JP2022046069A - Muscular atrophy inhibitor - Google Patents

Abstract

To provide a muscular atrophy inhibitor capable of rectifying (preventing or treating) sarcopenia by suppressing the appearance of MuRF1 or Atogin-1.SOLUTION: This muscular atrophy inhibitor includes at least one kind of a crushed article or an extract selected from the group consisting of a coriander, a cardamom, a long pepper, and cinnamon.SELECTED DRAWING: None

Description

The present invention relates to a muscular atrophy inhibitor.

Sarcopenia is a syndrome associated with age-related loss of skeletal muscle mass and strength. It has been confirmed that high-intensity training is effective for the prevention and improvement of sarcopenia. However, in the elderly, it is physically burdensome to actively carry out high-intensity exercise. Therefore, a nutritional approach that can be easily implemented is desired for the prevention and improvement of sarcopenia.
For example, as an ingredient having an effect of improving muscle function, a muscle atrophy inhibitor containing proanthocyanidin as an active ingredient (Patent Document 1), and a transition of muscle fiber type during disuse muscle atrophy using fruit-derived polyphenol as an active ingredient. A muscle fiber type transition inhibitor (Patent Document 2) and a muscle function decline inhibitor containing catechins as an active ingredient (Patent Document 3) have been reported.

Japanese Unexamined Patent Publication No. 2002-338464 Japanese Unexamined Patent Publication No. 2006-328031 Japanese Unexamined Patent Publication No. 2008-13473

MuRF1 and Atogin-1 have been reported as genes involved in muscle loss. An object of the present invention is to provide a muscular atrophy inhibitor capable of suppressing the expression of MuRF1 or Atogin-1 and improving (preventing or treating) sarcopenia.

As a result of diligent research on a muscle atrophy inhibitor capable of improving (preventing or treating) sarcopenia, the present inventor surprisingly found that coriander, cardamom, long pepper, or cinnamon are involved in muscle loss in MuRF1 and Atogin-1. It was found to suppress.
The present invention is based on these findings.
Therefore, the present invention
[1] A muscular atrophy inhibitor containing at least one crushed product or extract selected from the group consisting of coriander, cardamom, long pepper, and cinnamon as an active ingredient.
[2] The muscular atrophy inhibitor according to [1], which suppresses the expression of MuRF1 and / or Atologin-1.
[3] A food composition for suppressing muscular atrophy containing the muscular atrophy inhibitor according to [1] or [2], and the muscular atrophy inhibitor or [3] according to [4] [1] or [2]. A method for suppressing muscular atrophy (excluding medical practice for humans), which comprises a step of ingesting an effective amount of the food composition for suppressing muscular atrophy described in 1.
Regarding.

According to the muscular atrophy inhibitor of the present invention, skeletal muscle atrophy can be suppressed and sarcopenia can be prevented or ameliorated.

It is a graph which showed the effect on the expression of the MuRF1 gene of the distilled water extract. It is a graph which showed the effect on the expression of the Antrogin-1 gene of the distilled water extract. It is a graph which showed the effect on the expression of MuRF1 gene of a hot water extract. It is a graph which showed the effect on the expression of the Antrogin-1 gene of the hot water extract. It is a graph which showed the effect on the expression of MuRF1 gene of an ethanol extract. It is a graph which showed the effect on the expression of the Antrogin-1 gene of the ethanol extract. It is a graph which showed the effect on the expression of the Antrogin-1 gene of the hexane extract.

《Muscle atrophy inhibitor》
The muscular atrophy inhibitor of the present invention contains at least one crushed product or extract selected from the group consisting of coriander, cardamom, long pepper, and cinnamon as an active ingredient. The muscular atrophy inhibitor of the present invention preferably suppresses muscular atrophy by suppressing the expression of MuRF1 and / or Atrogin-1.

《Sarcopenia》
Sarcopenia is a decrease in muscle strength and physical function associated with a decrease in muscle mass due to aging, illness, lack of exercise, malnutrition, and the like. Sarcopenia can be prevented and improved with moderate exercise. However, if an elderly person is hospitalized for a long period of time due to a fracture or the like, muscle mass may decrease, exercise may become impossible, and a virtuous cycle may occur in which sarcopenia progresses.
As used herein, amelioration of sarcopenia includes prevention or treatment.

(coriander)
Coriander used as an active ingredient of the muscular atrophy inhibitor of the present invention is a kind of Umbelliferae and is used as a spice. As spices, the whole plant is used, the fruit and seed part are used as coriander seed, and the leaf and stem part are used as coriander leaf and pakuchi. Further, the coriander of the present invention may have any shape, such as the state as it is harvested or the state of a crushed powder or paste. The site to be used is not particularly limited, and may be only a specific site or a mixture of two or more types of sites. Preferred sites are coriander seeds or coriander leaves. Further, as the active ingredient of the muscular atrophy inhibitor of the present invention, a raw state or a dry state may be used.

(cardamom)
The cardamom used as the active ingredient of the muscular atrophy inhibitor of the present invention may be green cardamom, black cardamom, raw or dried. Further, it may have any shape such as the state as it is harvested or the state of a crushed powder or paste. The preferred moiety is the fruit used as a spice, but it may be another moiety or a mixture of the fruit and the other moiety. As the active ingredient of the muscular atrophy inhibitor of the present invention, a dried product or a raw site may be used.

(Long pepper)
The long pepper used as an active ingredient of the muscular atrophy inhibitor of the present invention is a vine woody plant of the family Pepper family, also called long pepper (piper longum L.). The long pepper of the present invention may have any shape, such as as it is harvested or as a crushed powder or paste. The site to be used is not particularly limited, and may be only a specific site or a mixture of two or more types of sites. A preferred site is the fruit used as a spice. Further, as the active ingredient of the muscular atrophy inhibitor of the present invention, a raw state or a dry state may be used.

(cinnamon)
The cinnamon used as the active ingredient of the muscle atrophy inhibitor of the present invention is a tree of the genus Cinnamomum, and may be Chinese cinnamon, Ceylon cinnamon, Indonesian cinnamon, or Indian cinnamon, as it is harvested, or as a powder. Alternatively, it may have any shape, such as the state of crushed paste. The site to be used is not particularly limited, and may be only a specific site or a mixture of two or more types of sites. A preferred site is the bark used as a spice. Further, as the active ingredient of the muscular atrophy inhibitor of the present invention, a raw state or a dry state may be used.

《Crushed material》
In the present invention, the crushed product is a state in which coriander, cardamom, long pepper or cinnamon (hereinafter, when referred to as coriander or the like, means coriander, cardamom, long pepper, cinnamon) is crushed. It can be powdery, granular, or pasty, for example. The pulverized product is preferably a powder. The process for processing into a crushed product or powder is not particularly limited, but by any method usually used by those skilled in the art, using a crushing device or instrument such as a crusher, a mill, a blender, a mixer, and a stone mill. Examples include the process of crushing a plant. Prior to pulverization, the plant may be dried by any method usually used by those skilled in the art, for example, hot air of 100 ° C. or higher, warm air that does not affect the active ingredient, freeze-drying, etc., and the water content is 10% by weight or less. The dried plant may be stored for a certain period of time.

When the coriander or the like contained in the muscular atrophy inhibitor of the present invention is a pulverized product, the average longest diameter of the pulverized product such as coriander is 0.01 to 4 mm, preferably 0. Those having a diameter of 02 to 1 mm, more preferably 0.02 to 0.5 mm can be used. Further, it is possible to use a crushed product such as coriander having a maximum diameter of 0.01 to 4 mm, preferably 0.02 to 1 mm, more preferably 0.02 to 0.5 mm in an amount of 90% by weight or more. can. In addition, 90% by weight or more of crushed material such as coriander passes through 4.7 mesh (4 mm) 16 mesh (1 mm) or 30 mesh (0.5 mm) in the JIS test sieve mesh conversion table. can do. When the longest diameter of a pulverized product such as coriander is 1 mm or less, the expression of MuRF1 or Atrogin-1 of the present invention is improved. Therefore, it is preferable to use a crushed product having a maximum diameter of 4 mm or less.
The average longest diameter of coriander and the like can be measured using a known device for measuring the particle size.

《Extract》
The muscular atrophy inhibitor of the present invention can contain an extract such as coriander. The muscular atrophy inhibitor of the present invention can also contain both a pulverized product such as coriander and an extract. As the coriander or the like used for extracting the active ingredient of the muscular atrophy inhibitor of the present invention, the fruit may be used raw or dried. Further, it may be processed into a crushed product or a powder so that the extraction efficiency is improved.

Extraction of the active ingredient is a conventional extraction method used for extraction of a component derived from a plant, for example, but not limited to, a steam distillation method, a solvent extraction method, and a squeezing method (direct, high temperature, or low temperature). ), Or a supercritical extraction method can be used. A combination of these extraction methods, for example, a method of extracting with a solvent after squeezing may be used.

The extraction solvent used for extraction by the solvent extraction method is not limited as long as the active ingredient can be sufficiently extracted in the extract, and for example, an organic solvent, an aqueous solvent, or an organic solvent and an aqueous solvent. Mixtures can be used.

Extracts such as coriander contained in the muscle atrophy inhibitor of the present invention can be extracted with organic solvents such as alcohol, acetone, benzene, esters, ethyl acetate, hexane, chloroform, and diethyl ether. The alcohol is not limited as long as the active ingredient can be sufficiently extracted in the extract, but for example, a monohydric alcohol having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, isopropyl alcohol, and butyl alcohol. Alcohol can be used. Polyhydric alcohols having 2 to 5 carbon atoms such as 1,3-butylene glycol, propylene glycol, dipropylene glycol, and glycerin can also be used. Preferred alcohols are monohydric alcohols having 1 to 5 carbon atoms, more preferred alcohols are methanol and ethanol, and most preferred alcohols are ethanol.

The extract such as coriander contained in the muscular atrophy inhibitor of the present invention can be extracted with an aqueous solvent. The aqueous solvent is not limited as long as it contains water, and for example, water, physiological saline, a buffer solution, or the like can be used. Examples of the buffer solution include phosphate buffer solution, sodium phosphate buffer solution, sodium carbonate buffer solution, citrate buffer solution, citrate phosphate buffer solution, acetate buffer solution, Tris buffer solution and the like. The pH of the aqueous solvent is not particularly limited.

The extract such as coriander contained in the muscular atrophy inhibitor of the present invention can be extracted by a mixture of an organic solvent and an aqueous solvent. The amount of the aqueous solvent contained in the extraction solvent is not limited as long as the active ingredient can be sufficiently extracted in the extract, but is, for example, 50% by weight or more, 70, based on the total amount of the extraction solvent. It can be% by weight or more, or 90% by weight or more.

When an extract such as coriander contained in the muscular atrophy inhibitor of the present invention is extracted by a solvent extraction method, the extraction temperature is particularly limited as long as the active ingredient can be sufficiently extracted into the extract. However, it is preferably −50 ° C. to 100 ° C., more preferably 0 ° C. to 100 ° C., and most preferably 5 ° C. to 100 ° C.

The extraction time of the extract such as coriander contained in the muscular atrophy inhibitor of the present invention is, for example, when it is in the state of coriander or the like, that is, when it is raw or dried, or when it is processed into a crushed product or powder. It can be appropriately determined according to the processing state. Further, the extraction time can be appropriately determined according to the extraction conditions such as, for example, the temperature of the extract or the presence or absence of stirring or shaking. The extraction time is preferably 5 seconds to 1 week, more preferably 5 seconds to 1 day, and most preferably 10 minutes to 12 hours. Further, when the extract such as coriander contained in the muscular atrophy inhibitor of the present invention is extracted by the solvent extraction method, it is preferable to carry out the extraction while stirring or shaking so as to improve the extraction efficiency.

The squeezing method is a method of extracting an extract of coriander or the like by physically applying pressure to coriander or the like. There are a direct pressing method performed at room temperature, a high temperature pressing method performed at high temperature, and a low temperature pressing method performed at low temperature. The extract contained in the muscular atrophy inhibitor of the present invention can be extracted by using any squeezing method.

The extract contained in the muscular atrophy inhibitor of the present invention can be extracted by using a supercritical extraction method. The supercritical extraction method is a method of extracting an extract from a specific plant using a substance in a supercritical state. As a substance in a supercritical state, for example, carbon dioxide has a strong dissolving power, and is therefore generally used for decaffeination of coffee or extraction of flavors and pharmaceutical components from natural raw materials such as plants.

(MuRF1)
MuRF1 (Muscle RING-Finger Protein-1) is a RING-type ubiquitin ligase with a molecular weight of 40 kDa that is specifically expressed in skeletal muscle and myocardium. It is known that the expression level is increased during muscular atrophy. On the other hand, it has been reported that muscle atrophy does not occur in MuRF1 knockout mice, and it is considered to be a protein related to muscle atrophy.

(Atrogin-1)
Atrogin-1, also referred to as MAFbx-1, is an SCF complex ubiquitin ligase having a molecular weight of 40 kDa. In the SCF complex ubiquitin ligase, cullin is used as a platform protein, E2 binds to the RING domain of Rbx1 (Roc1), and the F-box protein bound via Skp1 recognizes the substrate protein and ubiquitinates it. Atrogin-1 is also known to have an increased expression level during muscle atrophy, while it has been reported that muscle atrophy does not occur in Atrogin-1 knockout mice.

The active ingredient in the present invention, a ground product such as coriander or an extract, is a subject [animal] alone or, preferably with a conventional carrier or diluent which is pharmaceutically or veterinarily acceptable. , Preferably mammals (particularly humans)] in effective amounts. It is also possible to feed animals other than humans in the form of food and drink as feed.

The muscular atrophy inhibitor of the present invention may consist of a pulverized product or an extract such as coriander, or may contain a pulverized product or an extract such as coriander. When the muscular atrophy inhibitor of the present invention contains a pulverized product such as coriander or an extract, other additives can be contained.

The dosage form of the muscular atrophy inhibitor of the present invention is not particularly limited, and is a powder, a fine granule, a granule, a tablet, a capsule, a suspension, an emulsion, a syrup, an extract, or a pill. Oral preparations such as, or parenteral preparations can be mentioned.

The oral preparation includes, for example, gelatin, sodium alginate, starch, corn starch, sucrose, lactose, glucose, mannit, carboxymethyl cellulose, dextrin, polyvinylpyrrolidone, crystalline cellulose, soy lecithin, sucrose, fatty acid ester, talc, magnesium stearate, and the like. Excipients such as polyethylene glycol, magnesium silicate, silicic anhydride, or synthetic aluminum silicate, binders, disintegrants, surfactants, lubricants, fluidity promoters, diluents, preservatives, colorants, It can be produced according to a conventional method using a fragrance, a flavoring agent, a stabilizer, a moisturizing agent, a preservative, an antioxidant or the like.

Examples of parenteral preparations include injections. In the preparation of injections, in addition to the active ingredient, for example, a water-soluble solvent such as physiological saline or Ringer's solution, a water-insoluble solvent such as vegetable oil or fatty acid ester, an tonicity agent such as glucose or sodium chloride, and a solubilizing agent. , Stabilizers, preservatives, suspending agents, emulsifiers and the like can be optionally used.

The muscular atrophy inhibitor of the present invention is, but is not limited to, a dry solid product obtained by drying an extract such as coriander, for example, 0.01 to 500 mg / mL, preferably 0. It can be contained in an amount of 1 to 400 mg / mL, more preferably 0.5 to 300 mg / mL, still more preferably 1 to 200 mg / mL, still more preferably 2 to 150 mg / mL, and most preferably 1 to 100 mg / mL.

The dose of coriander or the like of the present invention can be appropriately determined, and for example, as a dry solid obtained by drying the extract, 1 to 3000 mg / kg body weight / day, preferably 5 to 2000 mg / day. kg body weight / day, more preferably 10 to 1500 mg / kg body weight / day, still more preferably 15 to 1000 mg / kg body weight / day, still more preferably 20 to 800 mg / kg body weight / day, most preferably 25 to 400 mg / kg body weight. / Can be a day.

When the muscular atrophy inhibitor of the present invention is used, the dose as a dry solid (or freeze-dried powder) of the obtained extract is 0.1 to 1000 mg / kg body weight / day, preferably 0.5. ~ 500 mg / kg body weight / day, more preferably 1 to 250 mg / kg body weight / day, still more preferably 3 to 100 mg / kg body weight / day, still more preferably 5 to 50 mg / kg body weight / day, or most preferably 8 to. It can be 30 mg / kg body weight / day.

Of course, the above administration method is an example, and other administration methods may be used. It is desirable that the method, dose, administration period, administration interval, etc. of the muscular atrophy inhibitor to humans be determined by a controlled clinical trial.

The muscle atrophy inhibitor of the present invention can be administered to humans, but the administration target may be animals other than humans, and pets such as dogs, cats, rabbits, hamsters, guinea pigs, and squirrels; cows. And domestic animals such as pigs; experimental animals such as mice and rats; and animals bred in zoos and the like.

<< Food composition >>
The food composition of the present invention contains the muscular atrophy inhibitor of the present invention. The food composition of the present invention may contain a food material. In the present specification, the food material can be eaten as a finished food or beverage such as curry or beer, a spice as a raw material such as curry or beer, or as a food or beverage as it is such as miso or sake. And also includes those that can be used as raw materials for cooking.

As foods, specifically, fresh cooked foods such as salads; cooked foods such as croquettes, fried white fish, pizza, and hamburgers; fried cooked foods such as fried vegetables; tomatoes, peppers, celery, biting melons, carrots, and potatoes. , And vegetables such as asparagus and cooked products processed from these vegetables; sweets such as cookies, bread, biscuits, dried bread, cakes, rice cakes, sheep rice cakes, puddings, jellies, ice creams, chewing gums, crackers, chips, chocolates and candy. Kind; noodles such as udon, pasta, and buckwheat; fish paste products such as kamaboko, ham, and fish sausage; dairy products such as cheese, cream, and butter; miso, soy sauce, sauce, dressing, ketchup, mayonnaise, soup base , Noodle soup, curry flour, salt and pepper, sesame salt, mirin, oden no moto, ru, seasoning spices and other seasonings; soy foods such as tofu; konjac; and supplements. The food is preferably a sauce (eg, for curry, stew, gratin, or pasta), ru (eg, for curry or stew), mixed spices, seasoning spices, or supplements.

Beverages include, for example, coffee beverages; cocoa beverages; vegetable juices obtained from the above vegetables; fruit juice beverages such as grapefruit juice, orange juice, grape juice, and lemon juice; tea beverages such as green tea, tea, roasted tea, and oolong tea; Alcoholic beverages such as beer, wine (red wine, white wine, or sparkling wine, etc.), sake, plum wine, sparkling liquor, whiskey, brandy, shochu, lamb, gin, and liqueurs; milk beverages; soy milk beverages; liquid foods; and Sports drinks and the like can be mentioned. The beverage is preferably spice tea (eg herbal tea or chai).

The food composition of the present invention can be, for example, powdery, granular, solid, liquid, capsule, paste, gel or tablet.

These foods or beverages may include antioxidants, fragrances, acidulants, colorants, emulsifiers, preservatives, seasonings, sweeteners, spices, pH regulators, stabilizers, antioxidants, vegetable oils, animal oils, if desired. , Sugar and sugar alcohols, vitamins, organic acids, fruit juice extracts, vegetable extracts, grains, beans, vegetables, meats, seafood and other food additives and food ingredients alone or in combination of two or more. be able to. The blending amount of these food materials and food additives can be appropriately determined within a range that does not impair the object of the present invention.

These foods or beverages are commonly sterilized, for example, heat and pressure sterilization such as retort and autoclave, batch sterilization, plate sterilization, energization heat sterilization, microwave heat sterilization, and steam sterilization such as injection and infusion. Processing can be performed.

Foods and beverages include functional foods (beverages) and health foods (beverages). In the present specification, the "health food (beverage)" means a food or beverage that has or can be expected to have some effect on health, and the "functional food (beverage)" is described above. Among "health foods (beverages)", it means foods or beverages designed and processed so as to sufficiently express a bioregulatory function (that is, an immunostimulatory function). Functional foods and health foods can be in the form of granules, solids, liquids, capsules, gels or tablets.

Food or beverages include feed for animals. Examples of the target animal include primates such as humans, cows, pigs, sheep, goats, horses, dogs, cats, rabbits, rats, mice and the like.

<< Method of suppressing muscle atrophy >>
The method for suppressing muscular atrophy of the present invention includes a step of ingesting an effective amount of the muscular atrophy inhibitor or the food composition for suppressing muscular atrophy. However, the method for suppressing muscular atrophy of the present invention does not include medical treatment for humans. Specifically, it includes ingestion of a muscular atrophy inhibitor in a training gym, a nursing facility, etc., and ingestion of a food composition for suppressing muscular atrophy.
The "muscle atrophy inhibitor" and the "food composition for suppressing muscular atrophy" in the method for suppressing muscular atrophy of the present invention are described in the respective sections.

Hereinafter, the present invention will be specifically described with reference to Examples, but these do not limit the scope of the present invention.

<< Example 1 >>
In this example, distilled water extracts were prepared from coriander seeds (dried fruit parts) and cardamom (dried fruit parts).
10 times the amount of distilled water (200 g) was added to 20 g of coriander seeds or cardamom and mixed. The mixture was stirred at 4 ° C. for 12 hours using a stirrer. The extract was centrifuged at 3200 rpm for 10 minutes. The supernatant was collected and suction filtered through a filter having a maximum pore size of 20 to 25 μm. The obtained supernatant was freeze-dried to obtain a dry solid product of a water extract.

<< Example 2 >>
In this example, hot water extracts were prepared from cinnamon (dried bark), coriander seeds (dried fruit part) and cardamom (dried fruit part / seed part).
To 20 g of cinnamon, coriander seed or cardamom was added 10 times the amount of distilled water (200 g) and mixed. Using a stirrer, the mixture was stirred at 95 ° C. for 1 hour. The extract was centrifuged at 3200 rpm for 10 minutes. The supernatant was collected and suction filtered through a filter having a maximum pore size of 20 to 25 μm. The obtained supernatant was freeze-dried to obtain a dry solid product of a hot water extract.

<< Example 3 >>
In this example, ethanol extracts were prepared from long pepper (dried fruit) and cinnamon (dried bark).
Extraction was performed by suspending long pepper and cinnamon powder in 100% ethanol so as to be 1 g / mL, and shaking for 15 minutes twice. After extraction, the supernatant was collected by centrifugation. The solvent was dried on an evaporator, the weight of the residue was weighed, and then dissolved in a mixed solution of dimethyl sulfoxide (DMSO) and ethanol to a concentration of 200 mg / mL to obtain a test sample.

<< Example 4 >>
In this example, a hexane extract was prepared from cinnamon (dried bark).
Extraction was performed by suspending the powder of long pepper (dried fruit part) in 100% n-hexane so as to be 200 g / mL, and shaking it for 15 minutes twice. After extraction, the supernatant was collected by centrifugation. The solvent was dried on an evaporator, the weight of the residue was weighed, and then dissolved in dimethyl sulfoxide (DMSO) to 200 mg / mL to obtain a test sample.

<< Example 5 >>
Using mouse myoblast lines, the effects of the spice extracts obtained in Examples 1 to 4 on the expression of MuRF1 and Atrogin-1 were investigated.
Mouse myoblast line C2C12 was seeded on a 12-well plate to a 2 × 10 ⁴ cells / mL. After culturing in 10% FBS-DMEM for 24 hours, the medium was replaced with 0.5% FBS-DMEM (differentiation-inducing medium) (the differentiation-inducing medium was replaced once 48 hours after the start of induction). After differentiation, it was replaced with a differentiation-inducing medium containing each spice extract (final concentration 0.01 mg / mL), and after treatment for 24 hours, it was replaced with a differentiation-inducing medium containing each spice extract and dexamethasone (Dex: 1 mM). Time processed. The addition of dexamethasone enhances the expression of the MuRF1 gene and the Atrogin-1 gene, and serves as a model for muscle atrophy.

The expression of MuRF1 gene and Atrogin-1 gene in the collected cells was measured by Real-time RT PCR.
The precultured cell supernatant was removed and the cell surface was washed with PBS (-). 1 mL of Sepasol RAN I Super G (Nacalai Tesque) was added to the cells and the cells were collected by pipetting. Further, 200 μL of chloroform was added, and the mixture was vigorously inverted and mixed for 2 minutes. After mixing, the mixture was centrifuged at 12,000 × g for 15 minutes. After centrifugation, 498 μL of the aqueous solution layer was recovered.
An equal amount of propanol was added to the recovered aqueous solution layer, and after inversion miscibility, the mixture was allowed to stand on ice for 10 minutes. Then, it was centrifuged at 12,000 × g for 10 minutes. After centrifugation, the supernatant was removed, 75% ethanol was added, the mixture was inverted and mixed, and the mixture was centrifuged at 12,000 × g for 5 minutes. After centrifugation, the supernatant was removed, and after air-drying, ultrapure water was added and the mixture was allowed to stand on ice for 15 minutes. The absorbance of the mRNA solution was measured with a spectrophotometer (BioSpec-nano, SHIMADZU) to determine the concentration of mRNA in the solution.
An aqueous RNA solution containing 100 ng of RNA in a microtube was filled up to 13.5 μL with sterile ultrapure water, and 0.5 μL of oligo dT primer was added. The mixture of mRNA and oligo dT primer was heated at 70 ° C. for 5 minutes and then cooled at 4 ° C. Further, 5 μL of dNTP, 5 μL of 5 × reverse transcription reaction buffer, 0.5 μL of RNase inhibitor (Invitrogen), 0.2 μL of reverse transcriptase (Nippon Gene), and 0.3 μL of ultrapure water were added at 42 ° C. Was heated for 60 minutes. Then, it cooled at 4 degreeC. After completion of the reaction, the mixture was diluted 40-fold with ultrapure water.
To the PCR96well plate, add 1 μL of cDNA, 1 μL of MuRF1, or 1 μL of forward primer and reverse primer of Atrogin-1, 3 μL of ultrapure water, and 5 μL of DNA detection solution, and add 95 ° C-1 minute, (95 ° C-3 seconds, The reaction was carried out under the conditions of 60 ° C.-30 seconds) × 40 cycles. The analysis used the comparative Ct method. β-actin was used as an endogenous control and was used to correct gene expression of interest. The device used was StepOne Plus (manufactured by Life Technologies). Each primer is as follows.
MuRF1 forward primer: 5'-TGAGGTGCTACTTGCTCCT-3'; SEQ ID NO: 1)
MuRF1 reverse primer: 5'-TCACCTGGTGGCTATTCTCC-3'; SEQ ID NO: 2)
Atrogin-1 forward primer: 5'-ATGCACACTGGTGCAGAGAG-3'; SEQ ID NO: 3)
Atrogin-1 reverse primer: 5'-TGTAAGCACAGAGGCAGGTC-3'; SEQ ID NO: 4)

Distilled and hot water extracts of cardamom suppressed the expression of MuRF1 and Atrogin-1 (FIGS. 1-4). The distilled water extract of Korean seed suppressed the expression of MuRF1 and Atrogin-1 (Figs. 1 and 2), and the hot water extract suppressed the expression of Atrogin-1 (Fig. 4). Hot water extract and ethanol extract of cinnamon suppressed the expression of Atrogin-1 (FIGS. 4 and 6). The ethanol extract of Long Pepper suppressed the expression of MuRF1 (Fig. 5), and the hexane extract suppressed the expression of Atrogin-1 (Fig. 7).

The muscular atrophy inhibitor of the present invention can suppress the expression of MuRF1 and Atrogin-1, whose expression is enhanced during muscular atrophy, and can improve the symptoms of sarcopenia.

Claims (4)

An muscular atrophy inhibitor containing at least one crushed product or extract selected from the group consisting of coriander, cardamom, long pepper, and cinnamon as an active ingredient. The muscle atrophy inhibitor according to claim 1, which suppresses the expression of MuRF1 and / or Atologin-1. A food composition for suppressing muscular atrophy, which comprises the muscular atrophy inhibitor according to claim 1 or 2. A method for suppressing muscular atrophy (provided, medical practice for humans), which comprises a step of ingesting an effective amount of the muscular atrophy inhibitor according to claim 1 or 2 or the food composition for muscular atrophy according to claim 3 to a subject. except for).

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