JP2014024776A - Prophylactic and therapeutic agent for nash - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide safe and effective prophylactic/therapeutic agent for NASH.SOLUTION: The agent contains as an active component lactobacillus cells that can reduce the serum ALT value by 65% or more when tested as follows: mice are fed to uptake 3 g/day of a high fat diet containing lactobacillus cells (1×10/g) for 12 weeks; 9-12 weeks after the start of the diet, carbon tetrachloride was subcutaneously administered to the mice twice a week (initial dose: 0.05 ml/kg BW (body weight), then 0.1 ml/kg BW) in total 8 administrations; then the serum ALT values are compared with those of the control mice to which lactobacillus cells were not fed.

Description

  The present invention relates to a prophylactic / therapeutic agent for nonalcoholic steatohepatitis (NASH).

  In recent years, an increasing number of obese people have no drinking habits but become fatty liver due to cirrhosis or liver cancer. Among fatty liver diseases, a case in which a fatty liver disorder similar to alcoholic liver disorder is observed despite no apparent alcohol consumption is called nonalcoholic fatty liver disease (NAFLD), of which hepatocytes in liver tissue A case in which steatohepatitis with balloon-like swelling is observed is particularly referred to as Nonalcoholic steatohepatitis (NASH). Since the NASH has a high risk of becoming cirrhosis or liver cancer, development of a preventive and therapeutic means is desired.

  Streptococcus thermophilus SSP Sarivarius (CD8 strain) (DSM 14667) is known as a lactic acid bacterium that exhibits NASH therapeutic effects (Patent Document 1).

JP 2005-512590 A

However, the lactic acid bacterium Streptococcus thermophilus S.P. Sarivarius (CD8 strain) has a higher serum concentration than that before administration when humans are administered 1,800 billion bacteria per day for 4 months. The value of ALT (alanine transaminase) was reduced by about 50%, and the effect was low. There is no description of reduction of inflammation in the lobule of the liver or improvement of fibrosis.
Therefore, an object of the present invention is to provide a NASH preventive / therapeutic agent that can be safely taken on a daily basis and has a high NASH preventive / therapeutic effect.

Therefore, the present inventor first made various studies to produce a model animal exhibiting a pathological condition peculiar to human NASH. Surprisingly, a drug that induces obesity by ingesting a high fat diet and causing liver damage. (Hereinafter referred to as a hepatic disorder agent) is administered to humans more than 4 times, which is a large fatty liver, inflammation around the central vein in the leaflet of the liver, and hepatocytes are swollen like a balloon. It has been found that a model animal having a pathological condition peculiar to NASH can be prepared, and that the NASH preventive and therapeutic effect of the test substance can be evaluated and a NASH preventive and therapeutic agent can be screened by using the model animal. The NASH model animal is used for screening using various components so that a high-fat diet containing lactic acid bacteria containing 1 × 10 ¹⁰ / g of lactic acid bacteria can be given to mice for 3 weeks. Carbon tetrachloride from the 9th to 12th week from the start of administration of the high-fat diet containing lactic acid bacteria (first time: 0.05 ml / kg BW (body weight), second time and later: 0.1 ml / kg BW) Is administered subcutaneously twice a week for a total of 8 times, lactic acid bacteria whose serum ALT value is reduced by 65% or more compared to mice not administered with lactic acid bacteria have the effect of improving NASH-specific symptoms, etc. The present invention was completed by finding it useful as a preventive and therapeutic agent.

Therefore, the present invention allows mice to ingest a high-fat diet containing lactic acid bacteria containing 1 × 10 ¹⁰ / g of lactic acid bacteria so that the daily intake is 3 g for 12 weeks, and the administration of the high-fat diet containing lactic acid bacteria is started. From the 9th to the 12th week from the start, carbon tetrachloride (first time: 0.05 ml / kg BW (body weight), second time and later: 0.1 ml / kg BW) was subcutaneously administered twice a week for a total of 8 times. The present invention provides a NASH prophylactic / therapeutic agent comprising as an active ingredient a lactic acid bacterium whose serum ALT value is reduced by 65% or more as compared with mice not administered with lactic acid bacterium.

The NASH prophylactic / therapeutic agent of the present invention allows a mouse to ingest a high-fat diet containing lactic acid bacteria containing 1 × 10 ¹⁰ / g of lactic acid bacteria so that the daily intake is 3 g for 12 weeks. When carbon tetrachloride (first time: 0.05 ml / kg BW, second time and later: 0.1 ml / kg BW) was subcutaneously administered twice a week for a total of 8 times from the 9th week to the 12th week from the start of administration of Lactic acid bacteria whose serum ALT value is reduced by 65% or more compared to mice not administered with lactic acid bacteria are effective ingredients, and are useful in that they significantly improve NASH-specific symptoms and have a high NASH preventive and therapeutic effect. Furthermore, it is highly effective and useful in that the serum AST (aspartate aminotransferase) value is reduced by 40% or more compared to mice not administered with lactic acid bacteria.

It is a figure which shows the histopathological change in the liver of a microbial cell test group. A, B: Normal diet group (normal liver), C, D: Control group (NASH), E, F: L. plantarum group (findings where inflammation in leaflets was slightly suppressed), G, H: L. salivarius group (findings in which inflammation in leaflets was slightly suppressed). A-H: x20. A, C, E, G: HE staining, B, D, F, H: MT staining.

The NASH prophylactic / therapeutic agent of the present invention allows a mouse to ingest a high-fat diet containing lactic acid bacteria containing 1 × 10 ¹⁰ / g of lactic acid bacteria so that the daily intake is 3 g for 12 weeks. When carbon tetrachloride (first time: 0.05 ml / kg BW, second time and later: 0.1 ml / kg BW) was subcutaneously administered twice a week for a total of 8 times from the 9th week to the 12th week from the start of administration of Lactic acid bacteria whose serum ALT value is reduced by 65% or more compared to mice not administered with lactic acid bacteria are effective ingredients. More specifically, the bacterial powder-containing high-fat diet described in Table 2 below (the number of bacteria per bacterial powder-containing high-fat diet: the number of bacteria in 1 g of bacterial powder-containing high-fat diet is determined by the following formula: weight of powder bacteria bacterial powder amount (g) × bacterial powder containing high-fat diet in 1g produced by viable count / lyophilized at the end of the culturing (g) .L.Crispatus: 2.47 × 10 12/1 .754 × 0.0005 = 7.04 × 10 ^{8 /} g, B. Bifidum: 2.74 × 10 ¹² /1.55×0.0005=8.84×10 ^{8 /} g, L. Plantarum: 7. ^{36 × 10 12 /2.737×0.0005=1.34×10 9 /g,L.Salivarius:2.4×10 12} /2.995×0.0005=4.01×10 8 / g, S. thermophilus: 4.6 × 10 ¹¹ /2.444×0.0005=9.41×10 ⁷ / G) for 12 weeks (average feed weight per mouse L. Crispatus: 2.4 g / day, B. Bifidum: 2.4 g / day, L. Plantarum: 2.4 g / day, L. Salivarius: 2.4 g / day, S. thermophilus: 2.6 g / day), carbon tetrachloride from the 9th to 12th week from the start of administration of the high-fat diet containing bacterial powder (initial: 0.05 ml) / Kg BW, second and subsequent times: 0.1 ml / kg BW) is administered subcutaneously twice a week for a total of 8 times. Blood is collected 24 hours after the final administration of carbon tetrachloride, and the ALT level in serum is measured using transaminase CII test Wako (Wako Pure Chemical Industries). It is a lactic acid bacterium whose ALT value is reduced by 65% or more compared to a mouse not administered with lactic acid bacteria (a mouse administered with the high fat diet shown in Table 2 for 12 weeks and administered with carbon tetrachloride eight times in the same manner as described above). . The average intake of feed per mouse in this lactic acid bacteria non-administered group is 2.5 g / day. The number of lactic acid bacteria ingested by the mouse at this time is less than 1 × 10 ^{10 /} g × 3 g × 7 × ¹² = 2.52 × 10 ¹² in any bacterial species, but the number of bacteria is smaller than this. Thus, when ALT is reduced by 65% or more compared to mice not administered with lactic acid bacteria, a high-fat diet containing 1 × 10 ¹⁰ / g lactic acid bacteria is given to the mice for 12 weeks, so that the daily intake is 3 g. Carbon tetrachloride (first time: 0.05 ml / kg BW, second time and later: 0.1 ml / kg BW) twice a week in the 9th to 12th weeks from the start of administration of the high-fat diet containing the lactic acid bacteria. Even when administered subcutaneously a total of 8 times, the value of serum ALT is reduced by 65% or more compared to mice not administered with lactic acid bacteria. Moreover, it is more preferable to use a lactic acid bacterium whose serum AST value is reduced by 40% or more when the AST is measured by the same measurement method as that of the ALT, compared to a mouse not administered with a lactic acid bacterium.

  The type of lactic acid bacteria having such properties is not particularly limited, and Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus salivalus, Bacillus gasseri, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus delbruxy subspecies. Bulgaricus (Lactobacillus delbrueckii subsp. Bulgaricus), Lactobacillus del bruecki subspecies. Lactobacillus pulstocils, Lactobacillus plantus, Lactobacillus johnsoniii, Lactobacillus johnsoniii, Lactobacillus johnsoniii, Lactobacillus johnsoniii Streptococcus bacteria such as Streptococcus salivarius), Lactococcus lactis subsp. Lactococcus lactis subsp. Lactis, Lactococcus lactis subspecies. Bacteria of Lactococcus such as Lactococcus lactis subsp. Cremoris, Lactococcus raffinolactis, Bacteria of Enterococcus faecalis, Enterococcus ecterus Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium infantitis, Bifidobacterium address sentis (Bififi) Bifidobacterium bifidum, Bifidobacterium catenuratum, Bifidobacterum bibium ), Bifidobacterium gallicum derived from human intestine, Bifidobacterium lactis used in foods, Bifidobac Potassium animalis (Bifidobacterium animalis) lactic acid bacteria and the like. Among these, at least one selected from the group consisting of Bifidobacterium bifidum, Lactobacillus plantarum, Lactobacillus salivarius, Streptococcus thermophilus is preferable, and Bifidobacterium bifidum YIT 10347 (FERM BP-10613), Lactobacillus plantarum YIT 0132 (FERM BP-11349), Lactobacillus salivarius YIT 0432 (FERM P-22129), Streptococcus thermophilus YIT2001 (FERM BP-7538) More than species are preferred. These strains are deposited at the National Institute of Advanced Industrial Science and Technology (NITE-IPOD) (1st, 1st, 1st East, 1-chome, Tsukuba, Ibaraki 305-8586). In addition, Streptococcus thermophilus SSP Sarivarius (CD8 strain) (DSM 14667) described in JP 2005-512590 A should be classified as Streptococcus salivarius, Streptococcus thermophilus is a different type of fungus.

In addition, the usage form of lactic acid bacteria can mention 1 or more chosen from the bacterial cell (live cell) and the processed material of the said bacterial cell, for example. The treated product is not particularly limited as long as it is a treated product by a conventional method. For example, the treated cells (dead cells), lyophilized products thereof, cultures containing them, bacterial ultrasonic waves, etc. Crushing liquid, bacterial enzyme treatment liquid, solid residue separated by solid-liquid separation means such as filtration or centrifugation; treatment liquid from which cell wall has been removed by enzyme or mechanical means, concentrate of the treatment liquid, Diluted product, dried product thereof, etc .; Nucleic acid-containing fraction obtained by dissolving bacteria with a surfactant and then precipitating with ethanol, etc .; On the other hand, for example, those subjected to separation / purification treatment such as separation by various chromatography.
The dead cells can be obtained, for example, by heat treatment, treatment with drugs such as antibiotics, treatment with chemical substances such as formalin, treatment with ultraviolet rays, treatment with radiation such as γ rays. As will be described later, the effectiveness can be expected regardless of whether it is a viable cell, dead cell, or other treated cell product, as long as it contains a certain number of viable cells at the end of cultivation of lactic acid bacteria.

In the present invention, “NASH preventive treatment” refers to prevention and / or treatment of NASH, and the lactic acid bacterium used in the present invention is a lactic acid bacterium containing 1 × 10 ¹⁰ / g lactic acid bacterium, as shown in Examples below. The mice were ingested with the high fat diet for 12 weeks so that the daily intake was 3 g, and carbon tetrachloride (first time: 9 weeks to 12 weeks from the start of administration of the high fat diet containing lactic acid bacteria) 0.05 ml / kg BW, second and subsequent times: 0.1 ml / kg BW) twice a week for a total of 8 subcutaneous administrations, the serum ALT value decreases by 65% or more compared to mice not administered with lactic acid bacteria It is preferably 70% or more, more preferably 80% or more. Further, the serum AST is preferably decreased by 40% or more, more preferably by 50% or more, and further preferably by 80% or more. In addition, inflammation and fibrosis in the lobule, which are preferably symptoms unique to NASH, are improved and have an excellent NASH preventive and therapeutic action. The lactic acid bacteria used in the present invention can also be used as an anti-inflammatory agent and anti-fibrotic agent in the leaflets.

  When the NASH prophylactic / therapeutic agent of the present invention is used as a pharmaceutical, examples of the dosage form of the oral administration preparation include tablets, capsules, granules, dragees, pills, fine granules, powders, powders, sustained release Formulations, suspensions, emulsions, syrups, emulsions, lyophilizers, solutions, elixirs, oral jellys and the like can be mentioned.

  Moreover, the formulation in the case of using as a pharmaceutical can be manufactured by a conventional method, The lactic acid bacteria of this invention may be used independently, and may be used in combination with a pharmaceutically acceptable carrier. Examples of the pharmaceutically acceptable carrier include excipients, binders, disintegrants, surfactants, lubricants, fluidity promoters, flavoring agents, coloring agents, flavoring agents, diluents, bactericides, Osmotic pressure adjuster, pH adjuster, emulsifier, preservative, stabilizer, absorption aid, antioxidant, UV absorber, wetting agent, thickener, brightener, activity enhancer, anti-inflammatory agent, isotonic Agents, soothing agents, flavoring agents and the like.

  Examples of the binder include starch, dextrin, gum arabic powder, gelatin, hydroxypropyl starch, methylcellulose, sodium carboxymethylcellulose, hydroxypropylcellulose, crystalline cellulose, ethylcellulose, polyvinylpyrrolidone, macrogol and the like.

  Examples of the disintegrant include starch, hydroxypropyl starch, carboxymethylcellulose sodium, carboxymethylcellulose calcium, carboxymethylcellulose, and low-substituted hydroxypropylcellulose.

Examples of the surfactant include sodium lauryl sulfate, soybean lecithin, sucrose fatty acid ester, polysorbate 80 and the like.
Examples of the lubricant include talc, waxes, hydrogenated vegetable oil, sucrose fatty acid ester, magnesium stearate, calcium stearate, aluminum stearate, polyethylene glycol and the like.
Examples of the fluidity promoter include light anhydrous silicic acid, dry aluminum hydroxide gel, synthetic aluminum silicate, magnesium silicate, and the like.

  Examples of the diluent include distilled water for injection, physiological saline, aqueous glucose solution, olive oil, sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol and the like.

In addition, the NASH preventive / therapeutic agent of the present invention can be used not only as a pharmaceutical as described above, but also as a food or drink, a quasi drug, a pet food, or the like.
In this case, the lactic acid bacteria may be contained in the food or drink as they are or with various nutritional components added. This food and drink can be used as a health food or food material useful for NASH prevention and treatment, and these foods and drinks or containers thereof may be marked with the effect described above. Specifically, when the NASH preventive / therapeutic agent of the present invention is blended in a food or drink, an additive that can be used as a food or drink is appropriately used, and a form suitable for food using conventional means, for example, granular or granular , Tablets, capsules, pastes, etc., and various foods such as processed meat products such as ham and sausage, fishery products such as kamaboko and chikuwa; bread, confectionery, butter, powdered milk, fermented food and drink Or added to beverages such as water, fruit juice, milk, soft drinks and tea beverages.

  Furthermore, fermented products such as fermented milk, lactic acid bacteria beverages, fermented soymilk, fermented fruit juice, and fermented plant fluid containing lactic acid bacteria that are active ingredients are suitably used as food and drink. These fermented foods and drinks can be produced according to conventional methods. For example, fermented milk is inoculated and cultured with lactic acid bacteria in a sterilized milk medium, and homogenized to obtain a fermented milk base. Subsequently, a separately prepared syrup solution is added and mixed, homogenized with a homogenizer or the like, and flavor is further added to obtain a final product. The fermented milk thus obtained can be a product of any form such as a plain type, a soft type, a fruit flavor type, a solid form, and a liquid form.

Moreover, there is no strict restriction | limiting in the dosage at the time of using the lactic acid bacteria which are the active ingredients of the NASH preventive-treatment agent of this invention. Since the effect obtained varies depending on various use modes such as the subject and applicable disease, it is desirable to set the dose appropriately. However, in terms of NASH preventive treatment, the number of lactic acid bacteria should be 1 × 10 ⁹ or more. The amount contained as a daily amount is preferable, the amount containing 5 × 10 ⁹ to 2 × 10 ¹² as a daily amount is more preferable, and the amount containing 1 × 10 ¹⁰ to 1 × 10 ¹² as a daily amount is further included. preferable. The number of bacteria in the present invention is calculated based on the number of living bacteria at the end of lactic acid bacteria culture.

  EXAMPLES Next, although an Example is given and this invention is demonstrated in detail, this invention is not limited to this at all.

Example 1
(1) Test group and test schedule The following experiment was performed using C57BL / 6N strain, 7-week-old mice. The normal diet group is a test group in which AIN93G, which is a general breeding feed, was ingested for 12 weeks. In the control group, for the onset of NASH, intake of a high fat diet for 12 weeks and carbon tetrachloride (initial: 0.05 ml / kg BW, second and subsequent: 0.1 ml / kg BW) from week 9 to week 12 Was administered subcutaneously a total of 8 times. Each lactic acid bacteria administration group was fed a diet containing 0.05% of each freeze-dried bacterial powder based on the high fat diet of the control group (the number of bacteria in the feed: the bacterial powder was freeze-dried from the culture solution) The number of bacteria in 1 g of feed is determined by the following formula: number of viable bacteria at the end of culture in culture medium / amount of bacterial powder produced by lyophilization (g) × weight of bacterial powder in 1 g of feed ( g) L. Crispatus: 2.47 × 10 ¹² /1.754×0.0005=7.04×10 ⁸ / g, B. Bifidum: 2.74 × 10 ¹² /1.55×0.0005= ^{8.84 × 10 8 /g,L.Plantarum:7.36×10 12 /2.737×0.0005=1.34×10 9} /g,L.Salivarius:2.4×10 12/2. ^{995 × 0.0005 = 4.01 × 10 8} /g,S.thermophilus ^{4.6 × 10 11 /2.444×0.0005=9.41×10 7 / g} ). The administration method and timing of carbon tetrachloride are the same as in the control group. The composition of the high fat diet containing AIN 93G, high fat diet and lactic acid bacteria powder is listed in Table 2. Feed and drinking water were freely consumed in each test (average feed weight per mouse normal diet: 2.4 g / day, control: 2.5 g / day, L. Crispatus: 2.4 g / day) B. Bifidum: 2.4 g / day, L. Plantarum: 2.4 g / day, L. Salvarius: 2.4 g / day, S. thermophilus: 2.6 g / day).
Necropsy was performed 24 hours after the final administration of carbon tetrachloride, and blood and organs were collected. Serum was prepared from blood and used to measure AST and ALT. The collected liver was immersed and stored in a 10% neutral buffered formalin solution for histopathological analysis.

(2) Measurement of AST and ALT levels in serum The AST and ALT levels in serum were measured using transaminase CII Test Wako (Wako Pure Chemical Industries). The operation was performed according to the manual.

(3) Histopathological analysis Each test group, 3 to 4 individual livers were used for histopathological analysis. The liver fixed with a formalin solution was cut out by a conventional method, embedded in paraffin, and then the paraffin block was sliced into a thickness of about 4 μm, stained with Hematoxylin-Eosin (HE), and observed with an optical microscope. In addition, Masson's trichrome (MT) staining was performed on paraffin sections to prove collagen fibers.
Intralobular inflammation and fibrosis were examined as diagnostic items for histopathology.
In the grading of the pathological condition, the entire visual field of the liver section is observed and no pathological condition is observed:-(negative), the pathological condition is 20% or less of the total visual field: ± (very mild), the pathological condition is 20 to 50% of the total visual field : + (Mild), 50-80% of all visual fields: ++ (moderate), 80% or more of all visual fields: ++ (severe).

(4) Results (a) Measurement of AST and ALT values in serum When AST values in serum were measured, L. plantarum, L .; A significant decrease was observed in the salivarius group (L. plantarum p = 0.026, L. sarivarius p = 0.021). B. In the bifidum group, a downward trend was observed (p = 0.07). S. A trend of decrease was also observed in the thermophilus group. In addition, when the ALT level in serum was measured, bifidum, L .; plantarum, L .; A significant decrease was observed in the salivarius group (B. bifidum p = 0.031, L. plantarum p = 0.012, L. sarivarius p = 0.008). S. In the thermophilus group, a tendency to decrease was observed (p = 0.054). However, L. In the crisptus group, neither AST nor ALT significantly decreased. The results of AST are shown in Table 3. Table 4 shows the results of ALT.

(B) Histopathological analysis The histopathological analysis of the liver was conducted, and the effect of the administration of the bacterial cells on the liver was examined. As a result, B.I. bifidum group, L. plantarum group, L. In some individuals in the salivarius group, inflammation in the lobule was milder than in the control group. L. plantarum group, S. p. In some thermophilus individuals, fibrosis was minor compared to the control group. The results are shown in FIG.

The number of bacteria that a mouse ingests per day is Bifidum: 2.12 × 10 ⁹ , L. Plantarum: 3.2 × 10 ⁹ , L. Salvarius: 9.62 × 10 ⁸ , S. thermophilus: 2.45 × 10 ⁸ . Since the weight of the mouse used was about 37 g, Bifidum: 5.73 × 10 ¹⁰ / kg BW / day, L. Plantarum: 8.65 × 10 ¹⁰ / kg BW / day, L.P. Sarivarius: 2.6 × 10 ¹⁰ / kg BW / day, S. var. thermophilus: 6.62 × 10 ⁹ / kg BW / day. According to the guidance of the US Food and Drug Administration (FDA), mouse intake is converted to human intake (calculation method: human dose (/ kgBW / day) = mouse dose (/ kgBW / day) × 3 (mouse body surface Conversion factor) / 37 (human body surface conversion factor)), Bifidum: 4.65 × 10 ⁹ / kg BW / day, L. Plantarum: 7.01 × 10 ⁹ / kg BW / day, L.P. Sarivarius: 2.11 × 10 ⁹ / kg BW / day, S. thermophilus: 5.37 × 10 ⁸ / kg BW / day. If the human body weight is 70 kg, in order to achieve the same effect as a mouse, Bifidum: 3.26 × 10 ¹¹ , L. Plantarum: 4.91 × 10 ¹¹ , L.C. Salivarius: 1.48 × 10 ¹¹ , S. thermophilus: 3.76 × 10 ¹⁰ . These are all less than 1 trillion 800 billion (1.8 × 10 ¹² ) described in JP-T-2005-512590. Therefore, the lactic acid bacterium of the present invention has an advantage that it has a higher effect with a smaller number of bacteria than the lactic acid bacterium described in JP-T-2005-512590.

  From the above results, it was shown that NASH model animals can be used to screen for NASH preventive and therapeutic agents, and that the lactic acid bacteria are useful as NASH preventive and therapeutic agents.

Claims (4)

The lactic acid bacteria-containing high-fat diet containing 1 × 10 ¹⁰ / g of lactic acid bacteria was ingested by the mice for 12 weeks so that the daily intake was 3 g. ˜12 weeks after carbon tetrachloride (initial: 0.05 ml / kg BW (body weight), second and later: 0.1 ml / kg BW) twice weekly for a total of 8 subcutaneous administrations A NASH preventive and therapeutic agent comprising, as an active ingredient, lactic acid bacteria whose serum ALT value is reduced by 65% or more.   The NASH prophylactic / therapeutic agent according to claim 1, wherein the lactic acid bacterium further lowers the serum AST value by 40% or more compared to a mouse not administered with the lactic acid bacterium.   The lactic acid bacteria are Bifidum, L.M. Plantarum, L.M. Salvarius and S. The NASH prophylactic / therapeutic agent according to claim 1 or 2, which is one or more selected from the group consisting of thermophilus.   The lactic acid bacteria are Bifidum YIT 10347, L.B. Plantarum YIT 0132, L. Salvarius YIT 0432 and S. The NASH prophylactic / therapeutic agent according to any one of claims 1 to 3, which is one or more selected from the group consisting of thermophilus YIT 2001.