WO2023063033A1 - Sarcopenic obesity inhibiting composition, and prophylactic and/or therapeutic composition containing the same for improving sarcopenic obesity induced by diabetes - Google Patents

Abstract

[Problem] To provide a novel sarcopenic obesity inhibiting composition, and a prophylactic and/or therapeutic composition containing the same for improving sarcopenic obesity induced by diabetes. [Solution] A sarcopenic obesity inhibiting composition containing (A) guar gum degradation products having an average molecular weight of 1.7×103 to 2.9×105, at least 70 mass% thereof being within this range, and (B) a guar bean protein, the viscosity of a 1 mass% aqueous solution of the composition being less than or equal to 50 mPa・s as measured at 25°C and 60 rpm using a B-type viscometer, wherein the guar gum degradation products are contained in guar-derived endosperm and obtained by using microbe-derived β-mannanase to hydrolyze and reduce the molecular weight of galactomannan polysaccharide that has a galactose to mannose content ratio (galactose:mannose) in the range 1:1.5 to 1:2.1, the dietary fiber content, defined by the enzymatic-HPLC method, is at least 70 mass%, the content of oligosaccharides in the guar gum degradation products 15 mass% or less, among the amino acid compositions in the contained proteins, (glutamic acid + glutamine + aspartic acid + asparagine) ≥ 100 mg/100g, (cystine + tyrosine + serine + threonine) ≥ 40mg/100g, and all amino acids ≥ 150mg/100g, and the ratio (A)/(B) is 1000 or less.

Description

Sarcopenic obesity inhibitory composition, and prophylactic and/or therapeutic composition containing the same for improving sarcopenic obesity induced by diabetes

The present invention relates to a composition for suppressing sarcopenia obesity, and a preventive and/or therapeutic composition containing the same for improving sarcopenia induced by diabetes.

Humans may experience a decrease in skeletal muscle mass and muscle strength due to aging or disease. This phenomenon is called "sarcopenia". Sarcopenia is a portmanteau of the Greek words sarco, meaning muscle, and penia, meaning decrease. Sarcopenia reduces resting energy expenditure and physical activity. However, as a result, body fat increases, which induces obesity, resulting in insulin resistance and an increase in tumor necrosis factor. This further promotes sarcopenia, resulting in a negative link between sarcopenia and obesity.
Thus, "sarcopenic obesity", which is a combination of sarcopenia and obesity, further increases the risk of lifestyle-related diseases beyond simple "obesity". Therefore, breaking the negative chain between sarcopenic obesity and muscle atrophy may extend healthy life expectancy. As such methods, enhancement of lipolysis, promotion of fat burning, maintenance/increase of muscle mass, etc. are conceivable.
As a prior art, there is a report that sarcopenic obesity can be prevented by ingesting fucoxanthin or fucoxanthinol, which is a type of polysaccharide (Patent Document 1).

JP 2020-132576 A

However, a satisfactory composition that can suppress sarcopenic obesity has not yet been obtained, leaving room for further research.
The present invention has been made in view of the above problems, and its object is to provide a novel composition for suppressing sarcopenic obesity, and a prophylactic and/or therapeutic composition containing the same for improving sarcopenic obesity induced by diabetes. It is to provide a composition.

The present inventor found that a composition containing a specific guar gum hydrolyzate and guar bean protein has sarcopenia obesity inhibitory activity, and basically completed the present invention.
Thus, the composition for suppressing sarcopenia obesity according to the present invention includes (A) a guar gum decomposition product having an average molecular weight of 1.8×10 ³ to 2.0×10 ⁵ and containing 70% by mass or more of those within the average molecular weight range. and (B) guar bean protein, and the viscosity of a 1% by mass aqueous solution when measured with a Brookfield viscometer at 25° C. and 60 rpm is 50 mPa s or less (more preferably 20 mPa s or less). wherein the guar gum decomposition product is contained in guar-derived endosperm, and the content ratio of galactose and mannose (galactose:mannose) is in the range of 1:1.5 to 1:2.1. It is obtained by hydrolysis using the derived β-mannanase to reduce the molecular weight, and has a dietary fiber content of at least 70% by mass and an oligosaccharide content of the guar gum hydrolyzate as determined by the enzyme-HPLC method. is 15% by mass or less, and among the amino acids contained in the protein (glutamic acid + glutamine + aspartic acid + asparagine) ≥ 100 mg/100 g, (cystine + tyrosine + serine + threonine) ≥ 40 mg/100 g, all It is characterized by amino acids≧150 mg/100 g and a ratio of (A)/(B) of 1000 or less.

A composition for preventing and/or treating sarcopenic obesity according to another invention is characterized in that it contains the sarcopenic obesity suppressing composition and is for improving sarcopenic obesity induced by diabetes. .
Further, food, drink and feed according to another invention are characterized by containing the composition for suppressing sarcopenia obesity. By ingesting this food and drink, it is possible to improve sarcopenic obesity induced by diabetes, so it is good for diabetic patients.
In the present invention, it is possible to provide foods, beverages and pharmaceuticals containing the composition of the present invention. Food and beverages include food and beverages, such as dietary supplements, health foods, foods for specified health uses, foods with function claims, diet therapy foods, general health foods, supplements, tea beverages, coffee beverages, and juices. , soft drinks, drinks, cooked rice, bread, noodles, dairy products, processed eggs, processed marine and livestock foods, confectionery, oils and fats and processed foods, seasonings, side dishes, and the like. Pharmaceuticals include pharmaceuticals or quasi-drugs, and are preferably oral formulations or enteral formulations, and can be in the form of liquids, tablets, granules, pills, syrups, and the like.
Feed in the present invention refers to food to be fed to organisms other than humans, and its form is not particularly limited. Organisms to which the feed can be applied are not particularly limited, but examples include farmed animals and pet animals. Farmed animals include livestock such as horses, cattle, pigs, sheep, goats, camels, and llamas, experimental animals such as mice, rats, guinea pigs, and rabbits, and poultry such as chickens, ducks, turkeys, and ostriches. be. Pet animals include, for example, dogs and cats.

According to the present invention, it is possible to provide a novel composition for suppressing sarcopenic obesity, and a sarcopenic obesity preventive and/or therapeutic composition containing the same for improving sarcopenic obesity induced by diabetes.

FIG. 10 is a graph showing body weight changes over time in Db/Db mice. FIG. In the graph, ● indicates the ND group, ▪ indicates the FFD group, and ▴ indicates the PHGG group (same in FIGS. 2 and 4). It is a graph which shows a blood glucose level change after (A) iPGTT test (intraperitoneal glucose tolerance test) and (B) ITT test (insulin tolerance test). FIG. 3 is a graph showing AUC (area under the curve) of blood glucose level graph after (A) iPGTT test and (B) ITT test. In the entire drawing, "*" indicates p<0.05, "**" indicates p<0.01, and "***" indicates p<0.001 (the same applies to other graphs). 2 is a graph showing changes over time in (A) oxygen partial pressure (VO ₂ ) and (B) carbon dioxide partial pressure (VCO ₂ ). 2 is a graph showing average values of (A) oxygen partial pressure (VO ₂ ) and (B) carbon dioxide partial pressure (VCO ₂ ) in each group in the light period (Light) and the dark period (Dark). Graph showing (A) grip strength measurements and (B) grip strength normalized to body weight. Graph showing (A) soleus weight and (B) soleus weight normalized to body weight. Graph showing (A) plantar muscle weight and (B) plantar muscle weight normalized to body weight. In each group, (A) a muscle tissue image of the soleus muscle and (B) a muscle tissue image of the plantar muscle are micrographs. It is a graph which shows the result of having calculated|required the cross-sectional area from (A) the microscope photograph figure of a soleus muscle, and (B) the microscope photograph figure of a soleus muscle. Graph showing (A) liver weight and (B) liver weight normalized to body weight. Graph showing (A) epididymal fat and (B) visceral fat mass normalized to body weight. Hematoxylin-eosin staining (HE), (B) Masson's triple staining (MT), and (C) Oil-red O staining (A) Hematoxylin-eosin staining (HE), (C) Micrographs of liver tissue taken from each group. be. FIG. 2 is a graph showing the results of examining (A) NAS (non-alcoholic fatty liver disease activity score) and (B) Oil Red O area. Graph showing the results of (A) ALT (alanine aminotransferase), (B) T-Chol (total cholesterol), (C) TG (triglycerides) and (D) NEFA (non-esterified fatty acids) in blood samples. is. 1 is a graph showing the results of examining the concentrations of (A) acetic acid, (B) propanoic acid, and (C) butanoic acid in feces. 1 is a graph showing the results of examining the concentrations of (A) acetic acid, (B) propionic acid and (C) butyric acid in serum. Among amino acids in muscle, (A) Valine, (B) Leucine, (C) Isoleucine, (D) Threonine, (E) Methionine, (F) 1 is a graph showing the results of examining the concentrations of phenylalanine, (G) lysine and (H) glutamic acid. Fig. 2 is a graph showing the results of examination of palmitic acid concentration in (A) feces, (B) serum and (C) muscle. Fig. 3 is a graph showing the results of expression analysis of each gene of (A) tnfa, (B) il6, (C) il1b, (D) cd36 and (E) il22 in the jejunum. FIG. 2 is a graph showing the expression analysis results of (A) trim63, (B) fbxo32, (C) tnfa, (D) il6, (E) foxo1 and (F) hdac4 genes in muscle. As inflammation-related cells in the small intestine, (A) ILC1, (B) ILC3, (C) ex-ILC3, (D) M1/CD45, (E) M2/CD45, and (F) M1/M2 ratio were investigated. It is a graph showing.

Next, embodiments of the present invention will be described with reference to the drawings. can be implemented in
Guar gum is a water-soluble natural polysaccharide obtained from the endosperm (more precisely, the cotyledons) of guar beans, and has two molecules of mannose linked in a straight chain and one molecule of galactose as a side chain. It is a polysaccharide and has an average molecular weight of about 2.0×10 ⁵ to 3.0×10 ⁵ . Guar gum is known to have physiological effects such as blood sugar level elevation inhibitory effect, cholesterol lowering effect, and bowel movement improving effect. In the present invention, the guar gum hydrolyzate is made from beans derived from the annual leguminous plant guar (scientific name: Cyanopsis tetragoloba), which is used for food in India, Pakistan, etc., and hydrolyzes the galactomannan polysaccharide contained in the endosperm. and water-soluble dietary fiber obtained by reducing the molecular weight. The method for hydrolyzing guar gum is not particularly limited and may be an enzymatic decomposition method, an acid decomposition method, or the like.

The enzyme used in the enzymatic decomposition method is not particularly limited as long as it is an enzyme that hydrolyzes linear mannose chains, but it is preferable to use β-mannanase derived from bacteria belonging to the genus Aspergillus or Rhizopus. The upper limit of the average molecular weight distribution of the guar gum decomposition product is 2×10 ⁵ or less, preferably 1.0×10 ⁵ or less, more preferably 2.5×10 ⁴ or less. The lower limit of the average molecular weight distribution of the guar gum decomposition product is 1.8×10 ² or more, preferably 3.0×10 ³ or more. When the average molecular weight distribution is smaller than 1.8×10 ² , it becomes difficult to sufficiently exhibit sarcopenia obesity inhibitory activity, and when the average molecular weight exceeds 2×10 ⁵ , the viscosity is too high to be included in food and drink. becomes difficult. The method for measuring the molecular weight ^{distribution is} not ^{particularly limited} . There is a method using a filtration chromatography method and the like.
The guar gum decomposition product of the present invention contains 70% by mass or more, preferably 80% by mass or more of those having the average molecular weight within the above range.

Galactose is a type of monosaccharide classified as aldohexose, and has a molecular formula of C ₆ H ₁₂ O ₆ and a molecular weight of 180 (both of which are the same as glucose). The configuration is 2-position (second from the top in the Fisher projection formula), -OH at 5-position is in the same direction, 3-position and 4-position are in the opposite direction, and the configuration at 5-position of D-galactose is D-glycerol Same as aldehyde.
Mannose is a type of monosaccharide classified as aldohexose, and has a molecular formula of C ₆ H ₁₂ O ₆ and a molecular weight of 180 (both of which are the same as glucose). The steric configuration is such that the --OH at the 2- and 3-positions are in the same direction and the 4- and 5-positions are in the opposite direction, and the steric configuration at the 5-position of D-mannose is the same as that of D-glyceraldehyde. Mannose is poorly metabolized in humans and hardly enters the glycolysis system when orally ingested.
The oligosaccharide content was 6 to 15% by mass (15% by mass or less) according to the gel filtration chromatograph analysis chart.
Amino acid composition in proteins can be measured by a known method using, for example, HPLC. Asparagine and glutamine are converted to aspartic acid and glutamic acid, respectively, during protein hydrolysis in pretreatment for amino acid analysis, and asparagine and aspartic acid, and glutamic acid and glutamine cannot be distinguished during measurement. Therefore, these are collectively quantified as aspartic acid and glutamic acid.

“Sarcopenia” means a condition (disease) in which muscles are reduced due to aging, lack of exercise, disease, or the like. If you don't use your muscles due to lack of exercise, your muscles will weaken and become thin, which not only invites further lack of exercise, but also causes bedridden. Sarcopenia increases the risk of diabetes (especially type 2 diabetes) and hypertension, and has other effects such as decreased walking speed, increased risk of falls, and increased mortality. .
“Sarcopenic obesity” means a state in which obesity occurs due to an increase in fat when calorie consumption is low relative to calorie intake in sarcopenia. Compared to before, if the height and weight are the same, it may be difficult to notice obesity just by looking at it, but there may be a significant decrease in muscle mass compared to before. Sarcopenic obesity is often seen in elderly people aged 65 and over, but progression begins around the age of 25 to 30 and progresses throughout life. This is one of the major causes of lowering the activity capacity of the elderly.
INDUSTRIAL APPLICABILITY The present invention has an improvement effect on sarcopenic obesity induced by diabetes (particularly elderly type 2 diabetes) among various diseases.
The composition for suppressing sarcopenia obesity of the present invention can be orally ingested as it is or mixed with foods, beverages, and the like. The dose of the composition for suppressing sarcopenia obesity when taken orally is not particularly limited, but is 0.5 g to 70 g (preferably 3 g to 30 g, more preferably 6 g to 18 g) per adult per day.

<Preparation of guar gum degradation product (PHGG)>
Example 1
After adjusting the pH to 4.5 by adding 0.1N hydrochloric acid to 900 g of water, 0.2 g of commercially available β-mannanase derived from bacteria belonging to the genus Aspergillus and 100 g of guar gum powder were added and mixed. The mixture was reacted at 40° C.-45° C. for 24 hours. After the reaction, the enzyme was deactivated by heating at 90°C for 15 minutes. The reaction solution was separated by suction filtration, and the clear solution obtained by removing insoluble matter was concentrated under reduced pressure (Yamato evaporator). A solid content of 20% by mass was obtained. This was dried with a spray dryer (manufactured by Okawara Kakoki Co., Ltd.) to obtain 65 g of a guar gum decomposition product as a powder.
A 0.5 (w/v) % concentration aqueous solution was obtained by dissolving the guar gum decomposition product in water. Using polyethylene glycol (average molecular weight: 2×10 ² , 2×10 ³ , 2×10 ⁴ and 1×10 ⁵ ) as molecular weight markers, gel filtration chromatography (column: YMC-Pack Diol-120, detector : Differential refractometer), the average molecular weight was about 20,000. Those having a molecular weight of 1.8×10 ³ to 2.0×10 ⁵ were contained in an amount of 85 mass % or more.

Moreover, when the viscosity of the 1 mass % aqueous solution was measured using a Brookfield viscometer at 25° C. and 60 rpm, it was 8 mPa·s.
When the content ratio of galactose and mannose (galactose:mannose) was measured, it was 1:1.7.
The dietary fiber content was measured by enzyme-HPLC method and found to be 90% by weight.
The oligosaccharide content was 7.3% by mass.
In addition, amino acid analysis of the protein contained in the guar gum decomposition product was performed, and the results were as shown in Table 1. Based on these results, (glutamic acid + glutamine + aspartic acid + asparagine), (cystine + tyrosine + serine + threonine), and total amino acids were determined to be 170 mg/100 g, 63 mg/100 g, and 359 mg/100 g, respectively. Met.
Further, the ratio of guar gum decomposition product to guar bean protein was found to be guar gum decomposition product/guar bean protein=279.

Example 2
After adjusting the pH to 3 by adding 0.1N hydrochloric acid to 900 g of water, 0.15 g of commercially available β-mannanase derived from bacteria belonging to the genus Aspergillus and 100 g of guar gum powder were added and mixed. The mixture was reacted at 40° C.-45° C. for 24 hours. After the reaction, the enzyme was deactivated by heating at 90°C for 15 minutes. The reaction solution was separated by suction filtration, and the clear solution obtained by removing insoluble matter was concentrated under reduced pressure (Yamato evaporator). A solid content of 20% by mass was obtained. This was dried with a spray dryer (manufactured by Okawara Kakoki Co., Ltd.) to obtain 68 g of guar gum decomposition product as a powder.
When the average molecular weight of the guar gum decomposition product was determined in the same manner as in Example 1, it was about 2.5×10 ⁴ . According to the results of the HPLC chart, 85% by mass or more of those having a molecular weight of 1.8×10 ³ to 2.0×10 ⁴ were contained.
Moreover, when the viscosity of the 1 mass % aqueous solution was measured using a Brookfield viscometer at 25° C. and 60 rpm, it was 10 mPa·s.
The content ratio of galactose and mannose (galactose:mannose) was measured to be 1:1.8.
The dietary fiber content was measured by enzyme-HPLC method and found to be 89% by weight.
The oligosaccharide content was 10% by mass.
In addition, when the amino acids in the protein contained in the guar gum decomposition product were analyzed, (glutamic acid + glutamine + aspartic acid + asparagine), (cystine + tyrosine + serine + threonine), and the total amino acids were determined, each was 134 mg / 100 g, 51 mg/100 g and 239 mg/100 g.
Further, the ratio of guar gum decomposition product to guar bean protein was found to be guar gum decomposition product/guar bean protein=418.

Example 3
After adjusting the pH to 4 by adding 0.1N hydrochloric acid to 900 g of water, 0.25 g of commercially available β-mannanase derived from bacteria belonging to the genus Aspergillus and 100 g of guar gum powder were added and mixed. The mixture was reacted at 50° C.-55° C. for 12 hours. After the reaction, the enzyme was deactivated by heating at 90°C for 15 minutes. The reaction solution was separated by suction filtration, and the clear solution obtained by removing insoluble matter was concentrated under reduced pressure (Yamato evaporator). A solid content of 20% by mass was obtained. This was dried with a spray dryer (manufactured by Okawara Kakoki Co., Ltd.) to obtain 70 g of a guar gum decomposition product as a powder.
When the average molecular weight of the guar gum decomposition product was determined in the same manner as in Example 1, it was about 1.5×10 ⁴ . According to the results of the HPLC chart, 86% by mass or more of those having a molecular weight of 1.8×10 ³ to 2.0×10 ⁵ were contained.

Moreover, when the viscosity of the 1 mass % aqueous solution was measured using a Brookfield viscometer at 25° C. and 60 rpm, it was 9 mPa·s.
When the content ratio of galactose and mannose (galactose:mannose) was measured, it was 1:2.0.
The dietary fiber content was measured by enzyme-HPLC method and found to be 88% by mass.
The oligosaccharide content was 9% by mass.
In addition, when the amino acids in the protein contained in the guar gum decomposition product were analyzed, (glutamic acid + glutamine + aspartic acid + asparagine), (cystine + tyrosine + serine + threonine), and the total amino acids were determined, each was 135 mg / 100 g, 62 mg/100 g and 280 mg/100 g.
Further, the ratio of guar gum hydrolyzate to guar bean protein was found to be guar gum hydrolyzate/guar bean protein=357.
Example 4
A guar gum hydrolyzate was prepared according to the examples of JP-A-5-117156 (page 4, line 3 to page 4, line 10). When the average molecular weight was determined according to Example 1, it was 5.5×10 ³ .

<Biological activity test>
(A) Test method 1. Test Animals Seven-week-old male diabetic Db/Db mice (manufactured by Shimizu Laboratory Surprise) were used. After one week of preliminary feeding, the 8-week-old mice were divided into the following three groups (n=6 for each group).
Either a normal diet group (ND group; 344.9 kcal/100 g (fat kcal) 4.6% (Clea)), a fiber-free diet group (FFD group) or a PHGG-added diet group (PHGG group) for 8 weeks Gave. For each group, pair feeding was performed so that the calorie intake was constant. Mice in each group were dissected at 16 weeks of age, and blood was collected and organs were excised.
2. Glucose and Insulin Tolerance Tests Weekly mice were fasted overnight for 14 hours and weighed. At 15 weeks of age, an intraperitoneal glucose tolerance test (iPGTT: 2 g/kg body weight) and an insulin tolerance test (ITT: 0.75 U/kg body weight) were performed after fasting for 14 hours and 5 hours, respectively. (manufactured by Sanwa Hong Kong Kenko Co., Ltd.)) was used to measure blood glucose. The area under the curve (AUC) of the iPGTT and ITT results were analyzed respectively.

3. Indirect Calorimetry In vivo indirect open circuit calorimetry was performed in a controlled ambient temperature (24±2° C.) metabolic chamber. A room temperature air flow rate (0.6 liter/min) was sucked into the room and monitored by a metabolic analyzer (O ₂ /CO ₂ analyzer MM202R (manufactured by Muromachi Kikai Co., Ltd.)). 24-hour _VO2 and _VCO2 were measured during a 12-hour light/dark cycle. Gas concentrations were monitored at the inlet and outlet of the closed chamber.
4. Measurement of Grip Strength At 16 weeks of age, grip strength was measured using a mouse grip dynamometer (Model DS2-50N (manufactured by IMADA)). Six consecutive measurements per day were performed at 1 minute intervals. The observer was blinded to which group each mouse belonged to. Grip strength was also normalized to body weight.

5. Weight Measurement of Soleus and Plantar Muscles The weights of the soleus (Soleus) and plantar (Planaris) muscles of each individual were measured. Each muscle weight was normalized to body weight.
6. Muscle Histology Soleus and plantar muscles were obtained, fixed in 10% buffered formaldehyde, and embedded in paraffin. Muscle sections were prepared and stained with hematoxylin and eosin staining. Images were taken with a BZ-X710 fluorescence microscope (manufactured by Keyence Corporation). Cross-sectional areas were measured with Image J (NIH).
7. Liver weight and visceral fat weight Liver weight and visceral fat weight (Epididymal Fat Weight) were measured. All were normalized to body weight.
8. Liver histology Livers were harvested, fixed in 10% buffered formaldehyde, and embedded in paraffin. Liver sections were prepared and stained with hematoxylin-eosin (HE), Masson's trichrome stain (MT) and Oil-red O. Images were taken with a BZ-X710 fluorescence microscope (manufactured by Keyence Corporation). In addition, the non-alcoholic fatty liver disease (NAFLD) activity score (NAS) was used to assess the severity of NAFLD.

9. Biochemical Testing Blood samples were taken from fasted mice to measure alanine aminotransferase (ALT) levels, total cholesterol (T-Chol), triglycerides (TG) and non-esterified fatty acids (NEFA). Biochemical tests were requested from Fujifilm Wako Pure Chemical Industries, Ltd.
10. Fecal and Serum Short Chain Fatty Acid (SCFA) Concentrations and Muscle Amino Acid Concentrations The SCFA composition of rat rectal feces and serum samples and the amino acid composition of rat plantar muscle were analyzed using an Agilent 7890B/7000D System (manufactured by Agilent Technologies). Analysis was performed using a gas chromatograph mass spectrometer (GC/MS). A sample of rectal feces (20 mg), serum (50 μL), and plantar muscle (20 mg) was added to 500 μL of acetonitrile and 500 μL of diluted water, ground in a ball mill at 4000 rpm for 2 minutes, and then shaken at 37° C. at 1000 rpm for 30 minutes. and centrifuged at 14000 rpm for 3 minutes at room temperature. Supernatant (500 μL) was added to 500 μL of acetonitrile and shaken at 1000 rpm at 37° C. for 3 minutes. Then, it was centrifuged at 14000 rpm for 3 minutes at room temperature, and the pH was adjusted to 8 with 0.1 mol/L NaOH to extract SCFA.
Next, SCFA and amino acid concentrations were automatically measured by an online solid phase extraction (SPE) method using an SPE-GC system SGI-M100 (manufactured by Icety Science).

11. Free Fatty Acids in Serum, Liver, Feces, and Soleus Muscle The composition of palmitic acid in mouse serum, liver, feces, and soleus muscle was measured using a gas chromatograph mass spectrometer (GC/MS), Agilent 7890B/7000D (manufactured by Agilent Technologies). was analyzed using 25 mL of serum and 15 µg of liver, feces and plantar muscle were methylated using a fatty acid methylation kit (manufactured by Nacalai Tesque). The final product was loaded onto a Varian capillary column (DB-FATWAX UI, Agilent Technologies). A CP-Sil88 for FAME (100 m×0.25 mm inner diameter×0.20 μm film thickness (manufactured by Agilent Technologies)) capillary column was used for the separation of fatty acids. The temperature of the column was maintained at 100° C. for 4 minutes, then increased at 3° C./min to 240° C. and held for 7 minutes. Samples were injected in split mode with a split ratio of 5:1. Each fatty acid methyl ester was detected with the selected ion monitoring mode. All results were normalized to the peak height of the C17:0 internal standard.

12. Gene expression analysis in jejunum and soleus muscle The jejunum and soleus muscle samples obtained by dissection were homogenized in an ice-cold QIAzol Lysis reagent (manufactured by Qiagen) to isolate total RNA. Total RNA (0.5 μg) was reverse transcribed using a high-capacity cDNA reverse transcription kit (manufactured by Applied Biosystems). The reverse transcription reaction was carried out at 37°C for 120 minutes and reverse transcription inactivation was carried out at 85°C for 5 minutes. Real-time reverse transcription-polymerase chain reaction (RT-PCT) was used to determine the mRNA expression levels of Trim63, Fbxo32, Tnfa, Il6, Foxo1 and Hdac4 for soleus muscle and Tnfa, Il6, Il1B, Cd36 and Il22 for jejunum. Expression levels were quantified. RT-PCT was performed using TaqMan Fast Advanced Master Mix (manufactured by Applied Biosystems).
The following method was used as PCT conditions. One cycle consisted of synthesis reaction at 50°C for 2 minutes and denaturation at 95°C for 20 seconds. Relative expression levels of each target gene were normalized to Gapdh threshold cycle (CT) values and quantified using the comparative threshold cycle 2-ΔΔCT method. Relative values were calculated with the signal of ND mouse set to 1.0.

13. Isolation and flow cytometry of mononuclear cells from the small intestine of mice After systemic perfusion with heparinized saline, intestinal lamina propria mononuclear cells were isolated using the lamina-specific dissociation kit (130-097-410 (Miltenyi, (manufactured by Biotech)). The cell pellet was resuspended in 40% Percoll and added to the top of a centrifuge tube containing 5 mL of 80% Percoll. The cells were washed twice with 2% FBS/PBS and subjected to post density gradient centrifugation at 420 xg for 20 minutes to obtain lamina propria mononuclear cells.
Stained cells were analyzed on a FACS CantoII and data were analyzed using FlowJo version 10 software (TreeStar). Gating for innate lymphoid cells included Biotin-CD3e (100304; clone: 145-2C11), Biotin-CD45R/B220 (103204; clone: RA3-6B2; 1/200), Biotin-Gr-1 (108404; clone: RB6-8C5; 1/200), Biotin-CD11c (117304; clone: N418; 1/200), Biotin-CD11b (101204; clone: M1/70; 1/200), Biotin-Ter119 (116204; clone : TER-119; 1/200), Biotin-FceRIa (134304; clone: MAR-1; 1/200), FITC-Streptavidin (405202; 1/500), PE-Cy7-CD127 (135014; clone: A7R34; 1/100), Pacific Blue-CD45 (103116; clone: 30-F11; 1/100), PE-GATA-3 (clone TWAJ, 1/50), ATC-RORγ (clone AFKJS-9, 1/50) , Fixable Viability Dye eFluor 780 (1/400) antibodies were used.
In addition, to gate M1 and M2 macrophages, APCT-CD45.2 (17045482; clone: 104, 1/50), PE-F4/80 (12480182; clone: BM8, 1/50), APC-Cy7-CD11b (47011282; clone: M1/70, 1/50), FITC-CD206 (MA516870; clone: MR5D3, 1/50), PE-CY7-CD11c (25011482; clone: N418, 1/50) antibodies board.
All the above antibodies were manufactured by eBioscience.

(B) Test results 1. Glucose and Insulin Tolerance Tests FIG. 1 shows changes in body weight of the ND group, FFD group and PHGG group. Fig. 2 shows changes in blood glucose level after each test of iPGTT and ITT performed at 15 weeks of age, and AUC is shown in Fig. 3, respectively. In the ND group, the blood glucose level increased in all tests, and it was considered that diabetes, which is characteristic of Db/Db mice, was developed (or was developing). On the other hand, in the PHGG group, the blood glucose level was significantly (p<0.01) lower than the ND group in all tests, and impaired glucose tolerance was improved.
2. Indirect Calorimetry FIG. 4 shows changes in oxygen partial pressure (VO ₂ ) and carbon dioxide partial pressure (VCO ₂ ) for each group over time. FIG. 5 shows the average values of _VO2 and _VCO2 during the light period (12 hours) and the dark period (12 hours). The PHGG group showed a significant (p<0.05) increase in basal metabolism compared to the ND and FFD groups.
3. Grip Strength Measurement FIG. 6(A) shows the grip strength measurement results, and FIG. 6(B) shows the results normalized to the body weight. The PHGG group had significantly stronger grip strength than the ND and FFD groups.

4. Weight Measurement of Soleus and Plantaris Muscles FIG. 7(A) shows the weight of the soleus muscle in each group, and FIG. 7(B) shows the results normalized to body weight. FIG. 8(A) shows the plantar muscle weight of each group, and FIG. 8(B) shows the results normalized to body weight.
The PHGG group had significantly heavier soleus and plantar muscle weights than the ND and FFD groups.
5. Muscle histology FIG. 9 shows histology of the soleus and plantar muscles in each group (representative micrographs). FIG. 10 shows the results of examining the cross-sectional area of each muscle.
The PHGG group had a significantly larger muscle cross-sectional area than the ND and FFD groups.
6. Liver weight and visceral fat content Fig. 11(A) shows liver weight, and Fig. 11(B) shows liver weight normalized to body weight. FIG. 12(A) shows visceral fat mass, and FIG. 12(B) shows visceral fat mass normalized to body weight.
The PHGG group had significantly larger liver weights than the ND and FFD groups. In addition, the PHGG group significantly decreased the normalized visceral fat mass compared to the ND group and the FFD group.

7. Liver histology FIG. 13 shows representative micrographs of liver tissues taken from each group stained with hematoxylin-eosin (HE), Masson's trichrome staining (MT) and Oil-red O. showed that. FIG. 14 shows the results of examining the NAS and Oil Red O areas for the three groups.
The PHGG group significantly improved fatty liver compared to the ND group and FFD group.
8. Biochemical Tests FIG. 15 shows the results of examining ALT, T-Chol, TG and NEFA in blood samples. For all test items, the PHGG group showed significantly lower values than the ND group or the FFD group. Thus, the PHGG group significantly improved lipid metabolism disorders.
9. SCFA and Amino Acid Concentrations in Feces and Serum FIG. 16 shows acetic acid, propanoic acid and butanoic acid concentrations in feces. FIG. 17 shows the concentration of each short-chain fatty acid in serum.
All short-chain fatty acids were significantly elevated in feces and serum in the PHGG group compared to the ND group or the FFD group.
FIG. 18 shows the concentrations of valine, leucine, isoleucine, threonine, methionine, phenylalanine, lysine and glutamic acid among amino acids in muscle. These amino acids are involved in muscle protein synthesis and inhibition of degradation.
All amino acids were significantly increased in the PHGG group compared to the ND group and the FFD group.

10. Palmitic Acid Concentrations in Feces, Serum, and Muscles FIG. 19 shows palmitic acid concentrations in feces, serum, and muscles.
The PHGG group showed a significant increase in feces and a significant decrease in serum and muscle compared to the ND and FFD groups.
11. Gene Expression Analysis in Jejunum and Soleus Muscle FIG. 20 shows the results of gene expression analysis in jejunum, and FIG. 21 shows the results of gene expression analysis in muscle.
In the jejunum, the expression of inflammation-related genes (tnfa, il6, il1b, cd36) was significantly reduced in the PHGG group compared to the ND group or FFD group. In muscle, the expression of muscle atrophy-related genes (trim63, fbxo32, tnfa, il6, foxo1, hdac4) was significantly reduced in the PHGG group compared to the ND group or FFD group.
12. Inflammatory Cells in the Small Intestine FIG. 22 shows the results of measuring ILC1, ILC3, ex-ILC3, M1/CD45, M2/CD45 and the M1/M2 ratio.
Inflammatory cells and factors related thereto (ILC1, ex-ILC3, M1/CD45) were significantly decreased in the PHGG group compared to the ND group and FFD group. In addition, the PHGG group significantly increased factors (IL22, IL3, M2 macrophages) that resolve inflammation.

As described above, significant sarcopenic obesity was observed in the ND group and FFD group due to overeating and lack of exercise. In contrast, PHGG was effective against both sarcopenia, diabetes and obesity. That is, improvement of impaired glucose tolerance, increased basal metabolism, improved grip strength/decreased muscle mass, decreased visceral fat content, improved liver fat, and improved lipid metabolism disorders were observed. The mechanism is increased short-chain fatty acids in feces and blood, increased branched-chain amino acids in muscles, decreased saturated fatty acids in blood, increased excretion of saturated fatty acids, and expression of muscle atrophy-related genes in skeletal muscle. decreased expression of inflammation-related genes in the small intestine, and decreased inflammatory cells in the lamina propria of the small intestine.
Thus, according to the present embodiment, a novel composition for suppressing sarcopenic obesity and a sarcopenic obesity preventive and/or therapeutic composition containing the same for improving sarcopenic obesity induced by diabetes could be provided. .

Claims (7)

(A) a guar gum decomposition product having an average molecular weight of 1.8×10 ³ to 2.0×10 ⁵ and containing 70% by mass or more of those within the average molecular weight range, and (B) guar bean protein, and B A sarcopenia obesity suppressing composition having a viscosity of 50 mPa s or less in a 1% by mass aqueous solution when measured using a type viscometer at 25° C. and 60 rpm,
The guar gum hydrolyzate is contained in the guar-derived endosperm, and the content ratio of galactose and mannose (galactose:mannose) is in the range of 1:1.5 to 1:2.1. and has a dietary fiber content of at least 70% by mass and an oligosaccharide content of 15% by mass of the guar gum hydrolyzate as determined by the enzyme-HPLC method. (Glutamic acid + glutamine + aspartic acid + asparagine) ≥ 100 mg/100 g, (cystine + tyrosine + serine + threonine) ≥ 40 mg/100 g, total amino acids ≥ 150 A composition for suppressing sarcopenia obesity, which is mg/100 g and has a ratio of (A)/(B) of 1000 or less. A composition for preventing and/or treating sarcopenic obesity, comprising the composition for suppressing sarcopenic obesity according to claim 1, for improving sarcopenic obesity induced by diabetes. A food or drink containing the composition for suppressing sarcopenia obesity according to claim 1 . A medicament containing the composition for suppressing sarcopenia obesity according to claim 1 . A medicament containing the preventive and/or therapeutic composition for sarcopenic obesity according to claim 2. A feed containing the composition for suppressing sarcopenia obesity according to claim 1 . A feed containing the composition for preventing and/or treating sarcopenic obesity according to claim 2.

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