Muscle enhancing agent, oral composition for muscle enhancement, and use of plasmalogen

Abstract

To provide a muscle enhancing agent and an oral composition for muscle enhancement that exhibit muscle satellite cell activation effects while ensuring high safety.SOLUTION: A muscle enhancing agent contains a plasmalogen as an active ingredient.SELECTED DRAWING: Figure 4

Description

[Technical field]

[0001] The present invention relates to a muscle-building agent, an oral composition for muscle building, and use of plasmalogen. [Background technology]

[0002] Skeletal muscles are the largest tissue in the human body and play important roles such as supporting the body and moving the limbs. If muscles are not used for a certain period of time due to lack of exercise, being bedridden, immobilization with a cast, etc., the muscles will become smaller and their function will decline. From the age of 40, human muscle mass decreases by more than 1% per year, and by the age of 80, it will have decreased to 50% of its maximum muscle mass. Muscle loss not only threatens the "healthy lifespan," the period during which one can live healthily and independently, but also increases arthritis, lower back pain, and chronic pain. As a result, the problem of musculoskeletal diseases such as age-related muscular fragility (sarcopenia), in which muscle mass or strength decreases with age, is becoming apparent in Japan, which has become a super-aging society.

[0003] Muscle satellite cells, which are skeletal muscle stem cells, are responsible for most of the regenerative ability of muscles. Normally, muscle satellite cells are in a quiescent state and do not proliferate, but when they are stimulated by skeletal muscle damage such as intense exercise, they become activated and express the muscle differentiation control factor MyoD, becoming precursor cells called myoblasts. Non-Patent Document 1 reports that a decrease in MyoD expression leads to a decrease in muscle mass and sarcopenia.

[0004] Muscles are made up of muscle fibers formed by myoblasts differentiating into myotubes, which then fuse together. Satellite cells play an essential role not only in muscle regeneration, but also in muscle growth during development and muscle hypertrophy through strength training. In addition, in intractable muscle diseases such as muscular dystrophy and the increasingly prevalent condition of sarcopenia, the number and function of satellite cells are reduced.

[0005] A method for activating satellite cells through high-intensity exercise has been reported. Non-Patent Document 2 reports that satellite cells and muscle fiber cross-sectional area increase with strength training at an intensity of 80% or more of the maximum lifting weight (1RM), lifting 8-12 times per set, 2-3 sets, 3 times a week for 3 months or more.

[0006] Furthermore, Patent Document 1 discloses a muscle activator that contains a bergamot plant extract as an active ingredient, with the intention of activating myoblasts or promoting muscle protein synthesis. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] JP 2023-029511 A [Non-patent literature]

[0008] [Non-Patent Document 1] Jin-A Kim, 6 others, “Sinensetin regulates age-related sarcopenia in cultured primary thigh and calf muscle cells”, BMC Complementary and Alternative Medicine, 2019, 19:287, 1-8 [Non-Patent Document 2] Lex B. Verdijk, et al., "Satellite cell content is specifically reduced in type II skeletal muscle fibers in the elderly", Am J Physiol Endocrinol Metab, 2007, 292, E151-E157 Summary of the Invention [Problem to be solved by the invention]

[0009] The high-stress exercise disclosed in Non-Patent Document 2 places a considerable burden on the exerciser and is not considered safe. High-stress exercise is particularly difficult for elderly people to perform. The effect of the muscle activator disclosed in Patent Document 1 on satellite cells is unclear. There is a demand for a safe muscle-building agent and oral composition for muscle building that have satellite cell activation effects.

[0010] The present invention has been made in consideration of the above circumstances, and aims to provide a highly safe muscle-building agent and oral composition for muscle building that have a muscle satellite cell activation effect. Another aim of the present invention is to provide use of plasmalogen for the production of the muscle-building agent or oral composition for muscle building. [Means for solving the problem]

[0011] The muscle-building agent according to the first aspect of the present invention comprises: Contains plasmalogen as an active ingredient.

[0012] The oral composition for muscle building according to the second aspect of the present invention comprises: Contains plasmalogen as an active ingredient.

[0013] The plasmalogen is It is a plasmalogen extracted from animal tissue. This may also be the case.

[0014] The animal tissue comprises: The tissue is an animal selected from the group consisting of shellfish, sea squirts and birds. This may also be the case.

[0015] The animal tissue comprises: Scallop tissue, This may also be the case.

[0016] The use according to the third aspect of the present invention comprises: The present invention relates to use of a plasmalogen for producing a muscle-building agent or an oral composition for muscle building. Effect of the Invention

[0017] According to the present invention, a highly safe muscle-building agent and an oral composition for muscle-building that have a muscle satellite cell activation effect are provided. Also, according to the present invention, there is provided use of plasmalogen for producing the muscle-building agent or the oral composition for muscle-building. [Brief description of the drawings]

[0018] [Figure 1] FIG. 1 shows a fluorescent microscope image of immunostained thigh muscle fibers. [Diagram 2] 1 shows the results of analyzing AMPK phosphorylation in soleus muscle, in which A shows the results of Western blotting for phosphorylated AMPK (pAMPK), and B shows the relative amount of pAMPK / AMPK based on the results of Western blotting. [Diagram 3] 1 shows the results of evaluating the expression of MyoD in soleus muscle, with A showing the results of Western blotting for MyoD and B showing the relative amount of MyoD based on the results of Western blotting. [Figure 4] Figure 1 shows the results of measuring the diameter of muscle fibers, A shows a bright field observation image of thigh muscle, and B shows the diameter of muscle fibers. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] The embodiments of the present invention will be described with reference to the drawings. Note that the present invention is not limited to the following embodiments and drawings. Note that in the following embodiments, the expressions "have", "include" and "contain" also include the meaning of "consisting of" or "consisting of".

[0020] The muscle-building agent according to the present embodiment contains plasmalogen as an active ingredient. Plasmalogen belongs to a subclass specific to glycerophospholipids, characterized by having a vinyl ether bond at the sn-1 position of the glycerol backbone and an ester bond at the sn-2 position. Plasmalogen is present in high concentrations in the cell membranes of many mammalian tissues.

[0021] The plasmalogen contained in the muscle-building agent is not particularly limited as long as it is generally classified as a plasmalogen. Examples of plasmalogens include Pls, PlsCho, inositol-type plasmalogens, and serine-type plasmalogens. Pls has a -CH group at the oxygen atom bonded to phosphorus at the sn-3 position of the glycerol skeleton. 2 CH 2 NH 2 PlsCho has a structure in which -CH is bonded to the oxygen atom attached to phosphorus at the sn-3 position of the glycerol backbone. 2 CH 2 N(CH 3 ) 3 It has a structure in which

[0022] Preferably, the plasmalogen is extracted from an animal tissue. The animal tissue is not particularly limited as long as it contains plasmalogen. For example, the animal tissue may be tissue of an animal selected from the group consisting of shellfish, sea squirts, and birds. More specifically, the tissue of aquatic animals such as shellfish, sea squirts, sea cucumbers, salmon, pacific saury, and bonito is preferred as the animal tissue. Preferably, the animal tissue is shellfish tissue. Among the animal tissues, edible parts are preferred. The animal tissue may be cut tissue, or may be crushed tissue to more efficiently extract plasmalogen.

[0023] Examples of shellfish include bivalve or snail shells such as scallops, mussels, and abalone. As shellfish, scallops are particularly preferred. Scallops are edible bivalve shells belonging to the Pectinidae family, and examples thereof include scallops belonging to the Mizuhopecten or Pecten genus. Specific examples of shellfish include scallops (Mizuhopectenyessoensis) harvested in Japan and European scallops (Pectenmaximus (Linnaeus)) harvested in Europe. Examples of edible parts of scallops include the adductor muscle and string.

[0024] Sea squirts are edible chordates belonging to the family Halocynthidae, and examples of such sea squirts include those belonging to the genus Halocynthia or Halocynthia genus. Specific examples of sea squirts include Halocynthia roretzi and Halocynthia aurantium. The edible part of the sea squirt is the flesh (fascia).

[0025] The bird is, for example, an edible bird. Examples of the bird include chickens, silkie chickens, ducks, etc. As for the tissue of the bird, breast meat is preferable because it is rich in plasmalogens.

[0026] Plasmalogens can be extracted using an organic solvent or a water-containing organic solvent, preferably in combination with an enzyme treatment. Examples of methods for extracting plasmalogens include ethanol extraction and hexane extraction.

[0027] In the ethanol extraction method, the animal tissue may be exposed to an extracting solution containing ethanol. The extracting solution containing ethanol used in the extraction step may be aqueous ethanol or a mixture of ethanol and another organic solvent. The ethanol concentration in the extracting solution is 50% by mass or more, preferably 80% by mass or more, more preferably 95% by mass or more, and may be 100% by mass. The amount of ethanol used is, for example, preferably 100 to 3000 mL, more preferably 200 to 2000 mL, and even more preferably 250 to 1000 mL per 100 g of animal tissue. The extraction using ethanol may be performed multiple times.

[0028] Preferably, the animal tissue exposed to the extract containing ethanol is an animal tissue treated with a protease. The protease may be, for example, a protease derived from a filamentous fungus such as Aspergillus or Rhizopus; a protease derived from a bacterium such as Bacillus; or a protease extracted from a plant such as papaya or pineapple. A commercially available protease may be used as the protease. In particular, a protease derived from a filamentous fungus is preferred, and a protease derived from Aspergillus oryzae is more preferred. In addition, since plasmalogen is unstable in acidic and alkaline conditions, a neutral protease is preferred.

[0029] The amount of protease used is, for example, 0.1 to 10.0 g, preferably 0.2 to 8.0 g, and more preferably 0.3 to 5.0 g, per 100 g of animal tissue.

[0030] After the extraction, the solid content is removed to recover the extract, and if necessary, the extract is dried and solidified to obtain an extract containing plasmalogen. The plasmalogen may be concentrated by subjecting the extract to a concentration treatment.

[0031] In the case of the hexane extraction method, the total lipids contained in the extract obtained by the ethanol extraction method are extracted with a mixed solvent of n-hexane and a water-soluble ketone solvent, and separated into an insoluble portion and a soluble portion. The water-soluble ketone solvent is, for example, acetone, methyl ethyl ketone, or a combination thereof, and is preferably acetone. When a mixture of n-hexane and acetone is used as the mixed solvent, the ratio is, for example, 4:6 to 6:4, preferably 4.5:5.5 to 5.5:4.5, in terms of volume ratio. The amount of the mixed solvent used is, for example, 10 to 30 mL per 1 g (dry mass) of total lipids.

[0032] The soluble portion obtained by the extraction process using the mixed solvent is dried and then extracted with a water-soluble ketone solvent, whereby the plasmalogen can be separated and recovered.

[0033] The amount of plasmalogen contained in the muscle-building agent according to the present embodiment is appropriately adjusted. For example, the muscle-building agent contains 0.000001 to 99.9 mass%, 0.00001 to 99.8 mass%, 0.0001 to 99.7 mass%, 0.001 to 99.6 mass%, 0.01 to 99.5 mass%, 0.1 to 99 mass%, 0.5 to 60 mass%, 1 to 50 mass%, or 1 to 2 mass% of plasmalogen as an active ingredient.

[0034] The muscle-building agent according to the present embodiment is administered to cells, living tissues, organisms, animals, etc. The muscle-building agent as a pharmaceutical is administered to humans and animals. The animals are preferably mammals, more specifically, dogs, cats, cows, pigs, horses, sheep, deer, etc.

[0035] The administration route of the muscle-building agent according to the present embodiment to humans is not particularly limited. The muscle-building agent is preferably used as an external preparation, an injection, or an oral preparation. The muscle-building agent may contain, for example, a plasmalogen and a pharmacologically acceptable carrier. The pharmacologically acceptable carrier is various organic carrier substances or inorganic carrier substances used as formulation materials. The pharmacologically acceptable carrier is, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring agent, etc. in a solid preparation, or a solvent, a dissolution aid, a suspending agent, an isotonic agent, a buffer, a flavoring agent, etc. in a liquid preparation. In addition, additives such as preservatives, antioxidants, colorants, and sweeteners may be blended as necessary.

[0036] Examples of excipients include lactose, sucrose, D-mannitol, D-sorbitol, starch, pregelatinized starch, dextrin, crystalline cellulose, low-substituted hydroxypropyl cellulose, sodium carboxymethylcellulose, gum arabic, pullulan, soft anhydrous silicic acid, synthetic aluminum silicate, magnesium aluminometasilicate, xylitol, sorbitol, and erythritol.

[0037] Examples of binders include, in addition to the above-mentioned excipients, pregelatinized starch, sucrose, gelatin, gum arabic, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, crystalline cellulose, sucrose, D-mannitol, trehalose, dextrin, pullulan, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, and the like.

[0038] Examples of disintegrants include, in addition to the above-mentioned excipients, lactose, sucrose, starch, carboxymethylcellulose, carboxymethylcellulose calcium, croscarmellose sodium, carboxymethylstarch sodium, low-substituted hydroxypropylcellulose, soft anhydrous silicic acid, calcium carbonate, and the like.

[0039] Examples of lubricants include magnesium stearate, calcium stearate, talc, colloidal silica, polyethylene glycol, and the like.

[0040] Examples of the stabilizer include paraoxybenzoic acid esters such as methylparaben and propylparaben, alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol, benzalkonium chloride, acetic anhydride, sorbic acid, etc. Examples of the flavoring agent include sweeteners, acidulants, fragrances, etc.

[0041] Examples of the solvent include water for injection, physiological saline, Ringer's solution, alcohol, propylene glycol, polyethylene glycol, sesame oil, corn oil, olive oil, cottonseed oil, etc. Examples of the solubilizing agent include polyethylene glycol, propylene glycol, D-mannitol, trehalose, benzyl benzoate, ethanol, tris(hydroxymethyl)aminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate, sodium salicylate, sodium acetate, etc.

[0042] Examples of suspending agents include surfactants such as stearyl triethanolamine, sodium lauryl sulfate, lauryl aminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, and glycerin monostearate; hydrophilic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose; polysorbates, and polyoxyethylene hydrogenated castor oil.

[0043] Examples of isotonicity agents include sodium chloride, glycerin, D-mannitol, D-sorbitol, glucose, xylitol, fructose, etc. Examples of buffering agents include buffer solutions such as phosphates, acetates, carbonates, citrates, etc.

[0044] Examples of preservatives include paraoxybenzoic acid esters, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid, etc. Examples of antioxidants include sulfites, ascorbates, etc. Examples of colorants include water-soluble colored tar dyes, lake dyes, natural dyes, etc. Examples of sweeteners include sodium saccharin, dipotassium glycyrrhizinate, aspartame, stevia, etc.

[0045] The muscle-building agent according to this embodiment is manufactured by a known method and is provided in the form of, for example, a liquid, cream, ointment, tablet, granule, fine granule, powder, tablet, capsule, or the like.

[0046] The dosage of the muscle-building agent according to the present embodiment is appropriately determined depending on the age, weight, symptoms, etc., of the human or animal to which it is administered. The muscle-building agent is administered so that the plasmalogen is in an effective amount. The effective amount is the amount of plasmalogen necessary to obtain the desired result, and is the amount necessary to delay, inhibit, prevent, reverse, or cure the progression of the condition to be treated or treated. Conditions or diseases that can be treated or prevented by muscle building include, for example, musculoskeletal diseases such as sarcopenia and intractable muscle diseases such as muscular dystrophy.

[0047] The dosage of the muscle-building agent according to the present embodiment is, for example, 0.000001 μg or more per day for an adult, preferably 1 μg or more or 500 μg or more, more preferably 1000 μg or more. The upper limit is, for example, 20000 μg per day, preferably 10000 μg. The muscle-building agent can be administered once or multiple times per day. The muscle-building agent may also be administered at various administration frequencies, such as every other day, once a week, every other week, or once a month. It should be noted that amounts outside the above ranges can also be used as necessary.

[0048] Plasmalogen, which is an active ingredient in the muscle-building agent according to the present embodiment, activates satellite cells and thickens muscle fibers, as shown in the following examples. The muscle-building agent is useful for repairing and strengthening muscles, as well as increasing muscle mass and strength. Since plasmalogen is a substance extracted from animal tissues, the muscle-building agent according to the present embodiment is highly safe.

[0049] The muscle-building agent according to this embodiment may be used as a reagent for the purpose of building muscle in in vitro and in vivo experiments.

[0050] In another aspect of the present embodiment, there is provided an oral composition for muscle building. Specific examples of the oral composition include supplements, food and drink compositions, functional foods, and food additives.

[0051] The form of the supplement is not particularly limited and may be any form such as tablets, powders, granules, capsules, sugar-coated tablets, films, lozenges, chewable tablets, solutions, emulsions, and suspensions. The supplement may contain any component normally used as a supplement.

[0052] The term "functional food" refers to food or beverage that is taken for the purpose of maintaining health, and includes foods with health claims, foods with functional claims, foods with nutrient functions, health foods, and nutritional supplements. As functional foods, foods with health claims or foods with nutrient functions are preferred. When commercializing the composition as a functional food, various additives used in foods, specifically colorants, preservatives, thickening stabilizers, antioxidants, bleaching agents, antibacterial and antifungal agents, acidulants, sweeteners, seasonings, emulsifiers, strengthening agents, manufacturing agents, and flavors, may be added to the oral composition for muscle building.

[0053] The functional food may be either a food or a beverage, and is not particularly limited as long as it can be taken orally. Examples of functional foods include beverages, confectionery, processed grain products, paste products, dairy products, seasonings, etc. Examples of beverages include nutritional drinks, soft drinks, black tea, green tea, etc. Examples of confectionery include candy, cookies, tablet candy, chewing gum, jelly, etc. Examples of processed grain products and noodles include bread, cooked rice, biscuits, etc. Examples of paste products include sausages, ham, kamaboko, etc. Examples of dairy products include butter, yogurt, etc.

[0054] The muscle-building oral composition may be added to food as a food additive. In this case, the food additive may be in the form of a paste, gel, powder, liquid, suspension, emulsion, granule, or the like, so as to be easily added to food.

[0055] The muscle-building oral composition according to the present embodiment may contain water, vitamins, minerals, organic acids, organic bases, fruit juice, flavors, functional ingredients, food additives, etc., within the range that maintains the effectiveness of muscle building. The muscle-building oral composition can be produced by a known method by adding ingredients other than the plasmalogen as necessary.

[0056] The above-mentioned oral composition for muscle building may be divided and stored in one or more containers so that the daily intake amount is the above-mentioned intake amount, in which case, preferably, one container contains one day's worth of oral composition for muscle building.

[0057] The above-mentioned oral composition for muscle building is provided in a form that can be distinguished from other products as a product in that it contains plasmalogen and is used for muscle building. For example, at least one of the packaging, instructions, and promotional materials of the oral composition for muscle building product indicates that it has a muscle building function.

[0058] When the muscle-building oral composition according to the present embodiment is provided as a food or beverage composition, it may be provided or sold as a food or beverage labeled with uses (including health uses) such as prevention of musculoskeletal diseases and intractable muscle diseases, risk reduction of these diseases, symptom relief of diseases, and treatment of diseases, in addition to muscle building. The act of "labeling" includes all acts for informing consumers of the above uses. As long as the expression can recall or infer the above uses, it all falls under the act of "labeling", regardless of the purpose of the labeling, the content of the labeling, the object to be displayed, the medium, etc.

[0059] It is preferable that the "labeling" be done in an expression that allows consumers to directly recognize the above-mentioned uses. Specifically, this includes the acts of transferring, delivering, displaying for the purpose of transferring or delivering, or importing food and beverage products or product packaging on which the above-mentioned uses are written, displaying or distributing advertisements, price lists, or transaction documents related to the products and writing the above-mentioned uses in information containing the above-mentioned uses and providing them by electromagnetic means (such as the Internet), etc.

[0060] Examples of "labeling" include labeling as health food, functional food, enteral nutritional food, special purpose food, health functional food, food for specified health uses, nutrient functional food, functional food, medical drug, etc. Among these, in particular, labeling approved under systems related to foods for specified health uses, foods with nutrient functions, or foods with functional claims, or similar systems, etc. Specific examples include labeling as foods for specified health uses, labeling as foods for conditional specified health uses, labeling that indicates an effect on the structure or function of the body, labeling that reduces the risk of disease, labeling of functionality based on scientific evidence, etc. More specifically, typical examples include labeling as foods for specified health uses (especially labeling of health uses) and similar labeling.

[0061] In another aspect of the present embodiment, there is provided a use of plasmalogen for the manufacture of a muscle-building agent or an oral composition for muscle building. In another aspect of the present embodiment, there is provided a muscle building method comprising a step of administering plasmalogen to a subject. In another aspect of the present embodiment, there is provided a plasmalogen for use in muscle building. Note that muscle building includes prevention, suppression, inhibition and recovery of a decrease in muscle mass or muscle strength.

[0062] The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples. EXAMPLES

[0063] [Preparation of sPls] sPls was prepared from scallops (Mizuhopecten yessoensis) in the following manner. Raw scallops were mixed with Coclase P (Mitsubishi Chemical Foods Corporation), ethanol was added, and the mixture was filtered under suction. The filtrate was dried in a rotary evaporator to obtain sPls.

[0064] [Administration of sPls] Four-week-old male c57BL6J mice (n=4 per group) were administered sPls at a dose of 1 mg / kg in drinking water for four weeks. Soleus and thigh muscles were collected from each mouse and analyzed as follows.

[0065] [Evaluation of AMPK activation by immunohistochemistry] Thigh muscles fixed in 4% paraformaldehyde (PFA) solution were stored in dHO. 2 The sections were then rinsed with 0.20 mL of 100 mL of 10% ethanol. The sections were then incubated at 37°C for 3 hours with 0.2% collagenase I solution to obtain samples. The 0.2% collagenase I solution was prepared using Buffer (50 mM Tris-HCl, 0.36 mM CaCl 2 , pH 7.4) containing collagenase type I (C0130, Sigma-Aldrich) at a concentration of 0.2%.

[0066] Wash the samples with phosphate-buffered saline (PBS) and dHO.2 After rinsing with O, the muscle fibers were disaggregated under a stereomicroscope, and single muscle fibers were placed on a slide glass and dried at 65°C and then at 37°C for 18 hours.

[0067] The samples were dried at 65°C for 15 minutes, then fixed with 4% PFA solution at room temperature for 40 minutes. After washing with PBS, they were treated with 0.5% Tx-100 / PBS at room temperature for 6 minutes. After washing with PBS, they were treated with blocking buffer (1% BSA-PBS) at room temperature for 20 minutes.

[0068] As primary antibodies, Phospho-AMPKα (Thr172) antibody (#2535, Cell Signaling Technology) and MyoD antibody (NB120-788, Novus Biologicals) were used, each diluted 2000-fold with 5% BSA-PBS. The primary antibodies were reacted with the samples for 18 hours at 4°C. After washing with PBS, the samples were reacted with a solution of Alexa Fluor 488 anti-rabbit IgG, Alexa Fluor 568 anti-mouse IgG (both from Invitrogen) and 3 mM DAPI diluted 2000-fold with 1% BSA-PBS for 1 hour at room temperature in the dark. After washing the samples with PBS, they were incubated in dH 2 The sections were rinsed with O, embedded in cover glasses, and observed using a BZ-X810 (Keyence Corporation).

[0069] [AMPK phosphorylation and MyoD expression analysis] Soleus muscles were homogenized in Buffer A (0.25 M sucrose, 10 mM Hepes-KOH (pH 7.5), 1 mM EDTA, protease inhibitor cocktail) using 18G to 25G needles, and protein was quantified and electrophoresed. The protein was then transferred to a PVDF membrane and detected by Western blotting using antibody solutions of Phospho-AMPKα (Thr172) antibody (#2535, Cell Signaling Technology) and AMPKα antibody (#5831, Cell Signaling Technology) diluted 2000-fold. Signals were quantified using Multi Gauge software version 3.0 software (Fujifilm). Signals obtained with Phospho-AMPKα (Thr172) antibody were divided by the signal obtained with AMPK antibody to standardize. Furthermore, the signal intensity in the cells treated with each treatment was expressed as a relative value, with the value obtained from untreated cells taken as 1. The trials were performed three or more times, and the results were shown as the average and standard deviation.

[0070] Samples similarly prepared from soleus muscle were detected and quantified using MyoD antibody (NB120-788, Novus Biologicals, diluted 2000-fold) and GAPDH antibody (M171-3, MBL, diluted 4000-fold).

[0071] [Measurement of muscle fiber diameter] The focal depth was adjusted so that the single muscle fibers prepared from the thigh muscle as described above were the thickest, and bright-field images were taken with a BZ-X810 (Keyence Corporation). The diameters of two or three points were then measured for each muscle fiber. A total of nine points were measured for muscle fibers not administered sPls, and a total of 29 points were measured for muscle fibers administered sPls.

[0072] [result] Figure 1 shows the results of evaluating AMPK activation by immunostaining. sPls increased the expression of pAMPK and the muscle differentiation marker MyoD in individual cells of thigh muscle, indicating that sPls increased the number of muscle satellite cells.

[0073] FIG. 2A shows the results of Western blotting for pAMPK. FIG. 2B shows the relative amount of pAMPK / AMPK based on the results of Western blotting. Activation of AMPK by sPls was observed in soleus muscle. FIG. 3A shows the results of Western blotting for MyoD. FIG. 3B shows the relative amount of MyoD based on the results of Western blotting. Increase in MyoD by sPls was observed in soleus muscle.

[0074] Figure 4A shows a bright field image of thigh muscle, and Figure 4B shows the diameter of muscle fibers. It was demonstrated that sPls activates APMK in muscle and thickens muscle fibers.

[0075] The present invention allows various embodiments and modifications without departing from the broad spirit and scope of the present invention. The above-described embodiments are for the purpose of explaining the present invention and do not limit the scope of the present invention. That is, the scope of the present invention is indicated by the claims, not the embodiments. Various modifications made within the scope of the claims and the scope of the invention equivalent thereto are considered to be within the scope of the present invention. [Industrial Applicability]

[0076] The present invention is useful as a muscle-building agent.

Claims

1. Contains plasmalogen as an active ingredient. Muscle-building agent.

2. Contains plasmalogen as an active ingredient. Oral composition for muscle building.

3. The plasmalogen is It is a plasmalogen extracted from animal tissue. The muscle-building agent according to claim 1 or the oral composition for muscle building according to claim 2.

4. The animal tissue comprises: The tissue is an animal selected from the group consisting of shellfish, sea squirts and birds. The muscle-building agent or oral composition for muscle-building according to claim 3.

5. The animal tissue comprises: Scallop tissue, The muscle-building agent or oral composition for muscle-building according to claim 3.

6. Use of plasmalogen for producing a muscle-building agent or an oral composition for muscle building.

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