KR20200076040A - Composition for preventing or treating gastritis and gastric ulcer comprising cabbage, broccoli sprout, broccoli leaf, kale as an active ingredient - Google Patents

Abstract

The present invention relates to a food composition which shows an effect of preventing or treating gastritis and gastric ulcer, and more particularly to a lactobacillus-fermented composition including cabbage, broccoli sprouts, broccoli leaves and kale as active ingredients to provide high efficiency of treating gastritis and gastric ulcer. The composition for preventing or treating gastritis and gastric ulcer uses four kinds of vegetables of Brassicaceae combined at an optimized ratio as active ingredients, and fermentation thereof is carried out with lactobacillus to produce an effective secondary metabolite, such as isothiocyanate. In this manner, antioxidation activity is reinforced and an effect of alleviating gastritis and gastric ulcer is improved through antibacterial and anti-inflammatory activities. The active ingredients are shown to have significantly improved blood test indexes related with gastritis and gastric ulcer, and thus are effective for producing functional foods for alleviating gastritis and gastric ulcer.

Description

Composition for preventing or treating gastritis and gastric ulcer comprising cabbage, broccoli sprout, broccoli leaf, kale as an active ingredient}

The present invention relates to a food composition showing gastroenteritis and gastric ulcer prevention or treatment effect, more specifically, the present invention is a fermentation composition consisting of cabbage, broccoli sprouts, broccoli leaves, kale to increase the efficiency of gastritis and gastric ulcer improvement It relates to a food composition comprising as an active ingredient.

Gastritis and gastric ulcer are gastrointestinal diseases caused by damage to the gastric mucosa due to exposure to various attack factors, both inside and outside. It is known as one of the most frequently occurring diseases worldwide. These gastrointestinal diseases are caused by psychological stress, hypersecretion of stomach acid, increased gastrointestinal motility, and Helicobacter pylori infection, which can be manifested in various forms, including bleeding, inflammation, ulceration, and mucosal damage, in addition to general pain. have. In particular, gastric ulcer is highly related to Helicobacter pylori infection, and it is known that more than 90% of adults are infected with this strain in developed countries as well as in Korea. In addition, nonsteroidal anti-inflammatory drugs (NSAIDs) such as aspirin and indomethacin are directly directed to the gastric mucosa through selective inhibition of cyclooxygenase-2, which facilitates local blood circulation. It is known to cause irritation and damage to the stomach, such as bleeding, inflammation, and ulcers.

In order to treat gastric ulcers and other gastric injury, hydrogen ion pump inhibitors that block acid secretion from gastric wall cells, antacids, histamine receptor blockers that promote acid secretion, gastric mucosal layer enhancer prostaglandin (PG) and its derivatives, In addition, antibiotics that inhibit the growth of Helicobacter bacterial are used.

The above various gastrointestinal damage treatment or prophylactic agents have side effects and resistance problems when taken for a long time. Due to this, efforts to develop a preventive or therapeutic agent with superior gastric damage effect from natural materials including food materials are actively being conducted. In other words, there is an urgent need to discover and develop natural materials that have the effect of preventing gastric damage, which has no side effects even if ingested and does not cause resistance or stability problems even after prolonged use.

Cruciferous vegetables such as cabbage, broccoli, and kale contain large amounts of glucocinolates, including glucoraphanin, glucoberine, and glucerucine. Although glucosinolate is not biologically active, it is degraded by the enzyme myrosinase (present in many plant cells and intestinal microflora) to form active isothiocyanates. These isothiocyanates, including sulforaphane, appear to have a number of health promoting properties and, in some experiments, appear to exert a variety of anti-cancer effects.

In addition, when fermenting these types of vegetables using lactic acid bacteria, it can be confirmed that the existing components are strengthened or converted into an activated form due to the production of secondary metabolites resulting from plant-specific ingredients, and the efficacy is doubled. Recently, studies on a fermented composition of a whole form capable of increasing the recovery of the active ingredient and dietary fiber without juice of the finished composition have been actively conducted.

Accordingly, the present inventors have tried to develop a natural product processing product that can enhance gastritis and gastric ulcer improvement functions while being safe, and include cabbage, broccoli sprouts, broccoli leaves, and kale, which have been proven to be safe from the past, as active ingredients. In the case of common knowledge by preparing the fermentation composition, it was confirmed that the gastric mucosa protection and gastritis-related indicator values were markedly improved, showing an excellent effect on gastritis and gastric ulcer improvement, and the present invention was completed.

In order to achieve the above object, the present invention is characterized in that it contains cabbage, broccoli sprouts, broccoli leaves, kale as an active ingredient. The task is to protect the gastric mucosa and significantly improve gastritis-related indicator values when the fermentation composition containing cabbage, broccoli sprouts, broccoli leaves, and kale is administered to an experimental animal, and has an excellent effect of improving gastritis and gastric ulcer. By illuminating, it is to provide a food composition that helps prevent or treat gastritis and gastric ulcer by utilizing the above mixture.

The present invention provides a gas composition for preventing or treating gastritis and gastric ulcer, comprising a fermentation composition consisting of cabbage, broccoli sprouts, broccoli leaves, and kale. The composition for preventing or treating gastritis and gastric ulcer of the present invention is not only safe, but also improves gastritis and gastric ulcer improvement by protecting the gastric mucosa and remarkably improving gastritis-related indicator levels, thus causing gastritis and gastric ulcer, or various diseases caused by it It is expected to be effective in the prevention and improvement of. In addition, it is expected that commercialization of the above-mentioned healthy vegetable fermented beverages will be possible to expand the consumption of functional vegetable drinks.

1 is a graph measuring the content of sulfolapan and indole-3-carbinol contained in cabbage, broccoli sprouts, broccoli leaves, and kale.
Figure 2 is a graph measuring the change in antioxidant capacity and total polyphenol content by fermentation time of the composition for preventing or treating gastritis and gastric ulcer of the present invention.
Figure 3 is a picture of the state of the gastric mucosa by H&E staining when the composition for preventing or treating gastritis and gastric ulcer of the present invention is administered to an experimental animal.
Figure 4 is a graph measuring the gastric mucosal lesion index (Mucosal lesions score) by the full gastric mucosa reading of Figure 3.
5 is a graph measuring the activity of glutathione peroxidase, an antioxidant index, through serum of experimental animals.
Figure 6 is a graph measuring the concentration of C-reactive protein (C-reactive protein) as an anti-inflammatory index through the serum of experimental animals.

Hereinafter, the present invention will be described in detail by examples.

However, the following examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following examples.

Preparation of a composition for preventing or treating gastritis and gastric ulcer, including cabbage, broccoli sprout, broccoli leaf, and kale as active ingredients

Preparation of a composition for preventing or treating gastritis and gastric ulcer comprising the cabbage, broccoli sprouts, broccoli leaves, and kale as an active ingredient of the present invention was performed by the following procedure.

1) shredding cabbage, broccoli sprouts, broccoli leaves, kale,

2) mixing 4 kinds of shredded vegetables with cabbage juice, isomaltooligosaccharide and aloe vera juice,

3) Inoculating the lactic acid bacteria culture medium in which lactic acid bacteria growth was activated through pre-culture and fermenting at a fermentation temperature of 35 to 39° C. for 18 to 30 hours,

4) When the fermentation is finished, pulverizing to a fine particle form using a micro mill facility,

5) Sterilizing the tubular heat exchange method at 103℃ for 2 minutes using a tubular sterilizer,

6) When sterilization is finished, filtration through a 30 mesh filtration net and packaging in a bag in box,

7) low temperature aging the packaged composition at a refrigeration temperature of 4° C. or lower for 70 to 75 hours,

8) After aging, it was prepared by freezing at a temperature of -18°C or lower.

The cabbage, broccoli sprouts, broccoli leaves, and kale are sources of abundant glucosinolate components, which are converted into isothiocyanate components having strong antibacterial and anticancer or anti-inflammatory activity through enzymatic hydrolysis. This isothiocyanate is characterized by including a -N=C=S functional group. In particular, it is a component that can help prevent gastric ulcers by inhibiting ammonia production by inhibiting Helicobacter pylori or inhibiting urease activity.

In the present invention, the mechanism of activating the isothiocyanate component through hydrolysis by microbial action through lactic acid bacteria fermentation before enzymatic hydrolysis in the intestine was utilized, and the effect of another secondary metabolite produced through fermentation metabolism To enhance the antioxidant activity, and intended to induce gastritis and gastric ulcer improvement effect through antibacterial and anti-inflammatory action.

The cabbage, broccoli sprouts, broccoli leaves, and kale can maximize the content of isothiocyanate-based components, sulfolaphane and indol-3-carbinol, through inventors' raw material composition experiments. The compounding ratio was constructed on condition.

Accordingly, the content of sulfolapan and indole-3-carbinol of the above four vegetable raw materials was measured. The analysis of sulfolapan and indole-3-carbinol was performed by the Korea Basic Science Institute, and was measured using a Ttiple Quadrupole LC-Mass spectrometry (Finnigan TSQ Quantum Ultra EMR) instrument. The test of all items was repeated three times, and the significance test between treatments was tested at p<0.05 level by ANOVA test and Duncan's multiple range test. HPLC analysis conditions are shown in [Table 1].

Sulforaphane, indole-3-carbinol analysis conditions Items Conditions HPLC Conditions Column Extend-C18 (2.1 × 150mm, 5μ, Agilent) Column Temperature 30℃ Eluents A: 0.1% FA / B: 0.1% FA/Acetonitrile Flow Rate 200 μl/min Injection volume 10 μl Mass spectrometer condition Ion Source Type ESI (positive mode) Spray Voltage 4000 Sheath Gas Pressure 40 Aux Gas Pressure 10 Capillary Temperature 300 ℃ SRM [m/z] Sulforaphane: 178.100 → 114.295 (Collision Energy 10)
Indole-3-carbinol: 130.000 → 77.400 (Collision Energy 30)

As a result, as shown in [Figure 1], it was confirmed that the content of sulfolapan was excellent in broccoli sprouts (p<0.05), and the content of indole-3-carbinol was statistically the highest in cabbage (p <0.05), followed by high content in broccoli sprouts (p<0.05). It was confirmed that the sulfolapan was also contained in kale and broccoli leaves at 1.31 μg/mL, 2.50 μg/mL, and indole-3-carbinol, respectively, at 0.13 μg/mL and 0.24 μg/mL.

In consideration of the above analysis results and the sensory properties of the mixture, preferably 15 to 30% by weight of cabbage, 10 to 20% by weight of broccoli sprouts, 3 to 10% by weight of broccoli leaves, and 1 to 5% by weight of kale, More preferably, cabbage may be used in an amount of 18 to 25% by weight, 12 to 18% by weight of broccoli sprouts, 4 to 7% by weight of broccoli leaves, and 2 to 4% by weight of kale.

The isomaltoligosaccharide was used for the purpose of supplying a carbon source that promotes fermentation, and various carbon sources can be used according to the user's purpose such as glucose, fructose, sugar, fructooligosaccharide, isomaltoligosaccharide, but isomaltoligosaccharide is preferred. It may be used, more preferably, an oligosaccharide content of 50% or more based on anhydride may be used by adding isomaltooligosaccharide at 10 to 20% by weight.

The lactic acid bacteria culture solution is prepared through pre-cultivation in order to promote the growth of lactic acid bacteria, the pre-cultivation process is mixed with 10% by weight per purified grape as a carbon source, 2.5% by weight of soy protein separated as a nitrogen source, and 85.5% by weight of purified water at 121℃ After sterilization for 15 minutes at 37°C, the Lactobacillus plantarum strain can be inoculated by 2% by weight, and cultured at 37°C for 18 hours. It can be used as a fermentation seed and inoculated by 0.1 to 1% by weight of the mixture.

The Lactobacillus plantarum may preferably be a Lactobacillus plantarum PMO 08 (KFCC-11028) strain.

The Lactobacillus plantarum PMO 08 (KFCC-11028) strain of the present invention is a lactic acid bacterium described in Korean Patent No. 263661 (2000. 5. 30) and defoams bile acids to lower blood cholesterol in vivo It has an effect of exhibiting an effect, and it is known that the survival rate in the stomach is excellent and the intestinal adhesion ability is excellent.

The fermentation process can be terminated when a certain acidity is reached by measuring the acidity of the fermentation product and using it as a quality indicator, and preferably, when the acidity is 0.9% to 1.1%, the fermentation can be terminated.

The crushing process is to pulverize in the form of fine particles using a micro mill facility, and the size of the particles can be preferably crushed to a size that can be filtered through a 0.4Ø filter net.

The low temperature aging process is a process of purifying a unique flavor by storing at a low temperature for a period of time in order to improve the flavor of the fermentation composition, and the aging temperature and time can be low temperature aging at 70° C. for preferably 70 to 75 hours. .

Food composition for preventing or treating gastritis and gastric ulcer, comprising cabbage, broccoli sprouts, broccoli leaves, and kale as an active ingredient of the present invention is a functional food, a nutritional supplement, a health food, and a food. Includes all forms of food additives. Food compositions of this type can be prepared in various forms according to conventional methods known in the art.

Verification of gastritis and gastric ulcer improvement effect of composition containing cabbage, broccoli sprout, broccoli leaf, and kale as active ingredients

<2-1> Measurement of Antioxidant Activity and Active Ingredients of Fermentation Composition

In order to verify the effect of improving the gastritis and gastric ulcer of the fermentation composition prepared by the method in <Example 1>, first, the antioxidant capacity of the composition and the content of active ingredients effective for gastric health were measured.

Antioxidant activity was evaluated through antioxidant activity and total polyphenol content by fermentation time.

Antioxidant activity was measured by partially applying the Blois method (Blois MS 1958) to measure the free radical scavenging activity of a sample using 1,1-diphenyl-2-picryhydrazyl (DPPH). As an experimental method, 1 g of each test sample dissolved in ethanol was added to 1 mL of 0.2 mM DPPH dissolved in methanol, and reacted for 10 minutes at room temperature, followed by ELISA reader (Bio-Rad, Hercules, model 680, CA, USA). Absorbance was measured at 517 nm. The control (control) was not added to the test solution, the blank (blank) was corrected for the effect of the sample without adding DPPH. Free radical scavenging activity (EDA) was calculated by the following equation.

}]×100EDA(Electro Donating Ability, Inhibition %) = [1-{

}]×100

The polyphenol compound content was measured by partially modifying the Folin Denis method (Swain & Hillis 1959) using a phenolic substance reacting with phosphomolybdic acid and showing a blue color. That is, each sample was ground evenly, 10 g was taken, immersed in 100 mL of 80% ethanol for 24 hours, and then filtered and extracted. After adding 5 mL of distilled water to 0.1 mL of the extraction solution, 0.5 mL of Folin-Ciocalteu's phenol reagent and 1.5 mL of 20% sodium carbonate were added, and then 2.9 mL of distilled water was added and reacted at room temperature for 2 hours to measure absorbance at 765 nm. The content of the polyphenol compound was calculated from a standard calibration curve prepared using gallic acid.

All items were tested in triplicate. As a result, as shown in [Fig. 2], it was confirmed that the antioxidant activity and total polyphenol content increased the most at 24 to 30 hours of fermentation time, the antioxidant activity increased up to 144%, and the total polyphenol content up to 30%. The increased results were confirmed. Therefore, it was confirmed that it has a high antioxidant capacity due to the production of secondary metabolites through fermentation, and is formulated to be effective in suppressing ulcers and inflammation.

In addition, in the case of the completed fermentation composition, the content of isothiocyanate effective for inhibiting Helicobacter pylori was measured by having high antibacterial activity as an effective ingredient effective for gastric health.

Total Isothiocyanate analysis was performed by the Korea Basic Science Institute, and was measured using a Triple Quadrupole LC-Mass spectrometry (Finnigan TSQ Quantum Ultra EMR) instrument. The test was repeated three times, and HPLC analysis conditions are shown in [Table 2].

Total isothiocyanate analysis conditions Items Conditions Column Kinetex C18 (4.6 × 250 mm, 5 μm, Phenomenex) Guard column UHPLC C18 (AJ0-8768, Phenomenex) Column Temperature 25℃ Eluent and run time 80% MEOH, 10min Flow Rate 0.8 mL/min Injection volume 10 μl UV detect 365 nm

It was confirmed that the content of the total isothiocyanate of the fermentation composition of the present invention contained an amount of 15.3 mg/mL as shown in [Table 3].

Total isothiocyanate content of fermentation composition Total isothiocyanate content (mg/mL) 15.3

<2-2> Verification of improvement effect of gastritis and gastric ulcer in fermentation composition

In order to verify the effect of improving the gastritis and gastric ulcer of the fermentation composition prepared by the method in <Example 1>, gastric tissue imaging and professional reading, gastric ulcer index (Ulcer Index), etc. were performed through acute gastritis-induced animal model using HCl/EtOH. Evaluation was made to verify the effectiveness of the fermentation composition of the present invention. To this end, 6-week-old male SD-type rats were purchased from Orient Bio Co., Ltd. and adapted for 1 week under normal temperature (23±1℃) and humidity (50±5%) with rats as a regular solid feed (AIN-93G). Each was randomly placed and used in the experiment. The experimental group was composed of four treatment groups: the normal group (normal group), the Ulcer group (HCl/EtOH gastritis-causing group), the Omeprazole group (positive control group), and the fermentation composition-administered group (test group). 900 mg/kg b.w. The fermentation composition of the present invention in a dose is 20 mg/kg b.w. in the Omeprazole group. Omeprazole (Sigma, USA), a medicine for gastric ulcer, was administered orally once a day for 7 days, respectively, and purified water was orally administered to the normal group and the Ulcer group for the same period. 1.5 mL of 150 mM HCl/60% EtOH was administered to the Ulcer group, the Omeprazole group, and the fermentation composition administration group at the end of the administration to induce symptoms of gastritis and gastric ulcer, and one hour later, the animals were sacrificed and gastric tissue and serum were sacrificed. Separated and used in the experiment. The extracted gastric tissue was stored in 10% Formalin solution, and the separated serum was stored in a deep freezer set at -70°C until analysis.

First, histopathological examination of the gastric mucosa by H&E (Hematoxylin & Eosin) staining was performed. Precise reading and analysis were performed by Northers Inc. As a result, as shown in [Fig. 3], most of the areas where erosion occurs in the stomach tissues of rats are observed with congestion and hemorrhage, and when erosion is severely observed, the submucosal layer In some cases, inflammatory cell infiltration appeared. No lesions were observed in the normal group. In the Ulcer group, symptoms of gastric ulcer and gastritis were clearly confirmed in 3 out of 6 specimens, accompanied by severe erosion, hyperemia, and bleeding, whereas the lesions were observed in the Omeprazole and fermentation groups. Weak. In view of this, it is judged that the fermentation composition of the present invention has a level of improvement effect similar to that of omeprazole, an ulcer treatment agent.

In order to add the objectivity of the read result, the gastric mucosal lesion index (Mucosal lesions score) by an expert read was measured and plotted in [Fig. 4]. Measurement criteria were 0 (no abnormality detected), 1 (minimal, weak), 2 (mild, moderate), 3 (moderate, severe), 4 (severe, very) depending on the severity of the lesion. Severity), and the significance test between experimental groups was tested at p<0.05 level by Duncan's multiple range test after ANOVA test.

When the lesion grades were classified according to the degree of erosion, congestion, and hemorrhage, the Ulcer group (ulcer-causing group) averaged 3.0 points, but the lesions were severe, but the Omeprazole group and the fermentation composition group each had 1.0 , The lesion was weak with 0.8 points, and the score was significantly reduced (p<0.05) compared to the inducing group, thereby confirming the gastritis and gastric ulcer improvement effect of the fermentation composition of the present invention.

In addition, the antioxidant index (Glutathione peroxidase) and anti-inflammatory index (C-reactive protein), which are the basis of gastric mucosal protection, were analyzed through rat serum.

Glutathione peroxidase (GSH-px) activity was measured according to the modified method of the Lawrence and burk (1976) method. The reaction mixture consisted of a glutathione peroxidase assay buffer containing 50 mM calcium phosphate buffer at pH 8.0 and 0.5 mM EDTA, and a NADHP assay reagent containing 5 mM NADHP, 42 mM GSH, 10 unit/ml glutathione reductase. homogenate solution) and samples of supernatant fluid with 50 mM calcium phosphate buffer were prepared by centrifuging 1,000 g at 4° C. for 10 minutes. 900 μl of glutathione peroxidase assay buffer, 50 μl of NADHP assay reagent, and 50 μl of samples were continuously added to the cuvette and mixed well (total amount of cuvette was 1.00 ml). The reaction was started by adding 10 μl of 30 mM t-BHP (tert-butyl hydroperoxide) or CHP (cumene hydroperoxide) (80%). Absorbance was recorded and measured by Wavelength: 340 nm, Initial delay: 15 seconds, Interval: 10 seconds, Number of readings: 6. The total amount of glutathione peroxidase was determined by the sum of the values of each experimental result using 30 mM t-BHP and CHP (80%).

C-reactive protein (C-RP) concentration was measured using a RayBio ^® Human CRP ELISA Kit (Norcross, GA, USA).

The significance test between experimental groups was tested at p<0.05 level by ANOVA test and Duncan's multiple range test.

Glutathione peroxidase (Glutathione peroxidase) is an antioxidant that plays a role in decomposing and detoxifying hydrogen peroxide and lipid peroxide that cause damage to the cell membrane. As shown in [Fig. 5], the concentration of the Ulcer group decreased sharply compared to the normal group, but Omeprazole The group and the fermentation composition administration group were significantly increased again (p<0.05), and in particular, the fermentation composition administration group was found to be 96% higher than the Ulcer group, and the fermentation composition of the present invention was considered effective in protecting the gastric membrane.

C-reactive protein (C-reactive protein) is a protein that rapidly rises 2 to 3 hours after an acute inflammatory reaction occurs, and is an indicator mainly measured for early diagnosis of inflammation and tissue damage. As shown in FIG. 6, the concentration was significantly decreased in the Omeprazole group and the fermentation composition administration group compared to the Ulcer group (p<0.05), and in particular, in the fermentation composition administration group, it was shown to decrease by 61% compared to the Ulcer group, so that inflammation was caused against acute gastritis. It is believed to have excellent efficacy in preventing gastritis by effectively controlling the reaction.

In the end, the composition for preventing or treating gastritis and gastric ulcer containing cabbage, broccoli sprouts, broccoli leaves, and kale as an active ingredient of the present invention is isothiosia through the fermentation of 4 kinds of cruciferous vegetables composed of optimal combination based on the active ingredient Gastric gastritis is produced by effectively generating gastric ulcers through antibacterial and anti-inflammatory action by generating effective secondary metabolites such as Nate and enhancing antioxidant activity, thereby preserving gastric mucosa through preclinical experiments and significantly improving related blood index levels. And gastric ulcer improvement.

Accordingly, the present invention provides a gas composition for preventing or treating gastritis and gastric ulcer, comprising a fermentation composition consisting of cabbage, broccoli sprouts, broccoli leaves, and kale. The fermentation composition for preventing or treating gastritis and gastric ulcer of the present invention is highly effective in manufacturing functional foods for gastritis and gastric ulcer improvement by protecting the gastric mucosa and significantly improving the gastric index-related blood index.

Claims (3)

Gastritis and gastric ulcer prevention or treatment composition comprising cabbage, broccoli sprout, broccoli leaf, and kale as active ingredients.
According to claim 1, wherein the composition for preventing or treating gastritis and gastric ulcer,
1) shredding cabbage, broccoli sprouts, broccoli leaves, kale,
2) mixing 4 kinds of shredded vegetables with cabbage juice, isomaltooligosaccharide and aloe vera juice,
3) Inoculating the lactic acid bacteria culture medium in which lactic acid bacteria growth is activated through pre-culture and fermenting at a fermentation temperature of 35 to 39° C. for 18 to 30 hours
4) When the fermentation is finished, pulverizing to a fine particle form using a micro mill facility,
5) Sterilizing the tubular heat exchange method at 103℃ for 2 minutes using a tubular sterilizer,
6) When sterilization is finished, filtration through a 30 mesh filtration net to carry a plastic bag (Bag in box),
7) low temperature aging the packaged composition at a refrigeration temperature of 4° C. or lower for 70 to 75 hours,
8) After aging, the composition characterized in that it is prepared through a step of freezing at a temperature of -18°C or lower.
According to claim 1, The cabbage, broccoli sprouts, broccoli leaves, kale, 18 to 25% by weight of cabbage, 12 to 18% by weight of broccoli sprouts, 4 to 7% by weight of broccoli leaves, 2 to 4% by weight of kale Gastritis and gastric ulcer prevention or treatment composition, characterized in that.

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