KR20110124516A - Composition for treating inflammation-related diseases comprising extracts from culture media of lactic acid bacteria - Google Patents

Abstract

PURPOSE: A composition containing extract isolated from lactic acid bacteria culture liquid is provided to suppress IL-6 secretion and to treat inflammatory diseases and cancer. CONSTITUTION: A composition for treating inflammatory diseases contains extract of lactic acid bacteria culture liquid. The lactic acid bacteria are Lactobacillus, Lactococcus, Leukonostoc, Pediococcus, or Bifidobacteria. The extract contains supernatant of the culture liquid except for the bacteria. The supernatant contains heat resistant ingredients. A composition for treating cancer contains the extract. A composition for treating thrombokinesis-related diseases contains the extract. A method for treating inflammatory diseases or cancer comprises a step of administering the extract.

Description

Composition for treating inflammation-related diseases comprising extracts from culture media of lactic acid bacteria}

The present invention relates to a composition containing a lactobacillus culture extract having a therapeutic effect on inflammatory diseases, blood vessel formation and / or cancer.

Chronic inflammation is usually caused by immune dysfunction. Immune dysfunction can be divided into Th2-type inflammation, characterized by eosinophilic inflammation, and Th17-type or Th1-type inflammation, characterized by neutrophil inflammation [J Allergy Clin Immunol. 2010; 125: S33-40].

It is known that IL (Interleukin) -6, which is produced by a pathogenic bacterium or a toxin derived from bacteria, is important for the generation of Th17 immune response during naive T cell differentiation. Induce differentiation into Th17 cells (Cytokine. 2009; 47 (3): 149-56).

In addition, IL-6 is known to play an important role in the formation of blood clots in the blood vessels.In the case of acute coronary syndrome and stroke, blood vessels such as cardiovascular and cerebrovascular vessels are formed. This is a disease caused by blockage.

In addition, the fact that the inflammatory response is important in the development of cancer has been raised in the art before, there is a suggestion that cancer occurs due to the tissue damage caused by inflammation and the error of its healing process. Recently, the STAT3 signaling pathway is involved in cancer development and Th17-type inflammation has been reported to be important in colorectal cancer [Cell. 2010; 140 (6): 883-99].

On the other hand, lactobacillus is a bacterium that breaks down sugars such as glucose or lactose to make lactic acid, and it is known as a bacterium that decomposes proteins but does not decay and thus has a beneficial effect on the human body.

As a beneficial mechanism of action of lactic acid bacteria, sugars are used as an energy source to ferment them to produce a large amount of lactic acid, thereby lowering the intestinal pH and inhibiting the intestinal harmful bacteria. However, there is no known action related to the inflammatory response of lactic acid bacteria.

The inventors have discovered that lactic acid bacteria have the activity of inhibiting the production of IL-6, which is an important mediator of the inflammatory response, and based on this, the lactic acid bacteria culture extract is used to treat and / or prevent inflammatory diseases, blood vessel thrombus formation and cancer. To provide a way to.

As a means for achieving the above technical problem, the present invention provides a composition containing the extract of lactic acid bacteria culture medium, the composition has the use of treating or preventing inflammatory diseases, vascular thrombosis, and cancer.

According to one embodiment of the present invention, the lactic acid bacteria culture medium is Lactobacillus (Lactobacillus), Lactococcus (Lactococcus), Leukonostoc, Pdiococcus, or Bifidobacteria (Bifidobacteria) It may be selected from materials extracted from the medium and combinations thereof, but is not limited thereto.

According to one embodiment of the present invention, the lactic acid bacteria culture extract is characterized in that the material extracted from the culture medium for Lactobacillus (Lactobacillus).

According to one embodiment of the present invention, the lactic acid bacteria culture extract is characterized in that the material extracted from the culture medium of Lactobacillus rhamnosus (Lactobacillus rhamnosus).

According to one embodiment of the present invention, the lactic acid bacteria culture extract is characterized in that the material extracted from the culture medium cultured Lactobacillus rhamnosus (Lactobacillus rhamnosus) using MRS broth medium.

According to one embodiment of the present invention, the lactic acid bacteria culture solution extract is characterized in that it comprises a culture solution supernatant from the lactic acid bacteria culture medium.

According to one embodiment of the present invention, the lactic acid bacteria culture extract is characterized in that it comprises a component that resists heat less than 3kDa.

In another aspect, the present invention provides a method for treating and / or preventing inflammatory diseases comprising the step of administering the extract of lactic acid bacteria culture fluid.

According to one embodiment of the present invention, the lactic acid bacteria culture solution administration may be selected from oral administration, nasal administration, airway administration, skin administration, intravascular intraperitoneal administration, and combinations thereof, but is not limited thereto.

In another aspect, the present invention provides a method for treating and / or preventing thrombotic disease, comprising administering an extract of a lactic acid bacteria culture medium.

In another aspect, the present invention provides a method for the treatment and / or prevention of cancer comprising the step of administering the extract of lactic acid bacteria culture fluid.

The lactic acid bacteria culture extract of the present invention has an action of inhibiting IL-6 secretion, which is an important mediator of the inflammatory response. The IL-6 is known to be involved in the development of cancer through Th17-type inflammatory response as well as the STAT3 signaling pathway, and also plays an important role in the generation of blood clots in blood vessels. Therefore, it is expected that the lactic acid bacteria culture extract of the present invention (particularly, heat resistant components of less than 3 kDa) can be used to treat and / or prevent inflammatory diseases, thrombosis, and cancer.

Figure 1 shows the results of measuring the expression of the spontaneous, time-dependent inflammatory mediator IL-6 after the culture of colon 26, a mouse colon cancer cell line by ELISA method.
Figure 2 shows the degradation and IkB degradation of IkB, an intrinsic inhibitor of NF-kB, to demonstrate that the expression of IL-6, a spontaneous and time-dependent expression of inflammatory mediator IL-6, in colon 26 cells, a mouse colon cancer cell line. The result was confirmed by western blotting method of phosphor-IkB.
FIG. 3 shows that NF-kB inhibitors, BAY 11-7082 and NF-, are used to confirm that expression of IL-6, an inflammatory mediator that expresses spontaneously and time-dependently, in colon 26 cells, a mouse colon cancer cell line, is dependent on NF-kB signaling. The expression level of IL-6 was confirmed by ELISA method using LY 294002, an inhibitor of PI3K, an upper molecule of kB.
4 is a result of confirming the expression of IL-6 by ELISA method after treatment with Lactobacillus (LR) concentration-dependently to colon 26 cells.
5 is a result of confocal microscopy using a Cell Tracker dye staining live cells to confirm that the inhibition of the expression of the inflammatory mediator IL-6 by Lactobacillus is not caused by cell necrosis.
6 shows that the activity of inhibiting the expression of IL-6 spontaneously expressed in colon 26 cells is specific activity by Lactobacillus, IL-6 in colon 26 cell culture cultured with the same number of Lactobacillus and bacillus This is the result of confirming the expression.
7 is an experimental protocol for confirming the therapeutic effect of Lactobacillus in inflammatory bowel disease animal model.
Figure 8 is the result of showing the therapeutic effect of Lactobacillus to the disease by measuring the length of the large intestine and the weight of the individual which is a common symptom of inflammatory bowel disease.
Figure 9 shows the protocol for separating the Lactobacillus culture medium according to the size of the molecule.
10 is a result of confirming the IL-6 expression inhibitory activity of the water-soluble fractions separated according to the protocol of Figure 9 by ELISA method.
11 is a result confirming that IL-6 expression inhibitory activity of isolated water-soluble components less than 3 kDa occurs in a concentration-dependent manner.
12 is a result of confirming the expression of IL-6 in the cell culture by heating the isolated water-soluble components of less than 3 kDa for 20 minutes at 100 ° C and treated with colon 26 cells.

The inventors have discovered that lactic acid bacteria have an activity of inhibiting the secretion of the inflammatory cytokine IL-6 and have completed the present invention based thereon.

The present invention relates to a composition containing the extract of lactic acid bacteria culture medium, the composition can be used for the treatment and prevention of inflammatory diseases, cancer, and thrombosis-related diseases.

IL-6 is known to be involved in the Th17-type inflammatory response characterized by neutrophil inflammation in the inflammatory response, specifically IL-6 induces differentiation into Th17 cells through the STAT3 signaling pathway. Therefore, the lactic acid bacteria culture extract of the present invention having the activity of inhibiting IL-6 secretion can be used for the treatment and / or prevention of inflammatory diseases. The type of the inflammatory disease is not limited, and may be used, for example, in the treatment of inflammatory bowel disease.

On the other hand, there is a suggestion that cancer is caused by tissue damage caused by inflammation and its healing process in association with inflammatory response and cancer, and STAT3 signaling pathway is related to cancer development, and in the case of colorectal cancer, Th17- Type inflammation is reported to be important. Therefore, lactic acid bacteria culture extract of the present invention that inhibits the secretion of IL-6 involved in Th17-type inflammatory response as well as STAT3 signaling pathway is expected to be used for anticancer treatment and prevention. There is no restriction on the type of the treatable cancer, and for example, it can be used for the treatment and prevention of colorectal cancer.

In addition, since the importance of IL-6 in the production of blood clots in blood vessels is already known, the extract of the lactic acid bacteria culture medium of the present invention which inhibits the secretion of IL-6 can be used for the treatment and prevention of thrombosis-related diseases. The thrombosis-related diseases may include acute coronary syndrome, stroke, and the like.

Hereinafter, the present invention will be described in more detail with reference to embodiments and / or embodiments of the present invention so that those skilled in the art may easily implement the present invention.

As a result of experiments evaluating IL-6 secretion in cancer cells, the present inventors confirmed that the secretion of IL-6 increases in a time-dependent manner without treating cancer cells. The present inventors used colon 26, a mouse-derived colorectal cancer cell line, as cancer cells, but the type of cancer cells that can be used is not limited.

As a result of examining the mechanism of spontaneous secretion of IL-6 in the cancer cells, expression of IL-6 was found to be dependent on the NF-kB pathway (PI3K and NF-kB). Specifically, IL-6 expression was reduced in colorectal cancer cells after treatment with BAY 11-7082, an NF-kB signaling inhibitor, and LY 294002, an inhibitor of PI3K, an NF-kB upstream molecule.

The present inventors looked at the effect of lactic acid bacteria on inflammation-related sitcaine secretion of cancer cells. There is no restriction on the type of cancer cells and lactic acid bacteria used at this time. For example, colon 26, a colon cancer cell line, can be used. Also available lactic acid bacteria can be selected from Lactobacillus (Lactobacillus), Lactococcus, Leukonostoc, Pdiococcus, or Bifidobacteria. For example, Lactobacillus rhamnosus GG can be used as the lactic acid bacteria of the present invention.

The present inventors confirmed that spontaneous IL-6 expression was reduced in a concentration-dependent manner in the Lactobacillus rhamnosus-treated group of colon cancer cells. In addition, it was confirmed that the reduction of IL-6 expression was not caused by cell necrosis.

In addition, the present inventors, in order to determine that the inhibitory effect of lactic acid bacteria on the expression of IL-6 in cancer cells is due to the specific activity of lactic acid bacteria, bacillus subtilis, a Gram-positive bacterium in colon 26, a mouse-derived colon cancer cell line under the same conditions Was treated. As a result, it was confirmed that bacillus subtilis did not inhibit the expression of IL-6 in cancer cells.

In addition, the present inventors have isolated and identified the active ingredient in the lactic acid bacteria culture medium, it was confirmed that less than 3kDa water-soluble, heat-resistant components of the lactic acid bacteria culture medium extract has a significant IL-6 secretion inhibitory effect.

Hereinafter, preferred examples are provided to aid in understanding the present invention. However, the following examples are merely provided to more easily understand the present invention, and the contents of the present invention are not limited by the examples.

Example  One. Colon cancer in cell  Inflammatory cytokines ( IL -6) secretion analysis

Experiments were performed to evaluate the secretion of inflammatory cytokines in cancer cells. In this study, we evaluated the expression level of inflammatory mediator IL-6 in colon 26, a mouse-derived colorectal cancer cell line.

Specifically, Colon 26 cells were dispensed into 24 well plates to be 2 × 10 ⁵ and incubated for 12 hours. After 12 hours, the cells were washed with PBS (phosphate buffer saline), and 500 µl of MEM (Minimum Essential Media) alpha medium was dispensed, and the cell culture solution was collected in time. ELISA was performed to confirm the expression level of the inflammatory mediator IL-6 in each cell culture, and the results are shown in FIG. 1.

As shown in Figure 1, it was confirmed that the time-dependent increase in the expression of IL-6 in the absence of any stimulation for colorectal cancer cells.

Example  2. Colon cancer in cell IL -6 expression mechanism evaluation

The present inventors confirmed the time-dependent increase in time-dependent expression of IL-6 in colon 26 cells in the absence of any stimulation from the results of Example 1, and performed experiments to identify the expression mechanism.

In this regard, to assess the expression of NF-kB-dependent IL-6, the degree of degradation and the phosphorylation of IkB, a precursor of proteosomal degradation of IkB, were examined for IkB, which acts to inhibit NF-kB activity. Confirmation was evaluated by blotting.

Specifically, the cells were divided into 5 x 10 ⁵ cells in a 60 mm culture dish, and after 12 hours, the cells were harvested in time-dependent manner to separate the protein of the cells. 30 μg of the protein was loaded per sample to confirm and evaluate the expression level of IkB and phospo-IkB, and the results are shown in FIG. 2. At this time, β-actin was used as a control.

As shown in FIG. 2, it was confirmed that IkB, an intrinsic inhibitor of NF-kB, was degraded from 12 hours when IL-6 expression was increased, and phosphorylation of IkB, a precursor of IkB proteosomal degradation, was observed at 6 and 9 hours. It was confirmed that the increase.

In addition, an experiment was performed to confirm whether the expression of IL-6, which is spontaneously expressed in colon 26 cells, actually occurs depending on the NF-kB pathway. In this regard, the expression level of IL-6 was analyzed using BAY 11-7082, an NF-kB signaling inhibitor, and LY 294002, an inhibitor of PI3K, a higher molecule of NF-kB.

Specifically, according to the method of Example 1 after 12 hours of cell dispensing BAY 11-7082 and LY 294002 at a concentration of 10nmol and after 12 hours to determine the amount of IL-6 expression in the cell culture solution using the ELISA method ( ELISA) was performed and the results are shown in FIG. 3.

As shown in FIG. 3, the expression of IL-6 was reduced in colorectal cancer cells after treatment with BAY 11-7082, an NF-kB signaling inhibitor, and LY 294002, an inhibitor of PI3K, an NF-kB upstream molecule.

From these results, it is clear that the expression of inflammatory mediator IL-6 in colon 26, a mouse-derived colorectal cancer cell line, is dependent on the NF-kB pathway (PI3K and NF-kB).

Example  3. Colon cancer cell of  Effect of Lactic Acid Bacteria on Inflammation-Related Cytokine Secretion

The present inventors conducted the following experiment to evaluate the effect of lactic acid bacteria on inflammation-related sitcaine secretion. In this regard, we examined the effect of Lactobacillus rhamnosus GG on the expression of IL-6 in colon 26, a mouse-derived colorectal cancer cell line.

Specifically, 12 hours after dispensing Colon 26 cells according to the method of Example 1 after releasing Lactobacillus rhamnosus GG in 5 x 10 ⁷ , 5 x 10 ⁸ CFU (colony-forming unit) / ml to MEM alpha respectively 500 µl of each well was dispensed and cultured for 12 hours, and then the expression of IL-6 in the cell culture was confirmed by ELISA, and the results are shown in FIG. 4.

As shown in FIG. 4, spontaneous IL-6 expression was significantly reduced (almost completely suppressed in the group treated with 5 × 10 ⁸ CFU / ml Lactobacillus rhamnosus (LR 5 × 10 ⁸ ) in colon cancer cells (colon 26). Was confirmed.

In addition, an experiment was performed to confirm that the result of FIG. 4 was caused by lactic acid bacteria rather than cell necrosis.

Specifically, after incubating colon26 with Lactobacillus rhamnosus for 12 hours according to the above method, the cell culture medium was removed, washed with PBS, 500 μl of MEM alpha was added, and the cell tracker (CellTracker, Invitrogen, NO.C 2925) solution was 5 μm. After the addition, the cells were incubated for 30 minutes. After washing with PBS, 500 μl of MEM alpha was added and incubated for another 30 minutes. The cover glass was placed in 500 µl of 4% paraformaldehyde and fixed at room temperature for 10 minutes. Then, the host stain (Hoescht dye, Sigma, No. B2883) was added to 5 µm and dyed for 10 minutes. After attaching the cover glass to the slide glass, it was observed on a confocal microscope and the result is shown in FIG.

As shown in Figure 5, it can be seen that the cell death by lactic acid bacteria (Lactobacillus rhamnosus) does not occur.

From the above results, the expression of spontaneous IL-6 in the colon cancer cells (colon 26) treated with 5 x 10 ⁸ CFU / ml Lactobacillus rhamnosus (LR 5 x 10 ⁸ ) was significantly reduced (almost completely suppressed). It was confirmed that the action was not caused by cell necrosis but by the action of lactic acid bacteria.

In addition, the inventors conducted an experiment to confirm that the inhibition of IL-6 expression in the colorectal cancer cells (colon 26) is due to the specific activity of lactic acid bacteria. In this regard, the results of analyzing the expression of inhibited IL-6 in colon 26 cells by repeating the above experiment using the Gram-positive bacillus bacillus subtilis are shown in FIG. 6.

As shown in Figure 6, when treated with the same number (5 x 10 ⁸ CFU) Lactobacillus rhamnosus and bacillus subtilis, Lactobacillus rhamnosus GG showed the activity to inhibit the secretion of IL-6, bacillus subtilis IL-6 It did not inhibit the expression of.

From the above results, it is clear that lactic acid bacteria such as Lactobacillus rhamnosus and the like have a specific activity of inhibiting expression of inflammatory cytokine IL-6 in colon cancer cells.

Example  4. inflammatory bowel disease ( inflammatory bowel disease , IBD Treatment Effect of Lactic Acid Bacteria in Animal Models

The inventors conducted an experiment to confirm the therapeutic effect of lactic acid bacteria in inflammatory bowel disease animal model. In this regard, Lactobacillus rhamnosus GG was used to evaluate its inflammatory effect.

Specifically, a DSS model was used to induce inflammatory bowel disease by mixing dextran sodium sulfate (DSS) in drinking water at 5% concentration using BALB / c female mice.

Before starting the model using the DSS according to the protocol shown in Figure 7 was administered orally administered Lactobacillus rhamnosus of 1 x 10 ⁹ CFU for 7 days at intervals of 24 hours. After that, the drinking water of the positive control group (PC) and the experimental group except the negative control (NC) and the experimental group were replaced with 5% DSS and grown for 7 days, during which the experimental group was 1 × 10 ⁹ CFU of Lactobacillus. rhamnosus was orally administered at 24 hour intervals.

To determine the weight loss, which is a symptom of inflammatory bowel disease, during the 7 days of DSS treatment, the weight of the mouse (B.wt) was measured at 10 am daily, and the mouse was dissected 24 hours after the last administration. By measuring the length of the colon (colon length) expressing the development to determine the degree of inflammatory bowel disease and the results are shown in FIG.

As shown in Figure 8, Lactobacillus rhamnosus treated in the experimental group (Lactobacillus) compared to the positive control (PC) reduced the colon length (colon length) and weight loss (B.wt change), which is a symptom of inflammatory bowel disease It was confirmed.

From the above results, it is clear that Lactobacillus rhamnosus has a therapeutic effect against DSS-induced inflammatory bowel disease model.

Example  5. Lactobacillus ( Lactobacillus rhamnosus GG ) Isolation and Identification of Active Ingredients from Culture Media

The inventors carried out experiments to isolate and identify the active ingredient after confirming the anti-inflammatory effect of lactic acid bacteria.

(One) Fractionation

According to the fractionation method shown in Fig. 9, MRS broth (GIBCO, Cat.no 288130, component per 1 L: Proteose Peptone No. 3 10 g, Beef Extract 10 g, Yeast Extract 5) g, Dextrose 20 g, Polysorbate 80 1 g, Ammonium Citrate 2 g, Sodium Acetate 5 g, Magnesium Sulfate 0.1 g, Manganese Sulfate 0.05 g, Dipotassium Phosphate 2 g) in 1 L Lactobacillus rhamnosus (3.6 x 10 ¹² CFU / L) After incubation for 12 hours, high-speed centrifugation (1000 xg, 20min, 4 ℃) to obtain a supernatant except Lactobacillus rhamnosus. The supernatant was separated into a culture solution of 100 kDa or more (50 ml) and a culture medium of less than 100 kDa (950 ml) by using a 100 kDa filter membrane, and then again using a 3 kDa filter membrane to 100 ml of the culture medium of less than 100 kDa. Was separated into a culture solution (2.5 ml) and a culture solution of less than 3 kDa (97.5 ml).

As a result of the fractionation method, water-soluble components of 100 kDa or more, 3 kDa or more and less than 100 kDa and less than 3 kDa were fractionated.

(2) lactic acid bacteria ( Lactobacillus rhamnosus GG ) Evaluation of Activity of Water-Soluble Components in Culture Medium

Experiment to evaluate the anti-inflammatory effect of the water-soluble component fractions in the lactic acid bacteria culture medium isolated according to the method of Example 5 (1). In this regard, the expression level of IL-6 was evaluated when the fractions were treated for colon 26, a mouse-derived colorectal cancer cell line.

Specifically, in the same manner as in Example 1, the respective water-soluble component fractions separated according to the molecular size were treated in the cells at the same ratio (50 μl / ml) and incubated for 12 hours to express the expression of IL-6 in the cell culture. Confirmation by ELISA and the result is shown in FIG.

As shown in Figure 10, it was confirmed that the expression of IL-6 is significantly inhibited in colorectal cancer cells treated with less than 3 kDa water-soluble fractions.

In addition, in order to evaluate whether the inhibition of expression of IL-6 in the water-soluble component fraction of less than 3 kDa is concentration dependent, 20 µl / ml of each of the water-soluble components of less than 3 kDa was added to colon 26 according to the method of Example 1. , And after 12 hours of 100 μl / ml treatment, confirmed the expression of IL-6 in the cell culture and the results are shown in FIG. 11.

As shown in FIG. 11, it can be seen that the water-soluble component of less than 3 kDa in Lactobacillus rhamnosus GG culture medium inhibits the expression of IL-6 in colorectal cancer cells in a concentration-dependent manner.

(3) lactic acid bacteria ( Lactobacillus rhamnosus GG ) 3 in culture kDa  Water Soluble Ingredient Analysis

The present inventors further performed an experiment to determine whether the water-soluble component of less than 3 kDa is a heat resistant component.

After heating the aqueous solution component of less than 3 kDa for 20 minutes at 100 ℃ and immediately left for 5 minutes at 4 ℃, incubated for 12 hours by treatment in Colon 26 according to the method of Example 1 with the unheated control group The expression of IL-6 in the cell culture was confirmed and the results are shown in FIG. 12.

As shown in FIG. 12, in the effect of inhibiting the expression of IL-6 in colorectal cancer cells, there was a difference between a group treated with a less than 3 kDa water-soluble component heated at 100 ° C. for 20 minutes and a group treated with a non-heated control group. It did not represent. From the above results, the water-soluble component of less than 3 kDa in the lactic acid bacteria culture medium having the activity of inhibiting the expression of IL-6 was identified as a component resistant to heat.

From the above, it can be clearly seen that the lactic acid bacteria culture extract of the present invention (particularly less than 3 kDa of the water-soluble component in the lactic acid bacteria culture extract) has an activity of inhibiting the secretion of IL-6. IL-6 is also an important mediator of the inflammatory response and is known to play an important role in the Th17-type inflammatory response by inducing differentiation into Th17 cells through the STAT3 signaling pathway.

Therefore, it is possible to prevent and / or treat inflammatory diseases using the lactic acid bacteria culture extract of the present invention, in addition, the STAT3 signaling pathway is related to cancer development, and in the case of colorectal cancer, Th17-type inflammation has been reported to be important. Therefore, it is expected that the lactic acid bacteria culture extract of the present invention can be used for anticancer treatment and / or prevention. In addition, since the importance of IL-6 in the formation of blood clots is already known, the lactic acid bacteria culture extract of the present invention may be used for the treatment and prevention of thrombosis-related diseases such as acute arterial syndrome and stroke.

The above description is merely illustrative of the technical idea of the present invention, and various modifications, changes, and substitutions may be made by those skilled in the art without departing from the essential characteristics of the present invention. will be. Accordingly, the embodiments disclosed in the present invention and the accompanying drawings are not intended to limit the technical idea of the present invention, but to describe the present invention. The scope of the technical idea of the present invention is not limited by the embodiments and the accompanying drawings. Is apparent to those skilled in the art.

Claims (34)

Inflammatory disease treatment composition containing the lactic acid bacteria culture extract.
The method of claim 1,
The lactic acid bacterium is Lactobacillus (Lactobacillus), Lactococcus (Lactococcus), Leukonostoc, Pediococcus (Pdiococcus), and the composition characterized in that it is selected from the group consisting of Bifidobacteria (Bifidobacteria).
The method of claim 1,
The lactic acid bacteria culture extract is a composition, characterized in that the material extracted from the culture medium Lactobacillus (Lactobacillus).
The method of claim 1,
The lactic acid bacteria culture extract is a composition, characterized in that the material extracted from the culture medium for Lactobacillus rhamnosus (Lactobacillus rhamnosus).
The method of claim 1,
The Lactobacillus culture extract Lactobacillus rhamnosus (Lactobacillus rhamnosus) MRS broth medium components (Proteose Peptone, Beef Extract, Yeast Extract, Dextrose, Polysorbate 80, Ammonium Citrate, Sodium Acetate, Magnesium Sulfate, Manganese Sulfate, and Dipotassium Phosphate) A composition characterized in that the material is extracted from the culture solution cultured in the containing medium.
The method of claim 1,
The lactic acid bacteria culture extract is characterized in that it comprises a culture supernatant except lactic acid bacteria.
The method of claim 6,
The supernatant comprises a heat resistant component. The method of claim 7, wherein
Wherein said heat resistant component comprises a heat resistant component of less than 3 kDa in size.
Cancer treatment composition containing the extract of lactic acid bacteria culture.
10. The method of claim 9,
The lactic acid bacterium is Lactobacillus (Lactobacillus), Lactococcus (Lactococcus), Leukonostoc, Pediococcus (Pdiococcus), and the composition characterized in that it is selected from the group consisting of Bifidobacteria (Bifidobacteria).
10. The method of claim 9,
The lactic acid bacteria culture extract is a composition, characterized in that the material extracted from the culture medium Lactobacillus (Lactobacillus).
10. The method of claim 9,
The lactic acid bacteria culture extract is a composition, characterized in that the material extracted from the culture medium for Lactobacillus rhamnosus (Lactobacillus rhamnosus).
10. The method of claim 9,
The Lactobacillus culture extract extracts Lactobacillus rhamnosus from MRS broth medium components (Proteose Peptone, Beef Extract, Yeast Extract, Dextrose, Polysorbate 80, Ammonium Citrate, Sodium Acetate, Magnesium Sulfate, Manganese Sulfate, and Dipotassium Phosphate). A composition characterized in that the material is extracted from the culture solution cultured in the containing medium.
10. The method of claim 9,
The cancer is a composition, characterized in that the colorectal cancer.
10. The method of claim 9,
The lactic acid bacteria culture extract is characterized in that it comprises a culture supernatant except lactic acid bacteria.
16. The method of claim 15,
The supernatant comprises a heat resistant component. The method of claim 16,
Wherein said heat resistant component comprises a heat resistant component of less than 3 kDa in size.
A composition for treating thrombogenic disease, comprising the extract of a lactic acid bacteria culture medium.
The method of claim 18,
The lactic acid bacterium is Lactobacillus (Lactobacillus), Lactococcus (Lactococcus), Leukonostoc, Pediococcus (Pdiococcus), and the composition characterized in that it is selected from the group consisting of Bifidobacteria (Bifidobacteria).
The method of claim 18,
The lactic acid bacteria culture extract is a composition, characterized in that the material extracted from the culture medium Lactobacillus (Lactobacillus).
The method of claim 18,
The lactic acid bacteria culture extract is a composition, characterized in that the material extracted from the culture medium for Lactobacillus rhamnosus (Lactobacillus rhamnosus).
The method of claim 18,
The Lactobacillus culture extract extracts Lactobacillus rhamnosus from MRS broth medium components (Proteose Peptone, Beef Extract, Yeast Extract, Dextrose, Polysorbate 80, Ammonium Citrate, Sodium Acetate, Magnesium Sulfate, Manganese Sulfate, and Dipotassium Phosphate). A composition characterized in that the material is extracted from the culture solution cultured in the containing medium.
The method of claim 18,
The lactic acid bacteria culture extract is characterized in that it comprises a culture supernatant except lactic acid bacteria.
The method of claim 23, wherein
The supernatant comprises a heat resistant component.
25. The method of claim 24,
Wherein said heat resistant component comprises a component of size less than 3 kDa.
Inflammatory disease treatment method comprising administering a lactic acid bacteria culture extract.
The method of claim 26,
The administration is characterized in that the administration is selected from the group consisting of oral administration, nasal administration, airway administration, skin administration and vascular administration.
The method of claim 26,
The lactic acid bacteria culture medium extract is characterized in that it comprises a component that resists heat of less than 3kDa size.
A method of treating inflammatory bowel disease, the method comprising the step of orally administering a component containing lactic acid bacteria culture medium extract resistant to heat of less than 3kDa.
Cancer treatment method comprising the step of administering the lactic acid bacteria culture extract.
The method of claim 30,
The lactic acid bacteria culture medium extract is characterized in that it comprises a component that resists heat of less than 3kDa size.
The method of claim 30,
Said cancer is colorectal cancer.
A method for treating thrombogenic disease comprising administering an extract of a lactic acid bacteria culture medium.
The method of claim 33, wherein
The lactic acid bacteria culture medium extract is characterized in that it comprises a component that resists heat of less than 3kDa size.

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