KR20230166478A - Composition for protecting stomach and intestines comprising Wongam fermented product as effective component - Google Patents

Abstract

The present invention relates to a composition for gastrointestinal protection containing a fermented persimmon product as an active ingredient, wherein the fermented persimmon product has an excellent effect in inhibiting cell proliferation caused by Helicobacter pylori bacteria compared to a raw persimmon extract, and inhibits cell proliferation caused by Helicobacter pylori bacteria. Since it has the effect of suppressing the production of NO and IL-8, the composition of the present invention containing fermented raw persimmon as an active ingredient can be usefully used as a health functional food for gastrointestinal protection, a feed additive, or a treatment for gastrointestinal diseases. .

Description

Composition for protecting stomach and intestines comprising Wongam fermented product as effective component}

The present invention relates to a composition for gastrointestinal protection containing fermented persimmon as an active ingredient.

Gastritis is an acute inflammation accompanied by infiltration of polymorphonuclear leukocytes in the superficial layer of the mucous membrane and is caused by causes such as drinking, painkillers, viral infection, bacterial food poisoning, eating hot food, and irregular meals. In acute cases, ulcers can be treated with mental and physical stability, diet, and drug therapy, but the recurrence rate is high and cure is difficult, so they may progress to chronic ulcers and cause complications such as gastrointestinal bleeding, pyloric stenosis, and perforation.

Among the various causes of gastritis, Helicobacter pylori ( H. pylori ) causes chronic gastritis and is known to be a representative cause of gastric mucosal damage. It is known to be caused by virulence factors (cytotoxin, urease, ammonia, etc.) that act locally by Helicobacter pylori and by the involvement of phagocytes that produce reactive oxygen metabolites. It is known to cause not only gastritis but also gastritis. It causes stomach cancer.

In Korea, the Helicobacter pylori infection rate is relatively high at 60-70%, and Helicobacter pylori is associated with gastritis, gastric ulcers, and stomach cancer. The importance of treatment for Helicobacter pylori infection is being emphasized as it is related to prevention of recurrence of ulcers, possibility of curing lymphoma, and relief of abdominal pain.

Recently, resistant H. pylori that does not respond to standard antibiotic therapy has emerged, and research to treat it is continuing. Accordingly, there is an increasing need for research on improving gastric health, which can prevent H. pylori infection, which is involved in the development of various diseases, just by consuming agricultural products at the level of daily intake.

Meanwhile, original persimmon was newly developed by the Rural Development Administration to complement the shortcomings of Manchurian licorice ( G. uralensis ) and European licorice ( G. glabra ), which are plants of licorice origin, after collecting and evaluating the characteristics of domestic and foreign genetic resources over a period of about 9 years. As a variety, the stems are straighter and thicker than Manchurian licorice, making it resistant to falling. The root length, thickness, and number of roots are greater than Manchurian licorice, and the yield is 359kg/10a, which is 2.27 times more than Manchurian licorice.

Wongam, a new variety of licorice, is a variety with excellent yield and cultivation safety, so if research is conducted on its functionality, it is expected to contribute to increasing farm income.

Meanwhile, as a prior study related to gastrointestinal protection compositions, Korea Patent No. 1026524 discloses the gastrointestinal protection effect of a herbal medicine composition containing herbal extracts of Uiryeongseon, Cheonhwabun, and Hedgehog herb, and Korean Patent Publication No. 2014-0001394 discloses the turmeric extract. Although a composition for gastrointestinal diseases caused by Helicobacter pyroni as an active ingredient has been disclosed, there has been no disclosure at all regarding a composition for gastrointestinal protection containing the fermented persimmon product of the present invention as an active ingredient.

The present invention was developed in response to the above-mentioned needs, and provides a gastrointestinal protection composition containing fermented persimmon as an active ingredient, wherein the fermented persimmon inhibits cell proliferation caused by Helicobacter pylori bacteria and reduces inflammation-related factors. The present invention was completed by confirming that the production of was suppressed.

In order to solve the above problems, the present invention provides a health functional food composition for gastrointestinal protection containing fermented persimmon as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for preventing or treating gastrointestinal diseases containing fermented persimmon as an active ingredient.

In addition, the present invention provides a feed additive for gastrointestinal protection containing fermented raw persimmon as an active ingredient.

The present invention relates to a composition for gastrointestinal protection containing a fermented persimmon product as an active ingredient, wherein the fermented persimmon product has an excellent effect in inhibiting cell proliferation caused by Helicobacter pylori bacteria compared to a raw persimmon extract, and inhibits cell proliferation caused by Helicobacter pylori bacteria. It has the effect of suppressing the production of NO (Nitric Oxide) and IL-8.

Figure 1 shows the results of confirming the cell proliferation inhibition effect according to the treatment of the fermented persimmon product (WGBB) and the persimmon extract (WG) of the present invention. Cell proliferation was induced by treatment with Helicobacter pylori. * means that the cell survival rate of the Wongam fermentation treatment group was statistically significantly reduced compared to the control group inoculated with Helicobacter pylori to induce cell proliferation, p<0.05.
Figure 2 shows the results of confirming the NO production inhibition effect according to the treatment of the fermented persimmon product (WGBB) and the persimmon extract (WG) of the present invention. NO production was induced by treating Helicobacter pylori. * indicates a statistically significant decrease in NO production in the Wongam fermented product treatment group compared to the control group in which NO production was induced by inoculation with Helicobacter pylori, p<0.05.
Figure 3 shows the results confirming the effect of inhibiting IL-8 production according to the treatment of fermented raw persimmon (WGBB) of the present invention. IL-8 production was induced by treatment with Helicobacter pylori. # indicates a statistically significant increase in IL-8 production in the control group inoculated with Helicobacter pylori compared to the untreated control group, p<0.05. * indicates a statistically significant decrease in IL-8 production in the Wongam fermented product treatment group compared to the control group in which IL-8 production was induced by inoculation with Helicobacter pylori, p<0.05.

In order to achieve the object of the present invention, the present invention provides a health functional food composition for gastrointestinal protection containing fermented persimmon as an active ingredient.

In one embodiment of the present invention, the raw persimmon fermented product may be fermented by inoculating raw persimmon with Bifidobacterium, but is not limited thereto.

The gastrointestinal protection includes, but is not limited to, protecting the stomach from abnormal cell proliferation and inflammation caused by infection with Helicobacter pylori.

The composition is preferably manufactured in a dosage form selected from powder, granule, pill, tablet, capsule, candy, syrup and beverage, but is not limited thereto.

When using the health functional food composition of the present invention as a food additive, the fermented raw material can be added as is or used together with other foods or food ingredients, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on its purpose of use (prevention, health, or therapeutic treatment). Generally, when producing a food or beverage, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw materials. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range. There are no special restrictions on the types of foods above. Examples of foods to which the extract or its fraction can be added include meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, and tea. , drinks, alcoholic beverages, and vitamin complexes, etc., and includes all health functional foods in the conventional sense.

When the composition of the present invention is used as a health drink, it may contain various flavoring agents or natural carbohydrates as additional ingredients like conventional drinks. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as textrin and cyclotenstrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As a sweetener, natural sweeteners such as thaumatin and stevia extract or synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 g of the composition of the present invention. The composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal neutralizer, pH adjuster, stabilizer, preservative, glycerin, alcohol, carbonic acid. It may contain carbonating agents used in beverages. In addition, the composition of the present invention may contain pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These ingredients can be used independently or in combination. Although the ratio of these additives is not very important, the composition of the present invention is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight.

In addition, the present invention provides a pharmaceutical composition for preventing or treating gastrointestinal diseases containing fermented persimmon as an active ingredient.

The gastrointestinal disease may be caused by Helicobacter pylori, and is preferably one selected from gastritis, gastric ulcer, and stomach cancer caused by Helicobacter pylori, but is not limited thereto.

The composition of the present invention is preferably prepared in any one formulation selected from capsules, powders, granules, tablets, suspensions, emulsions, syrups, and aerosols, but is not limited thereto.

The composition of the present invention may further include pharmaceutically acceptable carriers, excipients, or diluents in addition to the above active ingredients, and may be in various oral or parenteral dosage forms. When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include capsules, powders, granules, tablets, pills, etc. These solid preparations contain one or more compounds and at least one excipient, such as starch, calcium carbonate, sucrose, or lactose ( It is prepared by mixing lactose, gelatin, etc. Additionally, in addition to simple excipients, lubricants such as magnesium stearate, talc, etc. are also used. Liquid preparations for oral administration include suspensions, emulsions, syrups, and aerosols. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspension solvents may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurel, glycero gelatin, etc. can be used. When administering parenterally, it is preferable to choose external dermal application or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine dura, or intracerebrovascular injection.

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat the disease with a reasonable benefit/risk ratio applicable to medical treatment, and the level of the effective amount is determined by the type, severity, and activity of the patient's disease. , can be determined based on factors including sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, drugs used simultaneously, and other factors well known in the field of medicine. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art.

The dosage of the composition of the present invention varies depending on the patient's weight, age, gender, health condition, diet, administration time, administration method, excretion rate, and severity of disease. The composition of the present invention can be used alone or in combination with surgery, radiation therapy, hormone therapy, chemotherapy, and methods using biological response modifiers.

In addition, the present invention provides a feed additive for gastrointestinal protection containing fermented raw persimmon as an active ingredient.

The feed additive of the present invention corresponds to supplementary feed under the Feed Management Act. In the present invention, the term 'feed' may mean any natural or artificial diet, meal, etc., or a component of the meal, for or suitable for eating, ingestion, and digestion by animals. The type of feed is not particularly limited, and feed commonly used in the art can be used. Non-limiting examples of the feed include plant feeds such as grains, roots and fruits, food processing by-products, algae, fiber, pharmaceutical by-products, oils and fats, starches, cucurbits or grain by-products; Examples include animal feeds such as proteins, inorganic substances, fats and oils, minerals, oils and fats, single-cell proteins, zooplanktons or food. These may be used alone or in combination of two or more types.

Hereinafter, the present invention will be described in more detail using manufacturing examples and examples. These manufacturing examples and examples are only for illustrating the present invention in more detail, and it is obvious to those skilled in the art that the scope of the present invention is not limited thereto.

Preparation Example 1. Preparation of raw persimmon extract

After adding 10 parts by weight of distilled water per 1 part by weight of dried raw material, it was extracted at 100°C for 4.5 hours, concentrated at 70°C for 3 hours, and dried at 60°C for 24 hours to be used as a sample for the experiment.

Preparation Example 2. Preparation of raw persimmon fermented product

1×10 ⁷ CPU of Bifidobacterium , 3 g of dried raw persimmon, and 1.5 g of glucose were added to 30 mL of distilled water, and then fermented for 24 hours at 37°C and 5% CO ₂ conditions. After fermentation, the fermented product was filtered using filter paper, then freeze-dried and used as a sample for the experiment.

Example 1. Inhibitory effect on gastric epithelial cell proliferation caused by Helicobacter pylori of Wongam fermented product

Human gastric epithelial cell line (AGS cell) was purchased from the Korea Cell Line Bank. Cells were cultured in RPMI 1640 medium containing 10% (v/v) FBS, infected with Helicobacter pylori bacteria at a ratio of 200:1, washed three times with PBS, and incubated for 24 hours. Raw persimmon extract or raw persimmon fermentation product was treated separately.

Cell viability was measured using the WST-1 analysis method. After culturing cells in a 96-well plate at a concentration of 2×10 ⁴ cells/well for 24 hours, WST-1 analysis solution was added and cultured for 1 hour. The number of surviving cells was confirmed by measuring absorbance at 450 nm.

As a result, as shown in Figure 1, it was confirmed that the proliferation of AGS cells was promoted by Helicobacter pylori bacteria and that cell proliferation was inhibited by treatment with raw persimmon extract or raw persimmon fermentation product. In addition, the cell proliferation inhibition effect of fermented raw persimmon product was superior to that of raw persimmon extract.

Example 2. Inhibitory effect on NO (Nitric Oxide) production caused by Helicobacter pylori in fermented raw persimmon products

To confirm the ability of raw persimmon extract and raw persimmon fermentation product to inhibit NO production, AGS cells were inoculated into a 6-well plate at a concentration of 1 × 10 ⁶ cells/well and cultured for 24 hours at 37°C and 5% CO ₂ conditions. Afterwards, the supernatant was removed, washed three times with PBS, and the raw persimmon extract or fermented persimmon was treated at different concentrations in RPMI 1640 medium without antibiotics, and then cultured at 37°C and 5% CO ₂ for 30 minutes.

AGS cells were inoculated with Helicobacter pylori at a ratio of 300:1, cultured at 37°C, 5% CO ₂ for 24 hours, treated with Griess reagent, and NaNO ₂ was used as a standard. The absorbance was measured at 540 nm.

As a result, as shown in Figure 2, when AGS cells induced to produce NO by Helicobacter pylori were treated with raw persimmon extract or raw persimmon fermented product, the effect of suppressing NO production was excellent, and the effect of suppressing NO production in the raw persimmon extract at 1 mg/mL was excellent. While the NO production reduction effect was about 18%, the same amount of raw persimmon fermented product showed a NO production reduction effect of about 21%, confirming that the NO production inhibition effect of the raw persimmon fermented product was superior to that of the original persimmon extract.

Example 3. Inhibitory effect on IL-8 production caused by Helicobacter pylori in fermented raw persimmon products

AGS cells were inoculated into a 24-well plate at a density of 2×10 ⁵ cells/well, cultured for 24 hours, and then the cells were washed with PBS, and then the raw fermented product was treated with the AGS cells at each concentration for 30 minutes. Afterwards, the cells were inoculated with Helicobacter pylori at a ratio of 200:1 and cultured for 24 hours. The supernatant was collected, centrifuged at 2,000 rpm for 10 minutes, and the supernatant was used to measure IL-8 concentration. IL-8 concentration was analyzed according to the protocol using an ELISA kit.

AGS cells infected only with Helicobacter pylori were used as a positive control, and standard IL-8 was serially diluted to the highest concentration of 250 pg/mL, and then a calibration curve was prepared.

As a result, as shown in Figure 3, when AGS cells induced to produce IL-8 by Helicobacter pylori are treated with the fermented persimmon product, there is an effect of suppressing IL-8 production.

Claims (7)

A health functional food composition for gastrointestinal protection containing fermented persimmon as an active ingredient. The health functional food composition for gastrointestinal protection according to claim 1, wherein the fermented raw persimmon product is fermented by inoculating the raw persimmon with Bifidobacterium. The health functional food composition for gastrointestinal protection according to claim 1, wherein the gastrointestinal protection protects the stomach from abnormal cell proliferation and inflammation caused by Helicobacter pylori infection. A pharmaceutical composition for preventing or treating gastrointestinal diseases containing fermented persimmon as an active ingredient. The pharmaceutical composition for preventing or treating gastrointestinal diseases according to claim 4, wherein the gastrointestinal diseases are caused by Helicobacter pylori. The pharmaceutical composition for preventing or treating gastrointestinal diseases according to claim 5, wherein the gastrointestinal disease is any one selected from gastritis, gastric ulcer, and gastric cancer caused by Helicobacter pylori. A feed additive for gastrointestinal protection containing fermented persimmon as an active ingredient.