JP6241644B2 - PPARγ expression improving agent, basal metabolism improving agent, muscle fatigue recovery improving agent, pharmaceutical composition for improving PPARγ expression, and food and drink for improving PPARγ expression - Google Patents

Description

  The present invention relates to a PPARγ expression improver containing agarooligosaccharide made from agar or agar-producing raw material, and a basal metabolism improver, fatigue recovery improver, pharmaceutical composition and food and drink containing the same.

  In recent years, metabolic syndrome, locomotive syndrome, sarcopenia, and the like have been regarded as problems, and as a result, health consciousness has increased and awareness regarding exercise has increased. Opportunities to move the body have decreased due to the development of transportation means and communication technology, resulting in lack of exercise, obesity, muscle weakness, and motor function decline. Exercise improves these problems, but it is difficult to get an effect in a short time. In a busy modern society, it is desired to exercise efficiently and recover quickly from fatigue. Furthermore, improving basal metabolism, making lipid burning natural, and being able to live an active life is one way to solve these problems. In order to solve these problems, it is necessary to develop a material that can improve basal metabolism by ingesting easily orally and accelerate recovery from fatigue due to exercise.

  Agar is dehydrated and dried mucilage extracted from red algae seaweeds such as prickly pear and ogonori. The structure is a polysaccharide composed of agarose and agaropectin. The structure of agarobiose, which is a repeating unit of agarose, consists of β-D-galactopyranose bonded at the 1,3-position and 3,6 anhydro-α-L-galactopyranose bonded at the 1,4-position. Agaropectin is a general term for ionic polysaccharides other than agarose in agar, and its structure is the same as that of agarose, but partially contains sulfate, methoxyl, pyruvate, and carboxyl groups. Contains a lot. The weight average molecular weight (Mw) is generally 200,000 to 400,000. Agaro-oligosaccharides (agarobiose, agarotetraose, agarohexaose, agarooctaose, etc.) produced by degrading this agar with acid or enzyme include systemic lupus erythematosus, immune-mediated glomerulonephritis, It is described that it is a prophylactic agent for various diseases such as multiple sclerosis, collagen disease, autoimmune disease, rheumatism, and it is further described that it is an antioxidant that suppresses active oxygen production (patent) Reference 1). It also describes that skin and bone aging is prevented by inhibiting metalloprotease production (Patent Document 2) and that there is a blood glucose lowering effect (Patent Document 3).

Patent No. 4007760 JP 2011-213707 A JP 2004-149471 A

  It is an object of the present invention to find that agarooligosaccharides have completely different actions from those conventionally known, and to provide a pharmaceutical composition and a food or drink using the same. That is, in the present invention, it is found that by ingesting agarooligosaccharide, the expression of PPARγ is improved, and as a result, basal metabolism is improved and a fatigue recovery effect can be obtained. Health improvement, slimming, and muscle strength improvement by improving basal metabolism It is intended to provide pharmaceutical compositions and foods and drinks for the purpose of improving health, slimming, improving muscular strength, etc., as a result of improvement in QOL, etc., and further fatigue recovery effect. .

  As a result of intensive studies to achieve the above object, the present inventors have found that, as an action not conventionally known for agarooligosaccharides, there are an action for improving basal metabolism and an action for improving fatigue recovery by improving the expression of PPARγ. The present invention has been reached.

  That is, the present invention relates to a PPARγ expression improver comprising as an active ingredient at least one sugar compound selected from the group consisting of agarobiose, agarotetraose, agarohexaose and agarooctaose.

  The present invention also relates to a basal metabolism improver or fatigue recovery improver characterized by containing the above PPARγ expression improver.

  Moreover, this invention relates to the pharmaceutical composition or food-drinks characterized by including said PPAR (gamma) expression improving agent.

  As described above, according to the present invention, by improving the expression of PPARγ, basal metabolism is improved and fatigue recovery effect is obtained. Health improvement, slimming, muscle strength improvement, QOL improvement, etc. by basal metabolism improvement, Furthermore, a fatigue recovery effect enables exercise to be continued, and as a result, a basal metabolism improver and fatigue recovery improver capable of improving health, slimming, and improving muscle strength, and a pharmaceutical composition and food and drink containing them. Can be provided.

It is the graph which showed the change of the expression level of PPARγ by changing the agarooligosaccharide concentration. It is the graph which showed the result of the basal metabolism improvement test by agarooligosaccharide intake. It is the graph which showed the result of the fatigue recovery improvement test by agarooligosaccharide intake. It is the graph which showed the accumulation | storage suppression effect of fat by addition of agarooligosaccharide.

Agaro-oligosaccharides are composed of repeating agarobiose (disaccharide), and are composed of agarotetraose (tetrasaccharide), agarohexaose (hexasaccharide), agarooctaose (octasaccharide), agarodecaose (10 sugars) and the like. ing. In the present invention, agarooligosaccharide, which is a sugar compound characterized by containing at least one of these, is used. The composition of agarooligosaccharide can be measured using high performance liquid chromatography. Measurement conditions are column TOSOH TSK-GEL ALPHA-2500 connected in series, elution under the conditions of solvent H ₂ O, flow rate 0.3 ml / min, temperature 60 ° C., detection can be examined by RI (differential refraction). .
The agarooligosaccharide can be produced using agar as a raw material, or can be produced using a solution obtained by hot water extraction of red algae such as Tengusa and Ogonori, which are raw materials for agar.

In the present invention, agarooligosaccharide can be produced, for example, by hydrolyzing agar. Specifically, as an acid decomposition method, the solid acid described in Japanese Patent No. 4796697 is used, a mineral acid such as sulfuric acid or hydrochloric acid is used, or an organic acid such as acetic acid or citric acid is used. However, any method may be used. A product produced by hydrolysis with an acid is called an agarooligosaccharide.
On the other hand, what is produced by enzymatic degradation with β-agarase is called neoagaro-oligosaccharide because of the difference in the cleavage site of the agar molecular main chain. Those produced by enzymatic degradation with α-agarase become agarooligosaccharides as in the case of hydrolysis with acid. However, in the present invention, both agarooligosaccharide and neoagarooligosaccharide exhibit the same effect. Therefore, in the present invention, both are hereinafter collectively referred to as agarooligosaccharides.

(Peroxisome proliferator-responsive receptor γ (PPARγ) expression improving effect)
The reason why basal metabolism is improved and the fatigue recovery effect is increased by ingesting agarooligosaccharide is as follows. Ingestion of agarooligosaccharide increases glucose uptake into cells in the metabolic system and promotes the TCA cycle in mitochondria. This can be explained by the fact that peroxisome proliferator-responsive receptor γ (PPARγ) gene is activated and its expression level increases. That is, the present invention can be said to be a PPARγ expression improver.
Here, peroxisome proliferator-responsive receptor γ (PPARγ) is a nuclear receptor protein belonging to the steroid receptor superfamily and is a γ subtype protein. PPARγ promotes glucose uptake from blood into cells in fat cells and peripheral tissues such as muscle and liver.
In the present invention, the effect of improving the expression of PPARγ can be evaluated not only in humans but also in mice. The expression of PPARγ can be quantitatively measured using RT-PCR or the like for mRNA complementary to the gene encoding PPARγ (PPARG). The expression level of PPARγ is preferably 1.5 times to 2000 times, and more preferably 2 times to 1000 times that when not taking agarooligosaccharide.

(Improvement of basal metabolism)
As described above, by improving glucose uptake into cells, sugar metabolism by the TCA cycle in mitochondria, which is an organ related to energy metabolism, becomes active. As a result, energy production increases and basal metabolism improves.
In the present invention, the effect of improving basal metabolism is that the oxygen consumption measured using metavaine (manufactured by Vine) is improved by 1.05 to 1.5 times compared to when not taking agarooligosaccharide. Say.
Moreover, in this invention, in order to exhibit the basal metabolism improvement effect most, it is preferable to ingest 50-5000 mg / day of agarooligosaccharide, and 100-5000 mg / day is more preferable. However, the intake is not limited to this amount.

(Fatigue recovery improvement effect)
Two pathways are known for sugar metabolism, one is a glycolytic system that does not require oxygen, and the other is a TCA cycle that requires oxygen. In the glycolytic system, an anaerobic reaction is performed using glycogen as a base, and it becomes a source of energy for rapid exercise, and the fatigue substance lactic acid accumulates in the muscle. On the other hand, in the TCA cycle, glucose is converted into carbon dioxide and water by an aerobic reaction to obtain energy, and lactic acid is not produced. This reaction takes place in the mitochondria.
Generally, muscle fatigue is said to be due to accumulation of lactic acid in the muscle by exercise. When glucose uptake into cells is increased by ingestion of agarooligosaccharide, energy production by TCA cycle is higher than glycolysis, and production of lactic acid by glycolysis is reduced, resulting in a fatigue recovery effect. As described above, the glycolysis system has the advantage that it can quickly supply the energy required for rapid exercise. However, when the production rate of pyruvic acid by the glycolytic system is fast and exceeds the treatment rate of pyruvic acid by mitochondria, pyruvic acid is converted to lactic acid. In this way, the glycolytic system produces lactic acid, which causes muscle fatigue and muscle pain.
In contrast, when the expression of PPARγ is improved, the ratio of glycolysis to sugar metabolism by the TCA cycle increases. This is also clear from non-patent prior literature (THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 286, No. 46, pp. 40013-40024, November 18, 2011). As a result, accumulation of lactic acid in the body is reduced, resulting in an effect of improving fatigue recovery.

In the present invention, the effect of improving fatigue recovery means that the ratio of blood lactic acid level is reduced by 5 to 50% as compared with the case of not taking agarooligosaccharide.
Moreover, in this invention, in order to exhibit the fatigue recovery improvement effect most, it is preferable to take agarooligosaccharide 50-5000 mg / day, and 100-5000 mg / day is more preferable. However, the intake is not limited to this amount.

(Fat burning effect)
L-carnitine is known to have a fat burning effect and a slimming effect, but it is not sufficient as an effect. According to the basal metabolism improving agent and the fatigue recovery improving agent of the present invention, the basal metabolism and the fatigue recovery effect are improved by the above-described mechanism, and as a result, a fat burning effect superior to L-carnitine can be achieved.

Examples of the pharmaceutical composition and food and drink according to the present invention include tablets, capsules, pills, powders, granules, fine granules, jellies, and liquids as pharmaceutical compositions. Examples of the food and drink include beverages, jellies, candies, confectionery, health foods, and the like, and there are no particular restrictions on the types of foods and beverages that can be used.
The pharmaceutical composition of the present invention only needs to contain the PPARγ expression improver showing the basal metabolism improving effect and fatigue recovery improving effect of the present invention, and other known excipients as long as the effects of the present invention are not hindered. Etc. can be appropriately contained.

  EXAMPLES Hereinafter, although this invention is demonstrated concretely based on an Example, these do not limit the objective of this invention.

(Experimental Example 1: Production of agarooligosaccharide)
Agaro-oligosaccharide 1
: Commercially available agarooligosaccharide (Agaoligo: manufactured by Takara Bio Inc.)
Constituent sugar Disaccharide 26.2
Tetrasaccharide 27.7
Hexasaccharide 23.1
Octasaccharide 18.9
10 sugars 4.1
Agaro-oligosaccharide 2
: 50 g of agar (Ultra Agar AX-30: manufactured by Ina Food Industry Co., Ltd.) was dissolved by heating in 1000 g of purified water, 2 g of concentrated sulfuric acid was added, and the mixture was stirred at 90 ° C. for 3 hours. After adjusting the pH to 3.5 with sodium hydroxide, the solution was treated with activated carbon, filtered through filter paper, and further filtered through a 1 μm filter to obtain a solution. This solution was pulverized by vacuum lyophilization.
Constituent sugar Disaccharide 31.5
Tetrasaccharide 30.1
6 sugars 21.2
Octa sugar 11.6
10 sugars 5.6
Agaro-oligosaccharide 3
: Neo agarooligosaccharide was produced by the method of Example 1 of JP-A-2-65789.
Constituent sugar Disaccharide 1.5
Tetrasaccharide 37.5
6 sugars 41.3
Octasaccharide 6.3
10 sugars 13.4

(Experimental example 2: PPARγ expression improvement by ingestion of agarooligosaccharide)
Muscles were removed from the hind limb thighs of 3-week-old C3H / HeN mice, minced with scissors, and treated with protease. Centrifugation was then performed and only the cells were collected. The cell concentration was adjusted, and the cells were cultured under conditions of 37 ° C. and 5% CO ₂ . DMEM containing 10% horse serum was used as the medium. After confirming that the cells had differentiated into myocytes after 1 week of culture, a system was prepared in which agarooligosaccharide 1 was added to the medium at final concentrations of 50, 100, and 1000 μg / ml. As a comparison, an additive-free system was also prepared. After the addition, the mixture was allowed to stand for 24 hours under conditions of 37 ° C. and 5% CO ₂ , and then the cells were recovered with a cell scraper. Cells were isolated from total RNA using TRIzol (guanidine thiocyanate / phenol / chloroform), chloroform and isopropanol. Next, cDNA was prepared using reverse transcriptase, and the expression level of PPARγ mRNA was measured by real-time PCR using the cDNA as a template. The results are shown in FIG.

  As shown in FIG. 1, it was found that the intake of agarooligosaccharide enhanced the activity of the gene that takes glucose into cells.

(Experimental example 3: Basic metabolism improvement test)
Eleven healthy adults ingested agarooligosaccharide, and a basal metabolism improvement test was conducted by the crossover method. Specifically, two commercially available rice balls were ingested as a basal diet, and 500 mg each of agarooligosaccharides 1 to 3 were simultaneously ingested to measure basal metabolism (Examples 1 to 3). As a comparative example, only basic food (Comparative Example 1), and L-carnitine (Lonza Japan) 500 mg instead of agarooligosaccharide (Comparative Example 2) were also tested. The measurement of basal metabolism was calculated from oxygen consumption using Metavaine (Vine). The results are shown in FIG. The results are shown as average values.

  As shown in FIG. 2, it was confirmed that agarooligosaccharides 1 to 3 (Examples 1 to 3) have improved energy consumption and improved basal metabolism of subjects compared to Comparative Examples 1 and 2.

(Experimental example 4: Fatigue recovery test)
A fatigue recovery test was conducted on 10 healthy adults. Specifically, after measuring the blood lactic acid level before exercise, 500 mg each of agarooligosaccharides 1 to 3 was ingested and walked for 30 minutes on a treadmill (manufactured by Nakatsugi Health). The blood lactic acid level was measured immediately after the end of walking (Examples 4 to 6). As a comparative example, the same test was performed when agarooligosaccharide was not ingested (Comparative Example 3) and when L-carnitine 500 mg was used instead of agarooligosaccharide (Comparative Example 4). The results are shown in FIG. The results are shown as the rate of change when pre-exercise is 1.

  As shown in FIG. 3, when agarooligosaccharides 1 to 3 were ingested (Examples 4 to 6), the ratio of blood lactic acid levels was lower than that of Comparative Examples 3 and 4.

(Experimental Example 5: Inhibition of fat accumulation in culture test using mouse adipocytes)
The effect of agarooligosaccharide 1 on fat accumulation was examined using mouse adipocytes. Specifically, mouse 3T3-L1 fibroblasts that reached a confluent state were treated at 37 ° C., 5% in DMEM containing 10% fetal bovine serum supplemented with 5 μM insulin, 0.5 mM isobutylmethylxanthine, and 0.25 μM dexamethasone. The cells were cultured under CO ₂ conditions to induce fat differentiation. Furthermore, agarooligosaccharide 1 was added to final concentrations of 10, 100 and 1000 μg / ml, and cultured at 37 ° C. under 5% CO ₂ for 72 hours (Example 7). As a comparative example, the same test was performed when no agarooligosaccharide was added (Comparative Example 5) and when L-carnitine was added instead of agarooligosaccharide (Comparative Example 6). The cultured 3T3-L1 cells were fixed with a formalin solution. Thereafter, intracellular lipid droplets were stained with 0.5% oil red O staining solution, and the absorbance at 492 nm was measured. The results are shown in FIG.

  As shown in FIG. 4, the cultured cells to which agarooligosaccharide 1 was added suppressed the accumulation of fat depending on the concentration of agarooligosaccharide. This is because the cells have increased sugar uptake, resulting in a decrease in the amount of sugar used for synthesis into lipids. On the other hand, Comparative Example 6 had no lipid accumulation suppressing effect.

(Experimental Example 6: Preparation of pharmaceutical composition and food and drink)
<Example 8>
A tablet (1 tablet 0.3 g) for the purpose of improving basal metabolism and recovering from fatigue was prepared with the following formulation.
Agaro-oligosaccharide 1 50.0%
Lactose 30.0%
Crystalline cellulose 19.5%
Magnesium stearate 0.5%
<Example 9>
A powder for the purpose of improving basal metabolism and recovering from fatigue was prepared with the following formulation.
Agaro-oligosaccharide 2 50.0%
Lactose 40.0%
Corn starch 9.7%
Magnesium stearate 0.3%
<Example 10>
Beverages aimed at improving basal metabolism and recovering from fatigue were prepared with the following composition.
Agaro-oligosaccharide 3 2.5g
50g sucrose
Citric acid 5g
Orange juice (concentrated 5 times) 200g
Orange fragrance

(Awareness survey)
Awareness survey was performed using the pharmaceutical compositions and beverages prepared in Examples 8-10. Specifically, 10 people who are running every day are selected, and after taking the pharmaceutical composition or beverage of any of Examples 8 to 10 so that the agarooligosaccharide is 500 mg, running is performed for 5 km. received. After running, we conducted a survey on fatigue. In addition, 10 test subjects were asked to perform 5 km running without taking the product of the present invention 5 days before this test, and compared with that time.

Ingested items Subjects 1-3: Example 8
Test subjects 4-6: Example 9
Test subjects 7 to 10: Example 10

Results (out of 10)
Less fatigue than when not ingesting 8/10 people Same as when not ingesting 2/10 people (1 in each of Examples 9 and 10)
0/10 people feel tired compared to when not taking

  As described above, it was thought that most people had little fatigue. Thus, fatigue recovery (resulting in slimming effect) was observed in the pharmaceutical composition or food and drink containing agarooligosaccharide.

Claims (5)

  A PPARγ expression improver comprising as an active ingredient at least one sugar compound selected from the group consisting of agarobiose, agarotetraose, agarohexaose and agarooctaose.   A basal metabolism improver comprising the PPARγ expression improver according to claim 1.   A muscle fatigue recovery improver comprising the PPARγ expression improver according to claim 1. Features that PPARγ expression increased for Drug sets Narubutsu by containing PPARγ expression enhancing agent according to claim 1, wherein. A PPARγ expression improving food or drink comprising the PPARγ expression improving agent according to claim 1.

Images