KR20220166593A - A composition for improving the intestinal microbial environment comprising the extract of herbal mixture - Google Patents

Abstract

The present invention relates to a prebiotics composition including a mixed extract of Phellodendron amurense Ruprecht, Schizonepeta tenuifolia, Asparagus cochinchinensis, Asparagus cochinchinensis, Asparagus cochinchinensis, and Asparagus cochinchinensis or a fraction thereof, and a food and feed composition for improving intestinal function including the composition. The prebiotics composition of the present invention can promote intestinal health by promoting the growth of beneficial bacteria in the intestine and inhibiting harmful bacteria in the intestine.

Description

Composition for improving the intestinal microbial environment of herbal medicine mixture extract {A composition for improving the intestinal microbial environment comprising the extract of herbal mixture}

The present invention is yellow baek, hyeonggae, gosam, licorice, maekmundong and It relates to a prebiotics composition containing a mixed extract or a fraction thereof of dry hwangsa, and a food and feed composition for improving intestinal function containing the composition.

Probiotics are living microorganisms that can beneficially function by maintaining the balance of intestinal flora, and are known to have a variety of functions. For example, probiotics are known to have beneficial functions to the human body, such as inhibiting the growth of harmful bacteria through competition for intestinal nutrients and barrier attachment sites, activating immunity, and reducing lactose intolerance.

In order to enhance these beneficial functions, studies on prebiotics, which are main components used to more effectively proliferate and maintain probiotics, have also been continued. As a representative example of prebiotics, fructo-oligosaccharides (FOS) are known. Fructooligosaccharide is an oligosaccharide in the form of β-1, 2-linked fructose molecules to sugar and can be fermented by Lactobacillus and Bifidobacteria, so ingestion of FOS can increase Bifidobacterium and Lactobacillus species. It is known that it can

However, it is known that adverse effects may be caused by harmful bacteria and non-probiotics capable of metabolizing FOS in the intestine, so the development of prebiotics compositions that can more effectively proliferate and maintain probiotics is needed. is being demanded Accordingly, prebiotics using plants and containing grape seed powder have been studied (Korean Patent Registration No. 10-1973514), but prebiotics using herbal complex extracts have not been developed.

Under this background, the present inventors have made diligent efforts to develop a more effective prebiotics composition. As a result, the present invention has been completed by discovering the herbal composite extract of the present invention as an active ingredient of the prebiotics composition and confirming its effect.

One object of the present invention is yellow baek, hyeonggae, gosam, licorice, maekmundong and It is to provide a prebiotics composition comprising a mixed extract or a fraction thereof of dry yellow.

Another object of the present invention is to provide a food composition for improving intestinal function comprising the prebiotics composition.

Another object of the present invention is to provide a feed composition for improving intestinal function comprising the prebiotics composition.

Hereinafter, the contents of the present invention will be described in detail. On the other hand, the description and embodiment of one aspect disclosed in the present invention can also be applied to the description and embodiment of another aspect with respect to common matters. Additionally, all combinations of the various elements disclosed herein fall within the scope of the present invention. In addition, it cannot be seen that the scope of the present invention is limited by the specific descriptions described below.

One aspect of the present invention is yellow baek, hyeonggae, gosam, licorice, macmundong and Provided is a prebiotics composition comprising a mixed extract of dry yellow or a fraction thereof.

In the present invention, the mixed extract of Hwangbaek, Hyeonggae, Gosam, Licorice, Maemundong and Geonjihwang may be mixed with "KAJD" or "KHU-ATO-JIN-D". The mixed extract may be an extract of a mixture of hwangbaek, hyeonggae, gosam, licorice, maekmundong, and geonhwang extract, and may also be a mixture of hwangbaek, hyeonggae, gosam, licorice, maemundong, or geonhwang extract.

The term of the present invention, "yellow baek" (黃柏) is a dried bark of the yellow baek tree, and has a crude drug name Phellodendri Cortex and a scientific name Phellodendron wilsonii . It is characterized by its bitter taste and cold nature, and is known to be used for nerves, cystoscopy, colonoscopy, and cardiomyoscopy.

The term of the present invention, "Hyeonggae" (荊芥) is a dried whole plant of Hyeonggae, a Lamiaceae plant, and has the herbal name Schizonepetae Spica and the scientific name Schizonepeta tenuifolia . It is characterized by its bitter taste and warm nature, and is known to act on menopause and liver cirrhosis, and to exhibit digestive and antibacterial effects.

The term of the present invention, "gosam" (苦蔘) is the root of the skin of gosam, and has the herbal name Sophorae Radix and the scientific name Sophora flavescens . It has a peculiar smell, is persistent, and is characterized by very bitter and cold weakness.

The term of the present invention, "licorice" (甘草) is dried roots or stems of licorice, and has the herbal name Glycyrrhizae Radix and the scientific name Glycyrrhiza glabra . It has a characteristic odor and a sweet taste. Licorice is known to harmonize the toxicity of all medicines so that the medicinal effects appear well, control the heat and morale of the books, improve the communication of all blood vessels, and strengthen muscles and bones.

The term of the present invention, "Geonjihwang" (乾地黃) is a dried rhizome of Rehmanniae, a plant of the ginseng family, and has the herbal name Radix Rehmanniae Exsiccat or Rehmanniae Radix, scientific name Rehmanniae Exsiccat or Rehmannia adj glutinosa . It is characterized by its sweet taste and cold nature, and has diuretic and hypoglycemic action, and is known to be used for uterine bleeding, menstrual disorders, constipation, and general weakness.

Mixed extracts or fractions of yellow baek, hyeonggae, gosam, licorice, licorice root, and geonhwang have never been used for prebiotics purposes, and it was first discovered by the present inventors that there is a use for prebiotics.

In the present invention, the hwangbaek, hyeonggae, goginseng, licorice, maekmundong, and geonhwang may be purchased and used commercially, or those collected or cultivated in nature may be used, but are not limited thereto.

The term "extract", as used herein, refers to an extract obtained by extracting a mixture of yellow baek, myeonggae, gosam, licorice, coriander, and dry yellow, a dilution or concentrate of the extract, a dried product obtained by drying the extract, and a crude product of the extract It includes extracts of all formulations that can be formed using extracts themselves and extracts, such as extracts, purified products, or mixtures thereof.

The extracts of hwangbaek, hyeonggae, sorghum, licorice, rhubarb, and dried yellow can be extracted from various organs of natural, hybrid, and mutant plants, for example, roots, aerial parts, stems, leaves, flowers, fruit bodies, and fruit It can be extracted from plant tissue culture as well as from the skin.

Extracts or fractions of hwangbaek, hyeonggae, sorghum, licorice, mungmundong, and geonhwang have never been used for prebiotics purposes, and it was first identified by the present inventors as having prebiotics uses.

Specifically, the mixture of yellow baek, hyeonggae, gosam, licorice, ginseng, and dried yellow is 0.5 to 1.5: 0.5 to 1.5: 0.5 to 1.5: 0.5 to 1.5: 0.5 to 1.5: 0.5 A weight ratio of 1.5:0.5 to 1.5, specifically a weight ratio of 0.7 to 1.3:0.7 to 1.3:0.7 to 1.3:0.7 to 1.3:0.7 to 1.3:0.7 to 1.3, more specifically 1:1:1:1:1:1 It may be included in a weight ratio of, but is not limited thereto.

The extract of the present invention may be extracted with at least one solvent selected from the group consisting of water, alcohol having 1 to 4 carbon atoms, and mixed solvents thereof by extracting a mixture of yellow baek, hyeonggae, gosam, licorice, coriander, and dried yellow, but is not limited thereto. don't

In the mixed extract of Hwangbaek, Hyeonggae, Gosam, Licorice, Maemundong and Geonjihwang, the method for extracting the mixture is not particularly limited, and can be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method include a hot water extraction method, an ultrasonic extraction method, a filtration method, a reflux extraction method, and the like, which may be performed alone or in combination of two or more methods.

In the present invention, the type of extraction solvent used to extract the mixture is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the extraction solvent may include water, alcohol, or a mixed solvent thereof, and these may be used alone or in combination of one or more, and specifically, water may be used. When alcohol is used as a solvent, alcohol having 1 to 4 carbon atoms may be specifically used.

As used herein, the term "fraction" refers to a product obtained by performing fractionation in order to separate a specific component or a specific component group from a mixture containing various components.

In the present invention, a fractionation method for obtaining the fraction is not particularly limited, and may be performed according to a method commonly used in the art. Non-limiting examples of the fractionation method include a solvent fractionation method performed by treating various solvents, an ultrafiltration fractionation method performed by passing an ultrafiltration membrane having a constant molecular weight cut-off value, and various chromatography (size, charge, hydrophobicity). or a chromatographic fractionation method that performs separation based on affinity), and a combination thereof. Specifically, a method of obtaining fractions from the extract by treating the extract obtained by extracting the maple leaves of the present invention with a predetermined solvent may be mentioned.

In the present invention, the type of fractionation solvent used to obtain the fraction is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the fractionation solvent include polar solvents such as water and alcohol having 1 to 4 carbon atoms; non-polar solvents such as hexane, ethyl acetate, chloroform, and dichloromethane; or mixed solvents thereof. These may be used alone or in combination of one or more, but are not limited thereto.

In addition, the extract or fraction may be prepared and used in the form of a dry powder after extraction, but is not limited thereto.

As used herein, the term "prebiotic or prebiotics" refers to a component that is used by microorganisms, including beneficial bacteria, to promote the growth or activity of microorganisms, thereby exhibiting a beneficial effect on the health of the host. can mean

In the present invention, the prebiotic may be one that improves the intestinal microflora to exhibit a beneficial effect on the host's health, and reduces the level of blood in the stool, reduces the level of mucus, reduces the length of the intestine, and improves the intestinal microbiome by improving the intestinal tissue. It may be to improve the environment, but is not limited thereto.

In the present invention, "improvement of intestinal flora" may promote the growth or growth of beneficial bacteria in the intestine and inhibit the growth or growth of harmful bacteria in the intestine, and may mean maintaining the balance between beneficial bacteria and harmful bacteria in the intestine.

The "intestinal beneficial bacteria" may collectively refer to microorganisms that have beneficial effects on the human body while inhabiting the intestines. For example, intestinal beneficial bacteria may include probiotics.

The "probiotics" may refer to microorganisms that have a good effect on health in the body, Lachnospiraceae , Rikenellaceae, Prevotellaceae , Acetobacter family ( Acetobacteraceae ) , Lactobacillaceae ( Lactobacillaceae ) ) , Lactobacillus , Firmicutes , Verrucomicrobia , Bifidobacterium , Akkermansia , and the like, but are not limited thereto.

The "intestinal harmful bacteria" may collectively refer to microorganisms that live in the intestine and have a detrimental effect on the human body , such as enteritis . Erysipelotrichaceae ) , Porphyromonasaceae ( Porphyromonadaceae ) , Bacteroides family ( Bacteroidaceae ) , Disulfo Vibrio family ( Desulfovibrionaceae ) , Pasteurella family ( Pasteurellaceae ) , Enterobacteriaceae ( Enterobacteriaceae ) Bacteroidetes ( Bacteroidetes ), Escherichia coli ( Escherichia coli ), Clostridium ( Clostridium ), Staphylococcus ( Staphylococcus ) and the like may be exemplified, but are not limited thereto.

The prebiotics composition of the present invention may include a mixed extract of hwangbaek, myeonggae, gosam, licorice, kimmundong, and geonhwang, and the mixed extract is to include 0.001 to 80% by weight based on the weight of the total prebiotics composition It can, but is not limited thereto.

For example, as a result of administering the KAJD oriental medicine mixed extract of the present invention to the intestinal microbial inhibition group and the enteric microbial inhibition deepening group, the bloody feces level, mucus level, and intestinal length were prolonged in the intestinal microbial inhibition group and the deep intestinal microbial inhibition group. It was confirmed that there was an effect of reducing, improving intestinal tissue, increasing beneficial bacteria in the intestine, and reducing harmful bacteria (FIGS. 4 to 11).

This suggests that the KAJD herbal mixture extract of the present invention improves the intestinal flora and the intestinal microbial environment.

Another aspect of the present invention provides a food composition for improving intestinal function comprising the prebiotics composition.

The prebiotics are as described in other embodiments.

The term "improvement of intestinal function" as used herein refers to any change in the state that improves or normalizes the overall function of the intestine, improves the intestinal flora, promotes the proliferation or growth of beneficial bacteria in the intestine, suppresses the proliferation or growth of harmful bacteria in the intestine, balances beneficial and harmful bacteria in the intestine It may contain any one or more of oils and fats.

[0025] Food-wise acceptable salts that can be included in the food composition of the present invention are useful acid addition salts formed from food-acceptable free acids or metal salts formed from bases. As an example, inorganic acids and organic acids may be used as free acids. Hydrochloric acid, sulfuric acid, hydrobromic acid, sulfurous acid, or phosphoric acid may be used as the inorganic acid, and citric acid, acetic acid, maleic acid, fumaric acid, gluconic acid, methanesulfonic acid, or the like may be used as the organic acid. In addition, as the metal salt, an alkali metal salt or an alkaline earth metal salt, sodium, potassium or calcium salt may be used. However, it is not limited thereto.

The food composition of the present invention includes forms such as pills, powders, granules, precipitates, tablets, capsules or liquids, and the food to which the composition can be added includes, for example, various foods, such as beverages, There are chewing gum, tea, vitamin complexes, and health supplements.

Ingredients that can be included in the food composition of the present invention, other than containing the prebiotic composition of the present invention as an essential ingredient, there are no particular restrictions on other ingredients, and, like conventional foods, various herbal extracts, food additives, or natural carbohydrates etc. can be contained as an additional component. The content of the active ingredient in the food composition may be appropriately determined depending on the purpose of use (prevention, improvement or therapeutic treatment). At this time, the content of the active ingredient included in the composition is not particularly limited thereto, but may include 0.0001% to 10% by weight, preferably 0.001% to 1% by weight, based on the total weight of the composition.

In addition, the food auxiliary additives may include food auxiliary additives common in the art, for example, flavoring agents, flavoring agents, coloring agents, fillers, stabilizers, and the like.

Examples of the natural carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrins, cyclodextrins, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. In addition to the above, natural flavors (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can advantageously be used as flavoring agents.

In addition to the above, the food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and fillers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like may be contained. In addition, it may contain fruit flesh for the manufacture of natural fruit juice, fruit juice beverages and vegetable beverages. These components may be used independently or in combination.

In the present invention, the health supplement may include health functional food and health food.

The health functional food (functional food) is the same term as food for special health use (FoSHU), and is a medicine processed to efficiently display bioregulatory functions in addition to nutritional supply, and has high medical effect. means food. Here, "function (sex)" means to obtain useful effects for health purposes such as regulating nutrients for the structure and function of the human body or physiological functions.

Food containing the food composition of the present invention can be prepared by a method commonly used in the art, and can be prepared by adding raw materials and components commonly added in the art during the preparation. In addition, the formulation of the food may also be prepared without limitation as long as the formulation is recognized as a food. The food composition of the present invention can be prepared in various types of formulations, and unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long time by using food as a raw material, and has excellent portability.

Another aspect of the present invention provides a feed composition for improving intestinal function comprising the prebiotics composition.

The prebiotics and intestinal function improvement are as described in other aspects.

In the present invention, the term "feed composition" may mean any natural or artificial diet, one meal, etc., or a component of the one meal for animals to eat, ingest, and digest, or suitable therefor, and is well known in the art. It can be manufactured in a variety of shapes.

The type of feed is not particularly limited, and feeds commonly used in the art may be used. Non-limiting examples of the feed include vegetable feeds such as grains, root fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal or grain by-products; Animal feed such as proteins, inorganic materials, oils, mineral oils, oils, single cell proteins, zooplankton, or food may be mentioned. These may be used alone or in combination of two or more. Here, the animal is a concept including livestock and pets.

The feed composition of the present invention may further include a binder, an emulsifier, a preservative, etc. added to prevent quality deterioration, and amino acids, vitamins, enzymes, probiotics, flavors, and non-protein nitrogen added to increase efficacy It may further include compounds, silicate agents, buffers, coloring agents, extractants, oligosaccharides, and the like, and may further include feed mixtures and the like, but is not limited thereto.

The prebiotics composition of the present invention can promote intestinal health by promoting the growth of beneficial bacteria in the intestine and inhibiting harmful bacteria in the intestine.

1 is a schematic diagram showing a method for constructing an animal model (ABX) inhibiting intestinal microorganisms and an animal model (ABX+DNCB) with more intensified inhibition of intestinal microorganisms.
Figure 2 is a diagram showing the change in body weight of the normal group, ABX group, DNCB group, ABX + DNCB group, and KAJD oriental composite extract intake group (ABX + DNCB + KAJD).
Figure 3 is a diagram showing the change in dietary intake of the normal group, ABX group, DNCB group, ABX + DNCB group, and ABX + DNCB + KAJD group.
4 is a diagram showing changes in blood levels in feces of the normal group, the ABX group, the DNCB group, the ABX+DNCB group, and the ABX+DNCB+KAJD group.
5 is a diagram showing changes in the level of mucus in feces of the normal group, the ABX group, the DNCB group, the ABX+DNCB group, and the ABX+DNCB+KAJD group.
6 is a diagram showing changes in blood and mucus levels in feces of the normal group, the ABX group, the DNCB group, the ABX+DNCB group, and the ABX+DNCB+KAJD group.
7 and 8 are diagrams showing changes in intestinal length in the normal group, the ABX group, the DNCB group, the ABX+DNCB group, and the ABX+DNCB+KAJD group.
9 is a diagram showing changes in the state of intestinal tissue in the normal group, the ABX group, the DNCB group, the ABX+DNCB group, and the ABX+DNCB+KAJD group.
10 and 11 are diagrams showing changes in intestinal microorganisms in the normal group, the ABX group, the DNCB group, the ABX+DNCB group, and the ABX+DNCB+KAJD group.

Hereinafter, the present invention will be described in more detail through examples. However, these Examples and Experimental Examples are intended to illustrate the present invention, and the scope of the present invention is not limited to these Examples and Experimental Examples.

Preparation Example: Preparation and administration method of KAJD herbal complex extract

KAJD was manufactured by Han Poong Pharmaceutical (Jeonju, Korea) according to good manufacturing practices (GMP). KAJD is Phellodendri Cortex , Schizonepetae Spica , Sophorae Radix , Licorice ( Glycyrrhizae Radix ), Liriopis Tuber ) and As a hot-water extract of a mixture containing dry sulfur ( Radix Rehmanniae Exsiccat ) in a weight ratio of 1: 1: 1: 1: 1: 1, after extracting from water and removing all solvents using a vacuum concentrator, freeze-drying to obtain an extract powder manufactured. The obtained powder was dissolved in negative water and used.

In the present invention, the effect of KAJD was confirmed by orally administering 200 mg/kg of KAJD herbal complex extract dissolved in drinking water to the group inhibiting intestinal microorganisms every day at the same time for 2 weeks.

Example 1: Intestinal microbial suppression animal model (ABX) and intestinal microbial suppression intensified animal model (ABX + DNCB) construction

In order to establish an animal model (ABX model) for inhibiting intestinal microbes, ampicillin, metronidazole, vancomycin, and neo Four types of antibiotics containing neomycin sulfate were mixed and fed for 7 days, and then fed with plain water every 3 days. Four types of antibiotics were induced to suppress intestinal microorganisms by self-supplying while gradually increasing the concentration for 4 weeks.

In order to construct an animal model (ABX+DNCB model) with enhanced inhibition of intestinal microbes, antibiotics and water were fed as in the above-described ABX model construction method, while the back skin of the mice was shaved and 100 μl of It was induced by applying 2% DNCB (2,4-dinitrochlorobenzene).

More specifically, the feeding and application methods for constructing the intestinal microbial suppression animal model (ABX model) and the intestinal microbial suppression intensified animal model (ABX + DNCB model) are schematically shown in FIG. 1.

Example 2: Toxicity evaluation of KAJD

In order to confirm the presence or absence of toxicity due to ingestion of the KAJD herbal composite extract, the body weight and food intake of the mice ingested the KAJD herbal composite extract of the above-described preparation were checked.

Specifically, KAJD herbal composite extract dissolved in drinking water was orally administered daily at the same time for 2 weeks (ABX+DNCB+KAJD) to the group with enhanced inhibition of intestinal microbes (ABX+DNCB). As a result, it was confirmed that there was no significant toxicity of KAJD, since there was only a slight difference in body weight and dietary changes within the appropriate error range (FIG. 2 and FIG. 3).

Example 3: Confirmation of the prebiotics effect of KAJD

Example 3-1: Confirmation of improvement in blood stool level and condition in the intestine of KAJD

In order to evaluate the prebiotics effect of the KAJD oriental medicine mixed extract, the level of blood in the intestine and whether the condition was improved were confirmed.

Specifically, feces were collected from the normal group, the intestinal microorganism suppression group (ABX), the intestinal microorganism suppression enhanced group (ABX+DNCB), and the KAJD oriental medicine mixed extract administration group (ABX+DNCB+KAJD). After fixing with formaldehyde, the tissue was planted in a paraffin block to prepare a slide with paraffin sections.

A Hemoccult kit was used to visually check the inflammation or blood levels of stool samples prepared as slides. At this time, when 2 drops of the Hemoccult kit solution were added to the stool sample, the higher the degree of blue color change, the more severe the degree of inflammation or bloody stool. In addition, stool consistency was used by more than 3 people to score the degree of viscosity and blood confirmation of mouse stool.

As a result, it was confirmed that blood and mucus levels were higher in the feces of the enhanced intestinal microbial suppression group (ABX+DNCB) than in the feces of the intestinal microbial suppression group (ABX), in which blood and mucus were confirmed. In addition, as a result of oral administration of KAJD oriental medicine mixed extract (ABX+DNCB+KAJD) targeting the group with enhanced inhibition of intestinal microorganisms (ABX+DNCB), blood and mucus levels were significantly reduced, confirming that the environment of intestinal microorganisms was improved. (Figs. 4 to 6).

Accordingly, it was confirmed that the mixed extract of KAJD oriental medicine decreased the level of blood and mucus in feces, which was increased by inhibition of intestinal microorganisms.

Example 3-2: Confirmation of the effect of KAJD on reducing intestinal length

Since it is known that the length of the intestine is extended due to the decrease in the function of the intestinal cells when the intestinal microorganisms are suppressed, the change in the intestinal length was confirmed to evaluate the prebiotics effect of the KAJD herbal mixture extract.

Specifically, the length of the intestines separated from the normal group (Normal), intestinal microbial suppression group (ABX), intestinal microbial suppression enhanced group (ABX + DNCB), and KAJD oriental medicine mixed extract administration group (ABX + DNCB + KAJD) were measured. did

As a result, compared to the normal group (Normal), the intestinal length was extended in both the intestinal microbial suppression group (ABX) and the intestinal microbial suppression enhanced group (ABX+DNCB), whereas when the KAJD herbal mixture extract was administered (ABX+ DNCB + KAJD), it was confirmed that the length of the intestine was reduced to a level similar to that of the normal group (FIGS. 7 and 8).

Accordingly, it was confirmed that the KAJD oriental medicine mixture extract reduced the length of the intestine, which was prolonged by inhibition of intestinal microorganisms.

Example 3-3: Confirmation of intestinal tissue improvement effect of KAJD

In order to evaluate the prebiotics effect of the KAJD oriental medicine mixed extract, it was confirmed whether or not the intestinal tissue was improved.

In order to observe the morphology and condition of the intestine, intestines were collected from the normal group (Normal), intestinal microbial suppression group (ABX), intestinal microbial suppression enhanced group (ABX+DNCB), and KAJD oriental medicine mixed extract administration group (ABX+DNCB+KAJD). A part of the tissue was separated, fixed with 4% paraformaldehyde (PFA), and then paraffin sections were prepared using paraffin (Paraplast High Melt, Surgipath, Leica, Biosystem, The Netherlands).

For the prepared intestinal samples, immunofluorescence staining using ZO-1 antibody and immunofluorescence staining using Occludin antibody were performed along with H&E (Hematoxylin and Eosin) staining, which is histological staining. Since ZO-1 and Occludin are proteins constituting tight junctions between intestinal cells, they were targeted to evaluate whether tight junctions are disrupted.

As a result, compared to the normal group (Normal), the suppression of intestinal microorganisms in both the intestinal microbial suppression group (ABX) and the intestinal microbial suppression group (ABX+DNCB) increased the number of villi and crypts in the intestinal tissue. It was confirmed that the form of villi and glands was restored to a level similar to that of the normal group when the KAJD herbal mixture extract was administered (ABX + DNCB + KAJD), while the morphology of cells was suppressed and the tight junction between enterocytes was suppressed. (FIG. 9).

Accordingly, it was confirmed that the KAJD herbal mixture extract restored the function of the intestinal tissue, which had been deteriorated by the suppression of intestinal microorganisms.

Example 3-4: Confirmation of the effects of KAJD on increasing the proportion of beneficial bacteria and reducing the proportion of harmful bacteria in the intestine

In order to evaluate the prebiotics effect of the KAJD oriental medicine mixed extract, changes in intestinal microflora were confirmed.

Specifically, feces collected from the normal group, the intestinal microbial suppression group (ABX), the intestinal microbial suppression enhanced group (ABX+DNCB), and the KAJD oriental medicine mixed extract administration group (ABX+DNCB+KAJD) were tested. Changes in the number of microorganisms were observed through 16s rRNA metagenome sequencing.

The number of microorganisms was evaluated for both beneficial ( Lactobaciilacea, Lachnospiraceae, Rikenellaceae, Prevotellaceae, Acetobacteraceae ) and harmful bacteria ( Clostridiaceae 1, Ruminococcaceae, Erysipelotrichaceae, Porphyromonadaceae, Bacteroidaceae, Desulfovibrionaceae, Pasteurellaceae, Enterobacteriaceae ).

As a result, it was confirmed that the total number of microorganisms was lower in both the intestinal microbial suppression group (ABX) and the intestinal microbial suppression enhanced group (ABX + DNCB) compared to the normal group (Normal). In particular, it was observed that the number of harmful bacteria, which are microorganisms related to inflammation, was higher in the group with enhanced inhibition of intestinal microorganisms (ABX+DNCB).

On the other hand, when the KAJD herbal mixture extract was administered (ABX + DNCB + KAJD), it was confirmed that the microbial flora appeared at a level similar to that of the normal group (FIGS. 10 and 11).

Accordingly, it was confirmed that the KAJD herbal mixture extract reduced the number of harmful bacteria in the intestine that had been increased by inhibiting intestinal microorganisms, increased the number of beneficial bacteria in the intestine, and restored the flora to a level similar to normal.

From the above results, it was confirmed that the mixed extract of KAJD oriental medicine has an effect as a prebiotic to improve the microbial environment in the intestine.

From the above description, those skilled in the art to which the present invention pertains will be able to understand that the present invention may be embodied in other specific forms without changing its technical spirit or essential features. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not limiting. The scope of the present invention should be construed as including all changes or modifications derived from the meaning and scope of the claims to be described later and equivalent concepts rather than the detailed description above are included in the scope of the present invention.

Claims (9)

Hwangbaek, Hyeonggae, Gosam, Licorice, Maekmundong and A prebiotics composition comprising a mixed extract of dry yellow or a fraction thereof.
The method of claim 1, wherein the extract is extracted with at least one solvent selected from the group consisting of water, alcohols having 1 to 4 carbon atoms, and mixed solvents thereof from a mixture of yellow baek, hyeonggae, gosam, licorice, licorice root, and dry yellow, Prebiotics composition.
The method of claim 1, wherein the fraction is prepared by mixing a mixture of yellow and white, hyeonggai, goginseng, licorice, coriander, and dry yellow in water, alcohol having 1 to 4 carbon atoms, hexane, ethyl acetate, chloroform, dichloroform A prebiotics composition prepared by fractionation with a solvent selected from the group consisting of methane (Dichloromethane) and mixed solvents thereof.
The method of claim 2, wherein the mixture comprises yellow baek, hyeonggae, gosam, licorice, coriander, and dry yellow in a weight ratio of 0.5 to 1.5: 0.5 to 1.5: 0.5 to 1.5: 0.5 to 1.5: 0.5 to 1.5: 0.5 to 1.5 Phosphorus, a prebiotics composition.
The prebiotics composition according to claim 1, wherein the prebiotics composition comprises a mixed extract of yellow baek, myeonggai, gosam, licorice, lycundong, and geonhwang in an amount of 0.001 to 80% by weight based on the weight of the total composition.
The prebiotics composition according to claim 1, wherein the prebiotics composition inhibits the growth of harmful bacteria in the intestine and promotes the growth of beneficial bacteria in the intestine.
The method of claim 6, wherein the intestinal harmful bacteria are Clostridiaceae 1 ( Clostridiaceae 1 ) , Rumino coccaceae ( Ruminococcaceae ) , Erysipelotrichaceae ( Erysipelotrichaceae ) , Porphyromonas family ( Porphyromonadaceae ) , Bacteroides family ( Bacteroidaceae ) , Disulfovibrionaceae ( Desulfovibrionaceae ) , Pasteurella ( Pasteurellaceae ) , and At least one selected from the group consisting of Enterobacteriaceae , and the intestinal beneficial bacteria are Lactobaciilacea , Lachnospiraceae , Rikenellaceae , Provotella ( Prevotellaceae ) , and Acetobacter family ( Acetobacteraceae) , which is at least one selected from the group consisting of, a prebiotics composition.
A food composition for improving intestinal function comprising the prebiotics composition of any one of claims 1 to 7.
A feed composition for improving intestinal function comprising the prebiotics composition of any one of claims 1 to 7.

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