JP2011246414A - Lipid metabolism promoter in muscular tissue - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a promoter of lipid metabolism and/or lipid transportation in muscular tissue.SOLUTION: The promoter of the lipid metabolism and/or the lipid transportation in the muscular tissue contains polysaccharides, and has activities for promoting the lipid metabolism and/or the lipid transportation in the muscular tissue.

Description

  The present invention relates to an agent for promoting lipid metabolism in muscle tissue, including polysaccharides such as dietary fiber.

In recent years, lifestyle diseases such as obesity, high blood pressure, and abnormal blood sugar levels have increased in Japan due to the westernization of dietary habits, and an increase in medical expenses has become a major social problem. In particular, obesity is a problem not only for the accumulation of subcutaneous fat, but also for the accumulation of visceral fat, which is an attracting material for hypertension, diabetes, etc., and triggers for serious diseases such as heart disease. .
In order to control obesity, human efforts such as instruction to make food intake of nutrients in an appropriate state and calorie consumption by exercise etc. are effective, but the burden on individuals may increase. is there. Therefore, in order to reduce the burden on the individual and obtain an obesity effect, it would be more desirable if there was a functional food having an absorption suppression of excess nutrients and a fat burning effect.
Among functional food ingredients, dietary fiber has been reported to have functions such as intestinal regulation, cholesterol and carbohydrate absorption suppression, and is known to significantly reduce the risk of heart disease. (Patent Documents 1, 2, and 3).

  For example, recently, the US Food and Drug Administration (USFDA) has taken daily intake of 7 grams of soluble polysaccharides obtained from psyllium husks or psyllium seed husks as part of a diet low in saturated fat and cholesterol. It was confirmed that the total cholesterol concentration and LDL cholesterol concentration in the blood of the dogs were gradually lowered to moderate hypercholesterolemia, which could be a risk of coronary heart disease (CHD). In view of this discovery, the USFDA approves a CHD healing statement for labeling foods that contain a minimum of 95% purified soluble polysaccharide per standard consumption (RACC, 7 grams divided by 4 meals per day) is doing. The USFDA adopted the term “palladium shell” or “palladium seed shell” to define soluble polysaccharides.

In general, dietary fiber is a general term for indigestible components contained in food that are not digested by human digestive enzymes or are difficult to digest. Chemically, it is often a polysaccharide, and can be roughly classified into two types, soluble polysaccharides and insoluble polysaccharides.
Examples of such soluble polysaccharides include agar, pectin, guar gum, gum arabic, xanthan gum, carrageenan, tamarind seed gum, alginic acid, pullulan, locust bean gum, carboxymethylcellulose, psyllium, curdlan, gellan gum and mannans (glucomannan) , Galactomannan, etc.) (Patent Document 4).
Examples of insoluble polysaccharides include cellulose, lignin, hemicellulose A, C, chitin, chitosan, and collagen.

  Traditionally, dietary fiber has not been well studied nutritionally because it is not digested or difficult to digest. However, based on recent epidemiological survey results, the mechanism of action centering on the cholesterol reduction effect has been energetically studied (Non-patent Document 1).

  Particularly viscous soluble polysaccharides prevent bile acid absorption in the lumen of the lower small intestine, and much bile acid is lost by feces. Since blood cholesterol is primarily used in the liver to synthesize more bile acids to compensate for the loss of bile acids, blood cholesterol is thought to decrease.

  In other words, the effects of lowering the amount of intestinal circulating cholesterol and the amount of cholesterol pooled in the liver, the effects of lowering the total cholesterol in the blood, lowering LDL cholesterol, metabolic changes of lipoproteins, etc. Is proposed to be demonstrated. However, the number of research examples up to now is small compared to other research areas, and a lot of knowledge that should be clarified in the future remains (Non-Patent Documents 1, 2, and 3).

Special table 2001-522614 Special Table 2004-519254 Special table 2007-510743 gazette JP 2005-330198 A

“Guar gum and similar soluble fibers in the regulation of cholesterol metabolism: Current understandings and future research priorities” Vascular Health and Risk Management 2008: 4 (5) 1023-1033 Fernandez ML. 2001. Soluble fiber and indigestible carbohydrate effects on plasma lipids and cardiovascular risks. Curr Opin in Lipidol, 12: 35-40. Roy S, Vega-Lopez S, Fernandez ML. 2000.Gender and hormonal status affect the hypolipidemic mechanisms of dietary soluble fiber in guinea pigs.J Nutr, 130: 600-7.

  The functions of polysaccharides such as dietary fiber, which have been known so far, include improvement of bowel movement and blood cholesterol lowering action. The purpose of the present invention is to clarify new functions other than those described above, To find new uses for dietary fiber.

The inventors of the present invention conducted extensive research to solve the above-mentioned problems. As a result, it is not a conventional method for measuring blood cholesterol concentration and triglyceride concentration, but in a tissue using a DNA chip. By monitoring gene expression changes, the effects of dietary fiber known in the past, i.e., improvement of stool quality, blood cholesterol lowering effect by preventing the absorption of bile acids, etc., are found to promote lipid metabolism in muscle tissue Succeeded.
That is, the present invention is a lipid metabolism and / or lipid transport promoter containing a polysaccharide.

  The promoter of the present invention has an action of promoting lipid metabolism and / or lipid transport in muscle tissue. Examples of the polysaccharide include at least one selected from the group consisting of psyllium, guar gum, tara gum, tamarind seed gum, and chitosan. Among these, psyllium is particularly preferable. In addition, the promoter of the present invention can vary the expression level of genes related to lipid metabolism and / or lipid transport in muscle tissue. Examples of genes related to lipid metabolism and / or lipid transport include at least one selected from the group consisting of Lpl, Cd36, Cact, Acsl1, Acss1, Acadl, Acadm, Acads, Hadh, Hadha, Hadhb, Cpt1b, and Slc2a1. Can be mentioned.

Furthermore, the present invention is a composition for suppressing fat accumulation, which contains a polysaccharide and has an action of promoting lipid metabolism and / or lipid transport in muscle tissue.
Furthermore, the present invention is a composition for suppressing fat accumulation, containing psyllium and having an action of promoting fat metabolism and / or lipid transport in muscle tissue.
Furthermore, the present invention is a composition for improving and / or preventing obesity comprising a polysaccharide and having an action of promoting lipid metabolism and / or lipid transport in muscle tissue.
Furthermore, the present invention is a composition for treating and / or preventing metabolic syndrome comprising a polysaccharide and having an action of promoting lipid metabolism and / or lipid transport in muscle tissue.
The composition can also be used for dieting.

  In the present invention, polysaccharides such as dietary fiber have an action of promoting lipid metabolism and / or lipid transport in muscle tissue, and the effect thereof is excellent in suppressing fat accumulation. Therefore, the polysaccharide can be expanded as a composition for inclusion in a food ingredient for promoting lipid metabolism and / or lipid transport, or for a purpose of suppressing fat accumulation in food. In addition, it is possible to provide a food component and a food intake method capable of preventing and improving obesity and, in turn, preventing and improving lifestyle-related diseases by an anti-obesity action utilizing the lipid metabolism promoting effect in muscle tissue.

It is a figure which shows the body weight gain inhibitory effect of the composition of this invention. It is a figure which shows the result of having measured total cholesterol concentration and blood triglyceride concentration. It is a figure which shows the gene expression change in a liver. It is a figure which shows the gene expression change in a muscle. It is a schematic diagram which shows the gene expression pathway in a cell. It is a figure which shows the result of having measured the epididymal fat mass and the retroperitoneal fat mass.

  Hereinafter, the present invention will be described in detail. The scope of the present invention is not limited to these descriptions, and other than the following examples, the scope of the present invention can be appropriately changed and implemented without departing from the spirit of the present invention.

1. The polysaccharide polysaccharide means a carbohydrate in which a large number of monosaccharide molecules are linked by glycosidic bonds. Secreted by plant cell walls (cellulose, pectin, etc.), arthropod or fungal exoskeleton (chitin, etc.), algae cells (agarose, agar, carrageenan, etc.), microorganisms There are those derived from gel-like substances (such as xanthan gum). Non-digestible or difficult to digest polysaccharides are treated as dietary fiber. Dietary fiber can be roughly classified into two types: soluble polysaccharides and insoluble polysaccharides. Soluble polysaccharides form a sticky gel when mixed with liquid, and insoluble polysaccharides are digestive pathways. Will pass through almost unchanged.

In the present invention, plant-derived polysaccharides or soluble polysaccharides derived from animals or microorganisms can be mainly used. Examples of such polysaccharides include soybean polysaccharides, agar, pectin, guar gum, gum arabic, xanthan gum, carrageenan, tamarind seed gum, alginic acid, pullulan, locust bean gum, carboxymethylcellulose, psyllium, curdlan, gellan gum and mannans. (Glucomannan, galactomannan, etc.) are included, but are not limited thereto.
In the present invention, insoluble polysaccharides can also be used. Insoluble polysaccharides include chitin, chitosan, cellulose, hemicellulose, etc. Among these, chitin and chitosan are more preferable.

As dietary fiber as listed above, commercially available fibers can also be used, and specific examples are given below.
The soybean polysaccharide is a water-soluble polysaccharide that can be extracted from soybean or a processed soybean product, and can be obtained, for example, by crushing soybean beans and then extracting a water-soluble fraction. Moreover, it is possible to use a commercial item (Fuji Oil Co., Ltd.).
Agar is a polysaccharide composed of mucus of red algae such as Tengusa, and can be obtained, for example, by freeze-drying mucus of Tengusa. Commercial products (Wako Pure Chemical Industries, Ltd.) can also be used.
Pectin is a component consisting of complex polysaccharides combined with other components such as cellulose as a cell wall constituent of many plants, and can be obtained, for example, by hot water extraction from citrus peel. Commercial products (Wako Pure Chemical Industries, Ltd.) can also be used.
Gua gum is a water-soluble dietary fiber obtained from legume guar seeds and endosperm, and can be obtained, for example, by extracting a water-soluble fraction from legume guar. Moreover, it is possible to use a commercial item (MRC polysaccharide company).

Gum arabic is a water-soluble dietary fiber mainly composed of a polysaccharide (arabinogalactan) obtained from the arabic gum. For example, it can be obtained by drying secretions from the bark of Arabidopsis. Moreover, it is possible to use a commercial item (San-Eigen FFI Co., Ltd.).
Xanthan gum is a polysaccharide obtained by treating starch obtained from corn or the like, and can be obtained, for example, by fermenting starch derived from corn. Moreover, it is possible to use a commercial item (MRC polysaccharide company).
Carrageenan is a linear sulfur-containing polysaccharide derived from red algae, and can be obtained, for example, by alkaline extraction from red algae. Moreover, it is possible to use a commercial item (MRC polysaccharide company).

Tamarind seed gum is a natural polysaccharide obtained from the endosperm of leguminous tamarind, and can be obtained, for example, by hot water or alkali extraction from a tamarind plant. Moreover, it is possible to use a commercial item (MRC polysaccharide company).
Alginic acid is a dietary fiber contained in brown algae and can be obtained, for example, by alkaline extraction from kombu or seaweed. Moreover, it is possible to use a commercial item (San-Eigen FFI Co., Ltd.).
Pullulan is a polysaccharide using starch as a raw material, and can be obtained, for example, by fermentation of aureobasidium pullulans, which is black yeast, using starch as a raw material. Commercial products (Hayashibara Shoji Co., Ltd.) can also be used.

Locust bean gum is a polysaccharide obtained from the seeds of evergreen carob trees, and can be obtained, for example, by pulverizing the endosperm portion of the seeds of carob trees. Moreover, it is possible to use a commercial item (MRC polysaccharide company).
Tara gum is a kind of nonionic natural polysaccharide obtained from the endosperm part of legume plant cod, and it is possible to use a commercially available product (MRC polysaccharide).

Carboxymethylcellulose is a derivative of cellulose and can be obtained by the reaction of cellulose and chloroacetic acid. Commercial products (Wako Pure Chemical Industries, Ltd.) can also be used.
Psyllium is a water-soluble dietary fiber purified from the outer skin of the plant family Psyllium, which can be obtained by extraction from the outer skin of psyllium. Moreover, it is possible to use a commercial item (MRC polysaccharide company).

Curdlan is a fermented polysaccharide produced by microorganisms and can be obtained by fermentation of bacteria such as Agrobacterium and Alcaligenes. Commercial products (Kirin Kyowa Foods Co., Ltd.) can also be used.
Gellan gum is a polysaccharide produced by microorganisms, and can be obtained, for example, by cultivation of Pseudomonas. Moreover, it is possible to use a commercial item (San-Eigen FFI Co., Ltd.).
Glucomannan is a water-soluble polysaccharide that is abundant in konjac koji. It accounts for about 40% of the dry weight of konjac koji and can be obtained by extracting from this. A commercial product (Mitsubishi Corporation Foodtech) can be used.
Chitosan is a polysaccharide obtained by deacetylating chitin (made of poly-β1 → 4-N-acetylglucosamine) obtained from crustacean exoskeletons such as crabs and shrimps by boiling treatment in concentrated alkali. Commercial products (Hokkaido Soda) can also be used.

  In the present invention, among the above dietary fibers, psyllium, guar gum, tara gum, tamarind seed gum, chitosan, or a combination thereof is preferable.

2. Functional food and drink and functional composition (pharmaceutical composition)
In the present invention, a polysaccharide such as dietary fiber is used as a composition for dietary food or drink or functional food, or for fat accumulation suppression, obesity improvement (and / or prevention), or treatment (and / or prevention) of metabolic syndrome. Can be used as a functional composition or a pharmaceutical composition. In particular, the composition of the present invention is preferably used as a diet food. When using the composition of this invention as food-drinks, it is preferable to set it as the form as a health food, for example, a low-calorie food. The health foods also include foods and drinks (for example, functional foods such as foods for specified health use) with a label indicating that they have an action to suppress fat accumulation and improve obesity on containers and instructions.

In the present invention, “diet” means to suppress weight gain due to intake of a high fat diet, and the composition of the present invention can be used for the suppression.
Many low-calorie foods such as konjac, agar, mushrooms and seaweed are known as diet foods. And it can standardize so that other nutrients (for example, protein, a vitamin, mineral, etc.) may fully be contained in these foodstuffs.

  Further, “inhibition of fat accumulation” means that an increase in visceral fat (posterior peritoneal fat and epididymal fat) due to intake of a high fat diet is suppressed. It can be determined that fat accumulation is suppressed when there is a difference in the weight of retroperitoneal fat and epididymal fat compared to the intake of a high fat diet. Therefore, the composition of the present invention can be used for the purpose of the suppression.

  In the present invention, when the composition of the present invention is orally administered for the purpose of inhibiting fat accumulation, improving obesity or treating metabolic syndrome, solid preparations such as tablets, granules, powders, pills and the like applied thereto Alternatively, liquid preparations such as liquids and syrups may be used. In particular, granules and powders can be made into unit dosage forms as capsules, and in the case of liquid preparations, they can be made into dry products (eg freeze-dried products) that are redissolved when used.

  Among these dosage forms, the oral solid preparation contains additives such as a binder, an excipient, a lubricant, a disintegrant, and a wetting agent that are generally used in the composition. The oral liquid preparation contains additives such as a stabilizer, a buffering agent, a corrigent, a preservative, a fragrance, and a coloring agent that are generally used in the composition.

  In the present invention, when the polysaccharide is used as a functional composition or a pharmaceutical composition, the dose of the composition of the present invention is 1 mg / kg to 15 g / kg, preferably 20 mg / kg to 5 g / kg per day. And can be administered from one day to several weeks, months or years.

3. Food, Beverage and Feed Since the composition of the present invention can be administered orally, it can also be used as a food, beverage or feed (preferably as a diet or functional food, beverage or feed). In this case, the composition of the present invention can be eaten as it is or contained in an enteric capsule for administration. Or it can melt | dissolve in liquid (for example, water) so that it may become a suitable density | concentration, and it can add to foodstuff etc. by methods, such as mixing, immersion, application | coating, and spraying.

In the food, beverage and feed of the present invention, the content ratio of the polysaccharide contained in the composition of the present invention is not limited and can be appropriately set according to various uses. Moreover, also when obtaining the foodstuff, drink, or feed of this invention, in the well-known manufacturing method according to various uses, a polysaccharide can be contained by arbitrary methods or timing.
The form of the polysaccharide to be contained in the composition is not limited and may be a slurry or a powder, and may be appropriately selected according to various uses.

  The food or beverage of the present invention (hereinafter referred to as “food”) is not particularly limited, and includes any food that can be eaten by humans. Examples include flour-based foods, meat (processed meat), marine products, vegetable protein-containing foods (soy foods and soy protein-containing foods), dairy products, confectionery, other foods, or foods obtained by instantly processing these foods It is done.

Examples of foods using flour include breads such as bread and rolls, cereals, crackers, biscuits, hot cakes, cakes such as castella, udon, soba, Chinese soba, raw noodles, various pasta (spaghetti, macaroni, penne, fittine) Etc.), noodles such as rice noodles, pizza dough, and naan.
Examples of meat include hamburger, meatballs, ham, and wiener.
Examples of marine products include seafood and seaweeds.
Examples of vegetable protein-containing foods include tofu and soy milk.
Examples of dairy products include milk, cheese, yogurt and the like.
Examples of confectionery include rice cake, pudding, jelly, jam, curry, ice cream, sorbet, and gelato.
Examples of other foods include diet foods such as konjac jelly, creams, seasonings (miso, soy sauce, sauce, mayonnaise, dressing, etc.), and various supplements (health foods, tablets, tablets, etc.).

Examples of the beverage include, but are not limited to, mineral water, vegetable juice, fruit juice, tea (green tea, tea, oolong tea, barley tea, etc.), soft drink, soup, coffee, milk drink, and nourishing tonic drink. .
Examples of feed include, but are not limited to, livestock feed, pet food, and other feeds. The pet food can target, for example, obese cats, obese dogs and the like. Examples of livestock food include foods such as cows, pigs, horses and birds.

  The amount of the composition of the present invention added to these foods or feeds can be selected according to the kind of foods or feeds to be added. However, if the amount added is too small, the expected fat accumulation suppression, obesity improvement, diet effect, metabolic syndrome treatment effect, and other lifestyle-related disease improvement effects cannot be fully exhibited. On the other hand, if the amount added is too large, the flavor inherent to the food to be added may be impaired. Therefore, it is preferable to determine the addition amount in consideration of these circumstances.

4). Lipid metabolism and / or lipid transport promotion effect in muscle tissue To measure the action of promoting lipid metabolism and / or lipid transport in muscle tissue, for example, homogenize skeletal muscle and measure β-oxidation activity using palmitic acid However, it can also be measured by comparing a change in gene expression level (increase or decrease in expression) in muscle tissue with a control group. For example, the number of genes whose expression level is to be examined (for example, 205 if the number of genes is 205) or less, the significance level is 5% or 1% in the repetition of the test, and at least 3 samples that are at least independent in one group In the comparison between the groups including, those that are statistically significant compared to the control group (control) can be determined as “changed”. As a technique for measuring gene expression changes, techniques known to those skilled in the art such as Northern blotting, real-time PCR, and DNA chip can be used. It is more preferable to use a DNA chip in view of capturing a large number of genes simultaneously.

  Lipid metabolism refers to the action of degrading triglycerides by lipase on the cell surface, the action of fatty acids released from triglycerides being taken into the cells and breaking them down, and the fatty acids being taken up by mitochondria and then degraded by β-oxidation To act.

The genes to be measured are preferably major enzymes related to lipid metabolism, and the following genes can be exemplified.
Acsm1 (acyl-CoA synthetase medium-chain family member 1) (Accession No.:NM_054094),
Acsl1 (acyl-CoA synthetase long-chain family member 1) (Accession No.:NM_007981),
Acss1 (acyl-CoA synthetase short-chain family member 1) (Accession No.:_NM_080575),
Mdh1 (malate dehydrogenase 1, NAD (soluble)) (Accession No.:NM_008618),
Idh2 (isocitrate dehydrogenase 2 (NADP +), mitochondrial) (Accession No.:NM_173011),
Hadh (hydroxyacyl-Coenzyme A dehydrogenase nuclear gene encoding mitochondrial protein) (Accession No.:NM_008212),
Hadha (hydroxyacyl-Coenzyme A dehydrogenase / 3-ketoacyl-Coenzyme A thiolase / enoyl-Coenzyme A hydratase (trifunctional protein), alpha subunit) (Accession No.:NM_178878),
Hadhb (hydroxyacyl-Coenzyme A dehydrogenase / 3-ketoacyl-Coenzyme A thiolase / enoyl-Coenzyme A hydratase (trifunctional protein), beta subunit, nuclear gene encoding mitochondrial protein) (Accession No.:NM_145558),
Acadm (acyl-Coenzyme A dehydrogenase, medium chain) (Accession No.:NM_007382),
Acadl (acyl-Coenzyme A dehydrogenase, long-chain) (Accession No .: NM_007381),
Acads (acyl-Coenzyme A dehydrogenase, short chain) (Accession No .: NM_007383), and
Lpl (lipoprotein lipase) (Accession No .: NM_008509).

  Furthermore, in the present invention, it is more preferable to monitor the following genes related to lipid transport together with monitoring of the enzyme-related genes.

  Lipid transport refers to an action of taking in fatty acids released from triglycerides decomposed by lipase on the cell surface into cells, or an action of taking up into mitochondria because the fatty acids taken into cells are oxidized.

Cpt1b (carnitine palmitoyltransferase 1b, muscle) (Accession No.:NM_009948),
Cpt2 (carnitine palmitoyltransferase 2) (Accession No.:NM_009949),
Slc2a1 (solute carrier family 2 (facilitated glucose transporter), member 1) (Accession No.:NM_011400),
Cd36 (CD36 antigen) (Accession No.:NM_007643), and
Cact (solute carrier family 25 (mitochondrial carnitine / acylcarnitine translocase), member 20, nuclear gene encoding mitochondrial protein) (Accession No .: NM_020520).

  In the present invention, when the expression of the following gene is increased as compared with the subject, it can be determined that lipid metabolism and / or lipid transport is promoted.

Lpl (lipoprotein lipase) (Accession No.:NM_008509), which has the action of degrading triglycerides on the cell surface,
Cd36 (CD36 antigen), which has an action of taking in fatty acids released from triglycerides decomposed by lipase on the cell surface into cells (Accession No.:NM_007643),
Cact (solute carrier family 25 (mitochondrial carnitine / acylcarnitine translocase), member 20, nuclear gene encoding mitochondrial protein) (Accession No .: NM_020520)
Acsl1 (Acyl-CoA synthetase long-chain family member 1) (Accession No.:NM_007981), which promotes the degradation of fatty acids by forming a double bond from methylene dehydrogenation of acyl group
Hadh (hydroxyacyl-Coenzyme A dehydrogenase nuclear gene encoding mitochondrial protein) (Accession No.:NM_008212), Hadha (hydroxyacyl-Coenzyme A dehydrogenase / 3-ketoacyl-Coenzyme A thiolase / enoyl-Coenzyme A hydratase (trifunctional protein), alpha subunit) (Accession No.:NM_178878), Hadhb (hydroxyacyl-Coenzyme A dehydrogenase / 3-ketoacyl-Coenzyme A thiolase / enoyl-Coenzyme A hydratase (trifunctional protein), beta subunit, nuclear gene encoding mitochondrial protein ) (Accession No .: NM_145558), Acadm (acyl-Coenzyme A dehydrogenase, medium chain) (Accession No .: NM_007382), Acads (acyl-Coenzyme A dehydrogenase, short chain) (Accession No .: NM_007383).

In the present invention, the following genes having functions similar to those of the above genes may be included.
Slc2a1 (solute carrier family 2 (facilitated glucose transporter), member 1) (Accession No.:NM_011400),
Cpt1b (carnitine palmitoyltransferase 1b, muscle) (Accession No.:NM_009948), Cpt2 (carnitine palmitoyltransferase 2) (Accession No.:NM_009949),
Acss1 (acyl-CoA synthetase short-chain family member 1) (Accession No.:_NM_080575),
Acadl (acyl-Coenzyme A dehydrogenase, long-chain) (Accession No .: NM_007381).

  In the present invention, among the genes listed above, one or more of Lpl, Cd36, Cact, Acsl1, Acss1, Acadl, Acadm, Acads, Hadh, Hadha, Hadhb, Cpt1b, and Slc2a1 are analyzed in combination Is preferred.

  Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to these examples.

Lipid metabolism promoting effect test in muscle tissue and confirmation of known cholesterol lowering effect A feed intake group was set as described in Table 1, and a lipid metabolism promoting effect experiment using mice was conducted.

  The experimental animal used for the experiment was a 5-week-old C57BL / 6J male mouse (purchased from Charles River, Japan). All of these mice were reared between groups in rat cages, and food and water were ad libitum on a 12 hour light / dark cycle.

First, after acclimatizing the above mice with CE-2 solid feed (purchased from Nippon Claire) for 7 days, the mice were divided into the groups shown in Table 1 so that the average body weights were equal. went.
For each group, a feed according to the formulation shown in Table 2 below was prepared and ingested over a period of 10 weeks to perform the main breeding. Pyrium was corrected with cellulose during feed preparation. Correction means that if psyllium is not used, instead of them, the control (control) and the whole group are adjusted to the same mass of feed. It should be noted that both drinks and feeds were taken freely, and body weight and feed intake were measured every other day.
Ten weeks after the main breeding, pentobarbital anesthesia was intraperitoneally administered at 0.13 ml / 100 g body weight, and then the mice were dissected. All mice were fasted for 16 hours before dissection.
During the dissection, blood was collected by collecting blood, and liver, skeletal muscle (thigh muscle, gastrocnemius), epididymal fat, and retroperitoneal fat were collected.

The following items were measured using the collected samples.
-Adipose tissue weight-Blood triglyceride concentration: Measured by an enzymatic method using a measurement kit of Triglyceride E-Test Wako (Wako Pure Chemical Industries).
-Total cholesterol concentration in blood: Measured with cholesterol E-Test Wako (Wako Pure Chemical Industries).

Weight gain inhibitory effect The group (Group I) ingested high fat diet + psyllium was found to have an effect of inhibiting weight gain compared to the case of ingesting a high fat diet (Group B). Due to this effect, the weight at the 10th week was almost the same as that of the normal diet group (Group A) despite the high fat diet in Group I (Table 3).

<Blood collection and blood component measurement results>
Using serum collected by performing the above-described blood sampling, blood cholesterol and triglyceride concentrations were immediately measured. The results are shown in Table 4 and FIG. These were analyzed using Tukey's multiple comparison method. The critical value for rejection was q (3,8; 0.05) ÷ √2 = 2.857. As a result, the total cholesterol concentration was significantly lower in Group I than in Group B, and the blood triglyceride concentration was also significantly lower in Group I than in Groups A and B. Was obtained (Table 5).
That is, as is conventionally known, cholesterol-lowering action and triglyceride-lowering action were observed in group I in which psyllium, a kind of dietary fiber, was added to the diet.

<Changes in gene expression with DNA chip>
Three samples were randomly extracted from 4 samples in each group. These were designated as A1 to A3, B1 to B2, and I1 to I3, and used for gene expression analysis.
Total RNA was extracted and purified from these samples by column extraction (using Qiagen RNeasy Mini column). Using 1 μg of the obtained total RNA, biotin labeling and amplification reaction were performed with MessageAmp-Biotin Enhanced Kit (Ambion) according to the instructions disclosed by the manufacturer.

Acquisition of expression level data by DNA microarray Using DNA chip Genopearl manufactured by Mitsubishi Rayon Co., Ltd., a DNA chip carrying 205 kinds of oligo DNA probes listed in Tables 6 to 10 was prepared.
The above-mentioned biotin-labeled amplified RNA (5 μg) was fragmented by heating at 94 ° C. for 7 minutes 30 seconds using the 5 × Fragmentation Buffer attached to the kit.
Various reagents were added to the fragmented biotin-labeled RNA solution so that the final composition was 0.12 M Tris-HCl / 0.12 M NaCl / 0.05% Tween 20, and 150 μl of a hybridization sample was obtained.

  The DNA chip and the hybridization sample were reacted at 65 ° C. for 16 hours, and then washed twice at 65 ° C. for 20 minutes with a washing solution (0.12M Tris-HCl / 0.12M NaCl / 0.05% Tween 20). . Further, the DNA chip was replaced with a storage solution (0.12M Tris-HCl / 0.12M NaCl) at 65 ° C. for 10 minutes.

  The DNA chip after washing was immersed in Streptavidin-Cy5 (GE Healthcare BioScience KK) dissolved in 2 μg / ml with a stock solution for 30 minutes at room temperature, and then washed with the washing solution for 5 minutes at room temperature four times. The solution was replaced with the preservation solution for 5 minutes.

The DNA chip after Cy5 labeling uses a dedicated DNA chip detector to detect fluorescence at a plurality of exposure times from 0 seconds to 40 seconds, and in each spot, among the images that do not contain saturated pixels in the spot, Using the image with the longest exposure time, it was quantified as the fluorescence signal intensity converted per unit time.
Tables 6-10 show the genes and probes mounted on the DNA chip. Note that numbers 1 to 205 at the left end of Tables 6 to 10 correspond to SEQ ID NOs 1 to 205, respectively.

<Data analysis method>
The detected data was analyzed as follows.
The median value of blank spots (spots not equipped with probes) was subtracted from the detected fluorescence intensity data. Hereinafter, this value is referred to as signal intensity. Moreover, the standard deviation of the blank spot was calculated. Data showing signal intensity below the standard deviation of the blank spot was excluded from the analysis.
The internal control gene was Arbp. A chip having the highest Arbp signal intensity between the chips to be compared was selected, and an Arbp signal intensity ratio between the chips was calculated to obtain a correction coefficient between the chips. The signal intensity was corrected by multiplying the spot signal intensity of each chip by a correction coefficient.

For the data after correction, the average value of the signal intensity of the control group (Group A) was taken, and the signal intensity of each chip was divided by this intensity, and a log with a base of 2 was taken for that value. This was taken as the expression ratio.
Based on the expression ratio, numerical analysis was performed. In particular, numerical analysis of changes in gene expression in muscle was performed using Dunnet's multiple comparison method with Group B as a control. Table 11 shows the statistically significant differences. In addition, clustering was performed for those judged to be significant. The distance function of clustering used Pearson's distance coefficient.

<Changes in gene expression in the liver>
As is known so far, the expression of Hmgcr and Sqle, which are major enzymes of cholesterol synthesis genes, was found to compensate for cholesterol lost due to the effects of dietary fiber. In addition, in order to avoid the loss of cholesterol from the liver, the expression of the Abcb11 gene, which is a bile excretion transporter from the liver to the intestinal tract, decreased, and suppression of cholesterol outflow from the liver to the intestinal tract was observed. Furthermore, the expression of Abcg1 and Abcg4 genes decreased, and the suppression of cholesterol withdrawal from the liver by HDL was observed. This result confirmed the effect of dietary fiber known so far (FIG. 3). FIG. 3 shows the results of analysis of expression fluctuations of the above five genes using Dunnett's multiple comparison method with Group B as a control. <0.01).

Effect of promoting lipid metabolism in muscle tissue <Change in gene expression in muscle>
As a result of comparing Group B and Group I, the genes determined to be significant are summarized in Table 11 and FIG. Moreover, based on the function of these genes, the pathway regarding the said function was shown in FIG.

As a result, the expression level of genes related to lipolysis was significantly increased in group I compared to the high fat diet group (group B), indicating that the utilization efficiency of lipid was promoted. (*: P <0.05, **: p <0.01)
By repeating the Dunnett's test for each of the 205 genes, an average of about 10 genes can be considered when the significance level is 0.05, and an average of about 2 genes when the significance level is 0.01. It was thought that lipid metabolism was promoted from the biological function of the gene group judged to be significant even after examining various circumstances.

<Fat accumulation inhibitory effect by promoting lipid metabolism in muscle tissue>
Intra-abdominal fat (epididymal fat, retroperitoneal fat) of dissected mice was collected and weighed, and the high-fat diet group (Group B) and the group fed with high-fat diet + psyllium (Group I) were compared. (Table 12, FIG. 6).

The comparative analysis used Tukey's multiple comparison method. The rejection limit value is q (3,9; 0.05) ÷ √2 = 2.791. As a result, both epididymal fat and posterior peritoneal fat were significantly different between Group B and Group I, and the effect of inhibiting fat accumulation was observed in Group I (Table 13). (*: P <0.05)
This effect shows that the accumulation of fat can be suppressed to the same level as the normal diet (Group A), despite the fact that the Group I ingested dietary fiber has a high fat diet. there were.

  From the results of this Example, it was found that the lipid metabolism promoting effect of dietary fiber in muscle tissue has the effect of simultaneously suppressing weight gain and fat accumulation. Therefore, it was found that the dietary fiber used in the present invention contributes to the suppression of obesity and lifestyle-related diseases such as metabolic syndrome in addition to the cholesterol lowering action.

  Since the lipid metabolism promoter in muscle tissue of the present invention has the effect of suppressing weight gain and fat accumulation, it can be used as a functional food, drink or feed, or as a diet food, drink or feed, and further, can suppress fat accumulation. It is useful as a composition for obesity, a composition for improving and / or preventing obesity, and a composition for treating and / or preventing metabolic syndrome.

SEQ ID NOs: 1 to 205: synthetic DNA

Claims (9)

  A lipid metabolism and / or lipid transport promoter containing a polysaccharide.   The promoter according to claim 1, which has an action of promoting lipid metabolism and / or lipid transport in muscle tissue.   The promoter according to claim 1 or 2, which can vary the expression level of a gene related to lipid metabolism and / or lipid transport in muscle tissue. The gene related to lipid metabolism and / or lipid transport is at least one selected from the group consisting of Lpl, Cd36, Cact, Acsl1, Acss1, Acadl, Acadm, Acads, Hadh, Hadha, Hadhb, Cpt1b and Slc2a1. The accelerator according to claim 3. The promoter according to claim 1 or 2, wherein the polysaccharide is at least one selected from the group consisting of psyllium, guar gum, tara gum, tamarind seed gum and chitosan.   A composition for suppressing fat accumulation, comprising a polysaccharide and having an action of promoting lipid metabolism and / or lipid transport in muscle tissue.   A composition for suppressing fat accumulation, comprising psyllium and having an action of promoting fat metabolism and / or lipid transport in muscle tissue.   A composition for improving and / or preventing obesity comprising a polysaccharide and having an action of promoting lipid metabolism and / or lipid transport in muscle tissue.   A composition for treating and / or preventing metabolic syndrome comprising a polysaccharide and having an action of promoting lipid metabolism and / or lipid transport in muscle tissue.