Composition for alleviating or treating gastrointestinal diseases, comprising quercus acuta extract or fraction thereof

Abstract

The present invention relates to a composition for alleviating or treating gastrointestinal diseases, the composition comprising a Quercus acuta extract or a fraction thereof. The composition according to the present invention, for preventing, alleviating or treating gastric hyperacidosis, gastric mucosal damage, gastric ulcer, gastritis or gastroesophageal reflux disease, inhibits or restores gastric mucosal damage caused by excessive gastric acid secretion and has excellent gastric protection efficacy, and thus can be safely used without toxicity or side effects.

Description

Composition for improving or treating gastrointestinal diseases comprising red oak extract or fraction thereof

[0001] The present invention relates to a composition for improving or treating gastrointestinal diseases, comprising a natural extract as an active ingredient.

[0002]

[0003] Approximately 10% of the world's population suffers from digestive diseases, which are caused by factors such as excessive stress, drinking, smoking, and poor eating habits. These diseases manifest in various ways, including gastritis, gastric ulcers, reflux esophagitis, and stomach damage. The stomach stores food that enters the esophagus, breaks it down into easily digestible pieces, and regulates its passage into the duodenum. This coordinates with the secretion of pancreatic enzymes to ensure efficient digestion and absorption. When food enters the stomach, it secretes gastric acid, a strong acid, to help digest it. During this process, the gastric mucosa protects the lining from damage by the acid.

[0004] Hyperacidity refers to the condition of stomach acidity, which occurs when the stomach secretes excessive amounts of acid, resulting in stomach burning and pain. Gastric acid is a component of the gastric juice secreted for digestion and is highly acidic. If excessive amounts are produced, it can damage the stomach lining and irritate stomach cells, causing symptoms such as heartburn, belching, and stomach pain.

[0005] Hyperacidity can be caused by irregular eating habits such as overeating, binge eating, and late-night snacking; consuming spicy or salty foods, alcohol, cigarettes, and carbonated beverages that irritate the stomach; and tension and stress that induce gastric acid secretion. Gastritis refers to a condition in which the gastric mucosa is infiltrated by inflammatory cells and is classified as acute gastritis and chronic gastritis. Acute gastritis occurs when excessive gastric acid is secreted, while chronic gastritis is known to be caused by excessive stress. Furthermore, hyperacidity can cause various diseases such as reflux esophagitis, gastric ulcers, and duodenal ulcers. Medications used to treat gastritis caused by hyperacidity mainly reduce gastric acid and stress, which are factors that promote and recur in gastritis and gastric ulcers. Among these, drugs that suppress excessive gastric acid secretion are the most commonly used.

[0006] Treatments for gastritis and peptic ulcers include antacids, which neutralize excessive gastric secretions; histamine H2 receptor antagonists and proton pump inhibitors, which suppress gastric acid secretion; prostaglandins, which increase the stomach lining's resistance to digestive juices and suppress acid secretion; and gastric mucosal protective agents. In addition, steroidal and non-steroidal anti-inflammatory drugs are used to relieve pain, which is the main symptom of gastritis and peptic ulcers. However, their use is limited due to side effects such as gastrointestinal upset, rash, and urticaria, and a 50-80% relapse rate within one year of discontinuing medication. In particular, non-steroidal anti-inflammatory drugs such as aspirin and ibuprofen can worsen peptic ulcers, so their use is severely restricted in patients with ulcers, including those with other conditions, or they must be taken together with antacids. Therefore, there is a need to develop a natural product-based preventive and therapeutic material that is safe and effective and can alleviate pain, a major symptom of gastritis and peptic ulcers, and protect the stomach.

[0007]

[0008] The present invention provides a pharmaceutical composition for preventing or treating gastric diseases, which has excellent effects in preventing and recovering gastric damage and protecting the stomach.

[0009] The present invention provides a food composition for preventing or improving gastric diseases, which has excellent gastric protection effects and prevents and recovers gastric damage.

[0010]

[0011] 1. A pharmaceutical composition for preventing or treating gastrointestinal diseases, comprising an extract of red thorn tree or a fraction thereof.

[0012] 2. A pharmaceutical composition for preventing or treating gastrointestinal diseases, wherein in the above 1, the red thorn extract is at least one extract selected from the group consisting of the fruit, flower, leaf, stem, whole plant, branch, root and branch of the red thorn tree.

[0013] 3. A pharmaceutical composition for preventing or treating gastrointestinal diseases, wherein the red oak extract in the above 1 is an extract available in at least one solvent selected from the group consisting of water, alcohols having 1 to 4 carbon atoms, propanol, isopropanol, and butanol.

[0014] 4. In the above 1, the fraction is a pharmaceutical composition for preventing or treating gastrointestinal diseases, wherein the fraction is obtained by fractionating the red oak extract with at least one solvent selected from the group consisting of dichloromethane, hexane, chloroform, ethyl acetate, and butanol.

[0015] 5. A pharmaceutical composition for preventing or treating at least one gastric disease selected from the group consisting of hyperacidity, gastric mucosal damage, gastric ulcer, gastritis, and gastroesophageal reflux disease in the above 1.

[0016] 6. A pharmaceutical composition for preventing or treating gastric disease caused by excessive secretion of gastric acid in the above 1.

[0017] 7. A food composition for preventing or improving gastrointestinal diseases, comprising an extract of red thorn tree or a fraction thereof.

[0018] 8. A food composition for preventing or improving gastrointestinal diseases, wherein in the above 7, the red thorn extract is at least one extract selected from the group consisting of the fruit, flower, leaf, stem, whole plant, root, and branch of the red thorn tree.

[0019] 9. In the above 7, the red oak extract is a food composition for preventing or improving gastrointestinal diseases, which is an extract available in at least one solvent selected from the group consisting of water, alcohols having 1 to 4 carbon atoms, propanol, isopropanol, and butanol.

[0020] 10. In the above 7, the fraction is a food composition for preventing or improving gastrointestinal diseases, wherein the fraction is obtained by fractionating the red oak extract with at least one solvent selected from the group consisting of dichloromethane, hexane, chloroform, ethyl acetate, and butanol.

[0021] 11. A food composition for preventing or improving at least one gastric disease selected from the group consisting of hyperacidity, gastric mucosal damage, gastric ulcer, gastritis, and gastroesophageal reflux disease in the above 7.

[0022] 12. A food composition for preventing or improving gastric disease caused by excessive secretion of gastric acid in the above 7.

[0023]

[0024] The pharmaceutical composition according to the present invention contains an extract obtained from a natural product, red oak, or a fraction thereof as an active ingredient, and has excellent preventive or therapeutic effects on gastric mucosal damage, hyperacidity, gastritis, gastric ulcer, or gastroesophageal reflux disease without toxicity or side effects.

[0025] The food composition according to the present invention contains an extract or a fraction thereof obtained from a natural product, red oak, as an active ingredient, and has excellent gastric protective efficacy in inhibiting or recovering gastric mucosal damage without toxicity or side effects.

[0026]

[0027] Figure 1 shows the results of analyzing the effect of red thorn fruit extract on gastric damage lesions in an HCl / ethanol-induced acute hyperacidity animal model.

[0028] Figure 2 shows the results of confirming the effect of red thorn fruit extract on the degree of gastric damage (top) and inhibition of gastric damage (bottom) in an HCl / ethanol-induced acute gastric acid animal model.

[0029] Figure 3 shows the results of analyzing the effect of red thorn fruit extract on PGE2 concentration in an HCl / Ethanol-induced acute gastric acid animal model.

[0030] Figure 4 shows the results of analyzing the effect of red thorn fruit extract on MDA concentration in an HCl / ethanol-induced acute gastric acid animal model.

[0031] Figure 5 shows the results of analyzing the effect of red thorn fruit extract on COX-2 protein expression in a Raw 264.7 cell model.

[0032] Figure 6 shows the results of analyzing the effect of solvent-dependent fractions of red thorn fruit on PGE2 concentration in the Raw 264.7 cell model.

[0033]

[0034] The present inventors completed the present invention by revealing that an extract of red oak or a fraction thereof has the effect of protecting the gastric mucosa and improving a damaged stomach while increasing the secretion of prostaglandin E2 (PGE2).

[0035] The present invention provides a pharmaceutical composition for preventing or treating gastrointestinal diseases, comprising an extract of red oak or a fraction thereof.

[0036] The above 'red oak' is indicated by its scientific name, Quercus acuta. Thunb., and is an evergreen broad-leaved tree of the genus Quercus in the family Fagaceae that grows abundantly in sunny mountain slopes and valleys at elevations of 170 to 500 m in Jeollanam-do, Gyeongsangnam-do, Gyeongsangbuk-do, and Jeju-do.

[0037] In the present invention, the red thorn extract may be at least one extract selected from the group consisting of the fruit, flower, leaf, stem, whole plant, branch, root, and branch of the red thorn tree. In one embodiment, it may be an extract of the fruit of the red thorn tree, but is not limited thereto.

[0038] In the present invention, the red oak extract can be extracted by adding a solvent in an amount of 5 to 20 times the weight of the red oak raw material, but is not limited thereto.

[0039] In the present invention, the red oak extract may include any one of an extract obtained by extraction treatment, a diluted or concentrated extract, a dried product obtained by drying the extract, or a modified or purified product thereof. The extraction method is not particularly limited, and extraction may be performed at room temperature or at a temperature that minimizes the destruction of the active ingredient.

[0040] In the present invention, the red oak extract may be extracted at 80 to 120°C for 1 to 5 hours, but is not limited thereto. If the extraction time is less than 1 hour, the effective activity of the extract may be reduced, and if it exceeds 5 hours, the effect of increasing the activity of the extract may be minimal compared to the extraction efficiency.

[0041] In the present invention, the red oak extract may be an extract soluble in at least one solvent selected from the group consisting of water, alcohols having 1 to 4 carbon atoms, propanol, isopropanol, and butanol, but is not limited thereto. In one embodiment, it may be a hot water extract obtained by adding 10 times the amount of water to the red oak raw material and heating it.

[0042] In the present invention, the fraction of the red oak extract may be obtained by fractionating the red oak extract with at least one solvent selected from the group consisting of dichloromethane, hexane, chloroform, ethyl acetate, and butanol, but is not limited thereto.

[0043] In one embodiment, the fraction may be a fraction of red thorn fruit.

[0044] In one embodiment, the red oak fruit fraction may be a red oak fruit hexane fraction obtained by a method including the steps of: 1) obtaining a dry powder of a hot water extract from red oak fruit; 2) dissolving the dry powder in distilled water; 3) adding hexane to the dissolved substance and fractionating it to separate it into a hexane layer and an aqueous layer; and 4) obtaining the hexane layer.

[0045] In one embodiment, the red oak fruit fraction may be a red oak fruit chloroform fraction obtained by a method including the steps of: 5) adding chloroform to the aqueous layer of step 3) and fractionating the mixture into a chloroform layer and an aqueous layer; and 6) obtaining the chloroform layer.

[0046] In one embodiment, the red oak fruit fraction may be a red oak fruit ethyl acetate fraction obtained by a method including the steps of: 6) adding ethyl acetate to the aqueous layer of step 5) and fractionating the same to separate it into an ethyl acetate layer and an aqueous layer; and 7) obtaining the ethyl acetate layer.

[0047] In one embodiment, the red thorn fruit fraction may be a red thorn fruit butanol fraction obtained by a method including the steps of 8) adding butanol to the aqueous layer of step 6) and fractionating the same to separate it into a butanol layer and an aqueous layer; and 9) obtaining the butanol layer.

[0048] In the present invention, the red oak extract or a fraction thereof has the effect of increasing the secretion of prostaglandin E2 (PGE2), which regulates the regeneration and function of the gastric mucosal cell layer. The prostaglandin is synthesized by the production of arachidonic acid from membrane phospholipid by phospholipase A2, and cyclic oxygenase-1 (COX-1) and cyclic oxygenase-2 (COX-2), which belong to cyclic oxygenase (COX), participate in this metabolic pathway, and the synthesized prostaglandin is known to be involved in platelet aggregation, gastric mucosal protection, renal function regulation, or inflammatory response. In particular, prostaglandin E2 secreted from above promotes mucus secretion through prostaglandin E2 receptor 2 (EP2) or prostaglandin E2 receptor 4 (EP4) and suppresses damage to the gastric mucosa caused by external stimuli.

[0049] In one embodiment, an animal model in which hyperacidity was induced had a sharp decrease in prostaglandin E2 concentration, but an animal model in which hyperacidity was induced after treatment with a red thorn extract or a fraction thereof had an increase in prostaglandin E2 concentration, confirming that the red thorn extract has an effect of preventing, improving, or treating gastric damage.

[0050] In the present invention, the red oak extract or its fractions have the effect of inhibiting malondialdehyde (MDA) in gastric tissue. Malondialdehyde is a lipid metabolism product produced by lipid oxidation by reactive oxygen species and serves as an indicator substance for the degree of lipid peroxidation due to oxidative stress.

[0051] In one example, in an animal model in which hyperacidity was induced, the concentration of malondialdehyde increased rapidly, but in an animal model in which hyperacidity was induced after treatment with a red thorn extract, the increase in malondialdehyde was suppressed, confirming that the red thorn extract has a protective and improving effect on gastric damage.

[0052] In the present invention, the effective concentration of the red oak extract or its fraction may be 5 to 200 μg / ml, specifically 5 to 150 μg / ml, and more specifically 5 to 100 μg / ml.

[0053] In the present invention, the effective dosage of the red oak extract or its fraction may be 30 to 300 mg / kg, specifically 50 to 250 mg / kg, and more specifically 70 to 220 mg / kg.

[0054] The pharmaceutical composition of the present invention can prevent or treat gastrointestinal diseases by including the red oak extract or a fraction thereof as an active ingredient. The gastrointestinal diseases may be, but are not limited to, diseases caused by excessive gastric acid.

[0055] Hyperchlorhydria (also known as hyperacidity) is a very common gastrointestinal condition that occurs when the level of hydrochloric acid in the stomach is higher than the standard range. It is caused by excessive hydrochloric acid in the stomach, which causes symptoms such as heartburn, belching, stomach pain, and reflux. Damage to the gastric mucosa can occur due to attacks by gastric acid and pepsin, or stimuli such as drugs, stress, alcohol, and smoking. If the damage progresses beyond the muscularis mucosa, which is the most superficial layer of the mucosa, it becomes a gastric ulcer. Gastritis is an inflammation of the gastric mucosa, the inner lining of the stomach that protects the stomach from gastric acid or external stimuli. Gastroesophageal reflux disease (GERD) occurs when stomach contents flow back into the esophagus, causing symptoms such as heartburn and acid reflux, or other complications.

[0056] The pharmaceutical composition of the present invention can prevent or treat at least one gastric disease selected from the group consisting of hyperacidity, gastric mucosal damage, gastric ulcer, gastritis, and gastroesophageal reflux disease.

[0057] The content of the red oak extract or its fraction in the pharmaceutical composition of the present invention should be adjusted according to the symptoms of the disease, the degree of progression of the symptoms, the condition of the patient, etc., and may be, for example, 0.001 to 99.9 wt %, 0.001 to 80 wt %, 0.001 to 70 wt %, 0.001 to 60 wt %, 0.001 to 50 wt %, 0.001 to 40 wt %, 0.001 to 30 wt %, 0.001 to 20 wt %, or 0.001 to 10 wt % of the total composition, but is not limited thereto. The above content is a value based on the dry amount after removing the solvent.

[0058] The pharmaceutical composition of the present invention can be formulated by including one or more pharmaceutically acceptable carriers in addition to a pharmaceutically effective amount of the active ingredient for administration.

[0059] The pharmaceutical composition of the present invention can be prepared using pharmaceutically suitable and physiologically acceptable adjuvants. These adjuvants may include excipients, disintegrants, sweeteners, binders, coating agents, bulking agents, lubricants, glidants, or flavoring agents. Furthermore, fillers, anticoagulants, lubricants, wetting agents, flavoring agents, emulsifiers, and preservatives may be additionally included.

[0060] The pharmaceutical composition of the present invention can be formulated in the form of an oral formulation such as a powder, granules, tablet, capsule, suspension, emulsion, syrup, aerosol, topical preparation, suppository or sterile injectable solution using a method known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to a subject in need of the pharmaceutical composition of the present invention, including a human. The formulation can be a powder, granule, tablet, emulsion, syrup, aerosol, soft or hard gelatin capsule, sterile injectable solution or sterile powder.

[0061] The pharmaceutical composition of the present invention may be administered alone or in combination with a component known to exhibit therapeutic and preventive effects on known gastrointestinal diseases. The effective dosage may vary depending on the patient's age, sex, condition, weight, absorption, inactivation rate, and excretion rate of the active ingredient in the body, type of disease, and concomitantly administered drug. Generally, 0.001 to 150 mg, preferably 0.01 to 100 mg per 1 kg of body weight may be administered daily or every other day, or administered once or several times a day in divided doses. However, since the dosage may increase or decrease depending on the route of administration, severity of obesity, sex, weight, age, etc., the above dosage does not limit the scope of the present invention in any way.

[0062] In addition, the present invention provides a food composition for preventing or improving gastrointestinal diseases, which comprises a red oak extract or a fraction thereof.

[0063] The above red oak extract or fraction thereof is as described above in the description of the pharmaceutical composition.

[0064] As described above, the red oak extract of the present invention or a fraction thereof increases the secretion of prostaglandin E2 (PGE2) and suppresses malondialdehyde (MDA) in gastric tissue, thereby exhibiting an effect of preventing and recovering gastric mucosal damage, and thus has excellent gastric protective efficacy.

[0065] The food composition of the present invention can be provided as a variety of functional foods with gastrointestinal protective effects by including the red oak extract or a fraction thereof as an active ingredient. Furthermore, the food composition can be used in the manufacture of health functional foods for preventing or improving at least one gastrointestinal disease selected from the group consisting of hyperacidity, gastric mucosal damage, gastric ulcers, gastritis, and gastroesophageal reflux disease.

[0066] The food composition of the present invention includes all forms such as functional foods, nutritional supplements, health functional foods, and food additives. The food composition of the above type can be manufactured in various forms by including additional ingredients such as minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickeners, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, nutrients, and vitamins, which are commonly added according to conventional methods known in the art.

[0067] The above health functional food is the same term as food for special health use (FoSHU), and refers to a food with high medical and healthcare effects that is processed to efficiently exhibit bioregulatory functions in addition to providing nutrition. In addition to the active ingredient, the red thorn extract or its fraction, it can be formulated into a food composition by including one or more food-related or pharmaceutically acceptable carriers. The formulation form of the food composition may be a tablet, capsule, powder, granule, liquid, pill, solution, syrup, juice, suspension, emulsion, or drop. For example, for formulation in the form of a tablet or capsule, the active ingredient may be combined with an oral, non-toxic, pharmaceutically acceptable inert carrier, such as ethanol, glycerol, or water.

[0068] The food composition of the present invention, formulated in the above manner, can be used as a functional food for protecting the stomach or preventing and improving gastrointestinal diseases, or can be added to various foods. Foods to which the composition of the present invention can be added include functional foods such as nutritional supplements, weight control foods, and health supplements, teas, beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gum, candy, ice cream, alcoholic beverages, and vitamin complexes.

[0069] The preferred content of the red oak extract or its fraction in the food composition of the present invention is not limited thereto, but may be, for example, 0.01 to 99 wt %, 0.01 to 80 wt %, 0.01 to 70 wt %, 0.01 to 60 wt %, or 0.01 to 50 wt % of the final manufactured food. For example, when included in a health drink, 0.01 to 50 g may be added based on 100 ml, but is not limited thereto.

[0070]

[0071] Hereinafter, the present invention will be described in detail with examples and experimental results.

[0072]

[0073] Example 1. Preparation of red oak extract

[0074] 100 g of Quercus acuta. Thunb. fruit was extracted by adding distilled water equivalent to 10 times the weight of the raw material and heating at 100°C for 3 hours. The obtained extract was concentrated under reduced pressure and freeze-dried for storage, and then dissolved in distilled water before use in experiments.

[0075]

[0076] Example 2. Preparation of fractions of red oak extract

[0077] The dried powder of the hot water extract of the red oak fruit was completely dissolved in 1 L of distilled water, placed in a fractional funnel, 1 L of hexane was added, mixed, and fractionated to separate the hexane layer, which is a hexane-soluble layer, and the water layer, which is a hexane-insoluble layer, thereby obtaining only the hexane layer, thereby preparing a hexane fraction. The same process for the hexane-insoluble layer and water layer was repeated three times to obtain a hexane-insoluble fraction and a soluble fraction.

[0078] 1 L of chloroform was added to the remaining solution (aqueous layer), mixed, and fractionated to separate the chloroform-soluble layer (chloroform layer) and the chloroform-insoluble layer (aqueous layer), thereby obtaining only the chloroform layer, thereby preparing a chloroform fraction. The same process was repeated three times.

[0079] 1 L of ethyl acetate was added to the remaining solution (aqueous layer), mixed, and fractionated to separate the ethyl acetate-soluble layer (ethyl acetate layer) and the ethyl acetate-insoluble layer (aqueous layer), thereby obtaining only the ethyl acetate layer, thereby preparing an ethyl acetate fraction. The same process was repeated three times.

[0080] 1 L of butanol was added to the remaining solution (aqueous layer), mixed, and fractionated to separate the butanol-soluble layer (butanol layer) and the butanol-insoluble layer (aqueous layer), thereby obtaining only the butanol layer, thereby preparing a butanol fraction. The same process was repeated three times.

[0081] After fractionation of the butanol-soluble layer, the remaining butanol-insoluble layer was concentrated to remove the remaining organic solvent, thereby producing an aqueous fraction. Each fraction thus obtained was filtered through a vacuum filtration device, concentrated, and then freeze-dried to completely remove the solvent before use in this experiment.

[0082]

[0083] Example 3. Production of an animal model induced by hyperacidity.

[0084] To observe the effects of the hot water extract of the red thorn fruit and its fractions on improving gastric mucosal damage, 6-week-old male ICR mice were purchased from Samtako (Korea) and acclimated to the laboratory environment for 7 days. The conditions of the animal breeding room were controlled as follows: temperature 22±2℃, humidity 50±5%, and a 12-hour light / dark cycle. Sterilized solid feed and water were supplied in sufficient quantities.

[0085] The experimental animals were divided into five groups of six animals each, and the experimental groups were composed as shown in Table 1 below. The groups, except for the normal group, were treated with HCl / Ethanol as a factor inducing hyperacidity to induce acute gastric mucosal damage. Prior to inducing acute hyperacidity, only water was provided and fasted for 12 hours. On the day of the experiment, the control group was orally administered distilled water, the positive control group (P.Control) was orally administered Gelfos 1 pouch (20 g) / day for an adult (60 kg), and the test substance group (QA 100 mg / kg and QA 200 mg / kg) was orally administered red thorn fruit extract at concentrations of 100 and 200 mg / kg for 3 days. Then, all experimental groups, except the normal group, were administered 500 ㎕ of 150 mM / 60% Ethanol 30 minutes later. After 30 minutes, the stomach was opened under ether anesthesia and the stomach tissue was removed.

[0086] Number Experimental group Treatment 1 Normal group · After saline solution administration, no treatment 2 Control group · 150 mM HCl / 60% Ethanol 500 ㎕ administration 3 Positive control group (P.Control) · Oral administration of Gelphos 1 packet / day (based on adults) followed by 150 mM HCl / 60% Ethanol 500 ㎕ administration 4 QA 100 mg / kg · Oral administration of red thorn fruit extract 100 mg / kg followed by 150 mM HCl / 60% Ethanol 500 ㎕ QA 200 mg / kg · Oral administration of red thorn fruit extract 200 mg / kg followed by 150 mM HCl / 60% Ethanol

[0087]

[0088] Experimental Example 1. Effect of red oak extract on gastric damage lesions.

[0089] After the gastric tissue of the experimental animals was removed, an incision was made along the greater trochanter of the stomach, the contents were removed, washed in saline solution, and fixed. The degree of damage to the gastric mucosa was observed with the naked eye, and photographs were taken using a Canon camera (Tokyo, Japan). The damaged area of ​​the gastric mucosa was measured using Image J (National Institute for Health, Bethesda, MD, USA). As a result, the control group induced with hyperacidity showed severe hemorrhagic mucosal damage throughout the gastric mucosa, and the test group treated with HCl / Ethanol after oral administration of 100 and 200 mg / kg concentrations of red thorn fruit extract showed a tendency for mucosal damage to be reduced compared to the control group (Fig. 1).

[0090]

[0091] Experimental Example 2. Effect of Red Oak Extract on the Degree of Gastric Damage and Damage Inhibition

[0092] In order to observe the improvement effect of gastric damage caused by hyperacidity, the gastric damage (or gastric damage improvement) score was scored according to the degree of damage as shown in Table 2 below, and the degree of gastric damage in each experimental group was indicated (Fig. 2, top).

[0093] Score of the above damage: No damage 0 1-3 small gastric lesions (<10 mm long) 1-3 large gastric lesions (≥10 mm long) 2 1-3 deep gastric lesions 3 3 or more small gastric lesions 4 3 or more large gastric lesions 5

[0094] In addition, the gastric damage inhibition rate of each experimental group was measured using the following mathematical formula 1. As a result, the test substance groups treated with HCl / Ethanol after orally administering red thorn fruit extract at concentrations of 100 and 200 mg / kg each inhibited gastric damage lesions by 74.7% and 56.2%, respectively (Figure 2 below).

[0095]

[0096]

[0097] Experimental Example 3. Effect of red oak extract on serum PGE2 concentration

[0098] Prostaglandin E2 (PGE2) produced in the gastric mucosa plays a role in regenerating and protecting the gastric mucosa, and an increase in PGE2 secretion indicates a gastric mucosal protective effect. To analyze the effect of red thorn fruit extract on PGE2 concentration in an acute hyperacid animal model, PGE2 concentration was measured using enzyme-linked immunosorbent assay (ELSIA). Specifically, on the day of the experiment, after sample processing, the animals were opened under ether anesthesia, blood was collected from the abdominal aorta, and the serum was separated by centrifugation at 1,200 rpm for 10 minutes. The separated serum was stored at -70℃ and then used in the experiment. Using the separated serum, the experiment was performed according to the protocol suggested by the manufacturer using a Prostaglandin E2ELISA kit (RnD System, USA).

[0099] As a result, compared to the control group (Control) and positive control group (P.Control) in which the PGE2 concentration decreased after inducing hyperacidity, the PGE2 concentration of the test substance group (QA 100 mg / kg and QA 200 mg / kg) showed a similar level to that of the normal group (Normal), confirming the gastric mucosa protective effect of the red thorn fruit extract (Fig. 3).

[0100]

[0101] Experimental Example 4. Effect of Red Oak Extract on MDA Concentration in Gastric Tissue

[0102] MDA (malondialdehyde) is an indicator substance that can indicate the degree of lipid peroxidation and causes cancer when produced in the body. To analyze the degree of lipid peroxidation in gastric tissue and to determine whether the extract of the red thorn fruit has a protective effect against gastric damage caused by excessive gastric acid, the concentration of MDA in gastric tissue was measured. Specifically, on the day of the experiment, a certain size of gastric tissue was excised from the removed stomach, and the tissue sample was homogenized on ice using PBS buffer containing 1X BHT. Centrifugation was performed and only the supernatant was analyzed. Using the separated supernatant, the experiment was performed according to the protocol suggested by the manufacturer using the Lipid Peroxidation Assay (TBARS) kit (Dogen bio., Korea). The experiment is based on the reaction of MDA with the chromogenic reagent TBA. The MDA-TBA adduct formed by the reaction of MDA and TBA was measured at 540 nm using a colorimetric method to determine the MDA concentration per tissue weight.

[0103] As a result, compared to the control group (Control) in which the MDA concentration increased after inducing hyperacidity, the MDA concentration of the test substance group (QA 100 mg / kg and QA 200 mg / kg) was reduced to a similar degree as the positive control group (P.Control), confirming the gastric damage inhibition and protective effect of the red thorn fruit extract (Fig. 4).

[0104]

[0105] Experimental Example 5. Effect of red oak extract on COX-2 expression

[0106] COX-2 (cyclic oxygenase-2) is an enzyme involved in the metabolic pathway of prostaglandin synthesis together with COX-1 (cyclic oxygenase-1). To analyze the expression of COX-2 protein by the extract of the fruit of the red thorn tree, mouse macrophage RAW 264.7 was cultured in DMEM (Dulbecco's modified Eagle's medium) medium with 10% FBS and 1% penicillin-streptomycin. The cells were seeded at 3.5 X 10 in a 60 mm cell culture dish. 5 After uniformly distributing the cells and stabilizing for 24 hours, the samples were treated as untreated group, LPS 1 ㎍ / ㎖ treatment group, LPS 1 ㎍ / ㎖ and red thorn extract (10, 50, 100, and 200 ㎍ / ㎖) treatment group. After 24 hours of sample treatment, the samples were washed once with PBS (Phosphate-Buffered Saline), dissolved in 1X RIPA Buffer (Radio-Immunoprecipitation assay buffer), centrifuged at 12,500 rpm for 15 minutes at 4 ℃, and the supernatant was separated and stored frozen at -80 ℃ until the experiment. To measure the expression level of COX-2, β-protein, 20 μg of protein was electrophoresed using 8-10% SDS polyacrylamide gel, and the acrylamide gel was transferred to a polyvinylidene difluoride (PVDF) membrane. After preparing the membrane, 5% skim milk was added and reacted for a time at room temperature, and each primary antibody was reacted at a ratio of 1:1000 at 4 °C. The next day, the membrane was washed three times with TBST (Tris-Buffered saline-Tween 20), and the secondary antibody was reacted for 1 hour at room temperature, and then washed three times with TBST every 10 minutes. Then, the membrane was detected for protein expression using an enhanced chemiluminescence (ECL) Western blotting substrate solution.

[0107] As a result, the LPS treatment group treated with red thorn extract showed a higher COX-2 concentration than the control group (Con) treated with LPS only (Fig. 5).

[0108]

[0109] Experimental Example 6. Effect of solvent on PGE2 concentration of red oak fractions

[0110] The effects of fractions prepared by different solvents from the extract of the fruit of the deciduous tree in Example 2 on the concentration of prostaglandin E2 (PGE2) were analyzed using mouse macrophage RAW 264.7 cells. Cells were cultured in DMEM (Dulbecco's modified Eagle's medium) medium supplemented with 10% FBS and 1% penicillin-streptomycin, and cultured in a CO2 incubator at 37°C. Cells were cultured until 80% of the cell culture dish was filled. The cells were dispensed into 48-well plates and cultured. The cells were divided into an untreated group, a group treated with 500 ng / ㎖ of LPS, and a group treated with 20 ㎍ / ㎖ of each of the different solvents and cultured for 24 hours. The culture medium was then centrifuged at 1,000 rpm for 5 minutes, and the supernatant was separated and analyzed using the PGE2ELISA method.

[0111] As a result, the PGE2 concentration in all groups treated with the red oak fruit extract (Water fr.) and its fractionated hexane fraction (Hexane fr.), chloroform fraction (Chloroform fr.), ethyl acetate fraction (Ethyl acetate fr.), and butanol fraction (Butanol fr.) increased compared to the untreated group, confirming that the red oak fruit fraction also has a gastric mucosal protective effect (Fig. 6).

Claims

1. A pharmaceutical composition for preventing or treating gastrointestinal diseases, comprising an extract of red thorn tree or a fraction thereof.

2. A pharmaceutical composition for preventing or treating gastrointestinal diseases, wherein the red thorn extract in claim 1 is at least one extract selected from the group consisting of the fruit, flower, leaf, stem, whole plant, branch, root, and branch of the red thorn tree.

3. A pharmaceutical composition for preventing or treating gastrointestinal diseases according to claim 1, wherein the red oak extract is an extract soluble in at least one solvent selected from the group consisting of water, alcohols having 1 to 4 carbon atoms, propanol, isopropanol, and butanol.

4. A pharmaceutical composition for preventing or treating gastrointestinal diseases, wherein the fraction is obtained by fractionating the red oak extract with at least one solvent selected from the group consisting of dichloromethane, hexane, chloroform, ethyl acetate, and butanol in claim 1.

5. A pharmaceutical composition for preventing or treating at least one gastrointestinal disease selected from the group consisting of hyperacidity, gastric mucosal damage, gastric ulcer, gastritis, and gastroesophageal reflux disease according to claim 1.

6. A pharmaceutical composition for preventing or treating gastric disease caused by excessive secretion of gastric acid according to claim 1.

7. A food composition for preventing or improving gastrointestinal diseases, comprising an extract of red thorn tree or a fraction thereof.

8. A food composition for preventing or improving gastrointestinal diseases, wherein the red thorn extract of claim 7 is at least one extract selected from the group consisting of the fruit, flower, leaf, stem, whole plant, root, and branch of the red thorn tree.

9. A food composition for preventing or improving gastrointestinal diseases according to claim 7, wherein the red oak extract is an extract available in at least one solvent selected from the group consisting of water, alcohols having 1 to 4 carbon atoms, propanol, isopropanol, and butanol.

10. A food composition for preventing or improving gastrointestinal diseases, wherein the fraction is obtained by fractionating the red oak extract with at least one solvent selected from the group consisting of dichloromethane, hexane, chloroform, ethyl acetate, and butanol, according to claim 7.

11. A food composition for preventing or improving at least one gastrointestinal disease selected from the group consisting of hyperacidity, gastric mucosal damage, gastric ulcer, gastritis, and gastroesophageal reflux disease according to claim 7.

12. A food composition for preventing or improving gastric disease caused by excessive secretion of gastric acid according to claim 7.

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