WO2018101798A1 - Artemisia capillaris water-soluble extract, use therof, and preparation method therefor - Google Patents
Artemisia capillaris water-soluble extract, use therof, and preparation method therefor Download PDFInfo
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- WO2018101798A1 WO2018101798A1 PCT/KR2017/014030 KR2017014030W WO2018101798A1 WO 2018101798 A1 WO2018101798 A1 WO 2018101798A1 KR 2017014030 W KR2017014030 W KR 2017014030W WO 2018101798 A1 WO2018101798 A1 WO 2018101798A1
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- gastric
- mucosal
- gastrointestinal
- gastric mucosal
- extract
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/282—Artemisia, e.g. wormwood or sagebrush
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
Definitions
- the present invention is a water-soluble extract of Artemisia capillaris , a composition for the prevention or improvement of gastrointestinal diseases containing the same, more specifically mucosal damage gastrointestinal caused by excessive gastric acid and stress through the maintenance, protection and regeneration of the gastrointestinal mucosa system
- the present invention relates to a food composition and a pharmaceutical composition containing a water-soluble phosphorus mugwort extract having an effect of preventing or ameliorating related diseases.
- Injin mugwort has been used for fatty liver, chronic hepatitis, gastrointestinal diseases, constipation, jaundice, neuralgia, etc., and is effective for alcohol drinking and fat accumulation inhibitory people and liver disease, and anti-inflammatory, diuretic, gallstone inhibitory effects are known.
- hemostatic effect is a blood bleeding (nose blood), bleeding (blood vomiting), bleeding (blood to the anus). It is used to treat bleeding from the womb, and as a first aid to reduce pain.
- leaf extracts have been reported to have antioxidant and NF- ⁇ B inhibitory effects and to suppress gastric damage and inflammation based on them.
- the prevention of gastric damage due to the antioxidant effects of PGE2 and GSH in NSAIDS gastric injury model (World J Gastroenterol 2005; 11 (47): 7450-7456), and the antioxidant activity of PGE2 and GSH in chronic reflux gastritis model Effect of prevention of gastric injury due to effects (World J Gastroenterol 2005; 11 (47): 7430-7435), increased secretion of Ghrelin, a hormone with gastric protective effect in acute alcoholic gastritis model (Gut and liver 2011; 5 1): 52-56), reducing IL-8 and CCL20 through p38 inhibition and NF- ⁇ B inhibition in a TNF- ⁇ mediated gastric epithelial inflammation model (World J Gastroenterol 2006; 12 (30): 4850-4858). ) has been reported.
- the leaf extracts and the extract of Injin mugwort have an effect of improving acute gastritis or bacterial gastritis through suppression of the inflammatory response.
- the effectiveness of the extracts of leaf or wormwood in preventing or improving gastrointestinal mucosa have been no reports about the effectiveness of the extracts of leaf or wormwood in preventing or improving gastrointestinal mucosa.
- Another object of the present invention to provide a food composition for the prevention or improvement of mucosal damage gastrointestinal diseases.
- Another object of the present invention to provide a pharmaceutical composition for the prevention or treatment of mucosal damage gastrointestinal diseases.
- composition for the prevention or improvement of gastric mucosal damaging gastrointestinal diseases containing Artemisia capillaris water-soluble extract.
- composition for the prevention or improvement of gastric mucosal damaging gastrointestinal disease of 3 above which prevents or inhibits mucosal damage caused by gastritis or gastric ulcer caused by gastric acid, ethanol, stress or a combination thereof.
- composition for the prevention or improvement of gastric mucosal damage gastrointestinal disease of 3 above which inhibits or improves gastric mucosal damage through increased expression of gastric mucosal proteins or mucosal immune factors.
- the gastric mucosal protein or mucosal immune factor is MUC5AC, MUC6, rPigr, TFF1, TFF2 and TFF3 composition for the prevention or improvement of gastric mucosal damaging gastrointestinal disease of the above 7, wherein at least one selected from the group consisting of.
- composition for the prevention or improvement of gastric mucosal damaging gastrointestinal disease of 3 above which inhibits or improves gastric mucosal damage by inhibiting inflammatory cytokines, inhibiting the production of lipid peroxides, or increasing antioxidant enzyme activity.
- a pharmaceutical composition for preventing or treating gastric mucosal damaging gastrointestinal diseases containing Artemisia capillaris water-soluble extract.
- gastric mucosal protein or mucosal immune factor is at least one selected from the group consisting of MUC5AC, MUC6, rPigr, TFF1, TFF2, and TFF3.
- Concentration of the concentrate from which the precipitate was removed was second concentrated until the water content was 55 ⁇ 5% to obtain an Artemisia capillaris aqueous extract;
- Injinssuk (Artemisia capillaris) method for producing an aqueous extract containing.
- Aqueous extract of Artemisia capillaris is a gastric mucosal mucin protein (MUC6, MUC5AC), including the major regulatory factors (TFF1, 2, 3) of the gastrointestinal mucosa system.
- MUC6, MUC5AC gastric mucosal mucin protein
- TNF1, 2, 3 the major regulatory factors of the gastrointestinal mucosa system.
- rPigr mucosal immune related factors
- SOD antioxidant enzymes
- MDA lipid peroxides
- Phosphorus erythema extracts have the effect of preventing and inhibiting mucosal damage to gastritis / gastric ulcer induced by hydrochloric acid-ethanol by increasing the expression of key regulatory factors of the gastrointestinal mucosal system, thus protecting the gastrointestinal tract through promoting mucosal production, It can be used as natural medicines, health functional foods, functional drinks, etc. to prevent gastric damage caused by stress or stomach acid. It also prevents and treats gastritis and gastric ulcers, which can be caused mainly by eating spicy, spicy foods, and excessive gastric acid, indigestion, and gas bloating caused by frequent drinking or decreased mucosa following NSAIDs.
- Injin mugwort extract is applicable to the prevention and treatment of gastric cancer and colon cancer through the increase of TFF3.
- Figure 1 is a result confirming the effect of the extract of A. erysipelas water soluble extract (AEAC) in mast cell degranulation in the mast cells induced by compound 48/80.
- AEAC erysipelas water soluble extract
- Figure 2 is a result confirming the inhibitory effect of inflammatory cytokine by the water soluble extract (AEAC) in mast cells induced by PMA and A23187.
- AEAC water soluble extract
- Figure 3 is a result confirming the inhibitory effect of inflammatory cytokine by L. aeruginosa water-soluble extract (AEAC) in LPS-induced macrophages.
- AEAC L. aeruginosa water-soluble extract
- Figure 4 is a result of confirming the neutralizing ability of calcium carbonate, phosphorus mugwort soluble extract (AEAC), licorice extract (AC) and leaf extract (AEAA).
- Figure 5 is a result confirming the effect of H. ethanol-induced acute gastritis / gastric ulcer animal model influx of the extract of AJC in the production or secretion of histamine in serum.
- Figure 6 is a result confirming the gastric mucosa protective effect by the water soluble extract (AEAC) in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
- AEAC water soluble extract
- Figure 7 shows the results of confirming the effect of increasing the amount of gastric mucus by the extract of AJC in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
- Figure 9 is a result confirming the effect of increasing the activity of the superoxide dismutase by AJC (AEAC) in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
- AEAC superoxide dismutase
- 11 is a result of confirming the effect of increasing the expression of gastric mucosa protein MUC5AC after a single administration of AJC (AEAC) in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
- AEAC AJC
- rPigr a mucosal immune-related factor after a single administration of AJC in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
- Figure 13 is a result of confirming the effect of increasing the expression of TFF1, 2 and 3 involved in gastric mucosa protection and treatment after a single administration of AJC in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
- 16 is a result of confirming the effect of increasing the expression of rPigr, gastric mucosal immunity-related factors after repeated administration of AJC in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
- Figure 17 shows the effect of increasing the expression of TFF1, 2 and 3 involved in gastric mucosa protection and treatment after repeated administration of AJC in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
- FIG. 18 is a method for protecting and treating gastric mucosa protein, gastric mucosal immunity related factors and gastric mucosa after repeated administration of AJC or licorice extract (AC) in HCl / ethanol-induced acute gastritis / gastric ulcer animal model This is the result of confirming the effect of increasing the expression of the factors involved.
- AJC licorice extract
- FIG. 19 is a photograph comparing the degree of gastric mucosal injury caused by dissecting the Taiwanese stomach after single administration of AJC or Licorice extract (AC) in acute gastritis / gastric ulcer animal model induced by HCl / ethanol. to be.
- FIG. 20 is a quantitative analysis of gastric mucosal injury length caused by dissection of the Taiwanese stomach after single administration of AJC or Licorice Extract (AC) in acute gastritis / gastric ulcer animal model induced by HCl / ethanol. The result is.
- Figure 21 is a photograph comparing the degree of gastric mucosal damage caused by dissection of the stomach part of the stomach after repeated administration of AJC or licorice extract (AC) in acute gastritis / gastric ulcer animal model induced by HCl / ethanol to be.
- AJC licorice extract
- Figure 23 is a photograph comparing the degree of damage caused by dissecting the stomach part of the stomach after a single administration of AJC or AEAA in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
- TFF1 is not only known as a cancer suppressor gene in connection with gastric cancer, but has been reported to protect the gastrointestinal mucosa from various injuries, to stabilize the mucosal layer and to regulate the healing process of gastrointestinal epithelial tissue.
- TFF2 has a function of inhibiting gastric acid secretion as well as a protective effect on the gastrointestinal mucosa of TFF1.
- TFF3 is found not only in the gastrointestinal mucosa but also in breast milk and has been reported to exhibit excellent anti-inflammatory effects.
- mucin proteins directly constitute the gastrointestinal mucosa and protect the gastrointestinal epithelial tissue
- rPigr a mucosal immune-related factor
- the Artemisia capillaris water-soluble extracts increase the expression of mucosal proteins or mucosal immune factors, specifically one or more mucosal proteins or mucosal immune factors selected from MUC5AC, MUC6, rPigr and TFF3, inflammatory Maintains and improves the homeostasis of the gastrointestinal mucosa system by inhibiting cytokine, inhibiting the production of lipid peroxides, or increasing antioxidant enzyme activity, and inhibiting inflammation and promoting mucosal production, thereby preventing or improving gastric mucosal damage.
- the present invention has been completed by finding out.
- the present invention is a water-soluble extract of Artemisia capillaris having an action of preventing or improving gastric mucosal gastrointestinal diseases, compositions and food compositions for preventing or ameliorating gastric mucosal gastrointestinal disorders, gastric mucosal gastrointestinal disorders It relates to a pharmaceutical composition for prophylaxis or treatment, and a method for preparing Artemisia capillaris water-soluble extract.
- the first aspect of the present invention relates to a water-soluble extract of Artemisia capillaris having a prophylactic or ameliorating action of gastric mucosal damaging gastrointestinal diseases.
- Artemisia capillaris water-soluble extract according to the present invention is extracted from hot water of Injin mugwort, mucosal proteins or mucosal immune factors, specifically MUC5AC, MUC6, rPigr, TFF1, TFF2 and TFF3 at least one protein or mucosal immunity It increases the expression of the factor, maintains and improves the homeostasis of the gastrointestinal tract mucosal system through inhibiting inflammatory cytokines, inhibiting the production of lipid peroxides or increasing antioxidant enzyme activity, and exhibits the effect of inhibiting inflammation or promoting mucosal production.
- mucosal proteins or mucosal immune factors specifically MUC5AC, MUC6, rPigr, TFF1, TFF2 and TFF3 at least one protein or mucosal immunity It increases the expression of the factor, maintains and improves the homeostasis of the gastrointestinal tract mucosal system through inhibiting inflammatory cytokines, inhibiting the production of lipid peroxides or increasing
- the Artemisia capillaris water-soluble extract of the present invention prevents or ameliorates mucosal damaging gastrointestinal disorders caused by, for example, gastric acid, ethanol, stress, or a combination thereof, gastric acid, ethanol, stress or a combination thereof. It can be used to prevent or inhibit the damage to the mucous membrane by gastritis or gastric ulcer caused by.
- the second aspect of the present invention relates to a composition for the prevention or improvement of gastric mucosal damaging gastrointestinal disease containing the water extract of Artemisia capillaris .
- a third aspect of the present invention relates to a food composition for preventing or ameliorating gastric mucosal damaging gastrointestinal disease containing the water extract of Artemisia capillaris .
- a fourth aspect of the present invention relates to a pharmaceutical composition for the prevention or treatment of gastric mucosal damage gastrointestinal diseases containing Artemisia capillaris water-soluble extract.
- the pharmaceutical composition for the prevention or treatment of gastric mucosal damaging gastrointestinal diseases contains a water-soluble extract of Artemisia capillaris as an active ingredient, specifically gastric mucosal proteins or mucosal immune factors, specifically MUC5AC, MUC6, rPigr, TFF1, Increases the expression of one or more gastric mucosal proteins or mucosal immune factors selected from TFF2 and TFF3, maintains and improves the homeostasis of the gastrointestinal mucosal system through inhibiting inflammatory cytokines, inhibiting the production of lipid peroxides or increasing antioxidant enzyme activity, Inhibit inflammation or promote mucosal production.
- the pharmaceutical composition for preventing or treating gastric mucosal damaging gastrointestinal diseases may further include appropriate carriers, excipients, diluents, and the like commonly used in the preparation of pharmaceutical compositions in addition to the water-soluble extract of Artemisia capillaris . .
- the pharmaceutical composition according to the present invention may contain the Artemisia capillaris water-soluble extract as it is or may contain a pharmaceutically acceptable salt of the components contained in the extract.
- the pharmaceutical composition of the present invention may be used alone or in combination with other pharmaceutically active compounds of Artemisia capillaris .
- compositions for the prevention or treatment of gastric mucosal injury gastrointestinal diseases according to the present invention, powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, external preparations, suppositories, according to conventional methods and needs It may be used in any form such as sterile injectable solution.
- the fifth aspect of the present invention relates to a method for producing an aqueous extract of Artemisia capillaris .
- step (1) 10-14 times of water, preferably purified water, is added to Artemisia capillaris , dried and cut to an appropriate size, preferably at 80 ⁇ 20 ° C, more preferably at 90 ⁇ 10 ° C. Extract for 3-5 hours. Therefore, the hydrothermal water of step (1) may be preferably 80 ⁇ 20 ° C., more preferably 90 ⁇ 10 ° C. water.
- step (2) the extract obtained in step (1) is cooled to room temperature and then filtered, and the filtrate is concentrated under reduced pressure at a temperature of less than 60 ° C. first, and then centrifuged at 4000 to 7000 rpm or more, and the precipitate is Remove by filtration.
- step (3) the concentrated solution from which the precipitate is removed is concentrated under a second reduced pressure until the water content is 55 ⁇ 5% at less than 60 ° C. to obtain a water-soluble extract of Insam.
- Artemisia capillaris used as a raw material can be used as long as it is native to Korea.
- RNA samples NaOH, methyl orange, calcium carbonate, hydrochloric acid, ethanol were purchased from Sigma-Aldrich (St. Louis, MO, USA).
- Oligo (dT), dNTP, RNase-free water and Superscript III First-Strand Synthesis used in Polymerase chain reaction (PCR) analysis were purchased from Invitrogen (Carlsbad, CA, USA).
- the RNeasy Mini Kit and Rotor-Gene SYBR Green PCR Kit were purchased from Qiagen (Valencia, CA, USA) and oligonucleotide primers were purchased from Bioneer (Daejeon, Korea).
- HMC-1 Human Mast Cells (HMC-1) were purchased from the Bank of Korea Cell Line, and 10% fetal bovine serum (FBS), 100 U / ml penicillin and 100 ⁇ g / ml streptomycin (GibcoBRL, USA) Cultured under 37 ° C., 5% CO 2 environment with added IMDM medium (GibcoBRL, USA).
- FBS fetal bovine serum
- IMDM medium GibcoBRL, USA
- RAW264.7 cells a mouse-derived macrophage line, were cultured in a 37 ° C., 5% CO 2 environment using DMEM medium supplemented with 10% fetal calf serum and 1% penicillin-streptomycin.
- the concentrated solution from which the precipitate was removed was concentrated under a second reduced pressure until the moisture content was 55 ⁇ 5% at less than 60 ° C to obtain a water-soluble extract of AJU (AEAC, 45 ⁇ 5% solids), and 30-60 minutes at 85-90 ° C. Sterilized during.
- the extract was packaged in a container and refrigerated and used as the final sample for this experiment (yield: 8-13% based on the solids content of AEAC relative to the weight of the raw material).
- the content of scopoletin an index component of water soluble extracts, was analyzed by Dionex Chromelon TM Chromatography Data System at a wavelength of 344nm using a C18 column (4.6 ⁇ 250mm, 100 ⁇ , Kinetex). At this time, 0.05% formic acid aqueous solution was used as the A solvent and acetonitrile was used as the B solvent as the mobile phase. Analysis was carried out in mobile phase conditions of A / B (90/10), 13 min A / B (5/95), 16 min (Gradient) -A / B (90/10). As a result of HPLC analysis, the scopoletin content of the water-soluble extracts of the extract was averaged at 3.502 mg per g of the extract.
- the antacid test is a test method for confirming the antacid power of an agent that reacts with gastric acid, and in the present invention, it was tested according to the antacid test method specified in the Korean Pharmacopoeia.
- a mixture of licorice extract (AC) and leafy water soluble extract (AEAA), and the extract of AJC (AEAC) according to the present invention were mixed in 100 ml of 0.1 N HCl, and reacted for 1 hour in a shaking incubator at 37 ° C. After titration, the mixture was titrated with 0.1N NaOH. Methyl orange was used as an indicator and calcium carbonate was used as a positive control.
- compound 48/80 (Sigma, USA), a potent drug that increases the infiltration of mast cells by increasing Ca 2+ influx into the cell membrane of mast cells, was used. After dispensing the cultured mast cells into a 24 well plate, each sample was pretreated for 2 hours at different concentrations, treated with Compound 48/80, and after 30 minutes, the supernatant was taken and secreted into the medium. The amount was measured.
- the cytokine secretion of mast cells was induced by a combination of phorbol myristate acetate (PMA) and A23187. After the cultured mast cells were dispensed into 24 well plates, each sample was pretreated for 1 hour, and then treated with PMA and A23187. After 8 hours, supernatants or cells were taken and used for cytokine measurement.
- PMA phorbol myristate acetate
- Raw264.7 cells which are macrophages, were treated with LPS at a concentration of 1 ⁇ g / ml after pretreatment of each sample for 1 hour at concentration, and supernatants or cells were taken 24 hours later and used for cytokine measurement.
- Cytokine measurements were performed according to the manufacturer's instructions by purchasing sandwich ELISA kits for IL-1 and IL-6.
- the standard curve was calculated from the absorbance of standard and the amount of cytokine was quantified from the absorbance of each sample.
- Qiagen's Rotor-Gene Q 2 plex real-time DNA amplifier and Qiagen's two step PCR SYBR kit were used to perform real-time polymerase chain reaction according to the manufacturer's instructions, and the amount of amplified DNA was quantitatively compared. Specifically, pre-denaturation was performed at 95 ° C. for 5 minutes based on a cycling protocol, followed by 40 cycles of 5 seconds at 95 ° C. and 10 seconds at 60 ° C. for 3 minutes from 60 ° C. to 95 ° C. It was performed in the order of the rising melting step. Products amplified by real-time polymerase chain reaction were quantified using the Delta delta Ct method and corrected by the expression level of GAPDH for each sample. The base sequence of the primers used in the experiment is as shown in the following [Table 1].
- control group In the control group, the general formula, the positive control group, ranitidine or licorice extract, and the experimental group were orally administered AEAC. After 30 minutes, 1 mL of HCl-EtOH (60% EtOH containing 150 mM HCl) was orally administered to each of the control, positive control, and experimental groups.
- HCl-EtOH 50% EtOH containing 150 mM HCl
- mice Six male rats with a body weight of about 200 g were divided into three groups: a control group, a positive control group, and an experimental group.
- control group was orally administered with the general formula, the positive control group with licorice extract, and the experimental group with AEAC twice a day for 1 week. Fasting for more than 24 hours after the last oral administration.
- control group was administered with the general formula, the positive control group with the licorice extract and the experimental group with oral AEAC, and after 30 minutes, 1 mL of HCl-EtOH (60% EtOH containing 150 mM HCl) was orally administered to the control group, the positive control group, and the experimental group. It was.
- Qiagen's Rotor-Gene Q 2 plex real-time DNA amplifier and Qiagen's two step PCR SYBR kit were used to perform real-time polymerase chain reaction according to the manufacturer's instructions, and the amount of amplified DNA was quantitatively compared. Specifically, pre-denaturation was performed at 95 ° C. for 5 minutes based on a cycling protocol, followed by 40 cycles of 5 seconds at 95 ° C. and 10 seconds at 60 ° C. for 3 minutes from 60 ° C. to 95 ° C. It was carried out in the order of the rising melting step. Products amplified by real-time polymerase chain reaction were quantified using Delta delta Ct method and corrected by the expression level of ⁇ -actin for each sample. The base sequence of the primers used in the experiment is as shown in the following [Table 2].
- Malondialdehyde (MDA), a lipid peroxide, was measured using thiobarbituric acid reactive substances (TBARS) assay according to the method of Emerit et al. (Presse Med. 1986 Jun 14; 15 (24): 1131-3). .
- Superoxide disproportionation enzyme (Superoxide dismutase; SOD) is a representative radical trapping the enzyme in the body, superoxide anion, which acts to return to;; (hydrogen peroxide H 2 O 2) ( ⁇ O2 superoxide anion) hydrogen peroxide It is known. Superoxide dismutase was measured using a SOD Assay Kit (Dojindo Molecular Technologies, USA).
- the secretion of histamine in the mast cells induced by compound 48/80 was effectively reduced by curcumin, a positive control, while licorice extract (AC) or A. eryngii water-soluble extract (AEAC). As little changes in histamine levels were observed, water extract of A. erythema (AEAC) had no effect on degranulation inhibition of mast cells.
- the secretion of inflammatory cytokines IL-1 ⁇ and IL-6 was significantly increased in LPS-induced macrophages, and the increase in inflammatory cytokines was markedly decreased by AJC. It confirmed that it became. In addition, it was confirmed that there is no difference in efficacy due to the change in formulation of the liquid or powder process.
- leaf extract exhibits an antacid like the calcium carbonate.
- Leafy leaf extract (AEAA) is thought to have the effect of temporarily preventing gastric damage by acid through neutralization reaction.
- no neutralization-related functions were observed in the water extract of AJU (AEAC) and licorice extract (AC), which means that there are no side effects such as excessive secretion of gastric acid due to a simple neutralization reaction.
- the licorice extract (AC) of 100 mg / kg was 98.7% compared to the HCl / ethanol-administered group.
- the same dose of A. eryngii water extract (AEAC) showed a 104.2% increase, showing a slightly higher gastric mucus increase effect than licorice extract (AC).
- the water extract of A. erythema (AEAC) showed the effect of increasing the amount of gastric mucus in a dose-dependent manner, and showed the maximum effect at the highest dose of 400 mg / kg. It was confirmed that the protective effect against / ethanol-induced acute gastritis / gastric ulcer.
- the treatment and defense effect of AUGC in the acute gastritis / gastric ulcer animal model shows the inflammatory treatment effect by restoring the gastric mucosa's defense function which is reduced by inflammation by restoring SOD activity, an antioxidant enzyme.
- gastric mucosa protein MUC6 when repeated administration of 200 mg / kg or 400 mg / kg of agar water soluble extract (AEAC), gastric mucosa protein MUC6 is 2.4 to 3.9 times, MUC5AC compared to the case of administration only HCl / ethanol was increased 3.0-4.6 fold, rPigr, a factor related to gastric mucosal immunity, 3.9-4.3 fold, TFF1 2.5-4.1 fold, TFF2 9.4-18.2 fold, TFF3 3.2- 4.2 fold.
- AEAC agar water soluble extract
- the water extract of A. eryngii (AEAC) according to the present invention directly promotes the production of mucosal proteins, and it can be seen that it has a continuous gastric mucosal protection effect.
- the mucosal protein system seems to have a protective effect on the gastrointestinal mucosa due to the increase of TFF2, as well as a symptom improvement effect by suppressing gastric acid secretion.
- the gastric injury inhibition rate was 100% inhibition in both 200 mg / kg and 400 mg / kg administration groups when HJ / ethanol was administered followed by administration of AJC.
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Abstract
The present invention relates to an Artemisia capillaris water-soluble extract having an effect of preventing or alleviating gastric mucosal injured gastrointestinal diseases, and a composition for preventing or alleviating mucosal injured gastrointestinal diseases, containing the same and, more specifically, to a composition, a food composition, and a pharmaceutical composition for preventing or alleviating, mucosal injured gastrointestinal diseases, all of the compositions having an effect of preventing or alleviating mucosal injured gastrointestinal diseases caused by excessive stomach acid, alcohol, stress, and the like through the maintenance, protection, and regeneration of the gastrointestinal mucosal system.
Description
본 발명은 인진쑥(Artemisia capillaris) 수용성 추출물, 이를 함유하는 위장 질환의 예방 또는 개선용 조성물, 더욱 상세하게는 위장관 점막 시스템의 유지, 보호 및 재생을 통하여 위산 과다 및 스트레스 등에 의해 유발되는 점막 손상성 위장관계 질환의 예방 또는 개선 효과를 가지는 수용성 인진쑥 추출물을 함유하는 식품 조성물 및 약학 조성물에 관한 것이다. The present invention is a water-soluble extract of Artemisia capillaris , a composition for the prevention or improvement of gastrointestinal diseases containing the same, more specifically mucosal damage gastrointestinal caused by excessive gastric acid and stress through the maintenance, protection and regeneration of the gastrointestinal mucosa system The present invention relates to a food composition and a pharmaceutical composition containing a water-soluble phosphorus mugwort extract having an effect of preventing or ameliorating related diseases.
한국의 자생 쑥은 30여 가지로, 식용과 약용으로 구분해서 사용하고 있다. 그 중에 대표적인 것이 인진쑥(Artemisia capillaris)과 애엽(Artemisia asiatica)이다. 이들은 서로 성질이 달라 전통 한방에서는 서로 다른 용도로 사용해 왔다. 인진쑥은 차가운 성질 때문에 주로 간질환이나 위장병 등에 사용해 온 반면, 애엽은 따뜻한 성질 때문에 뜸을 뜰 때 주로 사용한다.There are about 30 kinds of native mugwort in Korea, which are divided into edible and medicinal. The representative ones are Artemisia capillaris and Artemisia asiatica . They have different properties and have been used for different purposes in traditional Korean medicine. Injin mugwort has been used mainly for liver disease and gastrointestinal diseases because of its cold nature, while the lobe is mainly used for moxibustion due to its warm nature.
인진쑥은 예로부터 지방간, 만성 간염, 위장병, 변비, 황달, 신경통 등에 사용하였고, 알코올 분해와 지방 축적 억제 작용이 있어 음주가 많은 사람과 간 질환자에게 좋으며, 소염, 이뇨, 담석 억제 작용이 알려져 있다.Injin mugwort has been used for fatty liver, chronic hepatitis, gastrointestinal diseases, constipation, jaundice, neuralgia, etc., and is effective for alcohol drinking and fat accumulation inhibitory people and liver disease, and anti-inflammatory, diuretic, gallstone inhibitory effects are known.
애엽은 인진쑥과 달리 성질이 따뜻하여 아랫배가 차갑거나 이로 인해 생리가 고르지 못한 경우에 사용하며, 지혈 효과가 있어서 육혈(코피), 토혈(입으로 토하는 피), 하혈(항문으로 쏟아내는 피). 궁에서 나오는 출혈의 치료에 쓰이고, 통증을 억제하는 구급약으로도 쓰인다.Unlike the jinjin mugwort, warm nature of the lower abdomen is cold or because of this uneven menstruation is used, and hemostatic effect is a blood bleeding (nose blood), bleeding (blood vomiting), bleeding (blood to the anus). It is used to treat bleeding from the womb, and as a first aid to reduce pain.
종래에 애엽 추출물은 항산화 및 NF-κB 억제 효능을 가지며, 이를 기반으로 위 손상 및 염증 억제 작용을 나타낸다는 것이 보고된 바 있다. 그 예로서, NSAIDS로 인한 위 손상 모델에서 PGE2, GSH의 항산화 효과에 기인한 위 손상 방지 효과(World J Gastroenterol 2005; 11(47): 7450-7456), 만성 역류성 위염 모델에서 PGE2, GSH의 항산화 효과에 기인한 위 손상 방지 효과(World J Gastroenterol 2005; 11(47): 7430-7435), 급성 알코올성 위염 모델에서 위 보호 효과를 갖는 호르몬인 그렐린(Ghrelin) 분비 증가(Gut and liver 2011;5(1):52-56), TNF-α 매개 위 상피세포 염증 모델에서 p38 억제 및 NF-κB 억제를 통해 IL-8과 CCL20을 감소시킨다는 것(World J Gastroenterol 2006;12(30):4850-4858)이 보고되었다. Previously, leaf extracts have been reported to have antioxidant and NF-κB inhibitory effects and to suppress gastric damage and inflammation based on them. As an example, the prevention of gastric damage due to the antioxidant effects of PGE2 and GSH in NSAIDS gastric injury model (World J Gastroenterol 2005; 11 (47): 7450-7456), and the antioxidant activity of PGE2 and GSH in chronic reflux gastritis model Effect of prevention of gastric injury due to effects (World J Gastroenterol 2005; 11 (47): 7430-7435), increased secretion of Ghrelin, a hormone with gastric protective effect in acute alcoholic gastritis model (Gut and liver 2011; 5 1): 52-56), reducing IL-8 and CCL20 through p38 inhibition and NF-κB inhibition in a TNF-α mediated gastric epithelial inflammation model (World J Gastroenterol 2006; 12 (30): 4850-4858). ) Has been reported.
인진쑥(Artemisia capillaris) 추출물의 경우 LPS 매개 염증 모델에서 NF-κB 억제를 통해 iNOS, COX2, TNF-α 등을 감소시킨다는 것(INT J MOL MED 13: 717-720, 2004)과, 헬리코박터에 의한 위장 질환 치료 효과 (대한민국 공개특허공보 제10-2002-0046488호, 일본 공개특허 공보 H08-295632)가 보고되었다. Artemisia capillaris extract reduced iNOS, COX2, TNF-α through NF-κB inhibition in LPS-mediated inflammation models (INT J MOL MED 13: 717-720, 2004), and gastrointestinal tract by Helicobacter Disease treatment effects (Korean Unexamined Patent Publication No. 10-2002-0046488, Japanese Unexamined Patent Publication H08-295632) have been reported.
이와 같이 애엽 추출물이나 인진쑥 추출물은 염증 반응의 억제를 통해 급성 위염 또는 세균성 위염을 개선하는 효과를 나타내는 것으로 보인다. 그러나 위장관 점막의 손상을 예방하거나 개선하는 데에 애엽이나 인진쑥 추출물이 효과가 있는지에 대해서는 현재까지 보고된 바가 없다. As described above, the leaf extracts and the extract of Injin mugwort have an effect of improving acute gastritis or bacterial gastritis through suppression of the inflammatory response. However, there have been no reports about the effectiveness of the extracts of leaf or wormwood in preventing or improving gastrointestinal mucosa.
본 발명의 목적은 점막 손상성 위장 질환의 예방 또는 개선 작용을 나타내는 천연물 유래의 추출물을 제공하는 것이다. It is an object of the present invention to provide extracts derived from natural products that exhibit a prophylactic or ameliorating action of mucosal damaging gastrointestinal diseases.
본 발명의 다른 한 가지 목적은 위장관 점막 시스템의 개선 및 보호를 통하여 위산 과다 및/또는 스트레스 등에 의해 유발되는 점막 손상성 위장 질환에 대해 개선 효과를 가지는 위장 질환의 예방 또는 개선용 조성물을 제공하는 것이다. It is another object of the present invention to provide a composition for the prevention or improvement of gastrointestinal diseases having an improvement effect on mucosal damaging gastrointestinal diseases caused by excessive gastric acid and / or stress through improvement and protection of the gastrointestinal mucosa system. .
본 발명의 다른 한 가지 목적은 점막 손상성 위장 질환의 예방 또는 개선용 식품 조성물을 제공하는 것이다.Another object of the present invention to provide a food composition for the prevention or improvement of mucosal damage gastrointestinal diseases.
본 발명의 또 다른 한 가지 목적은 점막 손상성 위장 질환의 예방 또는 치료용 약학 조성물을 제공하는 것이다.Another object of the present invention to provide a pharmaceutical composition for the prevention or treatment of mucosal damage gastrointestinal diseases.
상기와 같은 본 발명의 기술적 과제를 해결하는 본 발명의 과제 해결 수단은 다음과 같다. The problem solving means of this invention which solves the technical subject of this invention as mentioned above is as follows.
1. 위 점막 손상성 위장 질환의 예방 또는 개선 작용을 가지는 인진쑥(Artemisia capillaris) 수용성 추출물.1. Water-soluble extract of Artemisia capillaris , which has the effect of preventing or ameliorating gastric mucosal injuries.
2. 인진쑥을 열수로 추출한 것인 상기 1의 인진쑥(Artemisia capillaris) 수용성 추출물. 2. Artemisia capillaris water-soluble extract of the above 1, which is extracted with hot water mugwort.
3. 인진쑥(Artemisia capillaris) 수용성 추출물을 함유하는 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물.3. Composition for the prevention or improvement of gastric mucosal damaging gastrointestinal diseases containing Artemisia capillaris water-soluble extract.
4. 위장관 점막 시스템의 항상성 유지 및 개선 효과를 통해 위 점막 손상성 위장 질환을 예방 또는 개선하는 것인 상기 3의 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물.4. The composition for the prevention or improvement of gastric mucosal damaging gastrointestinal disease of the above 3, which prevents or improves gastric mucosal damaging gastrointestinal diseases through the effect of maintaining and improving homeostasis of the gastrointestinal mucosa system.
5. 위산, 에탄올, 스트레스 또는 이들의 조합에 의해 유발되는 점막 손상성 위장관계 질환을 개선하는 것인 상기 3의 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물.5. The composition for the prevention or improvement of gastric mucosal damage gastrointestinal diseases of the above 3 to improve the mucosal damage gastrointestinal diseases caused by gastric acid, ethanol, stress or a combination thereof.
6. 위산, 에탄올, 스트레스 또는 이들의 조합에 의해 유발되는 위염 또는 위궤양에 의한 점막 손상을 예방 또는 억제하는 것인 상기 3의 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물.6. The composition for the prevention or improvement of gastric mucosal damaging gastrointestinal disease of 3 above, which prevents or inhibits mucosal damage caused by gastritis or gastric ulcer caused by gastric acid, ethanol, stress or a combination thereof.
7. 위 점막 단백질 또는 점막 면역 인자의 발현 증가를 통해 위 점막 손상을 억제 또는 개선하는 것인 상기 3의 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물.7. Composition for the prevention or improvement of gastric mucosal damage gastrointestinal disease of 3 above, which inhibits or improves gastric mucosal damage through increased expression of gastric mucosal proteins or mucosal immune factors.
8. 상기 위 점막 단백질 또는 점막 면역 인자는 MUC5AC, MUC6, rPigr, TFF1, TFF2 및 TFF3로 구성된 군에서 1종 이상 선택되는 것인 상기 7의 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물.8. The gastric mucosal protein or mucosal immune factor is MUC5AC, MUC6, rPigr, TFF1, TFF2 and TFF3 composition for the prevention or improvement of gastric mucosal damaging gastrointestinal disease of the above 7, wherein at least one selected from the group consisting of.
9. 염증성 사이토카인 억제, 지질 과산화물의 생성 억제 또는 항산화 효소 활성 증가를 통해 위 점막 손상을 억제 또는 개선하는 것인 상기 3의 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물.9. The composition for the prevention or improvement of gastric mucosal damaging gastrointestinal disease of 3 above, which inhibits or improves gastric mucosal damage by inhibiting inflammatory cytokines, inhibiting the production of lipid peroxides, or increasing antioxidant enzyme activity.
10. 인진쑥(Artemisia capillaris) 수용성 추출물을 함유하는 위 점막 손상성 위장 질환의 예방 또는 개선용 식품 조성물.10. Food composition for the prevention or improvement of gastric mucosal damaging gastrointestinal disease containing Artemisia capillaris water-soluble extract.
11. 인진쑥(Artemisia capillaris) 수용성 추출물을 함유하는 위 점막 손상성 위장 질환의 예방 또는 치료용 약학 조성물. 11. A pharmaceutical composition for preventing or treating gastric mucosal damaging gastrointestinal diseases containing Artemisia capillaris water-soluble extract.
12. 위 점막 단백질 또는 점막 면역 인자의 발현 증가를 통해 위 점막 손상을 억제 또는 개선하는 것인 상기 11의 위 점막 손상성 위장 질환의 예방 또는 치료용 약학 조성물.12. The pharmaceutical composition for preventing or treating gastric mucosal damaging gastrointestinal disease of 11, which inhibits or ameliorates gastric mucosal damage through increased expression of gastric mucosal proteins or mucosal immune factors.
13. 상기 위 점막 단백질 또는 점막 면역 인자는 MUC5AC, MUC6, rPigr, TFF1, TFF2 및 TFF3로 구성된 군에서 1종 이상 선택되는 것인 상기 12의 위 점막 손상성 위장 질환의 예방 또는 치료용 약학 조성물.13. The pharmaceutical composition for preventing or treating gastric mucosal damaging gastrointestinal disease of 12, wherein the gastric mucosal protein or mucosal immune factor is at least one selected from the group consisting of MUC5AC, MUC6, rPigr, TFF1, TFF2, and TFF3.
14. 염증성 사이토카인 억제, 지질 과산화물의 생성 억제 또는 항산화 효소 활성 증가를 통해 위 점막 손상을 억제 또는 개선하는 것인 상기 11의 위 점막 손상성 위장 질환의 예방 또는 치료용 약학 조성물.14. The pharmaceutical composition for preventing or treating gastric mucosal damaging gastrointestinal disease of 11, wherein the gastrointestinal mucosa is inhibited or improved by inhibiting inflammatory cytokines, inhibiting production of lipid peroxides, or increasing antioxidant enzyme activity.
15. 인진쑥(Artemisia capillaris)을 열수로 추출하는 단계;15. Extracting Artemisia capillaris with hot water;
추출액을 상온까지 냉각시켜 여과하고, 여액을 부피가 10±5%가 될 때까지 1차 농축 한 후 침전물을 제거하는 단계; 및Cooling the extract to room temperature and filtering the filtrate, and firstly concentrating the filtrate until the volume is 10 ± 5% to remove the precipitate; And
침전물이 제거된 농축액을 수분 함량이 55±5%가 될 때까지 2차 농축하여 인진쑥(Artemisia capillaris) 수용성 추출물을 얻는 단계;Concentration of the concentrate from which the precipitate was removed was second concentrated until the water content was 55 ± 5% to obtain an Artemisia capillaris aqueous extract;
를 포함하는, 인진쑥(Artemisia capillaris) 수용성 추출물의 제조 방법. , Injinssuk (Artemisia capillaris) method for producing an aqueous extract containing.
본 발명에 따르면, 인진쑥 수용성 추출물(aqueous extract of Artemisia capillaris, 이하, 'AEAC'로 지칭되기도 함)은 위장관 점막 시스템의 주요 조절 인자(TFF1, 2, 3)를 비롯하여 위점막 뮤신 단백질(MUC6, MUC5AC), 점막 면역 관련 인자(rPigr), 항산화 효소(SOD)의 활성을 증가시키며, 염증성 사이토카인(IL-1β, IL-6, TNF-α), 지질 과산화물(MDA)의 생성을 억제시킬 수 있다. 인진쑥 수용성 추출물은 위장관 점막 시스템의 주요 조절 인자들의 발현을 증가시킴으로써 염산-에탄올에 의해 유발된 위염/위궤양에 대한 점막 손상의 예방 및 억제 효과를 나타내므로, 점막 생성 촉진을 통한 위장관 보호 작용과, 이를 통한 스트레스 또는 위산에 의한 위 손상을 방지하기 위한 천연물 의약품, 건강 기능 식품, 기능성 음료 등으로 이용될 수 있다. 또한, 자극적인 맵고 짠 음식을 주로 섭취하는 것으로 인해 발생할 수 있는 위염이나 위궤양의 예방 및 치료, 잦은 음주나 NSAIDs 투약에 따른 점막의 생성 저하로 인한 위산 과다, 소화 불량, 가스 팽만 등의 예방 및 치료, 스트레스로 인한 위산 과다 분비 및 위염, 위궤양의 예방 및 치료에도 사용될 수 있을 것으로 기대된다. 뿐만 아니라 인진쑥 수용성 추출물은 TFF3의 증가를 통하여 위암, 대장암에 대한 예방 및 치료에도 응용이 가능하다.According to the present invention, Aqueous extract of Artemisia capillaris (hereinafter also referred to as 'AEAC') is a gastric mucosal mucin protein (MUC6, MUC5AC), including the major regulatory factors (TFF1, 2, 3) of the gastrointestinal mucosa system. Can increase mucosal immune related factors (rPigr), antioxidant enzymes (SOD), and inhibit the production of inflammatory cytokines (IL-1β, IL-6, TNF-α) and lipid peroxides (MDA). . Phosphorus erythema extracts have the effect of preventing and inhibiting mucosal damage to gastritis / gastric ulcer induced by hydrochloric acid-ethanol by increasing the expression of key regulatory factors of the gastrointestinal mucosal system, thus protecting the gastrointestinal tract through promoting mucosal production, It can be used as natural medicines, health functional foods, functional drinks, etc. to prevent gastric damage caused by stress or stomach acid. It also prevents and treats gastritis and gastric ulcers, which can be caused mainly by eating spicy, spicy foods, and excessive gastric acid, indigestion, and gas bloating caused by frequent drinking or decreased mucosa following NSAIDs. In addition, it is expected to be used for the prevention and treatment of gastric acid secretion and gastritis caused by stress and gastric ulcer. In addition, Injin mugwort extract is applicable to the prevention and treatment of gastric cancer and colon cancer through the increase of TFF3.
도 1은 compound 48/80으로 유도된 비만세포에서 인진쑥 수용성 추출물(AEAC)이 비만세포 탈과립화에 미치는 영향을 확인한 결과이다.Figure 1 is a result confirming the effect of the extract of A. erysipelas water soluble extract (AEAC) in mast cell degranulation in the mast cells induced by compound 48/80.
도 2는 PMA 및 A23187로 유도된 비만세포에서 인진쑥 수용성 추출물(AEAC)에 의한 염증성 사이토카인 억제 효과를 확인한 결과이다.Figure 2 is a result confirming the inhibitory effect of inflammatory cytokine by the water soluble extract (AEAC) in mast cells induced by PMA and A23187.
도 3은 LPS로 유도된 대식세포에서 인진쑥 수용성 추출물(AEAC)에 의한 염증성 사이토카인 억제 효과를 확인한 결과이다.Figure 3 is a result confirming the inhibitory effect of inflammatory cytokine by L. aeruginosa water-soluble extract (AEAC) in LPS-induced macrophages.
도 4는 탄산칼슘, 인진쑥 수용성 추출물(AEAC), 감초 추출물(AC) 및 애엽 추출물(AEAA)의 중화 능력을 확인한 결과이다.Figure 4 is a result of confirming the neutralizing ability of calcium carbonate, phosphorus mugwort soluble extract (AEAC), licorice extract (AC) and leaf extract (AEAA).
도 5는 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC)이 혈청 내 히스타민의 생성 또는 분비에 미치는 영향을 확인한 결과이다.Figure 5 is a result confirming the effect of H. ethanol-induced acute gastritis / gastric ulcer animal model influx of the extract of AJC in the production or secretion of histamine in serum.
도 6은 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC)에 의한 위 점막 보호 효과를 확인한 결과이다.Figure 6 is a result confirming the gastric mucosa protective effect by the water soluble extract (AEAC) in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
도 7은 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC)에 의한 위 점액량의 증가 효과를 확인한 결과이다.Figure 7 shows the results of confirming the effect of increasing the amount of gastric mucus by the extract of AJC in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
도 8은 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC)에 의한 말론디알데하이드의 억제 효과를 확인한 결과이다.8 is a result confirming the inhibitory effect of malondialdehyde by the water soluble extracts of AJ (AEAC) in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
도 9는 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC)에 의한 초과산화물 불균등화효소의 활성 증가 효과를 확인한 결과이다.Figure 9 is a result confirming the effect of increasing the activity of the superoxide dismutase by AJC (AEAC) in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
도 10은 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC)을 단회 투여한 후, 위 점막 단백질인 MUC6의 발현 증가 효과를 확인한 결과이다. 10 is a result of confirming the effect of increasing the expression of gastric mucosa protein MUC6 after a single administration of AJC in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
도 11은 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC)을 단회 투여한 후, 위 점막 단백질인 MUC5AC의 발현 증가 효과를 확인한 결과이다. 11 is a result of confirming the effect of increasing the expression of gastric mucosa protein MUC5AC after a single administration of AJC (AEAC) in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
도 12는 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC)을 단회 투여한 후, 위 점막 면역 관련 인자인 rPigr의 발현 증가 효과를 확인한 결과이다. 12 is a result of confirming the effect of increasing the expression of rPigr, a mucosal immune-related factor after a single administration of AJC in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
도 13은 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC)을 단회 투여한 후, 위 점막 보호 및 치료에 관여하는 TFF1, 2 및 3의 발현 증가 효과를 확인한 결과이다. Figure 13 is a result of confirming the effect of increasing the expression of TFF1, 2 and 3 involved in gastric mucosa protection and treatment after a single administration of AJC in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
도 14는 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC)을 반복 투여한 후, 위 점막 단백질인 MUC6의 발현 증가 효과를 확인한 결과이다. 14 is a result of confirming the effect of increasing the expression of gastric mucosa protein MUC6 after repeated administration of AJC in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
도 15는 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC)을 반복 투여한 후, 위 점막 단백질인 MUC5AC의 발현 증가 효과를 확인한 결과이다. 15 is a result of confirming the effect of increasing the expression of gastric mucosa protein MUC5AC after repeated administration of AJC in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
도 16은 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC)을 반복 투여한 후, 위 점막 면역 관련 인자인 rPigr의 발현 증가 효과를 확인한 결과이다. 16 is a result of confirming the effect of increasing the expression of rPigr, gastric mucosal immunity-related factors after repeated administration of AJC in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
도 17은 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC)을 반복 투여한 후, 위 점막 보호 및 치료에 관여하는 TFF1, 2 및 3의 발현 증가 효과를 확인한 결과이다. Figure 17 shows the effect of increasing the expression of TFF1, 2 and 3 involved in gastric mucosa protection and treatment after repeated administration of AJC in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
도 18은 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC) 또는 감초 추출물(AC)을 반복 투여한 후, 위 점막 단백질, 위 점막 면역 관련 인자 및 위 점막 보호 및 치료에 관여하는 인자들의 발현 증가 효과를 확인한 결과이다. FIG. 18 is a method for protecting and treating gastric mucosa protein, gastric mucosal immunity related factors and gastric mucosa after repeated administration of AJC or licorice extract (AC) in HCl / ethanol-induced acute gastritis / gastric ulcer animal model This is the result of confirming the effect of increasing the expression of the factors involved.
도 19는 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC) 또는 감초 추출물(AC)을 단회 투여한 후, 위의 대만부를 절개하여 발생된 위 점막 손상 정도를 비교한 사진이다. 19 is a photograph comparing the degree of gastric mucosal injury caused by dissecting the Taiwanese stomach after single administration of AJC or Licorice extract (AC) in acute gastritis / gastric ulcer animal model induced by HCl / ethanol. to be.
도 20은 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC) 또는 감초 추출물(AC)을 단회 투여한 후, 위의 대만부를 절개하여 발생된 위 점막 손상 길이를 정량 분석한 결과이다. FIG. 20 is a quantitative analysis of gastric mucosal injury length caused by dissection of the Taiwanese stomach after single administration of AJC or Licorice Extract (AC) in acute gastritis / gastric ulcer animal model induced by HCl / ethanol. The result is.
도 21은 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC) 또는 감초 추출물(AC)을 반복 투여한 후, 위의 대만부를 절개하여 발생된 위 점막 손상 정도를 비교한 사진이다. Figure 21 is a photograph comparing the degree of gastric mucosal damage caused by dissection of the stomach part of the stomach after repeated administration of AJC or licorice extract (AC) in acute gastritis / gastric ulcer animal model induced by HCl / ethanol to be.
도 22는 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC) 또는 감초 추출물(AC)을 반복 투여한 후, 위의 대만부를 절개하여 발생된 위 점막 손상 길이를 정량 분석한 결과이다. 22 is a quantitative analysis of the gastric mucosal injury length caused by dissection of the stomach part of the stomach after repeated administration of AJC or licorice extract (AC) in an acute gastritis / gastric ulcer animal model induced by HCl / ethanol. The result is.
도 23은 HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 인진쑥 수용성 추출물(AEAC) 또는 애엽 추출물(AEAA)을 단회 투여한 후, 위의 대만부를 절개하여 발생된 손상 정도를 비교한 사진이다. Figure 23 is a photograph comparing the degree of damage caused by dissecting the stomach part of the stomach after a single administration of AJC or AEAA in acute gastritis / gastric ulcer animal model induced by HCl / ethanol.
위장관 점막 시스템은 다양한 인자들에 의해 조절되는 것으로 알려져 있다. 그 중에 대표적인 조절인자는 TFF(Trefoil factor) 패밀리에 속하는 TFF1, TFF2 및 TFF3이다. TFF1은 위암과 관련하여 암 억제 유전자로 알려져 있을 뿐 아니라, 위장관 점막을 다양한 손상으로부터 보호하고, 점막층의 안정화 및 위장관 상피조직의 치유 과정을 조절하는 것으로 보고되었다. TFF2는 TFF1이 갖는 위장관 점막에 대한 보호 효과 뿐 아니라 위산 분비를 억제하는 기능을 갖는다. TFF3은 위장관 점막 뿐 아니라 모유에도 함유되어 있으며, 탁월한 염증 억제 작용을 나타내는 것으로 보고되었다. 이외에도 뮤신 단백질(MUC6 및 MUC5AC)은 직접적으로 위장관 점막을 구성하면서 위장관 상피조직을 보호하며, 점막 면역 관련 인자인 rPigr 등은 위장관 점막 시스템의 주요 인자로 여겨진다. 하지만, 이러한 위장관 점막 시스템의 기능을 조절하는 의약품이나 천연물 유래의 추출물은 현재까지 알려진 바가 없다. The gastrointestinal mucosa system is known to be regulated by various factors. Representative regulators are TFF1, TFF2 and TFF3 belonging to the Trefoil factor (TFF) family. TFF1 is not only known as a cancer suppressor gene in connection with gastric cancer, but has been reported to protect the gastrointestinal mucosa from various injuries, to stabilize the mucosal layer and to regulate the healing process of gastrointestinal epithelial tissue. TFF2 has a function of inhibiting gastric acid secretion as well as a protective effect on the gastrointestinal mucosa of TFF1. TFF3 is found not only in the gastrointestinal mucosa but also in breast milk and has been reported to exhibit excellent anti-inflammatory effects. In addition, mucin proteins (MUC6 and MUC5AC) directly constitute the gastrointestinal mucosa and protect the gastrointestinal epithelial tissue, and rPigr, a mucosal immune-related factor, is considered to be a major factor in the gastrointestinal mucosal system. However, extracts derived from medicines or natural products that regulate the function of such gastrointestinal mucosal systems have not been known to date.
이에 본 발명의 발명자들은 인진쑥(Artemisia capillaris) 수용성 추출물이 점막 단백질 또는 점막 면역 인자, 구체적으로는 MUC5AC, MUC6, rPigr 및 TFF3 중에서 선택되는 1종 이상의 점막 단백질 또는 점막 면역 인자의 발현을 증가시키고, 염증성 사이토카인 억제, 지질 과산화물의 생성 억제 또는 항산화 효소 활성 증가를 통해 위장관 점막 시스템의 항상성을 유지 및 개선시키고, 염증 억제 및 점막 생성 촉진 효과를 나타냄으로써, 위 점막 손상성 위장 질환을 예방 또는 개선한다는 것을 밝혀내며 본 발명을 완성하였다. Accordingly, the inventors of the present invention, the Artemisia capillaris water-soluble extracts increase the expression of mucosal proteins or mucosal immune factors, specifically one or more mucosal proteins or mucosal immune factors selected from MUC5AC, MUC6, rPigr and TFF3, inflammatory Maintains and improves the homeostasis of the gastrointestinal mucosa system by inhibiting cytokine, inhibiting the production of lipid peroxides, or increasing antioxidant enzyme activity, and inhibiting inflammation and promoting mucosal production, thereby preventing or improving gastric mucosal damage. The present invention has been completed by finding out.
따라서 본 발명은 위 점막 손상성 위장 질환의 예방 또는 개선 작용을 가지는 인진쑥(Artemisia capillaris) 수용성 추출물, 이를 포함하는 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물 및 식품 조성물, 위 점막 손상성 위장 질환 예방 또는 치료용 약학 조성물, 및 인진쑥(Artemisia capillaris) 수용성 추출물의 제조 방법에 관한 것이다. Therefore, the present invention is a water-soluble extract of Artemisia capillaris having an action of preventing or improving gastric mucosal gastrointestinal diseases, compositions and food compositions for preventing or ameliorating gastric mucosal gastrointestinal disorders, gastric mucosal gastrointestinal disorders It relates to a pharmaceutical composition for prophylaxis or treatment, and a method for preparing Artemisia capillaris water-soluble extract.
본 발명의 첫 번째 측면은 위 점막 손상성 위장 질환의 예방 또는 개선 작용을 가지는 인진쑥(Artemisia capillaris) 수용성 추출물에 관한 것이다.The first aspect of the present invention relates to a water-soluble extract of Artemisia capillaris having a prophylactic or ameliorating action of gastric mucosal damaging gastrointestinal diseases.
본 발명에 따른 인진쑥(Artemisia capillaris) 수용성 추출물은 인진쑥을 열수로 추출한 것으로, 점막 단백질 또는 점막 면역 인자, 구체적으로는 MUC5AC, MUC6, rPigr, TFF1, TFF2 및 TFF3 중에서 선택되는 1종 이상의 단백질 또는 점막 면역 인자의 발현을 증가시키고, 염증성 사이토카인 억제, 지질 과산화물의 생성 억제 또는 항산화 효소 활성 증가를 통해 위장관 점막 시스템의 항상성을 유지 및 개선시키고, 염증 억제 또는 점막 생성 촉진 효과를 나타낸다. Artemisia capillaris water-soluble extract according to the present invention is extracted from hot water of Injin mugwort, mucosal proteins or mucosal immune factors, specifically MUC5AC, MUC6, rPigr, TFF1, TFF2 and TFF3 at least one protein or mucosal immunity It increases the expression of the factor, maintains and improves the homeostasis of the gastrointestinal tract mucosal system through inhibiting inflammatory cytokines, inhibiting the production of lipid peroxides or increasing antioxidant enzyme activity, and exhibits the effect of inhibiting inflammation or promoting mucosal production.
따라서 본 발명의 인진쑥(Artemisia capillaris) 수용성 추출물은 예를 들면 위산, 에탄올, 스트레스, 또는 이들의 조합에 의해 유발되는 점막 손상성 위장관계 질환을 예방 또는 개선하거나, 위산, 에탄올, 스트레스 또는 이들의 조합에 의해 유발되는 위염 또는 위궤양에 의해 점막이 손상되는 것을 예방하거나 억제하는 용도로 사용될 수 있다. Thus, the Artemisia capillaris water-soluble extract of the present invention prevents or ameliorates mucosal damaging gastrointestinal disorders caused by, for example, gastric acid, ethanol, stress, or a combination thereof, gastric acid, ethanol, stress or a combination thereof. It can be used to prevent or inhibit the damage to the mucous membrane by gastritis or gastric ulcer caused by.
따라서 본 발명의 두 번째 측면은 상기 인진쑥(Artemisia capillaris) 수용성 추출물을 함유하는 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물에 관한 것이다.Therefore, the second aspect of the present invention relates to a composition for the prevention or improvement of gastric mucosal damaging gastrointestinal disease containing the water extract of Artemisia capillaris .
또한 본 발명의 세 번째 측면은 상기 인진쑥(Artemisia capillaris) 수용성 추출물을 함유하는 위 점막 손상성 위장 질환의 예방 또는 개선을 위한 식품 조성물에 관한 것이다. In addition, a third aspect of the present invention relates to a food composition for preventing or ameliorating gastric mucosal damaging gastrointestinal disease containing the water extract of Artemisia capillaris .
본 발명의 네 번째 측면은 인진쑥(Artemisia capillaris) 수용성 추출물을 함유하는 위 점막 손상성 위장 질환의 예방 또는 치료용 약학 조성물에 관한 것이다.A fourth aspect of the present invention relates to a pharmaceutical composition for the prevention or treatment of gastric mucosal damage gastrointestinal diseases containing Artemisia capillaris water-soluble extract.
본 발명에 따른 위 점막 손상성 위장 질환의 예방 또는 치료용 약학 조성물은 인진쑥(Artemisia capillaris) 수용성 추출물을 유효 성분으로 함유함으로써 위 점막 단백질 또는 점막 면역 인자, 구체적으로는 MUC5AC, MUC6, rPigr, TFF1, TFF2 및 TFF3 중에서 선택되는 1종 이상의 위 점막 단백질 또는 점막 면역 인자의 발현을 증가시키고, 염증성 사이토카인 억제, 지질 과산화물의 생성 억제 또는 항산화 효소 활성 증가를 통해 위장관 점막 시스템의 항상성을 유지 및 개선시키고, 염증 억제 또는 점막 생성 촉진 효과를 나타낸다. The pharmaceutical composition for the prevention or treatment of gastric mucosal damaging gastrointestinal diseases according to the present invention contains a water-soluble extract of Artemisia capillaris as an active ingredient, specifically gastric mucosal proteins or mucosal immune factors, specifically MUC5AC, MUC6, rPigr, TFF1, Increases the expression of one or more gastric mucosal proteins or mucosal immune factors selected from TFF2 and TFF3, maintains and improves the homeostasis of the gastrointestinal mucosal system through inhibiting inflammatory cytokines, inhibiting the production of lipid peroxides or increasing antioxidant enzyme activity, Inhibit inflammation or promote mucosal production.
본 발명에 따른 위 점막 손상성 위장 질환의 예방 또는 치료용 약학 조성물은 인진쑥(Artemisia capillaris) 수용성 추출물 이외에, 약학적 조성물의 제조에 통상적으로 사용되는 적절한 담체, 부형제, 희석제 등을 더 포함할 수 있다. The pharmaceutical composition for preventing or treating gastric mucosal damaging gastrointestinal diseases according to the present invention may further include appropriate carriers, excipients, diluents, and the like commonly used in the preparation of pharmaceutical compositions in addition to the water-soluble extract of Artemisia capillaris . .
본 발명에 따른 약학 조성물은 인진쑥(Artemisia capillaris) 수용성 추출물을 그대로 함유하거나 추출물 중에 함유되어 있는 성분들의 약학적으로 허용 가능한 염을 함유할 수도 있다. 본 발명의 약학 조성물은 상기 인진쑥(Artemisia capillaris) 수용성 추출물을 단독으로 또는 다른 약학적 활성 화합물과 함께 사용될 수도 있다. The pharmaceutical composition according to the present invention may contain the Artemisia capillaris water-soluble extract as it is or may contain a pharmaceutically acceptable salt of the components contained in the extract. The pharmaceutical composition of the present invention may be used alone or in combination with other pharmaceutically active compounds of Artemisia capillaris .
본 발명에 따른 위 점막 손상성 위장 질환의 예방 또는 치료용 약학 조성물은 통상의 방법 및 필요에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구 제형, 외용제, 좌제, 멸균 주사 용액 등 어떠한 형태로도 사용될 수 있다.Pharmaceutical compositions for the prevention or treatment of gastric mucosal injury gastrointestinal diseases according to the present invention, powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, external preparations, suppositories, according to conventional methods and needs It may be used in any form such as sterile injectable solution.
또한 본 발명 다섯 번째 측면은 인진쑥(Artemisia capillaris) 수용성 추출물의 제조 방법에 관한 것이다. In addition, the fifth aspect of the present invention relates to a method for producing an aqueous extract of Artemisia capillaris .
본 발명에 따른 인진쑥(Artemisia capillaris) 수용성 추출물의 제조 방법은, Method for producing a water-soluble extract of Artemisia capillaris according to the present invention,
(1) 인진쑥(Artemisia capillaris)을 열수로 추출하는 단계;(1) extracting Artemisia capillaris with hot water;
(2) 추출액을 상온까지 냉각시켜 여과하고, 여액을 부피가 10±5%가 될 때까지 1차 농축 한 후 침전물을 제거하는 단계; 및(2) filtering the extract by cooling to room temperature, and firstly concentrating the filtrate until the volume is 10 ± 5%, and then removing the precipitate; And
(3) 침전물이 제거된 농축액을 수분 함량이 55±5%가 될 때까지 2차 농축하여 인진쑥(Artemisia capillaris) 수용성 추출물을 얻는 단계;를 포함한다. (3) obtaining a water-soluble extract of Artemisia capillaris by secondary concentration of the concentrate from which the precipitate is removed until the water content reaches 55 ± 5%.
상기 단계 (1)에서는 건조 및 적절한 크기로 절단된 인진쑥(Artemisia capillaris)에 10-14배의 물, 바람직하게는 정제수를 가하고, 바람직하게는 80±20℃, 더욱 바람직하게는 90±10℃에서 3-5 시간 동안 추출한다. 따라서 상기 단계 (1)의 열수는 바람직하게는 80±20℃, 더욱 바람직하게는 90±10℃의 물일 수 있다. In step (1), 10-14 times of water, preferably purified water, is added to Artemisia capillaris , dried and cut to an appropriate size, preferably at 80 ± 20 ° C, more preferably at 90 ± 10 ° C. Extract for 3-5 hours. Therefore, the hydrothermal water of step (1) may be preferably 80 ± 20 ° C., more preferably 90 ± 10 ° C. water.
상기 단계 (2)에서는 단계 (1)에서 얻은 추출액을 상온까지 냉각되도록 한 다음 여과하고, 여액을 60℃ 미만의 온도에서 1차 감압 농축한 후, 4000 내지 7000 rpm 이상으로 원심분리하고, 침전물을 여과하여 제거한다. In step (2), the extract obtained in step (1) is cooled to room temperature and then filtered, and the filtrate is concentrated under reduced pressure at a temperature of less than 60 ° C. first, and then centrifuged at 4000 to 7000 rpm or more, and the precipitate is Remove by filtration.
단계 (3)에서는 침전물이 제거된 농축액을 60℃ 미만에서 수분 함량이 55±5%가 될 때까지 2차 감압 농축하여 인진쑥 수용성 추출물을 얻는다.In step (3), the concentrated solution from which the precipitate is removed is concentrated under a second reduced pressure until the water content is 55 ± 5% at less than 60 ° C. to obtain a water-soluble extract of Insam.
본 발명에 있어서, 원료로 사용되는 인진쑥(Artemisia capillaris)은 국내에서 자생하는 것이면 어떤 것이라도 사용 가능하다.In the present invention, Artemisia capillaris used as a raw material can be used as long as it is native to Korea.
이하에서는 실시예를 통해 본 발명을 더욱 상세히 설명한다. 그러나 하기 실시예는 본 발명의 예시일 뿐, 본 발명의 범위가 이에 의하여 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to Examples. However, the following examples are merely illustrative of the present invention, and the scope of the present invention is not limited thereto.
실시예Example
1. 시약, 시료, 세포 및 실험동물 준비1. Preparation of reagents, samples, cells and laboratory animals
1-1. 시약1-1. reagent
NaOH, 메틸 오렌지, 탄산칼슘, 염산, 에탄올은 Sigma-Aldrich(St. Louis, MO, USA)에서 구입하였다. 중합효소 연쇄반응(Polymerase chain reaction; PCR) 분석에서 이용된 oligo(dT), dNTP, RNase-free water 및 Superscript III First-Strand Synthesis는 Invitrogen(Carlsbad, CA, USA)에서 구입하였다. RNeasy Mini Kit, Rotor-Gene SYBR Green PCR Kit는 Qiagen(Valencia, CA, USA)에서, oligonucleotide primer는 Bioneer(Daejeon, Korea)에서 각각 구입하여 사용하였다.NaOH, methyl orange, calcium carbonate, hydrochloric acid, ethanol were purchased from Sigma-Aldrich (St. Louis, MO, USA). Oligo (dT), dNTP, RNase-free water and Superscript III First-Strand Synthesis used in Polymerase chain reaction (PCR) analysis were purchased from Invitrogen (Carlsbad, CA, USA). The RNeasy Mini Kit and Rotor-Gene SYBR Green PCR Kit were purchased from Qiagen (Valencia, CA, USA) and oligonucleotide primers were purchased from Bioneer (Daejeon, Korea).
1-2. 시료1-2. sample
쑥 원료로는 경북 청송산 인진쑥(Artemisia capillaris)을 ㈜두손애약초에서 구입하여 사용하였고, 대조물질인 감초 추출물(추출물 고형분 40% 함유 분말)은 (주)서흥으로부터, 애엽은 (주)지뉴인텍으로부터 제공받아 사용하였다. 시료는 인진쑥 수용성 추출물(AEAC) 또는 대조물질(감초 추출물; AC, 애엽 수용성 추출물; AEAA)을 물에 녹여 농도 별로 제조하여 사용하였다.As a mugwort, Artemisia capillaris of Cheongsong, Gyeongbuk was purchased from Duson Aegyo Herbal Co., Ltd. The licorice extract (powder containing 40% of extract solids) as a control material was obtained from Seoheung Co., Ltd. It was supplied from and used. Samples were prepared by dissolving indigo plant water extract (AEAC) or a control material (Licorice extract; AC, leafy water extract; AEAA) in water by concentration.
1-3. 세포1-3. cell
인간 비만세포(Human Mast Cell; HMC-1)는 한국 세포주 은행에서 구입하였으며, 10% 우태아혈청(fetal bovine serum; FBS), 100U/ml 페니실린 및 100㎍/ml 스트렙토마이신(GibcoBRL, USA)이 첨가된 IMDM 배지(GibcoBRL, USA)를 이용하여 37℃, 5% CO2 환경 하에서 배양하였다. 또한, 마우스 유래 대식세포주인 RAW264.7 세포는 10% 우태아혈청 및 1% 페니실린-스트렙토마이신이 첨가된 DMEM 배지를 이용하여 37℃, 5% CO2 환경 하에서 배양하였다. Human Mast Cells (HMC-1) were purchased from the Bank of Korea Cell Line, and 10% fetal bovine serum (FBS), 100 U / ml penicillin and 100 μg / ml streptomycin (GibcoBRL, USA) Cultured under 37 ° C., 5% CO 2 environment with added IMDM medium (GibcoBRL, USA). In addition, RAW264.7 cells, a mouse-derived macrophage line, were cultured in a 37 ° C., 5% CO 2 environment using DMEM medium supplemented with 10% fetal calf serum and 1% penicillin-streptomycin.
1-4. 실험동물1-4. Laboratory animals
실험동물로는 생후 7주령 내외의 수컷 Sprague Dawley계 흰 쥐((주)효창, 한국)를 구입하여 온도 23±1˚C, 습도 55±5%로 조정된 항온항습 사육장치(대종기기상사, 한국) 내에서 1주일간 적응시킨 후 실험에 사용하였다. 전 실험 기간 동안 사료(샘타코, 한국)와 음수는 자유 섭취시켰다.As experimental animals, male Sprague Dawley rats (Hyochang Co., Ltd., Hyochang, Korea), aged 7 weeks old, were purchased and adjusted to a temperature of 23 ± 1˚C and a humidity of 55 ± 5%. It was used for 1 week after adapting in Korea). Feed (Sampaco, Korea) and drinking water were freely consumed during the entire experiment.
2. 인진쑥 수용성 추출물(AEAC) 시료 제조2. Preparation of AJC Aqueous Extract (AEAC) Sample
건조 및 절단된 인진쑥(Artemisia capillaris, 경북 청송산)을 추출기에 넣고, 10-14배의 정제수를 가한 다음, 90±5℃에서 3-5시간 동안 추출하였다. 얻어진 추출액을 1 micron 폴리프로필렌 필터로 여과하고, 여과된 추출액을 60℃ 미만에서 1차 감압 농축한 후, 7000rpm 이상으로 연속식 원심분리하고, 침전물을 여과하여 제거하였다. Dried and cut Injin mugwort ( Artemisia capillaris , Cheongsong, Gyeongbuk) was added to the extractor, 10-14 times purified water was added, and extracted at 90 ± 5 ° C. for 3-5 hours. The obtained extract was filtered with a 1 micron polypropylene filter, and the filtered extract was concentrated under reduced pressure at less than 60 ° C. first, followed by continuous centrifugation at 7000 rpm or more, and the precipitate was filtered off.
침전물을 제거한 농축액을 60℃ 미만에서 수분 함량이 55±5%가 될 때까지 2차 감압 농축하여 인진쑥 수용성 추출물(AEAC, 고형분 함량 45±5%)을 얻고, 85-90℃에서 30-60분 동안 살균하였다. 이 추출물을 용기에 포장하여 냉장 보관하면서 본 실험을 위한 최종 시료로 사용하였다(수율: 원재료 중량 대비 AEAC의 고형분 함량 기준 8-13%). The concentrated solution from which the precipitate was removed was concentrated under a second reduced pressure until the moisture content was 55 ± 5% at less than 60 ° C to obtain a water-soluble extract of AJU (AEAC, 45 ± 5% solids), and 30-60 minutes at 85-90 ° C. Sterilized during. The extract was packaged in a container and refrigerated and used as the final sample for this experiment (yield: 8-13% based on the solids content of AEAC relative to the weight of the raw material).
C18 컬럼(4.6×250mm, 100Å, Kinetex)을 이용하여 344nm의 파장에서 Dionex ChromelonTM Chromatography Data System으로 인진쑥 수용성 추출물의 지표 성분인 스코폴레틴(Scopoletin)의 함량을 분석하였다. 이때 이동상으로는 0.05% 포름산 수용액을 A 용매로 하고, 아세토나이트릴을 B 용매로 사용하였다. A/B (90/10), 13min A/B (5/95), 16min (Gradient) - A/B (90/10)의 이동상 조건에서 분석을 실시하였다. HPLC 분석 결과, 인진쑥 수용성 추출물의 스코폴레틴(Scopoletin) 함량은 인진쑥 추출물 1g 당 평균 3.502 mg으로 확인되었다.The content of scopoletin, an index component of water soluble extracts, was analyzed by Dionex Chromelon TM Chromatography Data System at a wavelength of 344nm using a C18 column (4.6 × 250mm, 100Å, Kinetex). At this time, 0.05% formic acid aqueous solution was used as the A solvent and acetonitrile was used as the B solvent as the mobile phase. Analysis was carried out in mobile phase conditions of A / B (90/10), 13 min A / B (5/95), 16 min (Gradient) -A / B (90/10). As a result of HPLC analysis, the scopoletin content of the water-soluble extracts of the extract was averaged at 3.502 mg per g of the extract.
3. 제산력 시험3. Antacid test
제산력 시험은 위산과 반응하여 제산 작용을 나타내는 제제의 제산력을 확인하는 시험법으로, 본 발명에서는 대한 약전에 명시된 제산력 시험법에 따라 시험하였다. The antacid test is a test method for confirming the antacid power of an agent that reacts with gastric acid, and in the present invention, it was tested according to the antacid test method specified in the Korean Pharmacopoeia.
0.1N HCl 100ml에 대조물질인 감초 추출물(AC) 및 애엽 수용성 추출물(AEAA)과, 본 발명에 따른 인진쑥 수용성 추출물(AEAC) 시료를 각각 1mg씩 혼합하고, 37℃의 진탕 배양기에서 1시간 동안 반응시킨 후, 0.1N NaOH로 적정하였다. 지시약으로는 메틸 오렌지를, 양성 대조물질로는 탄산칼슘을 사용하였다.A mixture of licorice extract (AC) and leafy water soluble extract (AEAA), and the extract of AJC (AEAC) according to the present invention, were mixed in 100 ml of 0.1 N HCl, and reacted for 1 hour in a shaking incubator at 37 ° C. After titration, the mixture was titrated with 0.1N NaOH. Methyl orange was used as an indicator and calcium carbonate was used as a positive control.
4. 세포 실험4. Cell experiment
4-1. 비만세포 탈과립화 유도4-1. Induction of mast cell degranulation
비만세포의 탈과립화 유도를 위해, 비만세포의 세포막 내로 Ca2+의 유입을 증가시켜 비만세포의 탈과립을 증가시키는 강력한 약물인 compound 48/80(Sigma, USA)을 사용하였다. 배양된 비만세포를 24 웰 플레이트(24 well plate)에 분주한 후, 각 시료를 농도 별로 2시간 동안 전처리한 후에 compound 48/80을 처리하고 30분 뒤 상층액을 취하여 배지 내에 분비되어 있는 히스타민의 양을 측정하였다. To induce degranulation of mast cells, compound 48/80 (Sigma, USA), a potent drug that increases the infiltration of mast cells by increasing Ca 2+ influx into the cell membrane of mast cells, was used. After dispensing the cultured mast cells into a 24 well plate, each sample was pretreated for 2 hours at different concentrations, treated with Compound 48/80, and after 30 minutes, the supernatant was taken and secreted into the medium. The amount was measured.
4-2. 사이토카인 분비 유도4-2. Induce cytokine secretion
비만세포의 사이토카인 분비는 phorbol myristate acetate(PMA)와 A23187을 혼합 처리하여 유도하였다. 배양된 비만세포를 24 웰 플레이트에 분주한 후, 각 시료를 1시간 동안 전처리한 후에 PMA와 A23187을 처리하고 8시간 뒤 상층액 또는 세포를 취하여 사이토카인 측정에 사용하였다. The cytokine secretion of mast cells was induced by a combination of phorbol myristate acetate (PMA) and A23187. After the cultured mast cells were dispensed into 24 well plates, each sample was pretreated for 1 hour, and then treated with PMA and A23187. After 8 hours, supernatants or cells were taken and used for cytokine measurement.
대식세포인 Raw264.7 세포는 각 시료를 농도 별로 1시간 동안 전처리한 후에 1 μg/ml의 농도의 LPS를 처리하고 24시간 뒤 상층액 또는 세포를 취하여 사이토카인 측정에 사용하였다.Raw264.7 cells, which are macrophages, were treated with LPS at a concentration of 1 μg / ml after pretreatment of each sample for 1 hour at concentration, and supernatants or cells were taken 24 hours later and used for cytokine measurement.
4-3. 사이토카인 측정4-3. Cytokine measurement
사이토카인 측정은 IL-1 및 IL-6에 대한 sandwich ELISA kit를 구입하여 제조사의 지시에 따라 시험을 수행하였다. Standard의 흡광도로부터 표준곡선을 구하고 이를 이용하여 각 샘플의 흡광도로부터 사이토카인의 양을 정량하였다.Cytokine measurements were performed according to the manufacturer's instructions by purchasing sandwich ELISA kits for IL-1 and IL-6. The standard curve was calculated from the absorbance of standard and the amount of cytokine was quantified from the absorbance of each sample.
4-4. 유전자 발현 분석4-4. Gene expression analysis
(1) RNA 추출 및 역전사(1) RNA extraction and reverse transcription
Qiagen사(USA)의 RNeasy mini kit를 사용하여 제조사의 지시에 따라 세포로부터 RNA를 추출하였다. 분리된 RNA는 Qiagen사(USA)의 cDNA synthesis kit를 사용하여 cDNA로 합성한 후, 실시간 중합효소 연쇄반응(real-time PCR)에 사용하였다.RNA was extracted from the cells according to the manufacturer's instructions using the RNeasy mini kit from Qiagen (USA). The isolated RNA was synthesized into cDNA using a cDNA synthesis kit of Qiagen (USA), and then used for real-time PCR.
(2) 실시간 중합효소 연쇄반응(2) Real time polymerase chain reaction
Qiagen의 Rotor-Gene Q 2 plex 실시간 DNA 증폭기 및 Qiagen사의 two step PCR SYBR kit를 사용하여 제조사의 지시에 따라 실시간 중합효소 연쇄반응을 수행하고, 증폭된 DNA의 양을 정량적으로 비교하였다. 구체적으로 사이클링 프로토콜(cycling protocol)을 기준으로 95℃에서 5분 동안 pre-denaturation을 실시한 후, 95℃에서 5초, 60℃에서 10초씩 40 사이클을 반복하고, 60℃에서 95℃까지 3분 동안 상승하는 melting 단계의 순서로 수행하였다. 실시간 중합효소 연쇄반응으로 증폭시킨 산물을 Delta delta Ct method를 이용하여 정량하고, 각 시료에 대해 GAPDH의 발현량으로 보정하였다. 실험에 사용된 프라이머의 염기 배열은 하기 [표 1]에 나타낸 것과 같다.Qiagen's Rotor-Gene Q 2 plex real-time DNA amplifier and Qiagen's two step PCR SYBR kit were used to perform real-time polymerase chain reaction according to the manufacturer's instructions, and the amount of amplified DNA was quantitatively compared. Specifically, pre-denaturation was performed at 95 ° C. for 5 minutes based on a cycling protocol, followed by 40 cycles of 5 seconds at 95 ° C. and 10 seconds at 60 ° C. for 3 minutes from 60 ° C. to 95 ° C. It was performed in the order of the rising melting step. Products amplified by real-time polymerase chain reaction were quantified using the Delta delta Ct method and corrected by the expression level of GAPDH for each sample. The base sequence of the primers used in the experiment is as shown in the following [Table 1].
| NameName | orientationorientation | Sequences (5' → 3')Sequences (5 '→ 3') |
| IL-6IL-6 | forwardforward | ATGAACTCCTTCTCCACAAGCATGAACTCCTTCTCCACAAGC |
| IL-6IL-6 | everseeverse | GTTTTCTGCCAGTGCCTCTTTGGTTTTCTGCCAGTGCCTCTTTG |
| TNF-αTNF-α | forwardforward | GGCAGTCAGATCATCTTCTCGAACGGCAGTCAGATCATCTTCTCGAAC |
| TNF-αTNF-α | everseeverse | GAAGGCCTAAGGTCCACTTGTGTGAAGGCCTAAGGTCCACTTGTGT |
5. 급성 위염/위궤양 유발 동물모델5. Acute Gastritis / Gas Ulcer-Induced Animal Model
5-1. 단회 투여군5-1. Single dose group
체중 200g 내외의 수컷 래트(rat) 6마리를 1군으로 하여 대조군, 양성 대조군 및 실험군의 3군으로 나누고, 일반 환경 조건 하에서 24시간 이상 절식시켰다. Six male rats with a body weight of about 200 g were divided into three groups, a control group, a positive control group, and an experimental group, and fasted for 24 hours or more under general environmental conditions.
대조군에는 일반식을, 양성 대조군에는 라니티딘(ranitidine) 또는 감초 추출물을, 실험군에는 AEAC를 경구 투여하였다. 30분이 지난 후에 대조군, 양성 대조군, 실험군 각각에 HCl-EtOH(150mM HCl을 함유한 60% EtOH) 1mL를 경구 투여하였다. In the control group, the general formula, the positive control group, ranitidine or licorice extract, and the experimental group were orally administered AEAC. After 30 minutes, 1 mL of HCl-EtOH (60% EtOH containing 150 mM HCl) was orally administered to each of the control, positive control, and experimental groups.
절식 및 절수 상태로 1시간 방치한 후, 에테르(ether)로 치사시킨 다음, 위를 적출하였다. 적출한 위는 2% 포르말린 용액 10mL에 10분간 담가서 고정시켰다. 위의 대만부를 절개하여 발생된 손상 길이(mm) 측정 및 조직병리학적 분석을 실시하였다. 유전자 분석을 위한 위 조직은 고정하지 않은 상태로 분리한 후 분자생물학적 분석에 사용하였다. 혈청은 혈액을 채취하고 상온에서 30분간 방치한 후 원심분리하여 상층액을 취하였다.After leaving for 1 hour in a fasted and watered state, it was killed by ether (ether), and then the stomach was extracted. The extracted stomach was fixed by soaking in 10 mL of 2% formalin solution for 10 minutes. The dissection of the upper part of the incision was performed to determine the length of injury (mm) and histopathological analysis. The tissues for genetic analysis were separated from the unfixed state and used for molecular biological analysis. Serum was collected from blood, left at room temperature for 30 minutes, and centrifuged to obtain supernatant.
5-2. 반복 투여군5-2. Repeat dose group
체중 200g 내외의 수컷 랫트 6마리를 1군으로 하여 대조군, 양성 대조군 및 실험군의 3군으로 나누었다. Six male rats with a body weight of about 200 g were divided into three groups: a control group, a positive control group, and an experimental group.
대조군에는 일반식을, 양성 대조군에는 감초 추출물을, 실험군에는 AEAC를 각각 하루 2회씩 1주일간 경구 투여하였다. 마지막 경구 투여 후 24시간 이상 절식시켰다. 다시 대조군에는 일반식을, 양성 대조군에는 감초 추출물을, 실험군에 AEAC를 각각 경구 투여하고, 30분 후에 HCl-EtOH (150mM HCl을 함유한 60% EtOH) 1mL를 대조군, 양성 대조군, 실험군 모두 경구 투여하였다. The control group was orally administered with the general formula, the positive control group with licorice extract, and the experimental group with AEAC twice a day for 1 week. Fasting for more than 24 hours after the last oral administration. Again, the control group was administered with the general formula, the positive control group with the licorice extract and the experimental group with oral AEAC, and after 30 minutes, 1 mL of HCl-EtOH (60% EtOH containing 150 mM HCl) was orally administered to the control group, the positive control group, and the experimental group. It was.
절식 및 절수 상태로 1시간 방치한 후, 에테르(ether)로 치사시킨 다음, 위를 적출하였다. 적출한 위는 2% 포르말린 용액 10mL에 10분간 담가서 고정시켰다. 위의 대만부를 절개하여 발생된 손상 길이(mm) 측정 및 조직병리학적 분석을 실시하였다. 유전자 분석을 위한 위 조직은 고정하지 않은 상태로 분리한 후 분자생물학적 분석에 사용하였다. 혈청은 혈액을 채취하고 상온에서 30분간 방치한 후 원심분리하여 상층액을 취하였다.After leaving for 1 hour in a fasted and watered state, it was killed by ether (ether), and then the stomach was extracted. The extracted stomach was fixed by soaking in 10 mL of 2% formalin solution for 10 minutes. The dissection of the upper part of the incision was performed to determine the length of injury (mm) and histopathological analysis. The tissues for genetic analysis were separated from the unfixed state and used for molecular biological analysis. Serum was collected from blood, left at room temperature for 30 minutes, and centrifuged to obtain supernatant.
5-3. 유전자 발현 분석5-3. Gene expression analysis
(1) RNA 추출 및 역전사(1) RNA extraction and reverse transcription
Qiagen사(USA)의 RNeasy mini kit를 사용하여 제조사의 지시에 따라 랫트의 위 조직으로부터 RNA를 추출하였다. 분리된 RNA는 Qiagen사(USA)의 cDNA synthesis kit를 사용하여 cDNA로 합성한 후, 실시간 중합효소 연쇄반응에 사용하였다.RNA was extracted from the stomach tissue of rats using the RNeasy mini kit from Qiagen (USA) according to the manufacturer's instructions. The isolated RNA was synthesized into cDNA using a cDNA synthesis kit of Qiagen (USA), and then used for real-time polymerase chain reaction.
(2) 실시간 중합효소 연쇄반응(2) Real time polymerase chain reaction
Qiagen의 Rotor-Gene Q 2 plex 실시간 DNA 증폭기 및 Qiagen사의 two step PCR SYBR kit를 사용하여 제조사의 지시에 따라 실시간 중합효소 연쇄반응을 수행하고, 증폭된 DNA의 양을 정량적으로 비교하였다. 구체적으로 사이클링 프로토콜(cycling protocol)을 기준으로 95℃에서 5분 동안 pre-denaturation을 실시한 후, 95℃에서 5초, 60℃에서 10초씩 40 사이클을 반복하고, 60℃에서 95℃까지 3분 동안 상승하는 melting 단계의 순서로 수행하였다. 실시간 중합효소 연쇄반응으로 증폭시킨 산물을 Delta delta Ct method를 이용하여 정량하고, 각 시료에 대해 β-actin의 발현량으로 보정하였다. 실험에 사용된 프라이머의 염기 배열은 하기 [표 2]에 나타낸 것과 같다.Qiagen's Rotor-Gene Q 2 plex real-time DNA amplifier and Qiagen's two step PCR SYBR kit were used to perform real-time polymerase chain reaction according to the manufacturer's instructions, and the amount of amplified DNA was quantitatively compared. Specifically, pre-denaturation was performed at 95 ° C. for 5 minutes based on a cycling protocol, followed by 40 cycles of 5 seconds at 95 ° C. and 10 seconds at 60 ° C. for 3 minutes from 60 ° C. to 95 ° C. It was carried out in the order of the rising melting step. Products amplified by real-time polymerase chain reaction were quantified using Delta delta Ct method and corrected by the expression level of β-actin for each sample. The base sequence of the primers used in the experiment is as shown in the following [Table 2].
| NameName | orientationorientation | Sequences (5' → 3')Sequences (5 '→ 3') |
| MUC5ACMUC5AC | forwardforward | GGCCAATGCGGCACTTGTACCAATGGCCAATGCGGCACTTGTACCAAT |
| MUC5ACMUC5AC | reversereverse | GTCATCTGGACAGAAGCAGCCCTCGTCATCTGGACAGAAGCAGCCCTC |
| MUC6MUC6 | forwardforward | TCCTACTTGCCAGGTCTTCCAACTCCTACTTGCCAGGTCTTCCAAC |
| MUC6MUC6 | reversereverse | TTGTGGGTGTTGACTTCGGTATAGTTGTGGGTGTTGACTTCGGTATAG |
| TFF1TFF1 | forwardforward | CCATGGAGCACAAGGTGACCTGTGCCATGGAGCACAAGGTGACCTGTG |
| TFF1TFF1 | reversereverse | GGGAAGCCACAATTTATTCTCTCCGGGAAGCCACAATTTATTCTCTCC |
| TFF2TFF2 | forwardforward | CCCACTTCCAAACCAAGCGTCGCCCACTTCCAAACCAAGCGTCG |
| TFF2TFF2 | reversereverse | CAGCAGTGCCCTTCAGTAGTGACAGCAGTGCCCTTCAGTAGTGA |
| TFF3TFF3 | forwardforward | TGCCATGGAGACCAGAGCCTTCTGTGCCATGGAGACCAGAGCCTTCTG |
| TFF3TFF3 | reversereverse | TAGAACAGCCTTGTGCTGACTGTATAGAACAGCCTTGTGCTGACTGTA |
| PigrPigr | forwardforward | AAGAGACACCACCTCGTCCCAGATAAGAGACACCACCTCGTCCCAGAT |
| PigrPigr | reversereverse | TGGTGCCCTTATGGCTTGGGTATGTGGTGCCCTTATGGCTTGGGTATG |
5-4. 위 손상 억제율 분석5-4. Gastric damage inhibition rate analysis
위 손상 억제율을 확인하기 위해, 10% 중성 포르마린액에 고정된 위의 대만(great curvature)을 따라 종단면을 만들고, 발생된 손상 길이(mm)를 측정하였다. 일반적인 조직 처리 과정을 거쳐 파라핀으로 고정하였다. 위 손상 억제율은 하기 계산식을 이용하여 계산하였다.In order to confirm the inhibition of gastric damage, a longitudinal section was made along the great curvature of the stomach fixed in 10% neutral formarin solution, and the damage length (mm) generated was measured. It was fixed in paraffin through a general tissue treatment process. Gastric damage inhibition was calculated using the following formula.
[계산식 1][Calculation 1]
억제율(%) = (대조군의 지수 - 시료군의 지수)/대조군의 지수 X 100% Inhibition = (index of control-index of sample) / index of control X 100
5-5. 조직병리학적 분석 및 각종 지표 측정5-5. Histopathological analysis and measurement of various indicators
위 점착성 점액의 측정은 Kitagawa 등(Arzneimittelforschung. 1986 Aug;36(8):1240-4.)의 방법에 따라 위 점막 표면(mg/g 조직)에 붙은 알시안 블루(alcian blue)의 농도로 계산하였다.The measurement of gastric cohesive mucus was calculated by the concentration of alcian blue adhered to the gastric mucosal surface (mg / g tissue) according to the method of Kitagawa et al. (Arzneimittelforschung. 1986 Aug; 36 (8): 1240-4.). It was.
지질 과산화물인 말론디알데하이드(Malondialdehyde; MDA)의 측정은 Emerit 등(Presse Med. 1986 Jun 14;15(24):1131-3)의 방법에 따라 thiobarbituric acid reactive substances(TBARS) assay를 이용하여 측정하였다. Malondialdehyde (MDA), a lipid peroxide, was measured using thiobarbituric acid reactive substances (TBARS) assay according to the method of Emerit et al. (Presse Med. 1986 Jun 14; 15 (24): 1131-3). .
초과산화물 불균등화효소(Superoxide dismutase; SOD)는 체내의 대표적인 유해 산소 포집 효소로서, 초과산화물 음이온(˙O2-; superoxide anion)을 과산화 수소(H2O2; hydrogen peroxide)로 환원시키는 작용을 하는 것으로 알려져 있다. 초과산화물 불균등화효소는 SOD Assay Kit(Dojindo Molecular Technologies, USA)를 이용하여 측정하였다.Superoxide disproportionation enzyme (Superoxide dismutase; SOD) is a representative radical trapping the enzyme in the body, superoxide anion, which acts to return to;; (hydrogen peroxide H 2 O 2) (˙O2 superoxide anion) hydrogen peroxide It is known. Superoxide dismutase was measured using a SOD Assay Kit (Dojindo Molecular Technologies, USA).
6. 실험 결과6. Experimental Results
도 1을 참조하여 보면, compound 48/80으로 유도된 비만세포에서 히스타민의 분비는 양성 대조물질인 커큐민에 의해 효과적으로 감소된 반면, 대조물질인 감초추출물(AC) 또는 인진쑥 수용성 추출물(AEAC)에 의해서는 히스타민의 수치 변화가 거의 관찰되지 않음으로써, 인진쑥 수용성 추출물(AEAC)은 비만세포의 탈과립 억제에 영향을 미치지 않는 것을 확인하였다.Referring to FIG. 1, the secretion of histamine in the mast cells induced by compound 48/80 was effectively reduced by curcumin, a positive control, while licorice extract (AC) or A. eryngii water-soluble extract (AEAC). As little changes in histamine levels were observed, water extract of A. erythema (AEAC) had no effect on degranulation inhibition of mast cells.
도 2를 참조하여 보면, PMA 및 A23187로 유도된 비만세포에서 인진쑥 수용성 추출물(AEAC)에 의해 염증성 사이토카인인 IL-6 및 TNF-α의 분비가 효과적으로 억제되는 것을 확인하였다.Referring to Figure 2, it was confirmed that the secretion of the inflammatory cytokines IL-6 and TNF-α by the P. aeruginosa-induced mast cells in a mast cell soluble extract (AEAC).
도 3을 참조하여 보면, LPS로 유도된 대식세포에서 염증성 사이토카인인 IL-1β 및 IL-6의 분비가 크게 증가하였고, 상기 염증성 사이토카인의 증가는 인진쑥 수용성 추출물(AEAC)에 의해 현저하게 감소되는 것을 확인하였다. 또한, 액상 또는 분말의 공정상의 제형 변화에 따른 효능의 차이가 없는 것을 확인하였다. Referring to FIG. 3, the secretion of inflammatory cytokines IL-1β and IL-6 was significantly increased in LPS-induced macrophages, and the increase in inflammatory cytokines was markedly decreased by AJC. It confirmed that it became. In addition, it was confirmed that there is no difference in efficacy due to the change in formulation of the liquid or powder process.
도 4를 참조하여 보면, 애엽 추출물(AEAA)은 탄산칼슘과 마찬가지로 제산력을 나타내는 것을 확인하였다. 애엽 추출물(AEAA)은 중화 반응을 통해 산에 의한 위 손상을 일시적으로 방지하는 효과를 나타내는 것으로 사료된다. 그러나 인진쑥 수용성 추출물(AEAC) 및 감초 추출물(AC)은 중화 관련 기능이 전혀 관찰되지 않았는데, 이는 단순 중화 반응에 따른 위산의 과분비 등의 부작용이 전혀 없다는 것을 의미한다.Referring to Figure 4, it was confirmed that leaf extract (AEAA) exhibits an antacid like the calcium carbonate. Leafy leaf extract (AEAA) is thought to have the effect of temporarily preventing gastric damage by acid through neutralization reaction. However, no neutralization-related functions were observed in the water extract of AJU (AEAC) and licorice extract (AC), which means that there are no side effects such as excessive secretion of gastric acid due to a simple neutralization reaction.
도 5를 참조하여 보면, HCl/에탄올로 유도된 급성 위염/위궤양 동물모델의 경우, in vitro 세포주 실험 결과와 마찬가지로 혈청 내 히스타민 양에 있어서 인진쑥 수용성 추출물(AEAC)에 의한 유의적인 변화가 관찰되지 않음으로써, 인진쑥 수용성 추출물(AEAC)은 히스타민의 생성 또는 분비 억제 효과에 영향을 미치지 않는 것을 확인하였다.Referring to FIG. 5, in the case of HCl / ethanol-induced acute gastritis / gastric ulcer animal model, there was no significant change by AJC in the amount of histamine in serum as in the in vitro cell line test results. As a result, it was confirmed that the extract of A. aeruginosa (AEAC) did not affect the production or secretion inhibitory effect of histamine.
도 6을 참조하여 보면, HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 알시안 블루를 이용하여 위 점막의 변화를 분석한 결과, 정상 대조군에 비해 HCl/에탄올로 급성 위염/위궤양을 유도한 경우, 알시안 블루의 염색 정도가 크게 감소되었고, 대조물질인 감초 추출물(AC)의 경우, 위 점막이 부분적으로 남아있는 것으로 보였으나 전체적으로 점막 손상이 많이 관찰된 반면, 인진쑥 수용성 추출물(AEAC)의 경우, 200 mg/kg의 투여 용량으로도 정상 대조군만큼의 점막 보호층이 남아있는 것을 확인하였으며, 또한, 용량 의존적인 점막층의 증가가 관찰되었다. Referring to FIG. 6, as a result of analyzing the change of gastric mucosa using alcian blue in an acute gastritis / gastric ulcer animal model induced by HCl / ethanol, HCl / ethanol induced acute gastritis / gastric ulcer compared to a normal control group. In this case, the degree of staining of Alcian blue was greatly reduced, and in the case of control licorice extract (AC), gastric mucosa seemed to remain partially, but a lot of mucosal damage was observed. In this case, it was confirmed that the mucosal protective layer remained as much as the normal control even at a dose of 200 mg / kg, and a dose-dependent increase in the mucosal layer was observed.
도 7을 참조하여 보면, 위 점막 조직에 결합된 알시안 블루의 양을 상대값으로 환산하여 위 점액량을 분석한 결과, 100mg/kg의 감초 추출물(AC)은 HCl/에탄올 투여군에 비하여 98.7% 증가하였고, 동일 투여량의 인진쑥 수용성 추출물(AEAC)은 104.2% 증가를 보여 감초 추출물(AC) 보다 약간 우세한 위 점액량 증가 효능을 나타내었다. 또한, 인진쑥 수용성 추출물(AEAC)은 용량 의존적으로 위 점액량의 증가 효능을 나타내었고, 최고 용량인 400mg/kg에서 최대의 효과를 나타냄으로써, 인진쑥 수용성 추출물(AEAC)이 위 점액량을 증가시켜 HCl/에탄올로 유도된 급성 위염/위궤양에 대해 보호 효능을 나타내는 것을 확인하였다.Referring to FIG. 7, as a result of analyzing the amount of gastric mucus by converting the amount of alcian blue bound to gastric mucosal tissue as a relative value, the licorice extract (AC) of 100 mg / kg was 98.7% compared to the HCl / ethanol-administered group. Increasingly, the same dose of A. eryngii water extract (AEAC) showed a 104.2% increase, showing a slightly higher gastric mucus increase effect than licorice extract (AC). In addition, the water extract of A. erythema (AEAC) showed the effect of increasing the amount of gastric mucus in a dose-dependent manner, and showed the maximum effect at the highest dose of 400 mg / kg. It was confirmed that the protective effect against / ethanol-induced acute gastritis / gastric ulcer.
도 8을 참조하여 보면, HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 지질 과산화 억제 작용을 확인하기 위해, 말론디알데하이드(MDA)의 양을 측정한 결과, HCl/에탄올 투여에 의해 MDA가 2.051 nM에서 3.927 nM로 약 2배 증가되었으며, 200mg/kg의 인진쑥 수용성 추출물(AEAC)에 의해 MDA가 0.103 nM로 감소되는 것을 확인함으로써, 이는 정상 대조군보다 훨씬 낮은 수준으로 인진쑥 수용성 추출물(AEAC)이 지질 과산화를 효과적으로 억제하는 것을 확인하였다.Referring to Figure 8, in order to confirm the lipid peroxidation inhibitory effect in HCl / ethanol-induced acute gastritis / gastric ulcer animal model, the amount of malondialdehyde (MDA) was measured, the MDA by HCl / ethanol administration It was approximately doubled from 2.051 nM to 3.927 nM and confirmed that the MDA was reduced to 0.103 nM by 200 mg / kg of erythrocyte water soluble extract (AEAC), which was much lower than the normal control. It was confirmed to effectively inhibit lipid peroxidation.
도 9를 참조하여 보면, HCl/에탄올로 유도된 급성 위염/위궤양 동물모델에서 초과산화물 불균등화효소(SOD)의 활성을 측정한 결과, 정상 대조군에서 SOD 활성이 30.6%인 반면, HCl/에탄올 투여에 의해 SOD가 거의 측정되지 않을 만큼 뚜렷한 활성 감소를 나타내었다. 반면, 200 mg/kg, 400 mg/kg의 인진쑥 수용성 추출물(AEAC)에 의해 SOD가 각각 59.7%, 59.6%로 뚜렷한 활성 증가를 나타내었다. 상기 결과로부터 급성 위염/위궤양 동물모델에 있어서 인진쑥 수용서어 추출물(AEAC)의 치료 및 방어 효과는 항산화 효소인 SOD 활성을 회복시킴으로써 염증 유발로 감소된 위 점막의 방어기능을 회복시켜 염증 치료 효과를 나타냄을 확인할 수 있었다.Referring to FIG. 9, as a result of measuring the activity of superoxide dismutase (SOD) in an HCl / ethanol-induced acute gastritis / gastric ulcer animal model, the SOD activity was 30.6% in the normal control group, whereas HCl / ethanol administration Showed a marked decrease in activity so that SOD was hardly measured. On the other hand, 200 mg / kg and 400 mg / kg of AJC extract showed a marked increase in SOD of 59.7% and 59.6%, respectively. From the above results, the treatment and defense effect of AUGC in the acute gastritis / gastric ulcer animal model shows the inflammatory treatment effect by restoring the gastric mucosa's defense function which is reduced by inflammation by restoring SOD activity, an antioxidant enzyme. Could confirm.
본 발명에 따른 인진쑥 수용성 추출물(AEAC)을 단회 또는 반복 투여하고, 위점막 보호에 관련된 유전자들의 발현량 차이를 분석한 결과를 도 10 내지 18에 나타내었다.The results of analyzing the difference in the expression level of genes related to gastric mucosa protection after single or repeated administration of AJC water extract (AEAC) according to the present invention.
도 10 내지 13을 참조하여 보면, 인진쑥 수용성 추출물(AEAC)을 단회 투여한 경우, HCl/에탄올만을 투여한 경우에 비하여 위 점막 단백질인 MUC6은 1.3배, MUC5AC은 2.8배 발현량이 증가하는 것을 확인하였으며, 위 점막 면역과 관련된 인자인 rPigr은 1.5배, 위 점막의 보호 및 치료와 관련된 TFF1은 5.4배, TFF2는 5.5배, TFF3은 1.2배 증가하는 것을 확인하였다.Referring to Figures 10 to 13, when a single administration of AJC (AEAC), it was confirmed that the expression of gastric mucosa protein MUC6 1.3 times, MUC5AC 2.8 times increased compared to the case of administration of only HCl / ethanol. RPigr, a factor related to gastric mucosal immunity, increased 1.5 times, TFF1 increased 5.4 times, TFF2 5.5 times, and TFF3 1.2 times related to gastric mucosal protection and treatment.
도 14 내지 17을 참조하여 보면, 200mg/kg 또는 400mg/kg의 인진쑥 수용성 추출물(AEAC)을 반복 투여한 경우, HCl/에탄올만을 투여한 경우에 비하여 위 점막 단백질인 MUC6은 2.4 내지 3.9배, MUC5AC는 3.0 내지 4.6배 발현량이 증가하였으며, 위 점막 면역과 관련된 인자인 rPigr은 3.9 내지 4.3배, TFF1은 2.5 내지 4.1배, TFF2는 9.4 내지 18.2배, TFF3은 3.2 내지 4.2배 증가하는 것을 확인하였다.Referring to Figures 14 to 17, when repeated administration of 200 mg / kg or 400 mg / kg of agar water soluble extract (AEAC), gastric mucosa protein MUC6 is 2.4 to 3.9 times, MUC5AC compared to the case of administration only HCl / ethanol Was increased 3.0-4.6 fold, rPigr, a factor related to gastric mucosal immunity, 3.9-4.3 fold, TFF1 2.5-4.1 fold, TFF2 9.4-18.2 fold, TFF3 3.2- 4.2 fold.
또한, 도 18을 참조하여 보면, 감초 추출물(AC) 보다 인진쑥 수용성 추출물(AEAC) 투여에 의해 위 점막 단백질 및 위 점막 면역과 관련된 인자들이 더 효과적으로 증가하는 것을 확인할 수 있었다.In addition, referring to Figure 18, it was confirmed that the administration of the Agaric soluble extract (AEAC) than the licorice extract (AC) increased more effectively the factors related to gastric mucosa protein and gastric mucosal immunity.
상기와 같이, 본 발명에 따른 인진쑥 수용성 추출물(AEAC)을 단회 및 반복 투여한 경우, MUC5AC, MUC6, rPigr, TFF1, TFF2, TFF3 등의 위 점막 단백질의 생성이 크게 촉진되었고, 이러한 점막 생성 촉진 효과는 지속적으로 유지되었다. 특히 MUC5AC 유전자는 HCl/에탄올만을 투여한 경우에 비하여 인진쑥 수용성 추출물(AEAC)에 의해 4.6배까지 증가하였고 TFF2는 최고 18배까지 증가하였다. As described above, when single and repeated administration of the water extract of AJU (AEAC) according to the present invention, the production of gastric mucosa proteins such as MUC5AC, MUC6, rPigr, TFF1, TFF2, TFF3 was greatly promoted, and this mucosa-producing effect Was maintained continuously. In particular, MUC5AC gene was increased by 4.6-fold and TFF2 by 18-fold by AEAC compared with HCl / ethanol alone.
따라서 본 발명에 따른 인진쑥 수용성 추출물(AEAC)은 직접적으로 점막 단백질의 생성을 촉진하며, 이를 통해 지속적 위점막 보호 효과를 갖는 것을 알 수 있다. 특히 점막 단백질의 시스템에서 TFF2의 증가에 따른 위장관 점막에 대한 보호 효과 뿐 아니라 위산 분비 억제에 따른 증상 개선 효과가 나타나는 것으로 보인다.Therefore, the water extract of A. eryngii (AEAC) according to the present invention directly promotes the production of mucosal proteins, and it can be seen that it has a continuous gastric mucosal protection effect. In particular, the mucosal protein system seems to have a protective effect on the gastrointestinal mucosa due to the increase of TFF2, as well as a symptom improvement effect by suppressing gastric acid secretion.
도 19를 참조하여 보면, HCl/에탄올로 위염/위궤양을 유도한 경우, HCl/에탄올을 투여하지 않은 정상 대조군(Veh)에 비해 굵은 띠 모양으로 선상 출혈 소견을 나타내었다. 여기에 감초 추출물(AC)을 단회 투여한 경우, 위점막 전체에서 출혈성 점막 손상이 관찰되었으나, 인진쑥 수용성 추출물(AEAC)을 단회 투여한 경우, 용량 의존적으로 HCl/에탄올 투여에 의한 위점막 손상이 현저하게 억제되었다. 헤마톡실린 & 에오신(H&E) 염색을 통한 병리 조직검사 결과에서도 HCl/에탄올을 투여하고 인진쑥 수용성 추출물(AEAC)을 투여한 경우, HCl/에탄올을 투여하지 않은 정상 대조군과 비교하였을 때, 위 상피조직 및 분비 세포들의 형태와 배열에 큰 차이가 없는 것을 확인하였다.Referring to FIG. 19, when gastritis / gastric ulcers were induced with HCl / ethanol, glandular bleeding was observed in a thick band shape compared to the normal control group (Veh) not receiving HCl / ethanol. In the case of single administration of licorice extract (AC), hemorrhagic mucosal damage was observed in the entire gastric mucosa. However, in the case of single administration of AJC, the gastric mucosal damage was markedly dose-dependently. Suppressed. In histopathologic findings of hematoxylin & eosin (H & E) staining, gastric epithelial tissues when HCl / ethanol and H. aerugin water-soluble extract (AEAC) were administered compared to normal controls without HCl / ethanol And it was confirmed that there is no significant difference in the form and arrangement of secretory cells.
도 20 및 하기 표 3을 참조하여 보면, 위 손상 억제율은 HCl/에탄올 투여 후, 감초 추출물(AC)을 단회 투여한 경우, 100mg/kg 투여군에서 9.64%로 상대적으로 낮은 효능(potency)을 보였다. 이에 비해 HCl/에탄올 투여 후 인진쑥 수용성 추출물(AEAC)을 단회 투여한 경우, 25mg/kg 투여군에서 10.59%, 100mg/kg 투여군에서 76.59%의 억제율을 보였고, 최고 용량인 400mg/kg 투여군에서는 91.46%의 억제율을 나타내었다. Referring to Figure 20 and Table 3 below, the above damage inhibition rate is After the administration of HCl / ethanol, licorice extract (AC) was administered once, showing a relatively low potency of 9.64% in the 100 mg / kg administration group. In contrast, after HCl / ethanol administration A single dose of AJC showed 10.59% inhibition in the 25mg / kg group and 76.59% in the 100mg / kg group, and 91.46% in the 400mg / kg group.
| SampleSample | Dose(mg/kg)Dose (mg / kg) | lesion index(mm)lesion index (mm) | Inhibition(%)Inhibition (%) |
| -- | -- | 6.02 ± 2.806.02 ± 2.80 | -- |
|
AC |
100100 | 5.44 ± 1.095.44 ± 1.09 | 9.649.64 |
| AEACAEAC | 2525 | 5.38 ± 1.145.38 ± 1.14 | 10.5910.59 |
| 5050 | 5.48 ± 0.695.48 ± 0.69 | 8.858.85 | |
| 100100 | 1.41 ± 0.431.41 ± 0.43 | 76.5976.59 | |
| 200200 | 0.90 ± 0.190.90 ± 0.19 | 84.9784.97 | |
| 400400 | 0.51 ± 0.050.51 ± 0.05 | 91.4691.46 |
도 21을 참조하여 보면, HCl/에탄올로 위염/위궤양을 유도한 경우, HCl/에탄올을 투여하지 않은 정상 대조군(Veh)에 비해 굵은 띠 모양으로 선상 출혈 소견을 나타내었다. 여기에 감초 추출물(AC)을 반복 투여한 경우, 위점막 전체에서 출혈성 점막 손상이 관찰되었으나, 인진쑥 수용성 추출물(AEAC)을 반복 투여한 경우, 용량 의존적으로 HCl/에탄올 투여에 의한 위 점막 손상이 현저하게 억제되었다. Referring to FIG. 21, when gastritis / gastric ulcers were induced with HCl / ethanol, glandular bleeding was observed in a thick band shape compared to the normal control group (Veh) not receiving HCl / ethanol. When licorice extract (AC) was repeatedly administered, hemorrhagic mucosal damage was observed in the entire gastric mucosa. However, when repeated administration of AJC was significantly dose-dependent, gastric mucosal damage was significant. Suppressed.
도 22를 참조하여 보면, 위 손상 억제율은 HCl/에탄올 투여 후, 인진쑥 수용성 추출물(AEAC)을 투여한 경우, 200mg/kg 투여군과 400mg/kg 투여군에서 모두 100%의 억제율을 나타내었다.Referring to FIG. 22, the gastric injury inhibition rate was 100% inhibition in both 200 mg / kg and 400 mg / kg administration groups when HJ / ethanol was administered followed by administration of AJC.
도 23을 참조하여 보면, HCl/에탄올로 위염/위궤양을 유도한 경우, 굵은 띠 모양으로 선상 출혈 소견을 나타내었다. 여기에 애엽 추출물(AEAA)을 단회 투여한 경우에는 위점막 전체에서 출혈성 점막 손상이 여전히 관찰되었으나, 인진쑥 수용성 추출물(AEAC)을 단회 투여한 경우에는 HCl/에탄올 투여에 의한 위점막 손상이 현저하게 억제되는 것을 확인하였다.Referring to Figure 23, when the gastritis / gastric ulcer induction with HCl / ethanol, showed a linear bleeding in the form of a thick band. Hemorrhagic mucosal damage was still observed throughout the gastric mucosa with a single dose of AEAA, whereas gastric mucosal damage by HCl / ethanol was significantly inhibited with a single dose of AJC. It confirmed that it became.
이상과 같이 본 발명을 실시예를 통하여 설명하였다. 본 발명이 속하는 기술 분야의 통상의 기술자라면 본 발명이 그 기술적 사상이나 필수적 특징을 변경하지 않고 다른 구체적인 형태로 실시될 수 있다는 것을 이해할 것이다. 따라서 본 명세서와 도면에 개시된 본 발명의 실시예들은 본 발명의 기술 내용을 쉽게 설명하고 본 발명의 이해를 돕기 위해 특정 예를 제시한 것일 뿐이며, 본 발명의 범위를 한정하고자 하는 것은 아니다. 본 발명의 범위는 특허청구범위에 의하여 나타내어지며, 특허청구범위의 의미 및 범위, 그리고 등가 개념으로부터 도출되는 모든 변경 또는 변형된 형태는 본 발명의 범위에 포함되는 것으로 해석되어야 한다.As described above, the present invention has been described through examples. Those skilled in the art will understand that the present invention can be implemented in other specific forms without changing the technical spirit or essential features. Therefore, the embodiments of the present invention disclosed in the specification and the drawings are only specific examples to easily explain the technical contents of the present invention and aid the understanding of the present invention, and are not intended to limit the scope of the present invention. The scope of the invention is indicated by the claims, and all changes or modifications derived from the meaning and scope of the claims, and equivalent concepts should be construed as being included in the scope of the invention.
Claims (15)
- 위 점막 손상성 위장 질환의 예방 또는 개선 작용을 가지는 인진쑥(Artemisia capillaris) 수용성 추출물.A water-soluble extract of Artemisia capillaris , which has a preventive or ameliorating effect on gastric mucosal damaging gastrointestinal diseases.
- 제 1항에 있어서, The method of claim 1,인진쑥을 열수로 추출한 것인 인진쑥(Artemisia capillaris) 수용성 추출물.It would extract the injinssuk with hot water injinssuk (Artemisia capillaris) aqueous extract.
- 인진쑥(Artemisia capillaris) 수용성 추출물을 함유하는 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물.Injinssuk (Artemisia capillaris) composition for the prevention and improvement of gastric mucosal DAMAGE gastrointestinal disorders, containing a water-soluble extract.
- 제 3항에 있어서,The method of claim 3, wherein위장관 점막 시스템의 항상성 유지 및 개선 효과를 통해 위 점막 손상성 위장 질환을 예방 또는 개선하는 것인 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물.A composition for preventing or improving gastric mucosal damaging gastrointestinal disease, which is to prevent or improve gastric mucosal damaging gastrointestinal disease through the effect of maintaining and improving homeostasis of the gastrointestinal mucosa system.
- 제 3항에 있어서, The method of claim 3, wherein위산, 에탄올, 스트레스 또는 이들의 조합에 의해 유발되는 점막 손상성 위장관계 질환을 개선하는 것인 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물.A composition for preventing or improving gastric mucosal gastrointestinal disorders, which is to improve mucosal gastrointestinal disorders caused by gastric acid, ethanol, stress or a combination thereof.
- 제 3항에 있어서,The method of claim 3, wherein위산, 에탄올, 스트레스 또는 이들의 조합에 의해 유발되는 위염 또는 위궤양에 대한 점막 손상을 예방 또는 억제하는 것인 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물.A composition for preventing or improving gastric mucosal damaging gastrointestinal disease, which prevents or inhibits mucosal damage to gastritis or gastric ulcer caused by gastric acid, ethanol, stress or a combination thereof.
- 제 3항에 있어서,The method of claim 3, wherein위 점막 단백질 또는 점막 면역 인자의 발현 증가를 통해 위 점막 손상을 억제 또는 개선하는 것인 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물.A composition for the prevention or improvement of gastric mucosal damage gastrointestinal disease, which is to inhibit or improve gastric mucosal damage through increased expression of gastric mucosal proteins or mucosal immune factors.
- 제 7항에 있어서,The method of claim 7, wherein상기 위 점막 단백질 또는 점막 면역 인자는 MUC5AC, MUC6, rPigr, TFF1, TFF2 및 TFF3로 구성된 군에서 1종 이상 선택되는 것인 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물.The gastric mucosal protein or mucosal immune factor is MUC5AC, MUC6, rPigr, TFF1, TFF2 and TFF3 is one or more selected from the group consisting of the composition for the prevention or improvement of gastric mucosal damage gastrointestinal diseases.
- 제 3항에 있어서, The method of claim 3, wherein염증성 사이토카인 억제, 지질 과산화물의 생성 억제 또는 항산화 효소 활성 증가를 통해 위 점막 손상을 억제 또는 개선하는 것인 위 점막 손상성 위장 질환의 예방 또는 개선용 조성물.A composition for preventing or ameliorating gastric mucosal injury, wherein the gastrointestinal mucosa is inhibited or improved by inhibiting inflammatory cytokines, inhibiting the production of lipid peroxides, or increasing antioxidant enzyme activity.
- 인진쑥(Artemisia capillaris) 수용성 추출물을 함유하는 위 점막 손상성 위장 질환의 예방 또는 개선용 식품 조성물.Injinssuk (Artemisia capillaris) Gastric DAMAGE preventing or food compositions for the improvement of gastrointestinal disorders, containing an aqueous extract.
- 인진쑥(Artemisia capillaris) 수용성 추출물을 함유하는 위 점막 손상성 위장 질환의 예방 또는 치료용 약학 조성물.Pharmaceutical composition for the prevention or treatment of gastric mucosal damage gastrointestinal diseases containing Artemisia capillaris water-soluble extract.
- 제 11항에 있어서,The method of claim 11,위 점막 단백질 또는 점막 면역 인자의 발현 증가를 통해 위 점막 손상을 억제 또는 개선하는 것인 위 점막 손상성 위장 질환의 예방 또는 치료용 약학 조성물.Pharmaceutical composition for the prevention or treatment of gastric mucosal damage gastrointestinal disease that inhibits or ameliorates gastric mucosal damage through increased expression of gastric mucosal proteins or mucosal immune factors.
- 제 12항에 있어서,The method of claim 12,상기 위 점막 단백질 또는 점막 면역 인자는 MUC5AC, MUC6, rPigr, TFF1, TFF2 및 TFF3로 구성된 군에서 1종 이상 선택되는 것인 위 점막 손상성 위장 질환의 예방 또는 치료용 약학 조성물.The gastric mucosal protein or mucosal immune factors MUC5AC, MUC6, rPigr, TFF1, TFF2 and TFF3 is one or more selected from the group consisting of pharmaceutical composition for the prevention or treatment of gastric mucosal damage gastrointestinal diseases.
- 제 11항에 있어서, The method of claim 11,염증성 사이토카인 억제, 지질 과산화물의 생성 억제 또는 항산화 효소 활성 증가를 통해 위 점막 손상을 억제 또는 개선하는 것인 위 점막 손상성 위장 질환의 예방 또는 치료용 약학 조성물.A pharmaceutical composition for preventing or treating gastric mucosal damaging gastrointestinal diseases, wherein the gastrointestinal mucosa is inhibited or improved by inhibiting inflammatory cytokines, inhibiting production of lipid peroxides, or increasing antioxidant enzyme activity.
- 인진쑥(Artemisia capillaris)을 열수로 추출하는 단계;Extracting Artemisia capillaris with hot water;추출액을 상온까지 냉각시켜 여과하고, 여액을 부피가 10±5%가 될 때까지 1차 농축 한 후 침전물을 제거하는 단계; 및Cooling the extract to room temperature and filtering the filtrate, and firstly concentrating the filtrate until the volume is 10 ± 5% to remove the precipitate; And침전물이 제거된 농축액을 수분 함량이 55±5%가 될 때까지 2차 농축하여 인진쑥(Artemisia capillaris) 수용성 추출물을 얻는 단계;Concentration of the concentrate from which the precipitate was removed was second concentrated until the water content was 55 ± 5% to obtain an Artemisia capillaris aqueous extract;를 포함하는, 인진쑥(Artemisia capillaris) 수용성 추출물의 제조 방법. Including, Artemisia capillaris method of producing a water-soluble extract.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20020035322A (en) * | 2000-11-06 | 2002-05-11 | 안용찬 | Cosmetic composition fot atopic skin containing artemisia extract |
| KR20140068831A (en) * | 2014-05-09 | 2014-06-09 | 주식회사 엘지생활건강 | Anti-wrinkle cosmetic composition |
| KR20150055667A (en) * | 2013-11-13 | 2015-05-22 | 주식회사 케이스템셀 | Comoposition for Preventing or Treating Osteoporosis Comprising Extract Containing Scoparone Derived from Artemisia Capillaris |
| KR20160008100A (en) * | 2014-06-30 | 2016-01-21 | 명지대학교 산학협력단 | Composition comprising extracts or fractions of Artemisia capillaris as an effective ingredient |
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| JPH08295632A (en) | 1995-03-02 | 1996-11-12 | Takeda Chem Ind Ltd | Antibacterial agent |
| KR100379189B1 (en) | 2000-12-14 | 2003-04-08 | 학교법인고려중앙학원 | Agents for gastroenteric disorder by Helicobacter pylori and screening method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20020035322A (en) * | 2000-11-06 | 2002-05-11 | 안용찬 | Cosmetic composition fot atopic skin containing artemisia extract |
| KR20150055667A (en) * | 2013-11-13 | 2015-05-22 | 주식회사 케이스템셀 | Comoposition for Preventing or Treating Osteoporosis Comprising Extract Containing Scoparone Derived from Artemisia Capillaris |
| KR20140068831A (en) * | 2014-05-09 | 2014-06-09 | 주식회사 엘지생활건강 | Anti-wrinkle cosmetic composition |
| KR20160008100A (en) * | 2014-06-30 | 2016-01-21 | 명지대학교 산학협력단 | Composition comprising extracts or fractions of Artemisia capillaris as an effective ingredient |
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| Title |
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| PARK, J.-M. ET AL: "The anti-inflammatory effects of acidic polysaccharide from Artemisia capillaris on Helicobacter pylori infection", JOURNAL OF CANCER PREVENTION, vol. 18, no. 2, 2013, pages 161 - 168, XP055606894 * |
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