Composition for suppressing mitochondrial dysfunction

Abstract

To provide a novel composition that inhibits a deterioration of mitochondrial functions and treats diseases involving a deterioration of mitochondrial functions.SOLUTION: The present invention discloses a composition for inhibiting deterioration of mitochondrial functions, containing lactic acid bacteria belonging to Lactobacillus paracasei as an active ingredient, and a composition for treating, preventing or ameliorating diseases involving mitochondrial functions. Herein, the mitochondrial functions can be evaluated by using, as indexes, film potential, oxidative stress, oxygen consumption rate, basic respiration, ATP metabolic turnover and forms. The inventive composition can be provided as a food composition or a supplement.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a composition for suppressing mitochondrial dysfunction. The present invention also relates to a composition for treating, preventing or ameliorating a disease accompanied by mitochondrial dysfunction. [Background technology]

[0002] Mitochondria are intracellular organelles that produce most of the adenosine triphosphate (ATP) in eukaryotic cells, and are also called the respiratory apparatus of the cell because they synthesize ATP while consuming oxygen vigorously. Mitochondria also have various functions, such as regulating intracellular calcium concentration and controlling apoptosis, and are essential organelles for cell survival. In normal cells, mitochondria maintain homeostasis by constantly repeating fusion and fission. It has been reported that mitochondrial dysfunction is one of the typical characteristics of neurological disorders and is a major risk factor for the onset and progression of neurodegenerative disorders (Non-Patent Documents 1 and 2).

[0003] To date, there have been proposals for a therapeutic agent for neurodegenerative diseases that contains a lactic acid bacteria fermentation product as an active ingredient (Patent Document 1), and the use of a novel lactic acid bacterium in a pharmaceutical for treating neurodegenerative diseases (Patent Document 2). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2017-222601 A [Patent Document 2] Special Publication No. 2020-505031 [Non-patent literature]

[0005] [Non-Patent Document 1] Sarkar S et al., NPJ Parkinsons Dis., 3:30 (2017) [Non-Patent Document 2] Yuanbo Wu et. Al., Mitochondrion, 49:35-45 (2019) Summary of the Invention [Problem to be solved by the invention]

[0006] An object of the present invention is to provide a novel composition for use in suppressing mitochondrial dysfunction, and a novel composition for use in the treatment, prevention, or amelioration of diseases accompanied by mitochondrial dysfunction. [Means for solving the problem]

[0007] The present inventors have now added Lactobacillus paracasei to a mouse-derived macrophage cell line, induced inflammation in the cells, and evaluated the membrane potential, oxidative stress, and cellular respiration of mitochondria in the cells. They found that the decrease in membrane potential, the increase in oxidative stress, and the decrease in cellular respiration were suppressed compared to a group in which Lactobacillus paracasei was not added. The present inventors also added Lactobacillus paracasei and evaluated the morphology of mitochondria in a mouse-derived macrophage cell line in which inflammation was induced. They found that the abnormality in mitochondrial morphology was improved compared to a group in which Lactobacillus paracasei was not added. The present invention is based on these findings.

[0008] According to the present invention, the following inventions are provided. [1] Lactobacillus paracasei ( Lactobacillus paracasei A composition for suppressing mitochondrial dysfunction and an agent for suppressing mitochondrial dysfunction, comprising lactic acid bacteria belonging to the strain Lactobacillus subtilis L.) as an active ingredient. [2] The composition and agent described in [1] above, wherein the function of mitochondria is cellular respiration and / or ATP production. [3] The composition and agent described in [1] or [2] above, wherein the mitochondrial function is indicated by one or more indicators selected from the group consisting of membrane potential, oxidative stress, oxygen consumption rate, basal respiration, ATP metabolic turnover and morphology. [4] The composition and agent according to any one of [1] to [3] above, wherein the mitochondria are mitochondria in one or more types of cells selected from the group consisting of macrophages, dendritic cells, lymphocytes and microglia. [5] Lactobacillus paracasei ( Lactobacillus paracasei A composition for treating, preventing and / or ameliorating diseases accompanied by decreased mitochondrial function, and an agent for treating, preventing and / or ameliorating diseases accompanied by decreased mitochondrial function, comprising lactic acid bacteria belonging to the strain Lactobacillus subtilis (Lactobacillus casei) as an active ingredient. [6] The composition and agent described in [5] above, wherein the disease associated with impaired mitochondrial function is a neurodegenerative disease. [7] The composition or agent according to any one of [1] to [6] above, wherein the Lactobacillus paracasei is Lactobacillus paracasei KW3110 strain (FERM BP-08634). [8] The composition or agent according to any one of [1] to [7] above, which is a food composition or a pharmaceutical composition. [9] The composition and agent described in [8] above, wherein the food is a beverage, a dairy product, a fermented food, a health food, a functional food, a nutritional supplement, a supplement, a health functional food or a food for the sick.

[10] Lactobacillus paracasei ( Lactobacillus paracasei A composition for reducing the risk of the onset and progression of mitochondrial dysfunction and an agent for reducing the risk of the onset and progression of mitochondrial dysfunction, comprising lactic acid bacteria belonging to the strain Lactobacillus subtilis (Lactobacillus casei) as an active ingredient.

[11] Lactobacillus paracasei ( Lactobacillus paracasei A composition for reducing the risk of developing a disease caused by decreased mitochondrial function and an agent for reducing the risk of developing a disease caused by decreased mitochondrial function, comprising lactic acid bacteria belonging to the strain Lactobacillus subtilis (Lactobacillus casei) as an active ingredient.

[0009] The above compositions and agents [1] to

[11] may be referred to herein as "the composition of the present invention" and "the agent of the present invention", respectively.

[0010] The compositions and agents of the present invention are advantageous in that they can be used as functional foods that are useful for suppressing mitochondrial dysfunction and treating neurodegenerative diseases, and can also be used as functional foods that are safe for mammals, including humans. [Brief description of the drawings]

[0011] [Figure 1] FIG. 1 shows the results of evaluating the effect of the KW3110 strain on the mitochondrial membrane potential of a mouse-derived macrophage cell line (Example 1). FIG. 1A shows plots of each cell in the control group (control), LPS-treated group (LPS), and LPS-treated + KW3110 group (LPS + KW3110) in which the fluorescence intensity of MitoTracker (registered trademark) Red CMXRos is plotted on the vertical axis (MitoTracker Red) and the fluorescence intensity of MitoTracker (registered trademark) Green FM is plotted on the horizontal axis (MitoTracker Green). The cell population surrounded by the broken line indicates cells containing mitochondria with relatively low membrane potential. FIG. 1B is a graph showing the ratio of cells containing mitochondria with low membrane potential (cells surrounded by the broken line in FIG. 1A) to the total cells in the control group, LPS-treated group, and LPS-treated + KW3110 group. * indicates significant difference (p<0.05), ** indicates significant difference (p<0.01). [Diagram 2] FIG. 2 shows the results of evaluating the effect of the KW3110 strain on oxidative stress in mitochondria of a mouse-derived macrophage cell line (Example 2). FIG. 2A shows plots of MitoSOX® Red fluorescence intensity on the vertical axis (MitoSOX) and MitoTracker® Green FM fluorescence intensity on the horizontal axis (MitoTracker Green) in each of the control group (control), LPS-treated group (LPS), and LPS-treated + KW3110 group (LPS + KW3110). The cell population surrounded by a broken line indicates cells with a relatively high level of oxidative stress in mitochondria. FIG. 2B is a graph showing the ratio of cells with a high level of oxidative stress to the total cells. ** indicates significant difference (p < 0.01). [Diagram 3]Figure 3 shows the results of evaluating the effects of the KW3110 strain on changes in oxygen consumption rate, basal respiration, and ATP metabolic turnover in a mouse-derived macrophage cell line in a control group (control), an LPS-treated group (LPS), and an LPS-treated + KW3110 group (LPS + KW3110) (Example 3). Figure 3A shows the effect on changes in oxygen consumption rate, Figure 3B shows the effect on basal respiration, and Figure 3C shows the effect on ATP metabolic turnover. In each graph, the vertical axis shows the relative value when the average oxygen consumption rate in stage I in the control group is set to 1. ** indicates a significant difference (p < 0.01). [Figure 4] Figure 4 shows the results of evaluating the effect of the KW3110 strain on mitochondrial morphology in a mouse-derived macrophage cell line in a control group (control), an LPS-treated group (LPS), and an LPS-treated + KW3110 group (LPS + KW3110) (Example 4). The white line at the bottom right of each figure indicates the scale bar (total length 400 nm). Description of the Invention

[0012] The lactic acid bacteria used as an active ingredient in the present invention is Lactobacillus paracasei ( Lactobacillus paracasei ), preferably Lactobacillus paracasei KW3110 strain ( Lactobacillus paracasei , FERM BP-08634) (hereinafter, simply referred to as "KW3110 strain") can be used.

[0013] The lactic acid bacteria used in the present invention may be a strain equivalent to the Lactobacillus paracasei KW3110 strain. Here, the equivalent strain refers to a strain derived from the Lactobacillus paracasei KW3110 strain, a strain from which the Lactobacillus paracasei KW3110 strain is derived, or a descendant strain of the strain.

[0014] The above Lactobacillus paracasei KW3110 strain has been deposited at the National Institute of Technology and Evaluation, Biotechnology Center, Patent Organism Depositary (NITE-IPOD), an international depositary authority under the Budapest Treaty for the Deposit of Patent Microorganisms (Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818, Japan) under the accession number FERM BP-08634 (Deposit date: February 20, 2004). A derivative strain of Lactobacillus paracasei KW3110 has been deposited at the same Patent Organism Depositary under the accession number FERM BP-08635 (Deposit date: February 20, 2004).

[0015] The composition and agent of the present invention preferably contain Lactobacillus paracasei cells or a processed product thereof, or both, as an active ingredient. Lactobacillus paracasei cells may be in an isolated form or in a culture form.

[0016] The live bacteria of Lactobacillus paracasei can be prepared by culturing in a medium. The culture of Lactobacillus paracasei can be performed by a known method using a known medium. As the medium, MRS medium, GAM medium, or LM17 medium can be used, and inorganic salts, vitamins, amino acids, antibiotics, serum, etc. may be added as appropriate. The culture may be performed at 25 to 40°C for several hours to several days. After the culture, the Lactobacillus paracasei cells are collected by centrifugation or filtration.

[0017] Examples of the processed Lactobacillus paracasei cells include disrupted live or dead cells (e.g., ultrasonic disrupted), dried live or dead cells (e.g., freeze-dried), disrupted dried products, enzyme-treated live or dead cells, etc. The dead cells can be obtained by, for example, heat treatment, treatment with drugs such as antibiotics, treatment with chemicals such as formalin, treatment with ultraviolet light, or treatment with radiation such as gamma rays. The enzyme-treated products also include products in which the cell walls of lactic acid bacteria are removed by enzymes or mechanical means. Furthermore, nucleic acid-containing fractions (e.g., DNA, RNA) of Lactobacillus paracasei are also included in the processed Lactobacillus paracasei cells, and can be obtained by dissolving Lactobacillus paracasei cells with a surfactant or the like and then precipitating with ethanol or the like. In this specification, the term "lactic acid bacteria of the present invention" is used to include Lactobacillus paracasei cells and their processed products.

[0018] The compositions and agents of the present invention can be provided as the lactic acid bacteria of the present invention alone, or as a mixture of the lactic acid bacteria of the present invention with other components (e.g., food ingredients, food additives, supplement ingredients, pharmaceutical additives). The amount of the lactic acid bacteria of the present invention in the compositions and agents of the present invention can be determined arbitrarily depending on the purpose, application, form, dosage form, symptoms, body weight, etc., and the present invention is not limited thereto, but the content can be 0.01 to 90% by mass, more preferably 0.1 to 50% by mass, based on the total amount.

[0019] According to a first aspect of the present invention, there are provided a composition for suppressing mitochondrial dysfunction, which contains Lactobacillus paracasei as an active ingredient (hereinafter, may be referred to as the "suppressive composition of the present invention"), and an inhibitor of mitochondrial dysfunction, which contains Lactobacillus paracasei as an active ingredient (hereinafter, may be referred to as the "inhibitor of the present invention").

[0020] Mitochondria are eukaryotic organelles with a double membrane structure consisting of an outer membrane and an inner membrane. In normal cells, mitochondria maintain homeostasis by constantly repeating fusion and fission. The main function of mitochondria in cells is the production of ATP using the potential difference between the outer and inner membranes. ATP production uses oxygen taken in by the cell, and carbon dioxide is produced in the process, and mitochondria have a respiratory function in cells. The main parts that perform the functions of mitochondria are the matrix space and the inner membrane. The inner membrane is folded to form cristae (pleated structures), and this structure increases the surface area of ​​the inner membrane, which is particularly important for mitochondrial function (cellular respiration and ATP production).

[0021] Mitochondrial dysfunction and suppression of mitochondrial dysfunction can be evaluated by known in vitro or in vivo direct or indirect measurement methods, such as measuring mitochondrial membrane potential, oxidative stress, oxygen consumption rate, basal respiration and ATP metabolic turnover, or observing mitochondrial morphology or structure. Mitochondrial dysfunction and suppression of mitochondrial dysfunction can also be evaluated using the amount of intracellular ATP, electron transport chain enzyme complexes or respiratory chain complexes, damage, mutation or copy number of mitochondrial DNA (mtDNA), mitophagy, metal ion metabolism, etc.

[0022] As described in the Examples below, the morphology of mitochondria can be evaluated by observation with an electron microscope, and in this case, the morphology or structure can be evaluated as normal or abnormal using the characteristic cristae structure of mitochondria and the size of mitochondria as indicators. The structure of mitochondria can also be evaluated by measuring the balance between mitochondrial fusion and fission by measuring the rate of mitochondrial fragmentation. For example, mitochondria can be specifically stained using fluorescent reagents such as MitoTracker (registered trademark), CellLight (registered trademark), or fluorescent conjugates of mouse anti-Complex V d-subunit monoclonal antibodies (all from ThermoFisher Scientific), Mito Bright LT (Dojindo Laboratories), and the like, and the length, area, or fluorescence intensity of the resulting stained image can be measured for evaluation.

[0023] The mitochondrial membrane potential can be evaluated by measuring the potential difference between the outer and inner mitochondrial membranes used for ATP production. The membrane potential can be measured using a known reagent for detecting the mitochondrial membrane potential. As the reagent, for example, MitoTracker (registered trademark) Red CMXRos (ThermoFisher Scientific), MitoTracker (registered trademark) Green FM (ThermoFisher Scientific), and JC-1 (Dojindo Laboratories) can be used. As a detection method, a fluorescence microscope (BZ-X; Keyence), a high content confocal imaging system (Operetta CLS; Perkinelmer, Cell Voyger CV8000; Yokogawa Electric), flow cytometry (FACS (registered trademark); BD Bioscience), etc. can be used.

[0024] Oxidative stress in mitochondria can be measured using reagents that directly detect known causative substances such as superoxide, peroxyl radical, hydrogen peroxide, hydroxyl radical, or peroxynitrite, or reagents that detect substances resulting from causative substances such as oxidized / reduced glutathione, nitric oxide, or lipid peroxidation compounds. Examples of the reagent include fluorescent reagents and colorimetric reagents, such as MitoSOX (registered trademark) Red, CellROX (registered trademark) Reagents / Kit, Image-iT (registered trademark) Lipid Peroxidation Kit, Dihydroethidium, DAF-FM Diacetate, Premo (registered trademark) Cellular Hydrogen Peroxide (H2O2) Sensor, ThiolTracker (registered trademark) Violet, Monochloro-bimane, Monobromo-bimane, Premo (registered trademark) Cellular Redox Sensor, Grx-1-roGFP (all ThermoFisher Scientific), Cell Meter (registered trademark) Intracellular Total ROS Fluorescence Measurement Kit, Total ROS / Superoxide Detection Kit, Amplite (registered trademark) Intracellular Hydrogen Peroxide Fluorescence Measurement Kit, Amplite (registered trademark) Fluorescent Hydrogen Peroxide Assay Kit, Cell Meter (registered trademark) Intracellular Fluorescent Hydrogen Peroxide Assay Kit, Hydrogen Peroxide Detection Kit, and CELLestial (registered trademark) Red. Hydrogen peroxide quantification kit, OxiSelect (registered trademark) hydrogen peroxide / peroxidase assay kit, OxiSelect (registered trademark) TBARS assay, OxiSelect (registered trademark) protein radical measurement ELISA kit, nitric oxide (NO) detection kit (all from Cosmo Bio Co., Ltd.), ROS-Glo (registered trademark) H2O2 Assay, GSH / GSSG-Glo (registered trademark) Assay, GSH-Glo (registered trademark) Glutathione Assay or Griess Reagent System (all from Promega), MitoPeDPP, Si-DMA (all from Dojindo Laboratories) can be used.

[0025] The oxygen consumption rate or oxygen consumption amount can be evaluated by a measurement method using a known extracellular flux analyzer or a measurement method using a reagent such as an extracellular O2 center. As an extracellular flux analyzer, for example, XFe24 / XFe96 (Agilent Technologies) can be used. As a reagent, for example, Oxygen Consumption Rate Assay Kit / MitoXpress-Xtra (Funakoshi), Mito-ID (registered trademark) Extracellular O2 Center Kit (Cosmo Bio) or Extracellular Oxygen Consumption Assay (Abcam) can be used. Basal respiration and ATP metabolic turnover can be calculated based on the average oxygen consumption rate as described in Example 3.

[0026] The amount of intracellular ATP can be evaluated by measuring it by known chemiluminescence methods, high performance liquid chromatography (HPLC) methods, or ATP imaging using a fluorescent probe. Examples of chemiluminescence methods include a method in which luciferin and ATP are converted to oxyluciferin by luciferase and the light emitted is measured. Examples of reagents that can be used include Luminescent ATP Detection Assay Kit (Abcam), Intracellular ATP Measurement Kit Ver. 2, Rhodamine 123 (Cosmo Bio), and Mitochondrial ToxGlo (registered trademark) Assay (Promega).

[0027] The electron transport enzyme complex or respiratory chain complex can be evaluated by measuring the enzyme activity of complexes involved in ATP production present in mitochondria, such as Complex I (NADH oxidase / Co-enzyme Q reductase), Complex II (succinate / Co-enzyme Q reductase), Complex III (CoQ cytochrome C oxidoreductase), Complex IV (cytochrome C oxidase), or Complex V (F1FoATP synthase). As a reagent, for example, MitoCheck Complex Activity Assay Kit (Funakoshi Co., Ltd.) can be used.

[0028] Damage, mutation, or copy number of mitochondrial DNA (mtDNA) can be evaluated by a known DNA measurement method, such as real-time PCR. As a reagent, for example, a mitochondrial DNA damage analysis kit (Cosmo Bio Co., Ltd.) can be used.

[0029] Mitophagy can be evaluated by using a fluorescent reagent that accumulates in mitochondria, fuses with lysosomes, and develops color when placed under acidic conditions. Examples of the reagent that can be used include Mitophagy Detection Kit (Dojindo Laboratories), Keima-Red (MBL Life Sciences), etc. Detection methods that can be used include a fluorescent microscope (BZ-X; Keyence), a high-content confocal imaging system (Operetta CLS; Perkinelmer, Cell Voyger CV8000; Yokogawa Electric), and flow cytometry (FACS (registered trademark); BD Bioscience).

[0030] Metal ion metabolism can be evaluated by quantifying calcium ions, iron (II) ions, etc. taken up into mitochondria or cells. Reagents that can be used include Rhod 2-AM, Mito-FerroGreen, Calcium Kit - Fluo 4, Fura 2 (all from Dojindo Laboratories), Calcium Assay Kit (Colorimetric) (Abcam), etc.

[0031] It is known that mitochondrial dysfunction is caused by pathogens such as bacteria, viruses, and parasites; diseases such as neurodegenerative diseases; aging; and stresses such as environmental irritants. Therefore, in the present invention, the term "mitochondrial dysfunction" is used to mean dysfunction caused by the above-mentioned stresses. Examples of bacteria include, but are not limited to, gram-negative bacteria, and may be the whole or part of the bacterial body, and examples of parts of the bacterial body include lipopolysaccharide (LPS).

[0032] In the present invention, the term "suppression of mitochondrial dysfunction" is used to mean not only suppressing the progression of mitochondrial dysfunction, but also maintaining or improving mitochondrial function.

[0033] In the present invention, "mitochondria" refers to mitochondria in cells such as macrophages, dendritic cells, lymphocytes, and microglia.

[0034] The suppressive composition and the suppressor of the present invention can be ingested or administered to a subject with mitochondrial dysfunction. The subject can be, for example, an elderly person aged 60 years or older, 65 years or older, or 70 years or older. In addition, the degree of mitochondrial dysfunction in the subject can be evaluated before the ingestion or administration of the suppressive composition and the suppressor of the present invention. The evaluation of the degree of mitochondrial dysfunction can be carried out according to the above-mentioned method. In the present invention, the mitochondrial dysfunction can be mitochondrial dysfunction in macrophages.

[0035] According to a second aspect of the present invention, there are provided a composition for treating, preventing or ameliorating a neurodegenerative disease, which contains Lactobacillus paracasei as an active ingredient (hereinafter, may be referred to as "the therapeutic composition of the present invention"), and an agent for treating, preventing or ameliorating a neurodegenerative disease, which contains Lactobacillus paracasei as an active ingredient (hereinafter, may be referred to as "the therapeutic agent of the present invention").

[0036] As described in the Examples below, Lactobacillus paracasei has the effect of suppressing mitochondrial dysfunction in macrophage cells. Here, it is known that mitochondrial dysfunction in microglia, which is an immune cell in the brain and also has macrophage-like functions, causes brain inflammation, which leads to the onset or progression of neurodegenerative diseases and the like. That is, it has been reported that mitochondrial dysfunction is one of the typical characteristics of neurodegenerative diseases and is a major risk factor for the onset and progression of neurodegenerative diseases and the like (Non-Patent Documents 1 and 2). Therefore, neurodegenerative diseases can be treated, etc. by suppressing mitochondrial dysfunction. In addition to neurodegenerative diseases, it is known that mitochondrial dysfunction causes mitochondrial diseases. Mitochondrial diseases are a general term for diseases caused by mitochondrial dysfunction, and can be treated, etc. by suppressing mitochondrial dysfunction. Therefore, the lactic acid bacteria of the present invention can be used as an active ingredient for the treatment, prevention, or improvement of diseases accompanied by mitochondrial dysfunction.

[0037] In the present invention, "diseases accompanied by mitochondrial dysfunction" include neurodegenerative diseases, mitochondrial diseases, etc. Neurodegenerative diseases include, for example, Alzheimer's disease, Parkinson's disease, and dementia with Lewy bodies. Mitochondrial diseases include, for example, Leber's hereditary optic neuropathy, Leigh's syndrome, and mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episode syndrome.

[0038] In the present invention, "treatment of a disease accompanied by decreased mitochondrial function" is used to mean not only suppressing the progression of the disease or its symptoms, but also alleviating or recovering from the disease or its symptoms.

[0039] The therapeutic composition and therapeutic agent of the present invention can be ingested or administered to a subject with a disease accompanied by mitochondrial dysfunction. The subject can be, for example, an elderly person aged 60 years or older, 65 years or older, or 70 years or older. In addition, the degree of mitochondrial dysfunction in the subject can be evaluated before the ingestion or administration of the therapeutic composition and therapeutic agent of the present invention. The degree of mitochondrial dysfunction can be evaluated according to the above-mentioned method. In the present invention, the mitochondrial dysfunction can be mitochondrial dysfunction in macrophages.

[0040] As shown in the examples below, the lactic acid bacteria of the present invention have the effect of suppressing mitochondrial dysfunction. Therefore, the composition and agent of the present invention can be ingested or administered to a subject at risk of developing and progressing mitochondrial dysfunction or a subject at risk of developing a disease caused by mitochondrial dysfunction, thereby reducing the risk of developing and progressing mitochondrial dysfunction or the risk of developing a disease caused by mitochondrial dysfunction. Here, "subjects at risk of developing and progressing mitochondrial dysfunction" and "subjects at risk of developing a disease caused by mitochondrial dysfunction" refer to subjects who do not have subjective symptoms of developing and progressing mitochondrial dysfunction or a disease caused by mitochondrial dysfunction, but who may develop and progress mitochondrial dysfunction in the future, or who may develop a disease caused by mitochondrial dysfunction, such as elderly subjects aged 60 years or older, 65 years or older, or 70 years or older. In addition, "reducing the risk of developing and progressing mitochondrial dysfunction" means that the probability of developing and progressing mitochondrial dysfunction is reduced, and "reducing the risk of developing a disease caused by mitochondrial dysfunction" means that the probability of developing a disease caused by mitochondrial dysfunction is reduced. That is, according to the third aspect of the present invention, there is provided a composition for reducing the risk of onset and progression of mitochondrial dysfunction and an agent for reducing the risk of onset and progression of mitochondrial dysfunction, comprising the lactic acid bacteria of the present invention as an active ingredient. According to the third aspect of the present invention, there is also provided a composition for reducing the risk of onset of diseases caused by mitochondrial dysfunction and an agent for reducing the risk of onset of diseases caused by mitochondrial dysfunction, comprising the lactic acid bacteria of the present invention as an active ingredient. The composition and agent of the third aspect of the present invention can be carried out according to the description of the first and second aspects of the present invention.

[0041] The compositions and agents of the present invention can be provided in the form of pharmaceuticals (e.g., pharmaceutical compositions), quasi-drugs, foods (e.g., food compositions), feeds (including pet foods), etc., and can be implemented as described below.

[0042] The active ingredient of the present invention can be orally administered to humans and non-human animals. Oral preparations include granules, powders, tablets (including sugar-coated tablets), pills, capsules, syrups, emulsions, and suspensions. These preparations can be formulated using pharmaceutically acceptable carriers by methods commonly used in the art. Pharmaceutically acceptable carriers include excipients, binders, diluents, additives, flavors, buffers, thickeners, colorants, stabilizers, emulsifiers, dispersants, suspending agents, and preservatives.

[0043] The active ingredient of the present invention can be orally ingested by humans and non-human animals, and a typical form of ingestion is food. When the active ingredient of the present invention is provided as a food, the active ingredient can be provided as it is or can be provided by being contained in the food. The food provided in this way is a food containing an effective amount of the active ingredient of the present invention. In this specification, "containing an effective amount" of the active ingredient of the present invention refers to a content such that the active ingredient of the present invention is ingested in a range described below when the amount normally consumed in each food is ingested. In addition, "food" is used to mean health food, functional food, nutritional supplement, health functional food (e.g., specific health food, nutritional functional food, functional food), special purpose food (e.g., food for infants, food for pregnant women, food for sick people) and supplements. In addition, health functional food includes food labeled with disease risk reduction. It goes without saying that when the active ingredient of the present invention is ingested by animals other than humans, the food referred to in the present invention is used as feed.

[0044] Since the active ingredient of the present invention has the above-mentioned effect of suppressing mitochondrial dysfunction and the effect of treating diseases accompanied by mitochondrial dysfunction, it can be contained in foods that are taken daily, or can be provided as a supplement. In this case, the composition and agent of the present invention can be provided in a unit package form in which the amount to be taken per meal is predetermined. Examples of the unit package form per meal include a form in which a certain amount is specified by pack, package, can, bottle, etc. In order to better exert the various actions of the composition and agent of the present invention, the intake amount per meal can be determined according to the daily intake amount of the active ingredient of the present invention, which will be described later. The food of the present invention may be provided with an explanatory matter regarding the intake amount displayed on the package, or with a document or the like in which the explanatory matter is described.

[0045] The predetermined intake amount per meal in the unit package form may be the effective intake amount per day, or the effective intake amount per day divided into two or more times (preferably two or three times). Therefore, the unit package form of the composition and agent of the present invention may contain the active ingredient of the present invention at the daily intake amount per human described below, or may contain the active ingredient of the present invention at an amount of one-half to one-sixth of the daily intake amount per human described below. For convenience of intake, the composition and agent of the present invention are preferably provided in a unit package form per meal (i.e., a unit package form per day) in which the intake amount per meal is the effective intake amount per day.

[0046] The form of the "food" is not particularly limited, and may be, for example, a beverage, a semi-liquid or gel form, or a solid or powder form. In addition, examples of the "supplement" include tablets produced by adding excipients, binders, etc. to the active ingredient of the present invention, kneading the mixture, and then tableting; granules and orally disintegrating tablets produced by adding excipients, binders, etc. to the active ingredient of the present invention and granulating the mixture; and capsules in which the active ingredient of the present invention is enclosed in a capsule or the like. When providing the supplement, in addition to the above-mentioned unit packaging form per meal or per day, it is also suitable to provide the supplement in a unit packaging form per week, per two weeks, per month, or per two months. In addition, it is desirable to display the intake amount per meal or per day in the latter unit packaging form, so that the user can take an effective amount of the active ingredient of the present invention according to the display.

[0047] The foods provided by the present invention are not particularly limited as long as they contain the active ingredient of the present invention, and examples thereof include non-alcoholic beverages such as soft drinks, carbonated drinks, beverages containing fruit juice, beverages containing vegetable juice, beverages containing fruit juice and vegetable juice, livestock milk such as cow's milk, soy milk, dairy beverages, drink-type yogurt, drink-type or stick-type jellies, coffee, cocoa, tea beverages, nutritional drinks, energy drinks, sports drinks, mineral water, near-water, and non-alcoholic beer-flavored beverages; carbohydrate-containing foods and beverages such as rice, noodles, bread, and pasta; dairy products such as cheese, hard or soft yogurt, fresh cream made from livestock milk or other fats and oils, and ice cream. Examples of the food include Western confectionery such as cookies, cakes, and chocolates, Japanese confectionery such as manju and yokan, tablet confectionery (refreshing confectionery) such as ramune, candies, gums, gummy candies, frozen confectionery such as jelly and pudding, and snacks; alcoholic beverages such as whiskey, bourbon, spirits, liqueur, wine, fruit wine, sake, Chinese wine, shochu, beer, non-alcoholic beer with an alcohol content of 1% or less, sparkling wine, other miscellaneous alcohol, and chuhai; processed foods using eggs, processed foods (including delicacies) made from seafood and meat (including liver and other organ meats), and soups such as miso soup; seasonings such as miso, soy sauce, furikake, and other seasoning seasonings; and liquid foods such as thick liquid foods. Mineral water includes both sparkling and non-sparkling mineral water. The food provided by the present invention also includes both food manufacturing raw materials and food additives.

[0048] Tea beverages include all of fermented, semi-fermented, and non-fermented teas, such as black tea, green tea, barley tea, brown rice tea, sencha, gyokuro tea, roasted green tea, oolong tea, turmeric tea, pu-erh tea, rooibos tea, rose tea, chrysanthemum tea, ginkgo leaf tea, and herbal teas (e.g., mint tea, jasmine tea).

[0049] Examples of fruits used in fruit juice beverages and beverages containing fruit and vegetable juice include apples, mandarin oranges, grapes, bananas, pears, peaches, mangoes, acai, blueberries, and plums, while examples of vegetables used in vegetable juice beverages and beverages containing fruit and vegetable juice include tomatoes, carrots, celery, pumpkins, cucumbers, and watermelons.

[0050] The intake amount of the active ingredient of the present invention can be determined depending on the recipient's sex, age, weight, symptoms, intake time, dosage form, intake route, and drugs to be combined, etc. The daily intake amount of the active ingredient of the present invention for adults can be specified, for example, by the number of bacteria, and the lower limit is 1 × 10 7 pieces, 1×10 8 pieces, 1×10 9 The maximum number of pieces is 1×10 11 pieces, 1×10 12 pieces, 1×10 13 These upper and lower limits can be arbitrarily combined, and the range of intake can be, for example, 1×10 7 ~1×10 13 pieces, 1×10 8 ~1×10 12 pieces, 1×10 9 ~1×10 11 The number of lactic acid bacteria can be measured by, for example, a fluorescent staining method, flow cytometry, or a culture method.

[0051] The daily intake of the active ingredient of the present invention for an adult can also be specified by the dry cell mass, and the lower limit can be 1 mg, 10 mg, or 25 mg, and the upper limit can be 1000 mg, 500 mg, or 100 mg. These upper and lower limits can be arbitrarily combined, and the range of the intake can be, for example, 1 to 1000 mg, 10 to 500 mg, or 25 to 100 mg.

[0052] The above-mentioned intake amount of the active ingredient of the present invention and the following intake timing and intake period are applicable to both non-therapeutic and therapeutic uses of the active ingredient of the present invention, and in the case of therapeutic use, intake can be read as administration. The composition and agent of the present invention can also be administered to mammals other than humans (e.g., cows, horses, sheep, pigs, dogs, cats, etc.), and the intake amount, intake timing and intake period can be determined with reference to the description for humans.

[0053] It is preferable to continue taking the active ingredient of the present invention during the period during which the effect of suppressing mitochondrial dysfunction or the effect of treating diseases accompanied by mitochondrial dysfunction is expected. From the viewpoint of better exerting the effect of suppressing mitochondrial dysfunction or the effect of treating diseases accompanied by mitochondrial dysfunction, the period of taking the active ingredient of the present invention can be, for example, one week or more, two weeks or more, three weeks or more, preferably one month or more (four weeks or more) at the above-mentioned daily dose. The interval of taking the active ingredient of the present invention can be once every three days, once every two days, or once a day at the above-mentioned daily dose, and is preferably once a day.

[0054] The composition and agent, food and medicine of the present invention may be labeled with information on the effect of suppressing mitochondrial dysfunction and the effect of treating diseases accompanied by mitochondrial dysfunction. In this case, the information can be easily understood by consumers. Examples of information on these foods and drinks include, but are not limited to, all or part of the following: · To sustain, enhance, improve, enhance, support, or maintain memory · Sustain, enhance, enhance, improve, enhance, support, or maintain cognitive function Sustain, enhance, improve, enhance, support, and maintain attention · To sustain, increase, enhance, improve, enhance, support, or maintain concentration Helps maintain memory and prevents memory decline Prevents carelessness and forgetfulness Improve memory retention and memory accuracy · Suppresses age-related memory decline

[0055] According to another aspect of the present invention, there is provided a method for suppressing mitochondrial dysfunction, comprising ingesting or administering an effective amount of lactic acid bacteria belonging to Lactobacillus paracasei or a composition comprising the same to a subject in need thereof. According to the present invention, there is also provided a method for treating, preventing and / or improving a disease accompanied by mitochondrial dysfunction, comprising ingesting or administering an effective amount of lactic acid bacteria belonging to Lactobacillus paracasei or a composition comprising the same to a subject in need thereof. According to the present invention, there is also provided a method for reducing the risk of developing a disease caused by mitochondrial dysfunction, comprising ingesting or administering an effective amount of lactic acid bacteria belonging to Lactobacillus paracasei or a composition comprising the same to a subject in need thereof. The method of the present invention can be carried out according to the description of the composition and agent of the present invention.

[0056] According to another aspect of the present invention, there is provided a use of lactic acid bacteria belonging to Lactobacillus paracasei or a composition comprising the same for the manufacture of an inhibitor of mitochondrial dysfunction, as an inhibitor of mitochondrial dysfunction, or in a method for inhibiting mitochondrial dysfunction. According to the present invention, there is provided a use of lactic acid bacteria belonging to Lactobacillus paracasei or a composition comprising the same for the manufacture of an agent for treating, preventing, and / or improving a disease accompanied by mitochondrial dysfunction, as an agent for treating, preventing, and / or improving a disease accompanied by mitochondrial dysfunction, or in a method for treating, preventing, and / or improving a disease accompanied by mitochondrial dysfunction. According to the present invention, there is provided a use of lactic acid bacteria belonging to Lactobacillus paracasei or a composition comprising the same for the manufacture of an agent for reducing the risk of developing a disease caused by mitochondrial dysfunction, as an agent for reducing the risk of developing a disease caused by mitochondrial dysfunction, or in a method for reducing the risk of developing a disease caused by mitochondrial dysfunction. The use of the present invention can be carried out according to the description of the composition and agent of the present invention and the method of the present invention.

[0057] According to another aspect of the present invention, there is provided a lactic acid bacterium belonging to Lactobacillus paracasei for use in suppressing mitochondrial dysfunction, for use in treating, preventing and / or improving diseases accompanied by mitochondrial dysfunction, or for use in reducing the risk of developing diseases caused by mitochondrial dysfunction. The lactic acid bacterium belonging to Lactobacillus paracasei can be used according to the description of the composition and agent of the present invention.

[0058] The method and use of the present invention may be used in mammals, including humans, and both therapeutic and non-therapeutic uses are intended. As used herein, "non-therapeutic" does not include surgical, therapeutic or diagnostic procedures for humans (i.e., medical procedures for humans), and specifically does not include methods in which a physician or a person under the supervision of a physician performs surgical, therapeutic or diagnostic procedures on humans. EXAMPLES

[0059] The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples.

[0060] Example 1: Confirmation test of the effect of KW3110 strain on mitochondrial membrane potential In this example, the effect of the KW3110 strain on the mitochondrial membrane potential of a mouse-derived macrophage cell line subjected to inflammatory stimuli was evaluated.

[0061] (1) Test and evaluation methods (i) Method for detecting mitochondrial membrane potential Mitochondria have a double membrane structure consisting of an outer membrane and an inner membrane, and produce energy (ATP synthesis) by utilizing the potential difference between the outer membrane and the inner membrane. Normal mitochondria have a high membrane potential and produce energy, while mitochondria with impaired function have a low membrane potential and produce less energy. Mitochondria from living cells were stained with MitoTracker (registered trademark) Red CMXRos and MitoTracker (registered trademark) Green FM (ThermoFisher Scientific). MitoTracker (registered trademark) Red CMXRos is a red fluorescent reagent (excitation wavelength: 579 nm, fluorescence wavelength: 599 nm) whose degree of accumulation changes depending on the membrane potential, and MitoTracker (registered trademark) Green FM is a green fluorescent reagent (excitation wavelength: 490 nm, fluorescence wavelength: 516 nm) that accumulates regardless of the membrane potential, and the mitochondrial membrane potential can be evaluated by the ratio of the fluorescence intensities of both fluorescent reagents.

[0062] (ii) Test method In this study, mouse-derived macrophage cell line J774A.1 (obtained from ATCC (American Type Culture Collection) (catalog number: TIB-67), hereinafter the same) was used. The cells were cultured in Dulbecco's modified Eagle medium (DMEM) (containing 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin) (GIBCO) at a cell density of 2.5 × 10 5 The cells were seeded at 100 cells / well on a 24-well plate and cultured at 37°C for 1 day. This was designated the "control group." Next, Lactobacillus paracasei KW3110 strain heat-killed cells (FERM BP-08634) were added to the cultured cells in the well plate at 1 μg / mL, and after 24 hours, lipopolysaccharide (LPS) (Invivogen) was added at 0.1 μg / mL and cultured for 24 hours. This was designated the "LPS-treated + KW3110 group" (LPS+KW3110). In addition, the cells in the well plate were cultured for 24 hours after adding LPS in the same manner as above, without adding the KW3110 strain. This was designated the "LPS-treated group" (LPS). Next, the medium was removed from all groups, and a medium (FluoroBrite DMEM (GIBCO)) containing MitoTracker (registered trademark) Green FM (100 nM) and MitoTracker (registered trademark) Red CMXRos (100 nM) was added, followed by culturing at 37°C for 15 minutes. The medium was then replaced with FluoroBrite DMEM medium not containing the two fluorescent reagents, and flow cytometry analysis was performed using FACS Canto II (BD Biosciences). FlowJo software (BD Biosciences) was used as the analysis software. In this test, a plot of the cell population was created based on the fluorescence intensity of the two fluorescent reagents, and the cell population with a relatively low membrane potential was identified, and the percentage (%) of the cell population with a relatively low membrane potential to the total cells was calculated.

[0063] (iii) Statistical analysis The measured values ​​were expressed as mean ± standard error. One-way analysis of variance was performed followed by Tukey's test, and a p<0.05 was considered to indicate a significant difference between groups.

[0064] (2) Results The results are shown in Figure 1. Compared to the untreated cell group (control group), the percentage of cells containing mitochondria with low membrane potential was significantly increased in the LPS-treated group. On the other hand, in the LPS-treated + KW3110 group, the percentage of cells containing mitochondria with low membrane potential was significantly decreased compared to the LPS-treated group. These results demonstrated that the decrease in mitochondrial membrane potential caused by the addition of LPS in macrophage cells was improved by the addition of the KW3110 strain.

[0065] Example 2: Confirmation test of the effect of KW3110 strain on mitochondrial oxidative stress In this example, the effect of the KW3110 strain on oxidative stress in mitochondria of mouse-derived macrophage cells subjected to inflammatory stimuli was evaluated.

[0066] (1) Test and evaluation methods (i) Method for detecting mitochondrial oxidative stress Superoxide, peroxyl radical, hydrogen peroxide, hydroxyl radical and peroxynitrite are oxidative stresses that react with intracellular nucleic acids, proteins and lipids, causing damage to tissues. MitoSOX® Red (ThermoFisher Scientific) is a reagent that quickly localizes to mitochondria in cells and emits red fluorescence (excitation wavelength: 510 nm, fluorescence wavelength: 580 nm) in response to reactive oxygen. In the following test, the mitochondria of cells were stained with MitoSOX® Red (ThermoFisher Scientific). Oxidative stress can be evaluated by the ratio of the fluorescence intensity of MitoSOX® Red to that of MitoTracker® Green FM described in Example 1.

[0067] (ii) Test method In this test, the same mouse-derived macrophage cell line as in Example 1 was used. The cells were seeded and cultured in a 24-well plate in the same manner as in Example 1. This was designated as the "control group." Next, Lactobacillus paracasei KW3110 strain heat-killed bacteria (FERM BP-08634) was added to the cultured cells in the well plate at 1 μg / mL, and 24 hours later, LPS was added at 0.1 μg / mL and cultured for 24 hours. This was designated as the "LPS-treated + KW3110 group" (LPS + KW3110). In addition, the KW3110 strain was not added to the cultured cells in the well plate, and LPS was added in the same manner as above and cultured for 24 hours. This was designated as the "LPS-treated group" (LPS). Next, the medium was removed from all groups, and a medium containing MitoTracker (registered trademark) Green FM (100 nM) and MitoSOX (registered trademark) Red (5 μM) was added, and the cells were cultured at 37 ° C for 15 minutes. Next, the medium was replaced with FluoroBrite DMEM medium not containing the two fluorescent reagents, and flow cytometry analysis was performed using FACS Canto II (BD Biosciences). FlowJo software (BD Biosciences) was used as the analysis software. In this test, a plot of the cell population was created based on the fluorescence intensity of the two fluorescent reagents, and the cell population with relatively high oxidative stress was identified, and the percentage (%) of the cell population with relatively high oxidative stress to the total cells was calculated.

[0068] (iii) Statistical analysis The measured values ​​were expressed as mean ± standard error. One-way analysis of variance was performed followed by Tukey's test, and a p<0.05 was considered to indicate a significant difference between groups.

[0069] (2) Results The results are shown in Figure 2. Compared to the untreated cell group (control group), the proportion of cells with a high level of oxidative stress increased significantly in the LPS-treated group. On the other hand, in the LPS-treated + KW3110 group, the proportion of cells with a high level of oxidative stress decreased significantly compared to the LPS-treated group. These results demonstrated that the increase in mitochondrial oxidative stress caused by the addition of LPS in macrophage cells was improved by the addition of the KW3110 strain.

[0070] Example 3: Confirmation test of the effect of KW3110 strain on cellular respiration In this example, the effect of the KW3110 strain on cellular respiration of mouse-derived macrophage cells subjected to inflammatory stimuli was evaluated.

[0071] (1) Test and evaluation methods (i) Method for evaluating cellular respiration using an extracellular flux analyzer The extracellular flux analyzer is an instrument that evaluates the rate of oxygen consumption by cells in the medium, i.e., cellular respiration, by measuring the oxygen concentration and pH in the microspace around cultured cells over time. In addition, various mitochondrial functions related to respiration can be evaluated by continuously measuring the oxygen consumption rate while sequentially adding a mixture of an ATP synthase inhibitor, oligomycin, an uncoupler, carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP), and respiratory chain complex inhibitors, rotenone and antimycin A, to the medium.

[0072] (ii) Test method In this test, the same mouse-derived macrophage cell line as in Example 1 was used. The cells were cultured on an XFe24 cell culture plate (Agilent Technologies) at a cell count of 1.5 × 10 4The cells were seeded at 100 cells / well and cultured at 37°C for 1 day. This was designated the "control group." Next, Lactobacillus paracasei KW3110 strain heat-killed cells (FERM BP-08634) were added to the cultured cells in the plate at 1 μg / mL, and 24 hours later, LPS was added to 0.1 μg / mL and cultured for 24 hours. This was designated the "LPS-treated + KW3110 group" (LPS+KW3110). In addition, LPS was added to the cultured cells in the plate in the same manner as above, without adding KW3110 strain, and cultured for 24 hours. This was designated the "LPS-treated group" (LPS). Next, in all groups, the medium was replaced with XF DMEM medium (Agilent Technologies) containing 10 mM glucose, 1 mM pyruvate, and 2 mM glutamine, and the cells were cultured at 37°C in an incubator without CO2 control for 1 hour, after which the oxygen consumption rate was measured. Oligomycin (1 μM) was added 20 minutes after the start of measurement, FCCP (1 μM) was added 45 minutes after the start of measurement, and rotenone / antimycin A mixed solution (0.5 μM) (Agilent Technologies) was added 70 minutes after the start of measurement in that order. The oxygen consumption rate of the cells in the medium was measured using a flux analyzer (XFe24, Agilent Technologies) at 0, 10, and 18 minutes (stage I), 27, 35, and 44 minutes (stage II), 52, 61, and 69 minutes (stage III), and 78, 86, and 95 minutes (stage IV) after the start of measurement. The oxygen consumption rate was measured three times for each stage, and the average of the three measurements was calculated as the average oxygen consumption rate for each stage. Basal respiration and ATP turnover were calculated using the following formulas (I) and (II), respectively.

[0073] [Calculation formula] Basal respiration = mean oxygen consumption rate in stage I - mean oxygen consumption rate in stage IV (I) ATP turnover = mean oxygen consumption rate in stage I – mean oxygen consumption rate in stage II (II)

[0074] (2) Results The results are shown in Figure 3. Compared to the untreated cell group (control group), the LPS-treated group had a lower oxygen consumption rate at all measurement points, and showed a significant decrease in the oxygen consumption rate in basal respiration and ATP metabolic turnover. On the other hand, the LPS-treated + KW3110 group had an increased oxygen consumption rate at all measurement points compared to the LPS-treated group, and a tendency toward an increase in the oxygen consumption rate in basal respiration and ATP metabolic turnover was observed. These results demonstrated that the decrease in cellular respiration of macrophage cells caused by the addition of LPS was improved by the addition of the KW3110 strain.

[0075] Example 4: Confirmation test of the effect of KW3110 strain on mitochondrial morphology In this example, the effect of the KW3110 strain on the morphology of mouse-derived macrophage cells subjected to inflammatory stimuli was evaluated.

[0076] (1) Test and evaluation methods (i) Method for evaluating mitochondrial morphology using transmission electron microscopy To evaluate the morphology of mitochondria, intracellular mitochondria were observed using a transmission electron microscope. Chemical fixation was used to prepare samples for observation. Chemical fixation is one of the common fixation methods for biological samples, and involves solidifying and stabilizing proteins and lipids in cells and tissues using fixatives such as glutaraldehyde, paraformaldehyde, and osmium tetroxide.

[0077] (ii) Test method In this test, the same mouse-derived macrophage cell line as in Example 1 was used. The cells were plated in a 35 mm dish at a cell count of 5.0 × 10 5The cells were seeded so that each cell was 1 piece and cultured at 37℃ for 1 day. This was the "control group". Next, Lactobacillus paracasei KW3110 strain heat-killed cells (FERM BP-08634) were added to the cultured cells in the dish at 1μg / mL, and 24 hours later, LPS was added at 0.1μg / mL and cultured for 24 hours. This was the "LPS-treated + KW3110 group" (LPS+KW3110). In addition, the cells in the dish were cultured for 24 hours after adding LPS in the same manner as above, without adding KW3110 strain. This was the "LPS-treated group" (LPS). Next, the medium was removed from all groups, and 0.1M phosphate buffer containing 2% glutaraldehyde and 2% paraformaldehyde was added and cultured for 1 day at 4℃. The medium was then removed, and 0.1M phosphate buffer containing 2% osmium tetroxide was added and incubated at 4°C for 1 hour. The samples were dehydrated with ethanol, transferred to Quetol-812 embedding medium, and reacted at 60°C for 48 hours. After the reaction, 70-nm-thick sections were prepared from the samples using a diamond knife, mounted on copper grids, stained with 2% uranyl acetate at room temperature for 15 minutes, washed with sterile water, and then stained with lead stain (Sigma) at room temperature for 3 minutes. The samples were observed with a transmission electron microscope JEM-1400Plus (JEOL) at an accelerating voltage of 100 kV, and images were taken with a CCD camera EM-14830RUBY2 (JEOL).

[0078] (2) Results The results of photographing mitochondria in macrophage cells are shown in Figure 4. Mitochondria in the control group were oval and had a normal cristae structure, but mitochondrial morphological abnormalities and collapse of the cristae structure were observed in the LPS-treated group. On the other hand, the LPS-treated + KW3110 group did not show the abnormalities observed in the LPS-treated group. These results demonstrated that the addition of KW3110 improved mitochondrial morphological abnormalities caused by the addition of LPS in macrophage cells.

Claims

1. A composition for suppressing mitochondrial dysfunction, comprising Lactobacillus paracasei KW3110 strain (FERM BP-08634) as an active ingredient.

2. The composition of claim 1 , wherein the mitochondrial function is cellular respiration and / or ATP production.

3. The composition according to claim 1 or 2, wherein the mitochondrial function is measured using one or more indicators selected from the group consisting of membrane potential, oxidative stress, oxygen consumption rate, basal respiration, ATP turnover and morphology.

4. The composition according to any one of claims 1 to 3, wherein the mitochondria are mitochondria in one or more types of cells selected from the group consisting of macrophages, dendritic cells, lymphocytes, and microglia.

5. A composition for treating, preventing and / or improving a disease accompanied by decreased mitochondrial function, comprising Lactobacillus paracasei KW3110 strain (FERM BP-08634) as an active ingredient.

6. The composition according to claim 5 , wherein the disease associated with mitochondrial dysfunction is a neurodegenerative disease.

7. The composition according to any one of claims 1 to 6, which is a food composition or a pharmaceutical composition.

8. The composition according to claim 7, wherein the food is a beverage, a dairy product, a fermented food, a health food, a functional food, a nutritional supplement, a supplement, a health functional food or a food for the sick.

Images