JP5815105B2 - Neutral fat reducing agent - Google Patents

Description

  The present invention relates to a neutral fat reducing agent.

  Lactobacillus brevis (Lactobacillus brevis) SBC8803 strain (Accession number: FERM BP-10632) has an antiallergic action (Patent Document 1), an alcoholic liver disorder suppressing action (Patent Document 2), and an intestinal tract protecting action. (Patent Document 3) is known.

International Publication No. 2008/023663 International Publication No. 2009/090961 JP 2010-83881 A

  An object of the present invention is to provide a novel use of the Lactobacillus brevis SBC8803 strain.

  The present invention provides a neutral fat reducing agent containing a bacterial cell belonging to Lactobacillus brevis or a processed product thereof as an active ingredient.

  Neutral fat plays important roles such as the role of maintaining body temperature and keeping it constant, the role of protecting internal organs, and the role of an emergency energy source. On the other hand, if neutral fat increases too much, various lifestyle-related diseases may occur. Since the neutral fat reducing agent of the present invention contains a bacterial cell of a strain belonging to Lactobacillus brevis or a processed product thereof as an active ingredient, the neutral fat reducing agent increases through an effect of reducing neutral fat or an effect of suppressing an increase in neutral fat. Too much neutral fat can be reduced.

  There are four subspecies of Lactobacillus brevis: brevis, grevesensis, otakiensis, and coagulans. As a strain in the neutral fat reducing agent of the present invention, a strain belonging to subspecies brevis is preferable, and among strains belonging to subspecies brevis, for example, Lactobacillus brevis SBC8803 strain is particularly preferable.

  In addition, the Lactobacillus brevis SBC8803 strain was established on June 28, 2006 at the National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center (1st, 1st East, 1st Street, Tsukuba City, Ibaraki Prefecture, Japan, 6th postal code 305-8586). ), And the accession number is FERM BP-10632.

  Lactobacillus brevis is a type of lactic acid bacterium that has been used for fermented foods for a long time, and has been established to be safe for living organisms. Therefore, the neutral fat reducing agent of the present invention is highly safe to living bodies and can be taken continuously for a long period of time. Therefore, it can be used as a pharmaceutical ingredient, a food / beverage product component, a food / beverage product additive, and the like.

  Moreover, fermented milk is preferable as the food or drink. Since fermented milk originally contains lactic acid bacteria, the addition of the neutral fat reducing agent of the present invention does not deviate from sanitary standards such as the number of bacteria, and there is little adverse effect on the flavor. It is suitable as a food or drink to which a natural fat reducing agent is added.

  ADVANTAGE OF THE INVENTION According to this invention, the safety | security to a biological body is high, and the novel neutral fat reducing agent which can be used also as a component of food-drinks is provided. Moreover, the pharmaceutical, food / beverage products, food / beverage additive, etc. containing such a neutral fat reducing agent are provided.

6 is a graph showing changes in mouse body weight in Example 2. 3 is a graph showing plasma neutral fat concentration in mice in Example 2. 4 is a graph showing the visceral fat weight of a mouse in Example 2. 6 is a graph showing the human plasma triglyceride concentration in Example 4.

  The neutral fat reducing agent of the present invention contains a bacterial cell of a strain belonging to Lactobacillus brevis or a processed product thereof as an active ingredient. The neutral fat reducing agent of the present invention can reduce neutral fat based on the neutral fat reducing action of the bacterial cells of the strain belonging to Lactobacillus brevis or the processed product thereof.

  In the present specification, the “neutral fat reducing action” refers to an action of reducing neutral fat or an action of suppressing an increase in neutral fat.

  “Neutral fat” means a glycerin ester of a fatty acid. On the other hand, since most of the neutral fats exist in the blood as triglycerides (triacylglycerols), “triglyceride (or TG)” is synonymous with “neutral fat” in the present specification. May be used.

  The role of neutral fat in the body is, for example, the role as an energy source, the role of preventing heat from being taken away from the body as subcutaneous fat, the body temperature being kept constant, the organ being fixed at a certain position as visceral fat, It has important roles essential for survival, such as protecting the organs from external impacts. On the other hand, if the neutral fat increases too much beyond the aptitude value, it can cause various diseases. For example, if the neutral fat concentration in the blood increases too much, it can cause arteriosclerosis, and if too much visceral fat or subcutaneous fat increases, it can contribute to obesity and metabolic syndrome. Moreover, when neutral fat accumulates in the liver, it can become fatty liver. Neutral fat is synthesized in the liver using excess carbohydrates from eating too much as a material, and synthesis is also promoted by alcohol consumption. For example, the proper value of neutral fat in the blood is considered to be less than 150 mg / dl (fasting) (Japanese Arteriosclerosis Society Hyperlipidemia Guidelines Guideline: Arteriosclerosis 25: 1-34, 1997), if this value is exceeded, it is assumed that “neutral fat has increased too much”.

  The triglyceride reducing agent of the present invention can reduce the triglyceride that has been excessively increased by the action of reducing triglyceride, or the triglyceride can be excessively increased by the action of suppressing the increase of triglyceride. It can also be suppressed.

  The strain in the neutral fat reducing agent of the present invention only needs to belong to Lactobacillus brevis, and may be, for example, one that can be separated from the natural world or one that can be obtained from a cell bank such as ATCC.

  As a microbial cell of the above strain or a processed product thereof, even if only one type of microbial cell or a processed product thereof is contained, a microbial cell of two or more strains or a processed product thereof is also contained. Also good. Bacteria can be produced in large quantities by culturing viable bacteria.

  The above strain can be cultured according to a culture method commonly used for culturing strains belonging to Lactobacillus brevis. For example, as a culture method, live bacteria are inoculated in an MRS medium, LB medium or the like having a pH of about 4 to about 6 and then in an incubator at about 30 to about 38 ° C. (particularly about 37 ° C.) for about 12 to about 48 hours. (Especially about 12 hours, about 24 hours, about 36 hours or about 48 hours).

  The neutral fat reducing agent of the present invention may contain, as an active ingredient, a processed product of bacterial cells obtained by subjecting the bacterial cells to heat sterilization treatment, physical sterilization treatment, and the like. As the treated product of the microbial cells, for example, a treated product obtained by heating the microbial cells at 100 ° C. or more for several minutes or more (for example, the microbial cells are subjected to autoclave treatment at a temperature of 110 to 125 ° C. for 10 minutes or more. Processed product), a processed product obtained by freeze-drying, spray-drying, etc. on the cells, or a processed product obtained by physically destroying the cells with an ultrasonic wave, a French press or the like. The neutral fat reducing agent of the present invention exhibits the same effects as when the bacterial cells (viable bacteria) are used even when such a processed product of bacterial cells (dead bacteria) is used as an active ingredient.

  There are four subspecies in Lactobacillus brevis, which are brevis, grevesensis, otakiensis and coagulans, respectively. Among these subspecies, the strain preferably belongs to the subspecies brevis. Strains belonging to the subspecies brevis can be separated by using the base sequence of 16S ribosomal DNA and the difference in acid production from the sugar as an index, and can be classified as strains not belonging to subspecies grebsensis, subspecies otachysis and subspecies coagulance.

  Moreover, as a strain which belongs to Lactobacillus brevis subspecies brevis, it is preferable to use Lactobacillus brevis SBC8803 strain. Lactobacillus brevis SBC8803 strain is deposited at the Patent Organism Depositary, National Institute of Advanced Industrial Science and Technology under the accession number FERM BP-10632, and is available.

  The neutral fat reducing agent of the present invention can take the form of various preparations as described below. That is, the neutral fat reducing agent of the present invention may be in any form such as solid (for example, powder obtained by freeze-drying), liquid (water-soluble or fat-soluble solution or suspension), paste, etc. , Powders, pills, granules, tablets, syrups, troches, capsules and the like.

  The above-mentioned various preparations may consist only of bacterial cells belonging to the strain belonging to Lactobacillus brevis or processed products thereof, and by molding the bacterial cells belonging to the strain belonging to Lactobacillus brevis or processed products thereof into the above dosage form. Can be prepared. The above-mentioned various preparations also include bacterial cells belonging to Lactobacillus brevis or processed products thereof, and pharmaceutically acceptable additives (excipients, binders, lubricants, disintegrants, emulsifiers, surfactants). , Bases, solubilizers, suspending agents, etc.) and can be prepared by molding. In this case, the content of the active ingredient is 2 to 50% by mass based on the total amount of the preparation.

  For example, the excipient includes lactose, sucrose, starch, dextrin and the like. Examples of the binder include polyvinyl alcohol, gum arabic, tragacanth, gelatin, hydroxypropylmethylcellulose, hydroxypropylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone and the like. Examples of the lubricant include magnesium stearate, calcium stearate, talc and the like. Examples of the disintegrant include crystalline cellulose, agar, gelatin, calcium carbonate, sodium bicarbonate, dextrin and the like. Examples of the emulsifier or surfactant include Tween 60, Tween 80, Span 80, and glyceryl monostearate. Examples of the base include cetostearyl alcohol, lanolin, polyethylene glycol, rice bran oil, fish oil (DHA, EPA, etc.), olive oil and the like. Examples of the solubilizer include polyethylene glycol, propylene glycol, sodium carbonate, sodium citrate, Tween 80 and the like. Examples of the suspending agent include Tween 60, Tween 80, Span 80, glyceryl monostearate, polyvinyl alcohol, polyvinyl pyrrolidone, methyl cellulose, hydroxymethyl cellulose, sodium alginate and the like.

  The neutral fat reducing agent of the present invention may be administered to humans or non-human mammals. The dosage and administration method can be appropriately determined according to the condition, age, etc. of the individual to be administered. Suitable administration methods include, for example, oral administration. As an example of the dosage and administration method, the neutral fat reducing agent of the present invention can be given orally once a day in an amount of 5 mg of the active ingredient.

  The neutral fat reducing agent of the present invention can be used as a pharmaceutical ingredient, a food / beverage product component, a food / beverage product additive, a feed component, a feed additive, and the like.

  For example, the neutral fat reducing agent of the present invention includes water, soft drinks, fruit juice drinks, milk drinks, alcoholic drinks, breads, noodles, rice, tofu, dairy products, fermented foods, fermented milk, soy sauce, miso, It can be used as an additive to food and drink such as confectionery. These foods and drinks may further contain other additives usually used in the art, and examples of such additives include bitters, flavors, apple fibers, soybean fibers, meat extracts, blacks. Vinegar extract, gelatin, corn starch, honey, animal and vegetable oils; monosaccharides such as glucose and fructose; disaccharides such as sucrose; polysaccharides such as dextrose and starch; sugar alcohols such as erythritol, xylitol, sorbitol, and mannitol; vitamin C and the like Vitamins. The neutral fat reducing agent of the present invention can also be used as a component for foods for specified health use, foods for special uses, food supplements, health foods, functional foods, foods for the sick, and the like. The food or drink containing the neutral fat reducing agent of the present invention may be a fermented product obtained by fermenting milk, skim milk, soy milk, etc. with a strain belonging to Lactobacillus brevis.

  Moreover, it is preferable to use the neutral fat reducing agent of the present invention as an additive to fermented milk. The addition amount of the neutral fat reducing agent of the present invention in this case can be determined so that the content of the active ingredient is 0.001 to 0.5% by mass based on the total amount of fermented milk. Fermented milk is defined as “milk or milk containing non-fat milk solids equal to or higher than this, fermented with lactic acid bacteria or yeast, and pasty or liquid, or those frozen”. Examples include yogurt, cheese, butter, drinking yogurt, lactic acid bacteria beverages, and sour milk beverages. Among them, yogurt originally contains lactic acid bacteria (live bacteria), so even if the neutral fat reducing agent of the present invention is added, the strain belonging to Lactobacillus brevis contained as an active ingredient has an adverse effect on taste and flavor. There is almost no fear and it is particularly preferable. Furthermore, the component specification of yogurt is that the non-fat milk solid content is 8.0% or more and the number of lactic acid bacteria (or the number of yeasts) is 10 million / ml or more. The addition is preferable because there is almost no risk of exceeding the standard such as the hygiene standard for the number of lactic acid bacteria.

  Hereinafter, based on an Example, this invention is demonstrated more concretely. However, the present invention is not limited to the following examples.

[Example 1]
(Preparation of neutral fat reducing agent)
The Lactobacillus brevis SBC8803 strain was cultured at 30 ° C. for 24 hours in a medium mainly composed of yeast extract and liquid sugar, and the cells were collected by centrifugation. A part of the collected cells was suspended in distilled water to obtain a 100 mg / ml cell suspension. This was used as a neutral fat reducing agent for “live bacteria”. On the other hand, a part of the collected cells are sterilized by autoclaving (121 ° C., 20 minutes), then lyophilized, and suspended in distilled water to obtain a suspension of 100 mg / mL treated cells. Obtained. This was defined as a “dead bacteria” neutral fat reducing agent.

[Example 2]
(Administration test of neutral fat reducing agent in mice)
Thirty-two C57BL / 6J mice (male, Charles River Japan) were divided into 4 groups of 8 mice in each group so that the mean value and variance of the body weight at the start of the study did not vary between groups. In addition, the mouse | mouth was used for the test at the time of a 5-week-old after passing through the acclimatization breed | strain for 1 week.

(Preparation of test feed)
The test feed (diet) was prepared to have the composition shown in Table 1. In Table 1, the unit of each component amount is g. Moreover, AIN93M was used as a feed during the acclimatization breeding period. In Table 1, the component amount of the neutral fat reducing agent (live bacteria) and the component amount of the neutral fat reducing agent (dead bacteria) are the dry weight of the microbial cells and the dry weight of the processed microbial cells.

(Raising conditions)
After acclimation and breeding for 1 week, each group of mice was allowed to freely feed and drink a predetermined test feed and tap water for 6 weeks. Throughout the acclimatization period and subsequent test feed administration period, the mice were kept in a rearing device (temperature 23 ± 5 ° C., relative humidity 55 ± 10%, EBAC, Japan Claire), with regular ventilation, light / dark time 12 hours (light period: (8 o'clock to 20 o'clock) under the condition of 4 animals / 1 cage.

(Weight measurement)
Mice were weighed at the start of breeding and every other week during the breeding period. The average of the measured values of 8 animals in each group was taken as the weight of each group.

  FIG. 1 is a graph showing the weight transition of mice. There was no significant difference in body weight between the Ctrl group (Ctrl) that did not contain the Lactobacillus brevis SBC8803 strain in the diet, Sample Group 1 (live bacteria), and Sample Group 2 (dead bacteria). On the other hand, the normal diet group (AIN93M) was significantly lower in weight with a risk rate of 5% than the other three groups.

(Measurement of plasma neutral fat concentration and visceral fat weight)
Six weeks after the start of test feed administration, mice in each group were subjected to cardiac blood sampling and dissection under ether anesthesia, and plasma neutral fat concentration, fat weight around the epididymis, retroabdominal wall fat weight, and mesenteric fat weight were measured. did.

  FIG. 2 is a graph showing the plasma triglyceride concentration in mice. Compared with the normal diet group that had been administered the normal diet (AIN93M), the plasma group had a significantly increased plasma triglyceride concentration in the Ctrl group that had been administered the high-fat, high-sucrose, and high-protein diet (Fig. 2). On the other hand, increase in plasma triglyceride concentration was observed in sample groups 1 (live bacteria) and 2 (dead bacteria) that had been administered a diet obtained by adding Lactobacillus brevis SBC8803 strain to a high fat, high sucrose, high protein diet (Figure 2). That is, it was found that the Lactobacillus brevis SBC8803 strain can suppress an increase in plasma neutral fat concentration caused by a high-fat, high-sucrose, and high-protein diet, whether it is live or dead. .

  FIG. 3 is a graph showing the visceral fat weight of mice. The mesenteric fat weight per body weight was not significantly different among the 4 groups (FIG. 3). The retroabdominal wall fat weight per body weight and the fat weight around the accessory testicles per body weight were significantly increased in the Ctrl group, sample group 1 (live bacteria) and sample group 2 (dead bacteria) compared to the normal diet group (FIG. 3). ). In addition, the retroabdominal wall fat weight per body weight was not significantly different between the Ctrl group, the sample group 1 (live bacteria), and the sample group 2 (dead bacteria), whereas the fat weight around the accessory testicles per body weight was Ctrl. Compared with the group, it was significantly lower in the sample group 1 (live bacteria) and the sample group 2 (dead bacteria) (FIG. 3). Furthermore, reflecting these results, the total visceral fat weight of sample group 1 (live bacteria) and sample group 2 (dead bacteria) was significantly increased compared to the normal food group, but compared to the Ctrl group. Was significantly lower (FIG. 3). From these results, it was found that the Lactobacillus brevis SBC8803 strain can suppress the accumulation of visceral fat caused by a high-fat, high-sucrose, high-protein diet, whether it is live or dead. . It is considered that the suppression of plasma triglyceride concentration by the action of suppressing the increase in plasma triglyceride concentration described above is one factor that can suppress the accumulation of visceral fat.

Example 3
(Preparation of yogurt with added neutral fat reducing agent)
To the yogurt produced according to a conventional method, the neutral fat reducing agent of “live bacteria” of Example 1 was added and mixed so that the amount of bacterial cells per 5 g of yogurt was 5 mg, and the yogurt containing the neutral fat reducing agent was added. Prepared.

Example 4
(Administration test of neutral fat reducing agent in humans)
A plasma GOT (AST) value of 36 IU / L or higher, a plasma GPT (ALT) value of 36 IU / L or higher, or a plasma γGTP value of 61 IU / L or higher (may be duplicated) Human subjects were allowed to live as usual except for eating 100 g of yogurt prepared in Example 3 once a day for 2 weeks, and at the start of the test and the second week after the start of the test. The plasma triglyceride concentration was measured.

  FIG. 4 is a graph showing the measurement results of plasma neutral fat concentration. The plasma triglyceride concentration 2 weeks after the start of the study was significantly reduced at a risk rate of 5% in all subjects. In addition, the average value of all the subjects with plasma triglyceride concentration after 2 weeks decreased by 58.7 mg / dL with respect to the average value of all subjects with plasma triglyceride concentration before the start of the test.

  This result is considered to indicate that Lactobacillus brevis SBC8803 strain has an action of reducing plasma neutral fat. On the other hand, Lactobacillus brevis SBC8803 strain suppresses the increase in plasma triglyceride, since the results were obtained after living as usual except for the condition of eating yogurt prepared in Example 3 once a day. It is also considered to indicate that

Claims (1)

  Lactobacillus brevis (Lactobacillus brevis) SBC8803 (Accession number: FERM BP-10632) A cell neutral fat reducing agent which contains the microbial cell of a strain, or its processed material as an active ingredient.

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