KR102021883B1 - Lactobacillus fermentum lm1016 and composition for preventing or treating of inflammatory diseases and metabolic diseases comprising the same - Google Patents

Abstract

The present invention relates to a lactobacillus fermentum LM1016 strain (KFCC11756P), and a composition for preventing or treating inflammatory diseases or metabolic diseases comprising the same. More specifically, the lactobacillus fermentum LM1016 strain of the present invention alleviates cell damage and growth inhibition caused by fine dust in human hepatic cell lines (HepG2 cell) and reduces reactive oxygen species (ROS) in cell lines. In addition, the present invention recovers a level of SOD (peroxide removing enzymes) to normal and increases activation of HO-1 (Heme lyase) to reduce stress. Moreover, when a diesel exhaust particulate matter (DEPM), a standard matter of fine dust, is injected to a mouse model, the present invention reduces generation of inflammatory cytokine TNF-α, IL-6, and IL-1ß increased in blood. In addition, the present invention reduces levels of liver enzyme AST and ALT in blood increased by fine dust, and accordingly, prevents and treats liver cell damage or inflammation by fine dust.

Description

LACTOBACILLUS FERMENTUM LM1016 AND COMPOSITION FOR PREVENTING OR TREATING OF INFLAMMATORY DISEASES AND METABOLIC DISEASES COMPRISING THE SAME}

The present application relates to a Lactobacillus fermentum LM1016 ( Lactobacillus fermentum LM1016) strain (KFCC11756P), and a composition for preventing or treating inflammatory or metabolic diseases including the same.

Recently, due to the rapid industrialization of Southeast Asian countries, air pollutants caused by diesel exhaust particulate matter (DEPM) or microparticle structure air pollutants, especially inflammatory liver damage, etc. Is increasing. This inflammatory liver injury is a serious problem, such as progressing to non-alcoholic fat liver disease (NAFLD), liver fibrosis, liver cancer or Type-II diabetes, etc. It is reported in the media or through several reports.

Particulate matters in the air pass through the lung tissue (alveolar epithelium) through the respiratory tract, causing primary damage to the lung tissue, and the PMs passing through the lung tissue are secondarily transmitted through the human body to intestinal It has been reported to affect microbiome changes, hepatocytes and kidneys. Among them, diesel combustion particles have been reported to have different effects depending on particle size and water solubility or water solubility. Water-soluble PMs were found to accumulate in the liver, kidneys, and heart through the lung mucosa, while water-insoluble PMs could accumulate in the blood or blood vessel walls. In addition, PM or nano-sized carbon blacks produced during diesel combustion act as important inducers for liver inflammation and transcriptional inflammation, resulting in non-alcoholic steatohepatitis (NASH). It has been proven to act as a trigger for serious inflammatory conditions that can progress.

Tomaru, M et al. Observed that serum ALT (aspartate aminotransferase) or AST (aspartate aminotransferase) concentrations, which are important markers of liver damage in obese diabetic mouse models, were significantly increased by DEPM exposure.

Exposure to DEPM of lung or intestinal mucosal tissues leads to an increase in blood levels of lipo-poly-saccharides (LPS) or lipo-teichoic acids (LTA) derived from microorganisms (Endotoxemia), which may affect TLR-II, TLR-IV, etc. It is recognized and induces an increase in inflammatory cytokines, and induces an increase in multiple inflammation in several related tissues. It has been shown that the response of macrophages present in these lung tissues to the surface TLR-II or TLR-IV to DEPM induces an increase in inflammatory cytokines and is also closely related to the development of asthma. .

Accordingly, an object of the present invention is to develop probiotics that can prevent or alleviate liver damage caused by fine dust.

Li Xue et al. Alleviate endotoxemia caused by increased blood levels of LPS in the intestine and non-alcoholic fatty liver (NAFLD) induced by TLR-IV activation and reduce the concentration of endotoxemia in the blood. Intestinal microbiome control with probiotics has been shown to play an important role.

Jun Li et al. Also demonstrated that probiotics can be used to control the intestinal microbiome in mouse models and consequently reduce cancer cell growth in the liver, an intestinal tissue. Hepatocyte growth was reduced by 40% in the control group fed probiotics compared to the control group that did not receive probiotics and also induced a decrease in the concentration of IL-17 at the intestinal and tissue ends and a reduction in the production of Th-17 cell lines. It was.

[Preceding technical literature]

[1] Kim J.W., S. Park, C. W. Lim, K.H. Lee and B.S. Kim, 2014. Toxicol. Res., 30 (2); 65-70.

[2] Fonceca A.M., G. R. Zosky, E.M. Bozanich E.M. Sutanto, A. Kicic, P.S. McNamara, D. A. Knight, P.D. Sly, D.J. Turner and S.M. Stick, 2018, Respiratory Res. 19; 15--25.

[3] Tomaru M., H. Takano, K. Inoue, R. Yanagisawa, N. Osakabe, A. Yasuda, A. Shimada, Y. Kato and H. Uematsu, 2007, Int. J. Mol. Med., 19; 17-22.

[4] Susanne B., M.J. Fenton and J.M. Soukup, 2002, Am. J. Respir. Cell Mol. Biol. 27; 611-618.

[5] Li Xue, J. He, N. Gao, X. Lu, M. Li, X. Wu, Z. Liu, Y. Jun, J. Liu, J. Xu and Y. Geng, 2017, Scientific Report , 7; 45176 / DOI: 10.1038 / srer45176 / www.nature.com/scietificreports.

[6] Jun Li, C.Y.J. Sung, N. Lee, Y Ni, J. Pihlajamaki, G. Panagiotou, 2016, PNAS, E1306-E1315 / www.pnas.org / cgi / doi / 10.1073 / pnas.1518189113.

The present application is to provide a composition for the prevention or treatment of Lactobacillus fermentum LM1016 ( Lactobacillus fermentum LM1016) strain (KFCC11756P), and inflammatory or metabolic diseases including the same.

However, the problem to be solved by the present application is not limited to the above-mentioned problem, another task that is not mentioned will be clearly understood by those skilled in the art from the following description.

A first aspect of the present application provides a Lactobacillus fermentum LM1016 strain (KFCC11756P).

The second aspect of the present application, the pharmaceutical composition for the alleviation, prevention or treatment of inflammatory diseases or metabolic diseases, including live bacteria, microorganisms, cultures, lysates, extracts, or mixtures of Lactobacillus Permanent LM1016 (KFCC11756P) as an active ingredient To provide a composition.

The third aspect of the present application is a food composition for the prevention or alleviation of inflammatory diseases or metabolic diseases comprising live bacteria, microorganisms, cultures, lysates, extracts, or mixtures of Lactobacillus permanent LM1016 (KFCC11756P) as an active ingredient to provide.

According to the embodiments of the present invention, the Lactobacillus permanent LM1016 strain reduces cytotoxicity or toxicity of organ tissues, thereby alleviating, preventing or treating an inflammatory disease or metabolic disease caused by fine dust.

According to the embodiments of the present invention, the Lactobacillus permanent LM1016 strain may reduce cell damage and growth inhibition by microdust in HepG2 cells as well as reduce free radical species (ROS) in the cell line. . In addition, it increases the level of oxidative stress scavenging enzyme SOD (Superoxide dismutase) and the activity of HO-1 (heme oxygemase-1, Heme degrading enzyme). Infusion of particulate matter may reduce the production of elevated inflammatory cytokines TNF-α, IL-6 and IL-1ß in the blood. In addition, it is possible to reduce the level of blood liver enzymes AST and ALT increased by fine dust. Accordingly, as a result, the Lactobacillus latent LM1016 strain of the present invention exhibits a palliative, prophylactic or therapeutic effect on hepatocyte damage or inflammation caused by fine dust.

1, in one embodiment of the present application, Lactobacillus plantarum LM1001 (Accession No .: KCCM42959), Lactococcus lactis LM1009 (Lactococcus lactis subsp. Lactis LM1009) in human hepatocytes (HepG2 cells), respectively ), Lactobacillus rhamnosus LM1011, Streptococcus thermophilus LM1012, Streptococcus thermophilus LM1012, accession no. Accession no. longum LM1062), Lactobacillus ruteri LM1063) It is a graph showing the results of measuring the survival rate of human hepatocytes when treated with dust (100 μg / mL) for 24 hours by the MTT method.
Figure 2, in one embodiment of the present application, when the pre-treatment of each lactic acid bacteria in human hepatocytes and treated with fine dust (100 μg / mL) for 24 hours, the cytotoxic effect of the lactic acid bacteria under treatment LDH ( It is a graph showing the results measured by lactate dehydrogenase.
Figure 3, in one embodiment of the present application, pretreatment of each lactic acid bacteria in human hepatocytes and treated with fine dust (100 μg / mL) and incubated for 24 hours to measure the concentration of reactive oxygen species (ROS) in the cells One graph.
Figure 4, in one embodiment of the present application, the pre-treatment of each lactic acid bacteria in human hepatocytes and the fine dust (100 μg / mL) of 24 hours of the superoxide dismutase (SOD) of the enzyme It is a graph showing the result of measuring activity.
Figure 5, in one embodiment of the present application, after pre-treatment of each lactic acid bacteria in human hepatocytes and fine dust (100 μg / mL) for 24 hours, the oxidative stress removal enzyme HO- by Kutty and Maines method This is a graph measuring the activity of 1 (heme oxygemase-1).
6 (a) to 6 (c) show an inflammatory cytokine secreted in a medium after pretreatment of respective lactic acid bacteria and fine dust (100 μg / mL) in a human hepatocyte cell line in one embodiment of the present application. , TNF-α, IL-6, and IL-1ß concentration is a graph measured by the ELISA method.
Figure 7 (a) and (b), in one embodiment of the present application, 100 mg / kg of fine dust (DEPM), or 100 mg / kg of fine dust (DEPM) and lactic acid bacteria (1.0E + 09 cfu / kg) after oral administration to the mice for 14 days, the concentration of (a) Aspartate Aminotransferase (AST) and (b) Alanine Transferase (ALT), which are blood markers of liver damage, is measured.
8, in one embodiment of the present application, after oral administration of 100 mg / kg of fine dust or 100 mg / kg of fine dust and lactic acid bacteria to a mouse for 14 days, LDH (Lactate), which is an indicator of blood damage in liver Dehydrogenase) is a graph of measurement.
Figure 9, in one embodiment of the present application, after oral administration of 100 mg / kg of fine dust, or 100 mg / kg of fine dust and lactic acid bacteria to the mouse for 14 days, BUN (Blood) which is a blood index of kidney damage Urea Nitrogen) is a graph measuring the change.

DETAILED DESCRIPTION Hereinafter, exemplary embodiments of the present disclosure will be described in detail with reference to the accompanying drawings so that those skilled in the art may easily implement the present disclosure. As those skilled in the art would realize, the described embodiments may be modified in various different ways, all without departing from the spirit or scope of the present invention. In the drawings, parts irrelevant to the description are omitted for simplicity of explanation, and like reference numerals designate like parts throughout the specification.

Throughout this specification, when a part is said to be "connected" with another part, this includes not only the "directly connected" but also the "electrically connected" between other elements in between. do.

Throughout this specification, when a member is located “on” another member, this includes not only when one member is in contact with another member but also when another member exists between the two members.

Throughout this specification, when a part is said to "include" a certain component, it means that it can further include other components, without excluding the other components unless otherwise stated. As used throughout this specification, the terms “about”, “substantially”, and the like, are used at, or in close proximity to, numerical values when manufacturing and material tolerances inherent in the meanings indicated are provided and an understanding of the present application is intended. Accurate or absolute figures are used to assist in the prevention of unfair use by unscrupulous infringers. As used throughout this specification, the term “step of” or “step of” does not mean “step for”.

Throughout this specification, the term "combination (s) thereof" included in the expression of a makushi form refers to one or more mixtures or combinations selected from the group consisting of components described in the expression of makushi form, It means to include one or more selected from the group consisting of the above components.

Throughout this specification, the description of “A and / or B” means “A or B, or A and B”.

Hereinafter, with reference to the accompanying drawings will be described embodiments and embodiments of the present application; However, the present disclosure may not be limited to these embodiments, examples, and drawings.

A first aspect of the present application provides a Lactobacillus fermentum LM1016 strain (KFCC11756P).

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may comprise a 16S-rRNA sequence of SEQ ID NO: 1.

In one embodiment of the present application, by the fine dust may reduce the activity of the cell or increase the toxicity of the cell, accordingly, damage or inflammation may occur in the organ.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may be to reduce cytotoxicity or toxicity of organ tissues due to fine dust, but may not be limited thereto.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may be to increase the activity of the cells reduced due to fine dust, but may not be limited thereto.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may be to reduce the toxicity of the increased cells due to fine dust, but may not be limited thereto.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may control the amount of oxidative stress scavenging enzyme changed due to fine dust, but may not be limited thereto. For example, the oxidative stress releasing enzyme may include SOD (Superoxide Dismutase) and / or HO-1 (Heme Oxygenase-1, Heme Degrading Enzyme), but may not be limited thereto.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may increase the amount of reduced oxidative stress scavenging enzyme due to fine dust, but may not be limited thereto. For example, when the production amount of the oxidative stress scavenging enzyme decreases with exposure to the fine dust, the production amount of the oxidative stress scavenging enzyme may be recovered and / or increased again by the Lactobacillus permantum LM1016 strain, but is not limited thereto. It may not be.

Of the oxidative stress scavenging enzymes, HO-1 (Heme Oxygenase-1) is a heme degrading enzyme and tends to increase simultaneously for its inhibition when oxidative stress in the cells increases.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may further increase the production of HO-1 (Heme degrading enzyme) increased due to fine dust, but may not be limited thereto. For example, when the production amount of HO-1 increases with exposure to the fine dust, the production amount of HO-1 may be further increased by the Lactobacillus fermentum LM1016 strain, but may not be limited thereto.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may reduce the production of increased inflammatory cytokines due to fine dust, but may not be limited thereto. For example, when the amount of inflammatory cytokines increases with exposure to the fine dust, the amount of inflammatory cytokines may be reduced by the Lactobacillus fermentum LM1016 strain, but may not be limited thereto.

In one embodiment of the present application, the inflammatory cytokine may be one or more selected from the group consisting of TNF-α, IL-6, IL-1β, and combinations thereof, but may not be limited thereto. have.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may be to alleviate the cytotoxicity caused by fine dust. For example, the Lactobacillus permanent LM1016 strain may increase the activity of the cells, reduce the toxicity of the cells, increase the production of oxidative stress scavenging enzymes, or reduce the production of inflammatory cytokines. It may mitigate cytotoxicity caused by dust, but may not be limited thereto.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may be to alleviate damage to organ tissue caused by fine dust.

In one embodiment of the present application, the Lactobacillus fermentum LM1016 strain may be to reduce blood liver toxicity, but may not be limited thereto. For example, the hepatotoxicity can be confirmed by using alanine aminotransferase (ALT), aspartate aminotransferase (AST), or lactate dehydrogenase (LDH), which are indicators of hepatotoxicity, and by treating the Lactobacillus fermentum LM1016 strain to fine dust. By increasing the liver toxicity can be reduced, but may not be limited thereto.

In one embodiment of the present application, the Lactobacillus fermentum LM1016 strain may be to reduce the renal toxicity in blood, but may not be limited thereto. For example, the renal toxicity may be confirmed through Blood Urea Nitrogen (BUN), which is an indicator of renal toxicity, and may reduce the renal toxicity raised by fine dust as the Lactobacillus latent LM1016 strain is treated. It may not be limited.

The second aspect of the present application, the mitigation of inflammatory diseases or metabolic diseases, including live bacteria, microorganisms, cultures, lysates, extracts, or mixtures of Lactobacillus fermentum LM1016 strain (KFCC11756P) as an active ingredient It provides a pharmaceutical composition for prophylaxis or treatment.

With respect to the pharmaceutical composition for alleviating, preventing or treating an inflammatory disease or metabolic disease according to the second aspect of the present application, detailed descriptions of portions overlapping with the first aspect of the present application are omitted, but the description thereof is omitted. The content described in the first aspect may be equally applied to the second aspect of the present application.

In one embodiment of the present application, the pharmaceutical composition does not specifically limit the content if it contains the Lactobacillus Permantum LM1016 strain, for example, from about 10 ² ea / ml to about 10 ¹¹ ea / ml, about 10 ² ea / ml to about 10 ⁸ ea / ml, about 10 ² ea / ml to about 10 ⁶ ea / ml, about 10 ² ea / ml to about 10 ⁴ ea / ml, about 10 ⁴ ea / ml to about 10 ¹¹ ea / ml, about 10 ⁴ ea / ml to about 10 ⁸ ea / ml, about 10 ⁴ ea / ml to about 10 ⁶ ea / ml, about 10 ⁶ ea / ml to about 10 ¹¹ ea / ml, about 10 ⁶ ea / ml to about 10 ⁸ ea / ml, or about 10 ⁸ ea / ml to about 10 ¹¹ ea / ml, but may not be limited thereto.

In one embodiment of the present application, by the fine dust may reduce the activity of the cell or increase the toxicity of the cell, accordingly, damage or inflammation may occur in the organ.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may be to reduce cytotoxicity or toxicity of organ tissues due to fine dust, but may not be limited thereto.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may be to increase the activity of the cells reduced due to fine dust, but may not be limited thereto.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may be to reduce the toxicity of the increased cells due to fine dust, but may not be limited thereto.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may control the amount of oxidative stress scavenging enzyme changed due to fine dust, but may not be limited thereto. For example, the oxidative stress releasing enzyme may include SOD (Superoxide Dismutase) and / or HO-1 (Heme Oxygenase-1, Heme Degrading Enzyme), but may not be limited thereto.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may increase the amount of reduced oxidative stress scavenging enzyme due to fine dust, but may not be limited thereto. For example, when the production amount of the oxidative stress scavenging enzyme decreases with exposure to the fine dust, the production amount of the oxidative stress scavenging enzyme may be recovered and / or increased again by the Lactobacillus permantum LM1016 strain, but is not limited thereto. It may not be.

Of the oxidative stress scavenging enzymes, HO-1 (Heme Oxygenase-1) is a heme degrading enzyme and tends to increase simultaneously for its inhibition when oxidative stress in the cells increases.

In one embodiment of the present application, the Lactobacillus LM1016 strain may further increase the production amount of HO-1 (Heme degrading enzyme) increased due to fine dust, but may not be limited thereto. For example, when the production amount of HO-1 increases with exposure to the fine dust, the production amount of HO-1 may be further increased by the Lactobacillus fermentum LM1016 strain, but may not be limited thereto.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may reduce the production of increased inflammatory cytokines due to fine dust, but may not be limited thereto. For example, when the amount of inflammatory cytokines increases with exposure to the fine dust, the amount of inflammatory cytokines may be reduced by the Lactobacillus fermentum LM1016 strain, but may not be limited thereto.

In one embodiment of the present application, the inflammatory cytokine may be one or more selected from the group consisting of TNF-α, IL-6, IL-1β, and combinations thereof, but may not be limited thereto. have.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may be to alleviate the cytotoxicity caused by fine dust. For example, the Lactobacillus permanent LM1016 strain may increase cell activity, reduce cell toxicity, further increase the production of oxidative stress scavenging enzymes, or reduce the production of inflammatory cytokines, It may mitigate the cytotoxicity caused by the fine dust, but may not be limited thereto.

In one embodiment of the present application, the Lactobacillus permanent LM1016 strain may be to alleviate damage to organ tissue caused by fine dust.

In one embodiment of the present application, the Lactobacillus fermentum LM1016 strain may be to reduce blood liver toxicity, but may not be limited thereto. For example, the hepatotoxicity can be confirmed by using alanine aminotransferase (ALT), aspartate aminotransferase (AST), or lactate dehydrogenase (LDH), which are indicators of hepatotoxicity, and by treating the Lactobacillus fermentum LM1016 strain to fine dust. By increasing the liver toxicity can be reduced, but may not be limited thereto.

In one embodiment of the present application, the Lactobacillus fermentum LM1016 strain may be to reduce the renal toxicity in blood, but may not be limited thereto. For example, the renal toxicity may be confirmed through Blood Urea Nitrogen (BUN), which is an indicator of renal toxicity, and may reduce the renal toxicity raised by fine dust as the Lactobacillus latent LM1016 strain is treated. It may not be limited.

In one embodiment of the present application, furthermore, the pharmaceutical composition is not particularly limited as long as it is used in diseases caused by cytotoxicity due to fine dust, but may preferably be an inflammatory disease or a metabolic disease.

In one embodiment of the present invention, the inflammatory disease is enteritis, gastritis, hepatitis, bronchitis, pleurisy, arthritis, rhinitis, keratitis, nephritis, peritonitis, spondylitis, pancreatitis, inflammatory pain, urethritis, cystitis, burn inflammation, dermatitis, periodontitis, One or more selected from the group consisting of gingivitis, degenerative neuritis, inflammatory pain, and combinations thereof, but may not be limited thereto.

In one embodiment of the present application, the inflammatory disease may be more preferably liver inflammation due to hepatitis, for example, fine dust, but may not be limited thereto.

In one embodiment of the present disclosure, the metabolic disease is at least one selected from the group consisting of type 1 diabetes, type 2 diabetes, asthma, tuberculosis, hypertension, obesity, coronary atherosclerosis, arteriosclerosis, and combinations thereof It may be, but may not be limited thereto.

In one embodiment of the present application, the pharmaceutical composition comprising the Lactobacillus permanent LM1016 strain, respectively, oral formulations of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc. according to a conventional method, It may be used in the form of external preparations, suppositories or sterile injectable solutions, but may not be limited thereto.

In one embodiment of the present application, when formulating the pharmaceutical composition, it may be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, or surfactants commonly used, but is not limited thereto. You may not.

In one embodiment of the present application, the solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may include at least one excipient, for example, in the Lactobacillus permanent LM1016. , Starch, calcium carbonate (calcium carbonate), sucrose (sucrose), lactose (lactose), glucose (Glucose), maltodextrin (Malto-dextrin), or gelatin may be prepared by mixing. In addition, for example, in addition to simple excipients, lubricants such as magnesium stearate and talc may be used, but may not be limited thereto.

In one embodiment of the present application, liquid preparations for oral administration include suspensions, solvents, emulsions, syrups, etc. In addition to water, liquid paraffin, which is a commonly used simple diluent, various excipients such as wetting agents, Sweeteners, fragrances, preservatives and the like may be included, but may not be limited thereto.

In one embodiment of the present application, formulations for parenteral administration may include, but are not limited to, sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations and suppositories. For example, as the non-aqueous solvent or suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like may be used, but may not be limited thereto. For example, as the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, etc. may be used, but the present invention may not be limited thereto.

The third aspect of the present application, the prophylactic, microbial, culture, lysate, extract, or mixture of Lactobacillus fermentum LM1016 strain (KFCC11756P) as an active ingredient, prevention of inflammatory disease or metabolic disease Or it provides a food composition for relief.

With respect to the food composition for the prevention or alleviation of inflammatory or metabolic diseases according to the third aspect of the present application, detailed descriptions of portions overlapping with the first or second aspect of the present application have been omitted, even if the description is omitted The content described in the first or second aspect of can be equally applied to the third aspect of the present application.

In one embodiment of the present application, the food composition does not specifically limit the content if it comprises the Lactobacillus Permantum LM1016 strain, for example, from about 10 ² ea / ml to about 10 ¹¹ ea / ml, about 10 ² ea / ml to about 10 ⁸ ea / ml, about 10 ² ea / ml to about 10 ⁶ ea / ml, about 10 ² ea / ml to about 10 ⁴ ea / ml, about 10 ⁴ ea / ml to about 10 ¹¹ ea / ml, about 10 ⁴ ea / ml to about 10 ⁸ ea / ml, about 10 ⁴ ea / ml to about 10 ⁶ ea / ml, about 10 ⁶ ea / ml to about 10 ¹¹ ea / ml, about 10 ⁶ ea / ml to about 10 ⁸ ea / ml, or about 10 ⁸ ea / ml to about 10 ¹¹ ea / ml, but may not be limited thereto.

In one embodiment of the present application, the food composition may be used in dietary supplements, dairy products, fermented products, food additives, or feed additives, but may not be limited thereto.

In one embodiment of the present application, the type of the food composition is not particularly limited. The food composition that may include the Lactobacillus fermentum LM1016 strain is, for example, dairy products, including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, or ice cream , Various soups, beverages, tea, drinks, alcoholic beverages and vitamin complexes, but may not be limited thereto.

In one embodiment of the present application, the food composition may be used in the form of pills, powders, granules, acupuncture, tablets, capsules or beverages, but may not be limited thereto.

In one embodiment of the present application, in the food composition, the beverage composition, except for containing the live bacteria, microorganisms, cultures, shreds, extracts, or mixtures of the Lactobacillus Permantum LM1016 strain is not particularly limited to the liquid component No, it may contain various flavors or natural carbohydrates, etc. as an additional component, such as a conventional beverage, but may not be limited thereto.

In one embodiment of the present application, the dietary supplement is a natural flavoring agent, for example, tautin, stevia extract (for example rebaudioside A, glycyrrhizin, etc.), or a synthetic flavoring agent as a flavoring agent. (Saccharin, aspartame, etc.), but may not be limited thereto.

In one embodiment of the present application, the health functional food, various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (cheese, chocolate, etc.), pectic acid And salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like, but may not be limited thereto.

In one embodiment of the present application, the health functional food may include, but is not limited to, natural fruit juice and fruit flesh for the production of fruit juice drinks and vegetable drinks. For example, the components can be used independently or in combination.

In one embodiment of the present application, the subject animal to which the feed additive is added to the feed may be a mammal, and includes, for example, an animal selected from the group consisting of rats, cows, dogs, horses, and combinations thereof. It may be, but may not be limited thereto.

Hereinafter, the present invention will be described in more detail with reference to examples of the present application, but the following examples are merely illustrated to aid the understanding of the present application, and the content of the present application is not limited to the following examples.

[ Example ]

Sample Preparation

Lactobacillus strains used in this example are as follows: Lactobacillus plantarum LM1001 (Lactobacillus plantarum LM1001, Accession No .: KCCM42959), Lactococcus lactis LM1009 (Lactococcus lactis subsp. Lactis LM1009), Lactobacillus rhamnosus LM1011 Lactobacillus rhamnosus LM1011), Streptococcus thermophilus LM1012 (Streptococcus thermophilus LM1012, accession no. Sus LM1019 (Lactobacillus rhamnosus LM1019), Lactobacillus acidophilus LM1060, Bifidobacterium breve LM1061, Bifidobacterium breve LM1062, Bifidobacterium long lumbarbacterium LM1063 (Lactobacillus ruteri LM1063).

The 11 kinds of lactic acid bacteria strains were prepared in the dried raw materials by the method of preparing, incubating, cell recovery, mixing of a cryoprotectant, lyophilization, etc. of each seed in Lactomason. Raw materials of each of the prepared strains were diluted with appropriate concentration in tertiary distilled water and then sterilized (121 ° C., 30 minutes) to obtain a sample. In vivo samples were orally administered with diluted concentration of bacteria.

Treatment of Laboratory Animals

A 6-week-old male ICR mouse was used for sale at Samtaco Korea (Korea). Breeding environment was raised under the temperature of 22 ± 2 ℃, humidity 50 ± 20%, lighting for 12 hours, and the feed was fed solid feed for mouse and the drinking water was unlimited. The animals were acclimated in the experimental animal room for one week before performing the experiment. Eleven strains of lactic acid bacteria prepared were orally administered once daily for 2 weeks at a concentration of 10 mg / kg and diesel exhaust particulate matter (DEPM) at 100 mg / kg. After 24 hours after the last oral administration, blood samples were obtained from the experimental animals and stored at -80 ° C for use in the experiment.

Cell culture

A549 cells or HepG2 cell cultures were incubated (37 ° C humidified 5%) using RPMI1640 medium containing 10% fetal bovine serum (FBS), 2 mM L-glutamine, 100 units / ml penicillin, and 100 μg / ml streptomycin. Cultured in a CO ₂ incbator).

Splenocytes  Isolation and Cultivation

The spleens were aseptically extracted from rats, washed with RPMI 1640 solution and then ground to liberate cells. The separated cell suspension was passed through a 200 mesh stainless steel sieve and then centrifuged at 4 ° C. and 1,200 rpm for 3 minutes, and the cell pellet obtained was suspended in ACK buffer for 5 minutes to remove red blood cells. The obtained splenocytes were suspended in RPMI 1640 containing 10% FBS and 1% penicillin-streptomycin so that the concentration of cells was 1 × 10 ⁶ cells / ml, and 500 μl was respectively dispensed into 48 well plates. Treatment was done by concentration. LPS (lipopolysaccharide) was divided into samples as a positive control and treated to cells. Cell groups treated as above were incubated for 3 to 48 hours in 37 ℃, 5% CO ₂ incubator, the supernatant of the culture was stored at -20 ℃ was used for cytokine production and cytotoxicity measurement.

Cytotoxicity measurement ( MTT  Assay)

Hepatocytes were adjusted to a cell concentration of 5 × 10 ⁶ cells / ml in a 96-well microtiter plate, and 100 μl of the hepatocytes were treated in each well, followed by incubation for 1 hour. After exchanging the culture solution, the MTT labeling mixture prepared by adding an electron coupling reagent to the MTT labeling reagent was treated for 10 hours (final concentration 0.5 mg / ml) for 4 hours for each well. After treatment, the absorbance (550 nm) was used to measure the cytotoxicity of the prepared probiotic bacteria themselves.

In culture LDH (Lactate Dehydrogenase ) Measure

LDH activity was measured by examining the extent to which substrate pyruvate was reduced to lactate. This reduction is reduced NADH oxidized and shows maximum absorbance at 340 nm. 2.7 ml of 0.1 M potassium phosphate buffer (pH 7.5) was added to the cuvette, 0.1 ml culture medium was added, 0.1 ml of 0.02 M sodium pyruvate was added, and 0.1 ml of NADH (0.2 mg) was added and mixed well. After that, absorbance (340 nm) was measured for 2 minutes.

Within hepatocytes ROS Reactive Oxygen Species

The amount of ROS in hepatocytes was measured using a fluorescent probe, DCF-DA. DCF-DA (2 ', 7'-Dichlorofluorescein diacetate) in the culture solution was treated with 25 μM per well for 15 minutes, and then treated with lactic acid bacteria and fine dust. Intracellular peroxides formed after the reaction were measured for fluorescence at excitation wavelength (485 nm) and emission wavelength (530 nm).

Cytokine Measurements: TNF -alpha, IL-6, IL- 1beta

The cytokine concentration accumulated in the culture medium was isolated and measured by the enzyme-linked immunosorbent assay (ELISA) (ELISA kit, R & D system, USA). 100 μl of the supernatant sample was added to a well plate coated with an antibody against cytokines and allowed to react at room temperature for 2 hours. The supernatant was removed and washed five times or more with a washing buffer prepared by mixing PBS and Tween 20 (Sigma). . After detecting the reaction antibody (antibody) solution was added to react with the antibody, HRP (Horseradish Peroxidase) enzyme combined with avidin was added to react for 15 minutes at room temperature. Thereafter, the reaction was confirmed by changing the color by putting the TMB solution as a substrate for the HRP enzyme. The presence of cytokines in the sample results in a change in color, indicating the presence or absence of cytokines. After stopping the reaction of the HRP enzyme with the TMB substrate by adding a stop solution (H ₂ SO ₄ ), the absorbance (450 nm) was measured using a microplate reader (thermo).

Determination of Oxidative Stress Reductase Activity: SOD, HO-1

In order to measure the activity of HO-1 (Heme Oxygenase) and SOD (Super-oxide Dismutase) among oxidative stress-removing enzymes, after processing samples in hepatocytes, PBS solution was added and the cells were crushed. at g for 15 min. SOD activity in hepatocytes was measured by absorbance (560 nm) using an evaluated with SOD kit (AAT Bioquest). HO-1 activity was measured according to Kutty and Maines methods.

Assessment of liver and kidney toxicity indicators

Hepatocyte damage was confirmed by measuring the amount of ALT and AST released into the serum of experimental animals in vivo. The amount of ALT and AST was measured according to the Reitman-Frankel method. Lactate dehydrogenase (LDH) activity in serum was measured using the Roche Diagnostics LDH kit. Blood urea nitrogen (BUN), a measure of blood damage, was measured using a common commercial assay kit (Asan Pharmaceuticals).

Statistical processing

The results obtained in the above examples were analyzed by one way ANOVA test using Statistical Package for Social Sciences (SPSS, 21, IBM, Armonk, NY, USA), and the significance between samples was two-sided. (Student's two tailed t-test) was compared at the P <0.05 level.

1.Evaluation of Cytotoxicity Efficacy Due to Fine Dust (In vitro)

1-1. 16S- of strain rRNA  result

16s rRNA sequencing results of Lactobacillus fermentum LM1016 (Accession No. KFCC11756P) are shown in SEQ ID NO: 1.

1-2. Cytotoxicity measurement ( MTT  assay, LDH  Activity measurement)

In order to set the cell treatment concentration of lactic acid bacteria in human hepatocyte HepG2 cells, the cytotoxicity was tested after treating the lactic acid bacteria samples for up to 48 hours. Therefore, the cytotoxicity mitigation efficacy due to the fine dust of the lactic acid bacteria sample was measured at a concentration at which no cytotoxicity was observed (1 to 10 μg / ml or less). Cytotoxicity was measured by the activity of free LDH (Table 1).

HepG2 cells were incubated overnight in Serum-free medium and then treated with lactic acid bacteria at a concentration of 0.1-10 ug / ml for 48 hours. Cell activity was analyzed by MTT assay and cytotoxicity was analyzed by LDH (Lactate Dehydrogenase assay). All data were marked by calculating the standard deviation after 3 experiments.

Group Cell viability
(% of control) LHD leakage assay
(fold of control) 48 hr Normal 100.0 ± 4.2 1.00 ± 0.01 LM1001 1ug / ml 101.9 ± 2.8 1.01 ± 0.01 5 ug / ml 101.2 ± 1.4 1.07 ± 0.04 10ug / ml 98.7 ± 3.3 1.16 ± 0.02 LM1009 1ug / ml 100.1 ± 5.4 1.03 ± 0.02 5 ug / ml 101.2 ± 2.1 1.04 ± 0.01 10ug / ml 100.0 ± 3.0 1.10 ± 0.04 LM1011 1ug / ml 101.4 ± 2.0 1.02 ± 0.01 5 ug / ml 101.0 ± 3.2 0.99 ± 0.02 10ug / ml 99.5 ± 4.2 1.07 ± 0.03 LM1012 1ug / ml 101.3 ± 8.7 1.05 ± 0.04 5 ug / ml 100.7 ± 5.1 1.06 ± 0.02 10ug / ml 100.6 ± 8.0 1.07 ± 0.02 LM1014 1ug / ml 102.2 ± 5.3 1.01 ± 0.03 5 ug / ml 102.0 ± 3.3 1.03 ± 0.02 10ug / ml 101.9 ± 2.7 1.07 ± 0.01 LM1016 1ug / ml 101.0 ± 4.6 1.03 ± 0.02 5 ug / ml 100.6 ± 8.4 1.03 ± 0.01 10ug / ml 98.1 ± 5.6 1.10 ± 0.03 LM1019 1ug / ml 101.5 ± 10.4 1.04 ± 0.01 5 ug / ml 103.6 ± 10.2 1.04 ± 0.01 10ug / ml 98.0 ± 4.8 1.13 ± 0.02 LM1060 1ug / ml 100.9 ± 1.8 1.01 ± 0.02 5 ug / ml 99.1 ± 4.5 1.04 ± 0.02 10ug / ml 98.4 ± 3.9 1.11 ± 0.02 LM1061 1ug / ml 99.2 ± 2.5 1.02 ± 0.02 5 ug / ml 98.8 ± 7.6 1.05 ± 0.02 10ug / ml 98.8 ± 5.6 1.13 ± 0.02 LM1062 1ug / ml 100.8 ± 4.9 0.99 ± 0.02 5 ug / ml 100.7 ± 2.9 1.06 ± 0.01 10ug / ml 98.6 ± 3.7 1.12 ± 0.03 LM1063 1ug / ml 101.6 ± 6.7 1.02 ± 0.02 5 ug / ml 100.0 ± 4.1 1.09 ± 0.02 10ug / ml 99.1 ± 4.5 1.15 ± 0.01

To study the cytotoxicity-reducing effects of lactic acid bacteria, lactic acid bacteria (LM1001, LM1009, LM1011, LM1012, LM1016, LM1019, LM1060, LM1061, LM1062, and LM1063) in HepG2 cell lines were subjected to fine dust (diesel exhaust) after 1 hour pretreatment. particulate matter (DEPM) was treated for 24 hours (100 ug, DEPM / ml). After treatment, the survival rate of human hepatocytes (HepG2 cells) was measured by MTT test method and graph. Lactobacillus strains and the normal control (no control) was not represented as NA1, microdust but treated stress control was also represented as NA2.

In comparison between the normal control group (NA1) and the stress control group (NA2) of FIG. 1, the cell activity was reduced by the fine dust (DEPM), and the survival rate of the human hepatocyte (HepG2 cell) decreased by 20%. Compared with the stress control treated with fine dust, human hepatocytes pretreated with LM1001, LM1011, LM1012, LM1016, and LM1061 were found to maintain viability of 91% or more even after fine dust treatment.

In addition, as a result of measuring cytotoxicity, the amount of LDH liberated by cell damage (NA2) increased 1.4 times as compared to the normal control (NA1) when treated with fine dust in human hepatocytes. The amount of LDH released from human liver cell line HepG2 pretreated with LM1001, LM1011, LM1012, LM1016, LM1060, and LM1061 lactic acid bacteria was the same as that of the normal control group.

1-3. Due to fine dust Oxidative  Stress Reduction Efficacy Assessment

Evaluation of oxidative stress reduction efficacy due to fine dust was conducted by using HepG2 cell line through evaluation of the efficacy of ROS (Reactive Oxygen Species) generation and evaluation of SOD (Superoxide Dismutase) or HO-1 (Heme Oxygenase-1) activity. It became.

11 lactic acid bacteria including the lactic acid bacteria strain of the present invention treated with human liver cell line for 1 hour and then treated with fine dust and incubated for 24 hours, the degree of fluorescence generated depending on the concentration of ROS produced (DCF-DA) Intracellular reactive oxygen species (ROS) concentration was measured through.

As a result, it was confirmed that free radicals in human hepatocytes increased by 2.2 times by fine dust, and reduced free radicals in human hepatocytes by LM1001, LM1011, LM1012, LM1016, LM1060, or LM1061 lactic acid bacteria. 3).

After evaluation of SOD and HO-1, HepG2 cells were treated with microdust for 24 hours, and then, the cells were collected to measure the activity of SOD (FIG. 4) and HO-1 (FIG. 5) in the cells. Experimental results, it was confirmed that the fine dust treatment increased the activity of the HO-1 enzyme, and further increased by the treatment of lactic acid bacteria LM1001, LM1012, and LM1016 (Fig. 5). HO-1 enzyme increases as a response to reduce the stress as the stress on the HepG2 cells increases. On the contrary, the activity of SOD enzyme was relatively decreased with the fine dust treatment compared to the normal control (NA1) which did not process the fine dust. Recovery to the state was confirmed to increase (Fig. 4).

1-4. Evaluation of the Effect on Inflammatory Cytokines Caused by Fine Dust

In order to evaluate the effect on inflammatory cytokines (TNF-α, IL-6, IL-1ß, etc.) due to fine dust, lactic acid bacteria in rat splenocytes were treated with 3 ~ 3 hours of diesel exhaust particulate matter (DEPM) after 1 hour pretreatment. 24 hours treatment.

After treatment with fine dust, the amount of cytokines TNF-α, IL-6 and IL-1ß released into the culture medium was measured by ELISA assay. As a result, the microdust treatment increased the production of inflammatory cytokines TNF-α, IL-6 and IL-1ß in human hepatocytes, and the production of TNF-α was decreased by strains of LM1001, LM1012, and LM1016. (FIG. 6A), the production amount of IL-6 was reduced by the strains of LM1001, LM1011, LM1012, LM1014, LM1016, and LM1060 (FIG. 6B). In addition, IL-1ß production was reduced by the strains of LM1001, LM1012, and LM1016 (Fig. 6C).

2. Evaluation of Long-term Tissue Toxicity Efficacy Due to Fine Dust of Lactic Acid Bacteria (in vivo )

2-1. Weighing Weight, Liver, Spleen, and Kidney

In the present application, in order to study the efficacy of long-term tissue toxicity mitigation due to the fine dust of lactic acid bacteria, diesel exhaust particulate matter (100 mg / kg) and lactic acid bacteria (10 mg / kg) were orally administered for 14 days every day. . Body weight, liver, spleen and kidney weights were measured after sample administration. As a result, no change in organs was observed between the group administered with fine dust and the group administered with lactic acid bacteria and fine dust at the same time.

Mice were ingested for 14 days at a concentration of 100 mg, DEPM / kg, body weight and lactic acid bacteria 10 mg / kg, and body weight daily. After 14 days, mice were dissected to determine tissue abnormalities and weight changes, and the following data (n = 6) were displayed by calculating the standard deviation.

In addition, when measuring the weight of each tissue, a significant weight change in weight and each tissue was not observed in the control group treated with each lactic acid compared to the standard group treated with DEPM only without feeding the lactic acid bacteria (Table 2). Sudden weight change in experimental animals is one of the indicators of health abnormalities. In the results of this experiment, the absence of a sharp change in body weight is an indicator that no disease occurs during the experiment and does not affect the experiment.

Group Body weight (g) Liver weight (g) Spleen weight (mg) Kidney weight (mg) Normal 23.0 ± 0.9 1.22 ± 0.05 72.55 ± 0.80 276.6 ± 5.29 DEP (100 mg / kg) 23.1 ± 1.04 1.21 ± 0.05 70.73 ± 1.00 275.9 ± 5.14 DEP + LM1001 (10 mg / kg) 24.3 ± 0.94 1.28 ± 0.18 71.93 ± 1.06 278.4 ± 3.36 DEP + LM1009 (10 mg / kg) 24.9 ± 1.06 1.28 ± 0.20 71.62 ± 1.99 276.3 ± 4.98 DEP + LM1011 (10 mg / kg) 22.7 ± 0.74 1.27 ± 0.17 69.90 ± 1.70 281.4 ± 9.02 DEP + LM1012 (10 mg / kg) 23.3 ± 0.92 1.32 ± 0.17 71.98 ± 1.23 275.4 ± 5.16 DEP + LM1014 (10 mg / kg) 23.1 ± 0.86 1.28 ± 0.16 71.37 ± 1.02 278.4 ± 4.64 DEP + LM1016 (10 mg / kg) 23.4 ± 1.61 1.19 ± 0.07 71.63 ± 1.48 274.1 ± 4.19 DEP + LM1019 (10 mg / kg) 23.6 ± 0.89 1.30 ± 0.16 71.23 ± 2.01 274.5 ± 3.44 DEP + LM1060 (10 mg / kg) 22.6 ± 0.64 1.07 ± 0.31 72.58 ± 0.71 276.5 ± 4.31 DEP + LM1061 (10 mg / kg) 23.1 ± 1.08 1.25 ± 0.20 71.22 ± 0.79 277.7 ± 5.30 DEP + LM1062 (10 mg / kg) 22.5 ± 1.04 1.33 ± 0.17 71.70 ± 0.83 277.6 ± 7.00 DEP + LM1063 (10 mg / kg) 23.3 ± 0.79 1.25 ± 0.07 70.57 ± 2.23 280.2 ± 3.74

2-2. Assessment of hepatic and renal toxicity indicators

Herein, fine dust (DEPM, 100 mg / kg) and lactic acid bacteria (10 mg / kg) were orally administered daily for 14 days for hepatotoxicity indicator and renal toxicity index evaluation ALT, AST, LDH, BUN measurement.

Hepatic toxicity and renal toxicity indices (ALT, AST, LDH, BUN measurements) were measured after sample administration. In the experimental results, ALT, AST and LDH were increased in the group of fine dust, and blood ALT and AST were significantly decreased in the group of lactic acid bacteria LM1001, LM1012 and LM1016 (Fig. 7). It was confirmed that the significantly reduced in the group treated with LA1001, LM1012, LM1016, LM1062 (Fig. 8).

Blood Urea Nitrogen (BUN) value for renal toxicity was not significantly different between normal control (NA1) without microdust treatment and stress control (NA2) with microdust treatment. However, LM1001, LM1012, LM1014, and LM1016 in the control group treated with both fine dust and lactic acid bacterium showed significant decrease within the range of normal control group and stress control group, indicating a decrease in blood BUN production. It is considered that the concentration of BUN increased in the blood by renal function impaired by fine dust decreases the concentration of BUN in the blood and within the concentration range between the normal control group and the stress control group with the reduction of renal damage by the lactic acid bacteria.

The above description of the present application is intended for illustration, and it will be understood by those skilled in the art that the present invention may be easily modified in other specific forms without changing the technical spirit or essential features of the present application. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive. For example, each component described as a single type may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined form.

The scope of the present application is indicated by the following claims rather than the above description, and it should be construed that all changes or modifications derived from the meaning and scope of the claims and their equivalents are included in the scope of the present application.

Korea Microbial Conservation Center (Domestic) KFCC11756P 20171213

<110> JEONNAM BIOINDUSTRY FOUNDATION <120> LACTOBACILLUS FERMENTUM LM1016 AND COMPOSITION FOR PREVENTING OR          TREATING OF INFLAMMATORY DISEASES AND METABOLIC DISEASES          COMPRISING THE SAME <130> DP20180495KR <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1483 <212> RNA <213> Artificial Sequence <220> <223> Lactobacillus fermentum LM1016 16S-rRNA sequence <400> 1 ctcaggatga acgccggcgg tgtgcctaat acatgcaagt cgaacgcgtt ggcccaattg 60 attgatggtg cttgcacctg attgattttg gtcgccaacg agtggcggac gggtgagtaa 120 cacgtaggta acctgcccag aagcggggga caacatttgg aaacagatgc taataccgca 180 taacaacgtt gttcgcatga acaacgctta aaagatggct tctcgctatc acttctggat 240 ggacctgcgg tgcattagct tgttggtggg gtaacggcct accaaggcga tgatgcatag 300 ccgagttgag agactgatcg gccacaatgg gactgagaca cggcccatac tcctacggga 360 ggcagcagta gggaatcttc cacaatgggc gcaagcctga tggagcaaca ccgcgtgagt 420 gaagaagggt ttcggctcgt aaagctctgt tgttaaagaa gaacacgtat gagagtaact 480 gttcatacgt tgacggtatt taaccagaaa gtcacggcta actacgtgcc agcagccgcg 540 gtaatacgta ggtggcaagc gttatccgga tttattgggc gtaaagagag tgcaggcggt 600 tttctaagtc tgatgtgaaa gccttcggct taaccggaga agtgcatcgg aaactggata 660 acttgagtgc agaagagggt agtggaactc catgtgtagc ggtggaatgc gtagatatat 720 ggaagaacac cagtggcgaa ggcggctacc tggtctgcaa ctgacgctga gactcgaaag 780 catgggtagc gaacaggatt agataccctg gtagtccatg ccgtaaacga tgagtgctag 840 gtgttggagg gtttccgccc ttcagtgccg gagctaacgc attaagcact ccgcctgggg 900 agtacgaccg caaggttgaa actcaaagga attgacgggg gcccgcacaa gcggtggagc 960 atgtggttta attcgaagct acgcgaagaa ccttaccagg tcttgacatc ttgcgccaac 1020 cctagagata gggcgtttcc ttcgggaacg caatgacagg tggtgcatgg tcgtcgtcag 1080 ctcgtgtcgt gagatgttgg gttaagtccc gcaacgagcg caacccttgt tactagttgc 1140 cagcattaag ttgggcactc tagtgagact gccggtgaca aaccggagga aggtggggac 1200 gacgtcagat catcatgccc cttatgacct gggctacaca cgtgctacaa tggacggtac 1260 aacgagtcgc gaactcgcga gggcaagcaa atctcttaaa accgttctca gttcggactg 1320 caggctgcaa ctcgcctgca cgaagtcgga atcgctagta atcgcggatc agcatgccgc 1380 ggtggaatac gttcccgggg ccttgtacac accgccccgt cacaccatga gagtttgtaa 1440 cacccaaagt cggtggggta accttttagg agccagccgc cta 1483

Claims (12)

delete delete Comprising the live bacteria, microorganisms, cultures, lysates, extracts, or mixtures of Lactobacillus permanent LM1016 strain (KFCC11756P) as an active ingredient,
A pharmaceutical composition for alleviating, preventing or treating an inflammatory disease or metabolic disease,
The Lactobacillus fermentum LM1016 strain (KFCC11756P) is to reduce the toxicity of cytotoxicity or organ tissues caused by fine dust,
The inflammatory diseases include enteritis, gastritis, hepatitis, bronchitis, pleurisy, arthritis, rhinitis, keratitis, nephritis, peritonitis, spondylitis, pancreatitis, inflammatory pain, urethritis, cystitis, burn inflammation, dermatitis, periodontitis, gingivitis, degenerative neuritis, inflammatory pain, And combinations thereof, one or more selected from the group consisting of
The metabolic disease is one or more selected from the group consisting of type 1 diabetes, type 2 diabetes, asthma, tuberculosis, hypertension, obesity, coronary atherosclerosis, arteriosclerosis, and combinations thereof,
Pharmaceutical compositions for alleviating, preventing or treating inflammatory or metabolic diseases.
delete The method of claim 3, wherein
The inflammatory disease or metabolic disease is caused by cytotoxicity due to fine dust, pharmaceutical composition for alleviating, preventing or treating inflammatory disease or metabolic disease.
delete delete Comprising the live bacteria, microorganisms, cultures, lysates, extracts, or mixtures of Lactobacillus permanent LM1016 strain (KFCC11756P) as an active ingredient,
As a food composition for preventing or alleviating an inflammatory disease or metabolic disease,
The Lactobacillus permanent LM1016 strain (KFCC11756P) is to reduce the toxicity of cytotoxicity or organ tissues due to fine dust,
Food composition for the prevention or alleviation of inflammatory diseases or metabolic diseases.
delete The method of claim 8,
The inflammatory disease or metabolic disease is caused by cytotoxicity due to fine dust, food composition for the prevention or alleviation of inflammatory disease or metabolic disease.
The method of claim 10,
The inflammatory diseases include enteritis, gastritis, hepatitis, bronchitis, pleurisy, arthritis, rhinitis, keratitis, nephritis, peritonitis, spondylitis, pancreatitis, inflammatory pain, urethritis, cystitis, burn inflammation, dermatitis, periodontitis, gingivitis, degenerative neuritis, inflammatory pain, And one or more selected from the group consisting of combinations thereof,
Food composition for the prevention or alleviation of inflammatory diseases or metabolic diseases.
The method of claim 10,
The metabolic disease is one or more selected from the group consisting of type 1 diabetes, type 2 diabetes, asthma, tuberculosis, hypertension, obesity, coronary atherosclerosis, arteriosclerosis, and combinations thereof,
Food composition for the prevention or alleviation of inflammatory diseases or metabolic diseases.

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