fermentation, Fermentation is called anaerobic fermentation. That is, the fermentation for obtaining the bellflower fermented product includes not only fermentation at the above-mentioned 35 ° C. for 7 days but also fermentation at 30 to 40 ° C. for 6 to 8 days.

The term fermentation as used herein includes materials that have undergone fermentation by microorganisms. It includes not only those obtained by fermentation using microorganisms, but also those obtained by further application of other microorganisms thereto.

In the present invention, steaming and boiling for boiled platycodon fermentation products includes steaming at 110 to 130 DEG C for 15 to 25 minutes.

In the present invention, the fermentation for making the bellflower fermented product is referred to as fermentation, in which the microorganism decomposes the organic matter by using the enzyme of the microorganism. The microorganisms for fermentation include Bacillus amyloliquefaciens, Sararomyces cerevisiae ≪ / RTI > and Lactobacillus plantarum.

As used herein, the term natural drying is a process of drying using natural wind at normal temperature, and includes drying in a natural ventilation state at a temperature of 15 to 25 ° C.

In the present invention, the fermented broth is completely dried and then wrapped in Korean paper, and the fermented broth is kept at room temperature and aged for aging to make the fermented product.

Furthermore, the health tea composition of the present invention comprising the fermented Dorado fermented product as an active ingredient can prevent disease associated with the immune cell growth promoting effect, inhibition of adipocyte differentiation and carbohydrate inhibition activity.

Therefore, it can be usefully used as a health tea comprising an effective amount of a fermented broth for prevention of bronchial diseases, immunity enhancement and obesity diseases.

The composition of the present invention may be a food composition. The food composition may contain, as an additional component, various flavors or natural carbohydrates such as an ordinary food composition, in addition to the active ingredient, a bellflower fermented product.

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. The above-described flavors can be advantageously used as natural flavorings (tau martin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.).

The food composition of the present invention can be formulated in the same manner as the above pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolates, foods, confectionery, pizza, ram noodles, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes, .

In addition, the food composition may contain flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and aging agents (cheese, chocolate, etc.), pectic acid, Salts of alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks.

Since the edible fermented product of the present invention is a natural substance, it has little toxicity and side effects, and can be safely used for long-term use for the purpose of preventing bronchial diseases, immunity enhancement and anti-obesity.

The health functional food of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, rings and the like for the purpose of preventing bronchial diseases, immunity enhancement and anti-obesity.

In the present invention, the term "health functional food" refers to a food prepared and processed by using raw materials or ingredients having useful functions in accordance with Law No. 6727 on Health Functional Foods, and the nutritional control on the structure and function of the human body Or for the purpose of obtaining a beneficial effect for health use such as physiological action.

The health functional foods of the present invention may contain conventional food additives and, unless otherwise specified, whether or not they are suitable as food additives are classified according to the General Rules for Food Additives approved by the Food and Drug Administration, Standards and standards.

Examples of the food items included in the above food additives include chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as persimmon extract, licorice extract, crystalline cellulose, high color pigment and guar gum; L-glutamic acid sodium preparations, noodle-added alkalis, preservative preparations, tar coloring preparations and the like.

For example, the health functional food in the form of tablets may be prepared by granulating a mixture obtained by mixing the active ingredient of the present invention with the excipient, binder, disintegrant and other additives in a usual manner, Or the mixture can be directly compression molded. In addition, the health functional food of the tablet form may contain a mating agent or the like if necessary.

The hard capsule of the capsule type health functional food can be prepared by filling a normal hard capsule with a mixture of the fermented product of the present invention, which is an effective ingredient of the present invention, with an additive such as an excipient. The soft capsule is prepared by mixing the fermented product And filling the mixture with a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative and the like, if necessary.

The ring-shaped health functional food can be prepared by molding a mixture of the active ingredient of the present invention, which is an effective ingredient of the present invention, and a mixture of excipients, binders, disintegrators, etc., by a known method, Or it may be coated with a material such as starch, talc.

The granular health functional food may be prepared by granulating a mixture of the active ingredient of the present invention, which is the active ingredient of the present invention, with the excipient, binder, disintegrant, etc., And the like.

The health functional food may be a beverage, a meat, a chocolate, a food, a confectionery, a pizza, a ramen, a noodle, a gum, a candy, an ice cream, an alcoholic beverage, a vitamin complex and a health supplement food.

Hereinafter, the present invention will be further described by way of examples. These examples are for further illustrating the present invention, and the scope of the present invention is not limited to these examples.

≪ Example 1 > Fermentation Process of Borage Material

1-1. Microbial strain

Three strains of lactic acid bacteria, yeast, and bacillus were identified as excellent microorganism strains by bacteriological screening experiment in bellflower. The strains were cultivated in Korea Microorganism Resource Center (KCTC) and Korea Microorganism Conservation Center (KCCM). Lactobacillus plantarum KCCM 11322 isolated from kimchi was used as a lactic acid bacterium. Saccharomyces cerevisiae KCCM 11201, a baker's yeast strain, was used as a yeast strain. Bacillus amyloliquefaciens KCTC 1660 bacteria were used. Yeast and Bacillus were cultured in TSB (Tryptic Soy Broth) at 35 ℃ for more than 15 hours and lactic acid bacteria were cultured under the same conditions using MRS broth medium.

1-2. Process of bellflower fermentation for making bellflower fermented water

In order to remove dirt and foreign matter, the bellflower was washed several times with clean water and then the water was removed. After removing the moisture, the bellflower was cut into slices having a thickness of 2 to 3 mm, steamed at 121 ° C for 20 minutes, removed at 35 to 40 ° C, and inoculated with about 1% of the bacteria. Three fermenting bacteria, lactic acid bacteria, yeast, and Bacillus bacteria were mixed, either alone or in combination. The bellflower inoculated with the fermenting bacteria was put into a container and put into an incubator and fermented at 35 ° C for 7 days. After the fermentation, it was naturally dried in a well-ventilated place, completely dried, and wrapped in Korean paper, followed by aging at room temperature.

<Example 2> Antimicrobial activity of tea prototype for bronchial disease bacteria

(Meat extract 1%, tryptone 1%, NaCl 0.5%) as a sample to investigate the efficacy of Platycodon grandiflorum and Platycodon grandiflorum fermented tea on the bacteria causing bronchial diseases, , glucose 0.2%, yeast extract 0.3%, pepton 0.1%, pH 7.3). The bacterial culture medium was modified with blood agar base (Difco 0045) and the antimicrobial activity assay was measured by disk agar plate diffusion method. After culturing each test microorganism in a suitable medium at a concentration of 10 ⁷ CFU / mL, the seeds were placed in a Petri dish and placed on a paper disc. 40 μL of sample extracts were added and cultured to observe the formation of inhibitory rings. When the inhibitory ring was generated within 24 hours, the antimicrobial activity was judged to be positive, and the diameter of the inhibitory ring was measured and compared.

The strains used for the antimicrobial activity test and the list used for the culture conditions Strains Cultivarion conditions Corynebacterium diphtheriae (KCCM 40413)
Klebsiella pneumoniae subsp. pneumoniae (KCCM 41285)
Streptococcus pyogenes (KCCM 11817) 37 ° C, Trypticase Soy Agar (BBL211043) with 5% defibrinated Sheep blood
37 ° C, Nutrient Agar
37 ° C, Trypticase Soy Agar (BBL211043) with 5% defibrinated sheep blood

Klebsiella pneumoniae subsp. As a result of the test for antimicrobial activity against pneumoniae , the antifungal activities of the strains of the strains and the strains of the strains of the strains were higher than those of the strains of the strains of Saccharomyces cerevisiae, Bacillus amyloliquefaciens , The fermented tea strains of Bacillus amyloliquefaciens and Lactobacillus plantarum were found to be relatively active in fermented tea and fermented tea of lactic acid bacteria. Especially, (See Fig. 2).

Observation of inhibitory ring formation by agar diffusion method (Unit: cm) A medium (platelet fermented product) B medium (tea with fermented borage) Bacillus 1.1 Bacillus 1.1 Lactobacillus 1.3 Lactobacillus 1.2 yeast fungus 1.1 yeast fungus 1.2 Three mixed bacteria 1.5 Three mixed bacteria 1.4 Non fermentation 1.0 Non fermentation 1.0

<Example 3> Effect of enhancing the immune cell viability of the seedlings using the seedlings and the seedlings

The immune function enhancement effect on the fermented broth and fermented tea was verified using T cell and B cell (RPMI 8226, KCLB No.10155), which are human immune cells. The cells were cultured in RPMI 1640 medium supplemented with 10% FBS at 5% CO 2 at 37 ° C. After the cells were adjusted to a concentration of 2.5 × 10 ⁴ cells in a 24-well plate, (0.5 μg / μL) were added to each well after 24 hours and then re-cultured for 8 days. Cells were counted using a hemocytometer in each well while culturing for 8 days. Immunoreactivity was measured according to cell growth and number of cells compared with control without sample extracts.

As a result of the stimulation of T cell and B cell growth of the human immune cells, the growth of the T cell and the B cell was promoted in the treatment with the bellflower fermented product. Especially , the fermentation of Bacillus amyloliquefaciens and the fermentation of Bacillus amyloliquefaciens Saccharomyces cerevisiae, Bacillus amyloliquefaciens and &lt; RTI ID = 0.0 &gt; Showed higher growth promotion in the mixed fermentations of the three strains of Lactobacillus plantarum (see Figs. 3A and 4A).

In addition, the growth of the T. cell and the B cell was also promoted in the Doraji fermentation tea, in particular tea fermented with Bacillus amyloliquefaciens and Saccharomyces cerevisiae, Bacillus amyloliquequa It was confirmed that the mixed cultivation of three strains of Bacillus amyloliquefaciens and Lactobacillus plantarum showed higher growth promotion (see FIGS. 3B and 4B).

<Example 4> Anti-obesity effect of tea prototype using 3T3-L1 cell

4-1. Preparation of 3T3-L1 cell

3T3-L1 cells were cultured in KCLB No. 10092.1 from Korean Cell Line Bank. Cells were cultured in DMEM containing 10% FBS in a 5% CO ₂ , 37 ° C incubator in a 75T-flask. After 3 to 4 days, the cells were treated with 0.05% Trypsin / 0.53 mM EDTA to separate the cells. The cells were collected at a centrifuge (2000 rpm, 2 min), and the cell density was diluted to 1 × 10 ⁴ cells / ml. and cultured in 1 mL increments. The cells were changed into new DMEM culture medium containing 10% FBS once every 2 days. After 3 to 4 days, the cells were fused. After 2 days, DMEM culture medium was supplemented with 5 mg / mL insulin, 0.25 mM dexamethasone and 0.5 mM IBMX Differentiation was induced. After 2 days, the cells were replaced with 5 mg / mL insulin alone and changed into new culture once every 2 days (Mac Dougald OA, Hwang CS, Fan H., Lane MD 1995). On day 6, cells were cultured in the absence of insulin, and after 7 days of incubation, the samples were treated with Oil red O staining 24 hours later.

4-2. Cytotoxicity measurement

MTT (methylthiazolyldiphenyltetrazolium bromide) assay for measuring cell viability was performed according to Sladowski et al. The cultured 3T3-L1 cells were plated at 1 × 10 ⁵ cells / well on a 96-well plate and cultured for 16-18 hours. The diluted sample extracts were treated for 24 hours. MTT solution dissolved at 5mg / ml was added to 10uL / 990ul medium and incubated at 37 ° C for 3 hours in 5% CO ₂ . After the culture was completed, the culture solution was removed, and the mixture was added to 200 μL / well of DMSO, stirred for 20 minutes, and absorbance was measured at 595 nm using an ELISA reader.

3T3-L1 cells showed the highest cell survival rate (Fig. 5A) in the group treated with 400 μg / ml of lactic acid bacterium in the group treated with the platycodon fermentation product (Fig. 5A) The highest cell viability was obtained when the strain fermented at 800 μg / ml (FIG. 5B). Although the cell survival rate did not increase in a concentration-dependent manner depending on the treatment concentration of each sample, overall cytotoxicity tended to be low.

4-3. Oil red O dyeing

The cell culture medium was discarded and 50 μL of 10% formaldehyde was added to each well to fix the cells, followed by incubation at 4 ° C. for 1 hour. After that, the formaldehyde was discarded and washed three times with PBS. Then, a solution of 0.25 g of Oil red O staining solution (Oil red O dissolved in 50 mL of isopropyl alcohol was mixed with distilled water at a ratio of 3: 2, filtered through a 0.45 μm filter Solution) was added to each well, and the plate was stained again at room temperature for 1 hour and then washed three times with PBS. The stained cells were observed under a microscope. After observation, the stained dye in the adipocyte was extracted with 500 μL of isopropyl alcohol per well and the absorbance was measured at 520 nm with a spectrophotometer.

The oil red O staining of 3T3-L1 cells showed the lowest lipid accumulation rate in the fermented product of Saccharomyces cerevisiae in the bellflower fermented product (see FIG. 6A) Showed the lowest lipid accumulation rate in Bacillus amyloliquefaciens bacterial fermentation tea (see FIG. 6B).

In addition, as a result of staining of 3T3-L1 cells with Oil red O, it was confirmed that in the fermented broth of Saccharomyces cerevisiae , the production of fat globules was reduced and the differentiation into fat cells was suppressed in the fermented platycodon (See FIG. 7), it was confirmed that the production of Bacillus amyloliquefaciens and Lactobacillus plantarum fermentation tea was reduced and the differentiation into adipocytes was inhibited in the tea using the bellflower fermented product (See Fig. 8).

<Example 5> Fat and carbohydrate inhibitory activity assay

5-1. α-Amylase inhibitory activity

Α-Amylase inhibitory activity was evaluated in order to investigate the blood glucose lowering effect of the fermented platycodon. The α-Amylase inhibitory activity was determined by dissolving 5 g of agar and 5 g of soluble starch in 500 mL of distilled water and sterilizing the mixture. 20 μl of 20 mg / ml plant extract and α-amylase were placed on a paper disc and incubated at 37 ° C. for 24 hours. I2 / KI (5 mM I2 in 3% KI ) Was added for 5 minutes. After the color was developed for 15 minutes, the area was measured by measuring the radius of the clear zone and calculating the inhibition rate as πr ² . Acarbose was used as a positive control and water and methanol as a negative control were used.

As a result of the assay of α-amylase inhibitory activity on the berry cultivated and non-fermented tea, the berry cultivars showed relatively high inhibitory activity in Bacillus amyloliquefaciens and Lactobacillus plantarum (See FIG. 9A). The bellflower fermented tea showed relatively high inhibitory activity in Lactobacillus plantarum and non-fermented group (see FIG. 9B), and both of the bellflower fermented tea and the non- Showed the lowest inhibitory activity in Saccharomyces cerevisiae fermentation tea (see FIG. 9).

5-2. α-Glucosidase inhibitory activity

The inhibitory activity of α-glucosidase was evaluated in order to investigate the blood glucose lowering effect of the fermented platycodon. The α-glucosidase inhibitory activity was determined by measuring the absorbance at 405 nm by mixing the plant extract at a concentration of 20 mg / mL, 100 μL of 0.15 U / mL α-glucosidase enzyme and 720 μL of 0.2 M potassium phosphate buffer (pH 6.8) And 100 μL of 5 mM 4-nitrophenyl-α-D-glucopyranoside (pNPG) was added and reacted at room temperature for 10 min. The absorbance was measured at the same wavelength and enzyme inhibition activity was calculated from the change in absorbance.

As a result of assaying α-glucosidase inhibitory activity on the strains of fermentation broth and non-fermentation broth, there were no significant differences according to fermentation strains in both the broth fermentation broth and broth fermentation broth, It was confirmed that the inhibitory activity was relatively high in the fermented tea (see Fig. 10).

5-3. Pancreatic lipase inhibitory activity

In order to investigate the anti - obesity effect of the fermented Doraji, lipase inhibitory activity, which plays an important role in fat absorption, was evaluated. Porcine pancreatic lipase (PPL, type II) activity was measured using p-nitrophenyl butyrate (p-NPB) as a substrate. First, the PPL stock solution was prepared to a concentration of 1 mg / mL in 0.1 mM potassium phosphate (pH 6.0), and stored at -20 ° C. To measure lipase inhibitory activity, 25 μL of extract (20 mg / mL) and 25 μL of PPL solution were added to 950 μL of 0.1 mM potassium phosphate (pH 7.2, 0.1% Tween 80) and incubated at 30 ° C. for 1 hour. NPB was added and reacted at 30 ° C for 5 minutes, and the absorbance was measured at 405 nm.

     Inhibition activity (%) = [1- (B-C) / A] X100)

A: Absorbance without added sample

B: Absorbance with addition of sample

C: Absorbance without addition of enzyme

The results of the porcine pancreatic lipase inhibition test for the fermented tea and the non fermented tea showed the highest inhibitory activity in the fermented tea without the fermentation and the relatively high inhibitory activity in the mixed fermented tea in the fermented tea. . In particular, Bacillus amyloliquefaciens fermentation tea showed the lowest inhibitory activity, indicating that there was a difference in the inhibitory activity depending on the fermentation strain (see FIG. 11).

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (13)

A composition for preventing or treating a bronchial disease, comprising a bellflower fermented product as an active ingredient. The method according to claim 1,
Wherein said bronchial disease is selected from the group consisting of cold, flu, tonsillitis, acute bronchitis, chronic bronchitis, pneumonia, bronchial asthma, nasopharyngitis, otitis media, tonsillitis and adenoid disease. A composition for enhancing immunity comprising a bellflower fermented product as an active ingredient. An anti-obesity composition comprising a bellflower fermented product as an active ingredient. The method of any one of claims 1, 3, and 4,
The above-mentioned bellflower fermented product is fermented using one or more microorganisms selected from the group consisting of Saccharomyces cerevisiae, Bacillus amyloliquefaciens and Lactobacillus plantarum. &Lt; / RTI &gt; The method of any one of claims 1, 3, and 4,
Wherein the bellflower fermented product is fermented using Saccharomyces cerevisiae, Bacillus amyloliquefaciens, and Lactobacillus plantarum . Heat-treating the cut bellflower to mellow the bellflower;
Inoculating the fermented broth to the cooked broth; And
And fermenting the inoculated bellflower. Heat-treating the cut bellflower to mellow the bellflower;
Inoculating the fermented broth to the cooked broth;
Fermenting the inoculated platycodon;
Naturally drying the fermented broth; And
And aging the naturally dried bellflower. 9. The method according to claim 7 or 8,
Wherein the bellflower mashing step is heat-treated at 115 to 125 DEG C for 15 to 25 minutes in a magnet. 9. The method according to claim 7 or 8,
Wherein the step of inoculating the fermenting microorganism is carried out after cooling the cooked broth to 35 to 40 占 폚. 9. The method according to claim 7 or 8,
Characterized in that the fermenting bacteria are at least one microorganism selected from the group consisting of Saccharomyces cerevisiae, Bacillus amyloliquefaciens and Lactobacillus plantarum. Way. 9. The method according to claim 7 or 8,
Wherein the inoculated bellflower is fermented in an incubator at 33 to 37 DEG C for 6 to 8 days. 9. The method of claim 8,
The natural drying step is naturally dried at room temperature for 1 to 3 days,
Wherein the aging step is carried out at room temperature for 6 to 8 days.

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