KR20130018621A - Composition for preventing, treating or improving gastrointestinal disease comprising plant originated lactic acid bacteria, lactobacillus plantarum pmo08 as an active ingredient - Google Patents

Abstract

PURPOSE: A composition containing Lactobacillus plantarum PMO08 for preventing and treating/relieving gastrointestinal diseases is provided to effectively suppress gastric and duodenal inflammation. CONSTITUTION: A pharmaceutical composition for preventing and treating gastrointestinal diseases contains a mixture of Lactobacillus plantarum PMO08(KFCC-11028) and a Dioscoreaceae plant, as an active ingredient. The gastrointestinal diseases include gastric ulcer, duodenal ulcer, acute gastritis, chronic gastritis, cardialgia, reflux esophagitis, duodenitis, dyspepsia, gastrointestinal hemorrhage, or gastric cancer. Dioscoreaceae plants include Dioscorea Batatas, Dioscorea Japonica, Dioscorea Bulbifera, Dioscorea nipponica, Dioscorea Tokoro, Dioscorea Tenuipes, Dioscorea Septembloba, or Dioscorea Quinqueoloba. A food composition for preventing and treating gastrointestinal diseases contains the mixture as an active ingredient. A pharmaceutical composition for preventing and treating gastrointestinal diseases contains Lactobacillus plantarum PMO08(KFCC-11028) as an active ingredient.

Description

[0001] The present invention relates to a composition for preventing, treating or ameliorating a gastrointestinal disorder, which comprises lactic acid bacteria Lactobacillus plantarum pimelo 08 as an active ingredient. [0002]

The present invention relates to a novel use of plant lactobacillus Lactobacillus plantarum pimae 08, and more particularly to a novel use of plant lactobacillus plantarum pumio 08 or lactobacillus plantarum pumio 08, As an active ingredient, to a composition for preventing and treating gastrointestinal diseases.

Lactic acid bacteria, which are abundant in fermented foods, play a role in decomposing fibrous and complex proteins into important nutritional components in a symbiotic relationship with the digestive system of the human body. Thus, the intestinal microbial environment of the host is improved in the gastrointestinal tract of animals including humans Living microorganisms that have a beneficial effect on the health of the host are collectively referred to as probiotics.

Lactic acid bacteria are lactic acid bacteria that produce carbohydrates and make lactic acid by using them. They are tuberous anaerobes or organisms that grow well in low oxygen. Lactic acid bacteria can be divided into five genus, Streptococcus, Lactobacillus, Leuconostoc, Bifidobacteria and Pediococcus.

Microorganisms of Streptococcus, a Streptococcus, are homozygous and are known to inhibit rot and pathogens by fermenting milk to produce lactic acid. The microorganisms of the genus Lactobacillus are lactic acid bacilli that are homozygous or heterozygous, and are commonly found in the fermentation process of dairy products or vegetables, and the microorganisms of the genus ruconosstock, which are heterozygous, are mainly involved in fermentation of vegetables. In addition, Bifidobacteria genus microorganisms are organized anaerobic bacteria that do not grow in the presence of oxygen, ferment sugar to produce L (+) type lactic acid that can be usefully metabolized in infants. Finally, the microorganism of Pediococcus is a homozygous fermenting microorganism in the form of sycamore, which is present in kimchi or pickled food and is involved in the fermentation of meat such as sausage.

Among them, microorganisms of the genus Lactobacillus are lactic acid bacterium that is homozygous or heterozygous fermenting, and are commonly found in the fermentation process of intestines, dairy products and vegetables of animals including humans. Lactobacillus microorganisms maintain intestinal pH acidic and inhibit the growth of harmful microorganisms such as E. coli and Clostridium, and improve diarrhea and constipation, as well as vitamin synthesis, anti-cancer effects, serum cholesterol lowering And so on. Acidophillin produced by lactic acid bacilli is known to inhibit the growth of dysentery, Salmonella, Staphylococcus aureus, and Escherichia coli. It inhibits the growth of diarrheal pathogens and normalizes intestinal microflora to stop diarrhea.

On the other hand, in order for the lactic acid bacteria to have a useful effect as a probiotic agent, it must be able to form a community by attaching it to the host tissue after ingestion and to exhibit lactic acid bacterial inherent functions. In Koreans, the length of digestion is longer than that of Westerners because of the long time required for digestion due to the vegetable-based diet, which has a high fiber content from ancient times. As the survival rate of lactic acid bacteria is lower than that of Westerners, I need a way to be able.

Recently, as the function of Lactobacillus plantarum isolated from fermented foods such as vegetables, fruits such as vegetables, or fruit and vegetables such as kimchi, it is emerging as a new functional material. Lactobacillus Planta Room Acid resistance, biliary and antibiotic resistance effects of Jay 124 (Korean Patent Application No. 2002-0049220), Lactobacillus Planta Room Jay 9 (Korean Patent Application No. 2003-0067073), Listeria Monocytogenes and Enterococcus Anticancer Activity against Lactobacillus , Lactobacillus Planta Room CJ LP 243 enhances the intestinal disease treatment and immunity (Republic of Korea Patent Application No. 2009-0102822 No.) effects, Lactobacillus Planta Room The antioxidant and anti-inflammatory efficacy of Hewlett-Packard 7711 (Korean Patent Application No. 2009-0058180) has been disclosed. Recently, Lactobacillus plantarum pmi 08 (Korean Patent Application No. 2009-0130699) having anti-cancer or antimicrobial activity has been disclosed.

However, the above results have confirmed the physiological activity of the lactic acid bacteria, the therapeutic effect of the ulcer, the dysfunction / disease on the stomach or duodenum, and the method for promoting the intrinsic function of the lactic acid bacteria was not considered .

Accordingly, the inventors of the present invention have investigated the function of Lactobacillus plantarum pmi 08 (KFCC-11028), a kind of plant lactobacillus, and found that Lactobacillus plantarum pimio 08 (KFCC-11028) The present invention has been completed based on the discovery that a complex with a dead plant has excellent gastrointestinal disease prevention and treatment effects or gastrointestinal health promotion effects such as prevention / improvement of gastrointestinal diseases.

It is therefore an object of the present invention to provide a new use of a mixture of Lactobacillus plantarum pumio 08 (KFCC-11028) or Lactobacillus plantarum pumio 08 and a fungus plant.

In order to accomplish the object of the present invention as described above, the present invention provides a composition for preventing gastrointestinal diseases, which comprises a mixture of Lactobacillus plantarum pmi 08 (KFCC-11028) And a pharmaceutical composition for therapeutic use.

In order to achieve the another object of the present invention, the present invention comprises a food composition for preventing and improving gastrointestinal diseases, comprising a mixture of Lactobacillus plantarum POM 08 (KFCC-11028) and an asteraceae as an active ingredient to provide.

In order to accomplish still another object of the present invention, the present invention provides a pharmaceutical composition for the prevention and treatment of gastrointestinal diseases, which comprises Lactobacillus plantarum pmi 08 (KFCC-11028) as an active ingredient.

In order to accomplish still another object of the present invention, there is provided a food composition for prevention and improvement of gastrointestinal diseases, which comprises Lactobacillus plantarum pmi 08 (KFCC-11028) as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for the prevention and treatment of gastrointestinal diseases, which comprises a mixture of Lactobacillus plantarum pyromo 08 (KFCC-11028) and a dead plant as an active ingredient.

The lactobacillus plantarum pimio 08 is present in any one or more form selected from the group consisting of live cells, dried germs, dead cells, and culture dried products.

In one embodiment of the present invention, the presence of duodenal puncture in the cysteamine-treated group was confirmed. As a result, the number and size of perforations of the lactic acid bacteria or the lactic acid bacteria / hemp mixture of the present invention were reduced compared to the cysteamine groups, and in particular, the lactic acid bacteria / hemp mixture did not cause perforation, &Lt; 1-2 > and < 1-3 >).

In another embodiment of the present invention, confirmation of such results can be performed by examining histopathological examination of the duodenum and histopathological findings through a blind test. The histopathological changes of inflammation include inflammation findings, depth and width of erosion or ulcer, And scored for the loss of chordae and villi. As a result, the lactic acid bacteria of the present invention or the lactic acid bacteria and horsetail of the present invention showed less tissue damage and inflammation, and the lactic acid bacteria and horsetail of the present invention had the most excellent effects (Examples <1-4 >).

The lactic acid bacterium contained in the lactic acid bacteria / hemaglobin mixture contains only about 0.25 times of the lactic acid bacteria administered to the lactic acid bacteria-only test group (G4) and contains only 0.20 times of the hemoglobin group administered to the hemoglobin test group (G3) , The effect of the mixture is much better than that of lactic acid bacteria and hemp alone.

In addition, experiments were conducted in stomach. The amount of gastric juice secretion was similar to that of the normal control group (S1) in the case of lactic acid bacteria + lactic acid bacteria, lactic acid bacteria, and lactic acid bacteria. However, the control group (S6) injected with PPI showed a significantly lower amount of gastric juice than that of the normal control group, indicating that there is a possibility of causing another secondary gastrointestinal disorder (see Example <2-2>). Gastric intestinal biopsy and histopathological examination were also performed. As compared with the control group (S2), which showed gastric damage due to indomethacin, the experimental group containing Lactobacillus, Lactobacillus, Was about 50% (see Example <2-3>).

Cyclooxygenase (COX) is a major enzyme for the synthesis of prostanoids (prostaglandins, prostacyclins and thromboxanes). COX has two types of isoforms, COX-1, which is expressed as a constituent in most tissues and is known to produce prostaglandin (PG) for general physiological function. However, COX-2 is induced in response to a variety of stimuli such as lipopolysaccharide (LPS), tumor promoters, oncogenes, carcinogens, mitogens, hormones, cytokines and growth factors. The inhibition of COX-2 expression was examined by immunohistochemistry. As a result, the concentration of COX-2 expression was significantly higher in the experimental group of lactic acid bacteria, lactic acid bacteria, and lactic acid bacteria than in the control group (S2) , But the group containing PPI (S6) expressed COX-2 at a level similar to that of the S2 group (see Example <2-4>).

Proton pump inhibitor (PPI), used as a positive control, is an anti-ulcer drug that suppresses proton pump and strongly inhibits gastric acid secretion. Gastric acid is secreted when the stimulus is applied through the histamine receptor, acetylcholine receptor, and somatostatin receptor present in the parietal wall located on the upper surface. Since PPI inhibits the proton pump, which is the final step of gastric acid secretion, It is powerful. PPI includes omeprazole, rabeprazole, pantoprazole, etc. In the present invention, pantoprazole is used. Pantoprazole is INN (international non-proprietary name) for the compound 5-difluoromethoxy-2 - [(3,4-dimethoxy-2-pyridinyl) methylsulfinyl] -1H-benzimidazole. The most common side effects of PPI are headache, diarrhea, abdominal pain, and nausea. PPI decreases gastric acid secretion and raises gastric pH, thus inhibiting the absorption of griseofulvin, ketoconazole, itraconazole, iron and vitamin B12 . PPI is also metabolized in the liver mitochondria (CYP2C19). Therefore, metabolism such as diazepam (barium), phenytoin (diratin) and wafarin (coumadin) which are metabolized through the same path is competitively inhibited and accumulated in the body excessively, so that side effects of drugs such as coumadin may occur.

Therefore, in the present invention, it has been confirmed that Lactobacillus plantarum pmi 08 (KFCC-11028), which is a plant lactic acid bacterium, has an excellent effect on the improvement of gastrointestinal health. Therefore, it is possible to prevent gastrointestinal diseases Thereby providing a therapeutic composition. In particular, the composition of the present invention may contain, as an active ingredient, a mixture of Lactobacillus plantarum pyromo 08 (KFCC-11028) and a marigold plant. The concentration of the lactic acid bacteria in the stomach and duodenum is not suitable for the growth of lactic acid bacteria. Therefore, the number of lactic acid bacteria in the stomach and duodenum is not suitable for the growth of lactic acid bacteria. About 0.25 times, and the gastrointestinal experiment was about 0.03 times).

Dioscoreaceae plants include, but are not limited to, for example, Dioscorea Batatas, Dioscorea Japonica, Dioscorea Bulbifera, Dioscorea nipponica, Dioscorea Tokoro, Dioscorea Tenuipes, Dioscorea Septembloba or Dioscorea Quinqueoloba.

Although the whole plants can be used in all parts, Rhizome parts can be used. For example, Dioscorea Rhizoma may include all the powder products that have been dried or steamed after removing the Rhizome juniper of the above-mentioned endemic plants. Further, it is possible to use the root material itself of the above-mentioned dead plants or the dried root material itself or the dried root material. In addition, hot water or alcohol extract (or its concentrate, extracted powder thereof) of a dead plant or an enzyme-treated product, juice, or crushed product through enzyme treatment may be used for the active ingredient.

The diseases to which the composition of the present invention is applicable may be gastrointestinal diseases. More specifically, it can be gastrointestinal disease, gastric ulcer, duodenal ulcer, gastritis or heartburn, reflux esophagitis, duodenitis, indigestion, stomach cancer.

Meanwhile, the composition of the present invention may be a pharmaceutical composition or a food composition.

The pharmaceutical composition according to the present invention comprises a pharmaceutically effective amount of Lactobacillus plantarum pimero 08 or a mixture of Lactobacillus plantarum pimero 08 and a plant or a mixture of at least one pharmaceutically acceptable carrier May be further included. The pharmaceutical compositions of the present invention may be administered to a patient in a single dose and may be administered by a fractionated treatment protocol in which multiple doses are administered over a prolonged period of time. The pharmacologically effective amount in the above refers to an amount showing a reaction higher than that of the negative control, and preferably refers to an amount sufficient to treat or prevent a gastrointestinal disorder. The effective amount of the mixture of Lactobacillus plantarum pimio 08 or lactobacillus plantarum pimero 08 according to the present invention is between 0.001 and 1000 mg / day / kg body weight, but is not limited thereto. However, the pharmaceutically effective amount may be appropriately changed depending on various factors such as the disease and its severity, the patient's age, body weight, health condition, sex, administration route and treatment period.

The composition of the present invention may be formulated in various ways according to the route of administration by a method known in the art together with the pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable" is a non-toxic composition that, when administered physiologically to humans, does not inhibit the action of the active ingredient and typically does not cause an allergic reaction such as gastrointestinal disorders, dizziness, or the like. Say The composition of the present invention may be formulated in various ways according to the route of administration by a method known in the art together with the pharmaceutically acceptable carrier. Routes of administration include, but are not limited to, oral or parenteral administration.

When the pharmaceutical composition of the present invention is orally administered, the pharmaceutical composition of the present invention may be formulated into a powder, a granule, a tablet, a pill, a sugar, a tablet, a capsule, a liquid, a gel , Syrups, suspensions, wafers, and the like. Examples of suitable carriers include saccharides including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol, and starches including corn starch, wheat starch, rice starch and potato starch, Cellulose such as methylcellulose, sodium carboxymethylcellulose and hydroxypropylmethyl-cellulose and the like, fillers such as gelatin, polyvinylpyrrolidone and the like. In addition, crosslinked polyvinylpyrrolidone, agar, alginic acid or sodium alginate and the like may optionally be added as a disintegrant. Further, the pharmaceutical composition may further comprise an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent and an antiseptic agent.

In addition, when administered parenterally, the pharmaceutical compositions of the present invention may be formulated according to methods known in the art together with suitable parenteral carriers.

Other pharmaceutically acceptable carriers may be referred to those described in Remington's Pharmaceutical Sciences, 19th ed., 1995, Mack Publishing Company, Easton, PA.

Furthermore, the pharmaceutical composition of the present invention can be administered in combination with known compounds having an effect of preventing and treating gastrointestinal diseases.

In addition, the mixture of Lactobacillus plantarum pimio 08 or Lactobacillus plantarum pimero 08 according to the present invention may be provided in the form of a food composition for the purpose of preventing and / or ameliorating gastrointestinal diseases . The food composition of the present invention includes all forms of functional food, nutritional supplement, health food and food additives, lactic acid bacteria drink, fermented milk and the like. Food compositions of this type can be prepared in various forms according to conventional methods known in the art.

For example, as a health food, a mixture of Lactobacillus plantarum pumio 08 or Lactobacillus plantarum pimero 08 of the present invention and a fungus plant itself is prepared in the form of a lactic acid bacterium, a fermented milk, a tea, a juice and a drink Be ingested, ingested, granulated, encapsulated and powdered. In addition, a mixture of Lactobacillus plantarum pumio 08 or Lactobacillus plantarum pimero 08 of the present invention and a mixture of a plant and a known plant material or an active ingredient known to have an effect of improving gastrointestinal diseases is mixed with the plant to form a composition Can be manufactured.

Functional foods also include beverages (including alcoholic beverages), fruits and their processed foods (such as canned fruits, bottled, jam, marmalade, etc.), fish, meat and processed foods such as ham, sausage, Breads and noodles (eg udon, buckwheat noodles, ramen noodles, spaghetti, macaroni, etc.), juice, various drinks, cookies, sugar, dairy products such as butter, cheese, edible vegetable oil, margarine, vegetable protein, Frozen foods, various seasonings (for example, soybean paste, soy sauce, sauces, etc.) by adding a mixture of Lactobacillus plantarum pumio 08 or Lactobacillus plantarum pimero 08 of the present invention.

In addition, Lactobacillus plantarum pemo 08 can be used as a starter to produce beverages mixed with milk, cereals, fruits, vegetables, legumes, saccharides, etc., alone or in combination with fermented plants.

The preferred content of the mixture of Lactobacillus plantarum pumio 08 or Lactobacillus plantarum pimero 08 and the fungi in the food composition of the present invention is not particularly limited but is preferably 0.001 to 50 Weight%.

In addition, in order to use the mixture of Lactobacillus plantarum pimio 08 of the present invention or Lactobacillus plantarum pimero 08 and the fungus plant in the form of food additives, it may be used in the form of powder or concentrate.

Meanwhile, the present invention provides a lactic acid bacterium beverage comprising a mixture of Lactobacillus plantarum pimero 08 and a marine plant as an active ingredient. In addition, the present invention provides a lactic acid bacterium beverage comprising Lactobacillus plantarum pimae 08 as an active ingredient. The lactic acid bacteria, the dead plants and the usefulness thereof of the present invention are as described above.

As described above, the mixture of Lactobacillus plantarum pumio 08 or Lactobacillus plantarum pimero 08 of the present invention effectively inhibits stomach and duodenal inflammation and effectively prevents / treats gastrointestinal diseases such as ulcers. And is effective for prevention and treatment / improvement of gastrointestinal diseases. Therefore, the microorganism of the present invention can be used for prevention, treatment, and improvement of gastrointestinal diseases.

Figure 1 shows a representative duodenal photograph of each experimental group.
Figure 2 shows the puncture rate of the duodenum by each experimental group.
Figure 3 shows the perforation size of the duodenum by each experimental group.
FIG. 4 shows photographs of hematoxylin and eosin staining (H & E stain) of the duodenum of each experimental group.
FIG. 5 shows a mean score graph of the inflammation findings of each experimental group.
FIG. 6 shows a graph of average score of the lumen and villus of each experimental group.
FIG. 7 shows a mean score graph of ulcer size for each experimental group.
Figure 8 shows a mean score graph of total pathological findings.
9 is a graph showing a result of measurement of the amount of gastric juice secreted by each experimental group.
FIG. 10A shows gastric mucosa photographs for each experimental group.
FIG. 10B shows a mean score graph for the damage index according to each experimental group.
FIG. 11 shows photographs of COX-2 expression level observed by immunohistochemical staining for each experimental group.

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

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

&Lt; Example 1 >

Duodenal damage treatment efficacy

<1-1> Setting and processing of experiment group

As shown in Table 1, the experimental animals (SD rats, 7 weeks old) were divided into 6 groups from G1 to G6 in the experimental group, and all the experimental groups were sacrificed for one day, followed by gastric nourishment with lactic acid bacteria (Lactobacillus plantarum pM (KFCC-11028) alone or in combination with lactic acid bacteria / fungi were administered to the stomach at the same dose and cysteamine was administered in the same manner as described above. Clinical symptoms were investigated 24 hours after administration.

Ma was used as a green tea powder from Nongandong Nonghyup, and PPI (Sigma-aldrich, Pantoprazole) was used as a positive control.

Experimental group Experimental material Dosage Number of animals G1 Normal 10 G2 Cysteamine (500 mg / kg) 15 G3 Cysteamine (500 mg / kg) + hemoglobin 2 g / kg body weight 10 G4 Cysteamine (500 mg / kg) + lactic acid bacteria 2x10 ⁹ / rat 10 G5 Cysteamine (500 mg / kg) + lactic acid bacteria / mash mixture Lactobacillus
5.0x10 ⁸ / rat 10 hemp
393 mg / kg body weight G6 Cysteamine (500 mg / kg) + PPI (proton pump inhibitor, pantoprazole) 10 mg / kg body weight 10

<1-2> Measurement of duodenal puncture rate

After the experiment, the duodenum was removed from the experimental animals and the duodenal puncture rate was measured for the cysteamine-treated group (G2). The kidneys extracted for each experimental group are shown in Fig.

A total of 14 perforations were observed in the group treated with cysteamine only. Four perforations were observed in G3, two in G4, zero in G5, and one in G6. G2 was 100%, G3 was 62.9%, G4 was 31.4%, G5 was 0%, G6 was 100%, and the average perforation rate of cysteamine group (G2) was 100% 17.5%, indicating that the perforation does not occur only in the high-concentration treated group of the plant-derived lactic acid bacteria complex (see FIG. 2).

<1-3> Measure duodenal puncture size

The puncture size of the duodenum extracted in the above Example <1-2> was measured.

As a result, the mean size of perforations in the experimental group was 2.73 mm in the cysteamine group (G2), 1 mm in G3, 0.3 mm in G4, 0 mm in G5, and 0.22 mm in G6. G2 is 100%, G3 is 36.7%, G4 is 11%, G5 is 0%, and G6 is 8.2% when the ratio of the perforation size to the G2 group is taken as 100% of the cysteamine group (G2) (See FIG. 3). It can be seen that the size of the perforation in all experimental materials was reduced compared to the G2 group.

<1-4> Histopathological examination for duodenal inflammation - Scoring measurement

The histopathological examination of the duodenal inflammation extracted in the above Example <1-2> was performed.

A portion of the duodenum was fixed to a 10% buffered neutral formalin solution to be cut to a certain thickness, and then subjected to a general tissue treatment, followed by paraffin embedding to prepare a tissue slice of 4 to 5 μm. Then, hematoxylin And Eosin staining (Hematoxylin & Eosin, H & E stain). In addition, histopathologic changes of inflammation were scored by blind test by observing inflammation findings, depth and width of erosion or ulcer, and loss of joint and villus.

As a result, it can be confirmed morphologically that the histopathologic change (G2) of the inflammation induced by cysteamine is most improved in the high concentration group of plant lactic acid bacteria (G5), as shown in [Fig. 4] The mean scores for the inflammatory findings in Fig. 5, the mean score for the loss of the chordae and villi in Fig. 6, and the mean score for the depth and breadth of erosion or ulceration in Fig. 7, The results were as follows.

The total pathological scores of each pathological score in FIG. 8 were 1.7 ± 0.8 for G1, 42.5 ± 31.5 for G2, 22.1 ± 22.8 for G3, 29.7 ± 32.6 for G4, 9.4 ± 16.2 for G5, 26.2 ± 12.3, and the highest score of 9.4 ± 16.2 in the high concentration group of vegetable lactic acid bacteria complex (G5).

< Example  2>

Gastric damage treatment efficacy

<2-1> Setting and processing of experiment group

(SD rats, 7 weeks old) were divided into six groups from S1 to S6 in the experimental group, and the live bacteria and fungi of the lactic acid bacteria (KFCC-11028), PPI (proton pump inhibitor, lansoprazole) (Indomethacin) at a dose of 25 mg / kg was administered by gavage to all experimental groups except S1. The indomethacin was suspended in 0.5% CMC (3% sodium bicarbonate). Seven hours after the administration of indomethacin, the stomach was removed for clinical examination.

Lactobacillus plantarum pomo 08 (KFCC-11028) cells were cultured in the same manner as in Example 1-1, except that the culture broth-tyndalization- The method was used as a method of separating containing liquid - preservative mixture - preliminary freezing - freeze - milling - mixing of dead cells with excipient '.

Experimental group Experimental material Dosage Number of animals S1 Normal 10 S2 Indomethacin (25 mg / kg) 10 S3 Indomethacin (25 mg / kg) + lactic acid bacteria / Lactobacillus
3x10 ⁸ / rat 10 hemp
225 mg / kg body weight S4 Indomethacin (25 mg / kg) + live bacteria of lactic acid bacteria 1x10 ¹⁰ / rat 10 S5 Indomethacin (25 mg / kg) + bacterium of lactic acid bacteria 1x10 ¹⁰ / rat 10 S6 Indomethacin (25 mg / kg) + PPI (proton pump inhibitor, pantoprazole) 100 mg / kg body weight 10

In the above Table 2, the administration dose refers to the dose of another sample except indomethacin.

<2-2> Measurement of gastric juice

The gastric excess was used as a biomarker for gastric damage because it acts as a gastric lesion. The stomach was removed from the experimental rats of Example <2-1>, and the forceps were removed from the small intestine, and the pyloric portion was slightly cut out and the gastric juice was placed in the tube. The gastric juice contained in the tube was centrifuged (3,000 rpm, 10 minutes) to measure the volume of the supernatant.

The results of the above experiment are shown in Fig. In Fig. 9, the amount of gastric juice in the group (S2) administered with indomethacin alone was more than twice that in the normal group (S1), and the group of lactic acid bacteria + yeast (S3), live cells (S4) The amount of secretion is similar to that of normal group. On the other hand, in the PPI-treated group (S6), the amount of gastric juice secretion is less than that of the normal group, which may cause secondary gastrointestinal side effects due to low gastric juice secretion.

<2-3> Measurement of gastric damage

A part of the stomach tissues extracted in Example <2-1> were fixed in 10% neutral buffered formalin. After fixed tissue was cut to a certain thickness, paraffin embedding was performed through general tissue processing, and 4 to 5 tissue sections were prepared. Hematoxylin & Eosin staining (H & E staining) The depth, depth, and degree of bleeding were scored. Scoring criteria are listed in Table 3 below.

score Degree of lesion 0 No inflammatory response in mucosa and submucosa One Mild infiltration of neutrophils in submucosa and lower mucosa but no gastric epithelial change 2 Moderate infiltration of neutrophils and macrophages in mucosa as well as submucosa and focal necrosis of gastric epithelia 3 Severe infiltration of neutrophils and macrophages in mucosa and submucosa and multifocal necrosis of gastric epithelia 4 Severe infiltration of neutrophils and macrophages in mucosa and submucosa and massive necrosis of gastric epithelia 5 Severe infiltration of neutrophils and macrophages in mucosa, submucosa and muscle layer, massive necrosis of all parts of gastric epithelium and presence of edema, erosinm and ulcer

The higher the score, the greater the degree of damage.

The above damage measurement results are shown in Fig. In the group treated with indomethacin alone (S2), the gastric mucosal damage was severe whereas the group containing lactic acid bacteria + hemp (S3), lactic acid bacteria (S4) and lactic acid bacteria (S5) It can be confirmed that the damage reduction effect is about 50%.

<2-4> Immunohistochemical staining ( Immunohistochemistry )through COX -2 expression observation.

Indomethacin increases the expression of COX-2 (cyclooxygenase-2), which mediates inflammation. Therefore, the amount of COX-2 expression was confirmed by immunological staining.

Immunohistochemistry (IHC) was performed by selecting slides for each group of stomach tissues in which the lesion score was measured in Example <2-3>. After deparaffinization with xylene, the mixture was treated with anhydrous alcohol, 90%, 75%, and 50% ethanol for 2 minutes each. In order to inhibit the activity of endogenous hydrogen peroxide, the cells were treated with 0.3% hydrogen peroxide-methanol for 10 minutes, washed with distilled water, and then washed with 50 mMol Tris buffer (TBS, pH 7.5). To remove the nonspecific reaction, the cells were treated with anti-chlorine serum for 30 minutes, and the remaining solution on the slide was removed. The primary antibody, COX-2, was then allowed to react at room temperature for 2 hours. After the primary antibody reaction, the cells were washed with TBS three times for 5 minutes and then allowed to react with the biotin-conjugated antibody for 20 minutes, followed by staining with the conventional ABC (Avidin-biotin complex) method. 3-amino-9-ethylcarbazole (AEC) was used as a coloring agent, and the cells were stained with Mayer's hematoxylin and then observed with an optical microscope.

The brown portion, COX-2 expression site, is shown in green and is shown in Fig. It is confirmed that the group containing the lactic acid bacteria live cells, the lactic acid bacteria live cells, and the lactic acid bacteria cells has a smaller amount of COX-2 expression than the S2 group. Particularly, the lactic acid bacterium + hematite (S3) and dead cells (S5) groups are similar to the control group (S1). However, S6 containing PPI showed COX-2 expression similar to that of S2 group.

As described above, the mixture of Lactobacillus plantarum Lactobacillus plantarum pamoate 08 or Lactobacillus plantarum pimero 08 of the present invention effectively prevents stomach and duodenal inflammation and effectively prevents gastrointestinal diseases such as ulcers. / Is effective to prevent / treat / improve gastrointestinal diseases. Therefore, the microorganism of the present invention can be used for prevention, treatment, and improvement of gastrointestinal diseases.

Korea Microbial Conservation Center (Domestic) KFCC11028 19980401

Claims (8)

A pharmaceutical composition for preventing and treating gastrointestinal diseases, comprising a mixture of Lactobacillus plantarum PEMO 08 (KFCC-11028) and an asteraceae as an active ingredient.
The composition of claim 1, wherein the gastrointestinal disease is a disease selected from the group consisting of gastric ulcer, duodenal ulcer, acute gastritis, chronic gastritis, heartburn, reflux esophagitis, duodenitis, indigestion, gastrointestinal bleeding and gastric cancer.
2. The method according to claim 1, wherein the dead plants are selected from the group consisting of Dioscorea Batatas, Dioscorea Japonica, Dioscorea Bulbifera, Dioscorea nipponica, Dioscorea Tokoro, Dioscorea Tenuipes, Wherein the composition is selected from the group consisting of Dioscorea Septembloba and Dioscorea Quinqueoloba.
Food composition for the prevention and improvement of gastrointestinal diseases, comprising a mixture of Lactobacillus plantarum PEMO 08 (KFCC-11028) and asteraceae as an active ingredient.
A pharmaceutical composition for preventing and treating gastrointestinal diseases, comprising Lactobacillus plantarum POM08 (KFCC-11028) as an active ingredient.
Food composition for preventing and improving gastrointestinal diseases, comprising Lactobacillus plantarum POM 08 (KFCC-11028) as an active ingredient.
A lactic acid bacterium beverage comprising a mixture of Lactobacillus plantarum POM08 (KFCC-11028) and an asteraceae as an active ingredient.
Lactic acid bacteria beverage containing Lactobacillus plantarum POM08 (KFCC-11028) as an active ingredient.

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