KR100625508B1 - Compositions inhibiting a decrease in bone weight and a decrease in bone salt content in disuse muscle atrophy - Google Patents

Abstract

Compositions containing, as an active ingredient, polyphenols purified from fruit, for example, rose fruit, have the effect of inhibiting muscular atrophy, particularly lung soluble muscular atrophy, in mammals. Useful for inhibiting bone weight and bone salt decline.

Muscular atrophy inhibiting composition, soluble muscle atrophy, oxidized glutathione, fruit polyphenol, bone weight, bone salt, soleus muscle, oxidative stress

Description

Compositions inhibiting a decrease in bone weight and a decrease in bone salt content in disuse muscle atrophy}

The present invention relates to a muscle atrophy inhibiting composition and bone weight and bone salt reduction inhibitory composition, more specifically, muscle atrophy inhibiting composition containing fruit polyphenols as an active ingredient and lung soluble muscle atrophy It relates to a bone weight and bone salt reduction inhibitory composition.

Muscular atrophy can be largely classified into inactive (lung insoluble) disuse atrophy and progressive muscular atrophy.For progressive muscular atrophy, for example, muscular dystrophy ) Neurogenic atrophy such as sclerosis, progressive spinal muscular atrophy, Kugelberg-Belander disease, Bergnich-Hoffmann disease, and, for example, progressive muscular dystrophy [ducenen type, Becker Myogenic atrophy, such as type, zone type, facial thyroid type, and muscular tonic dystrophy.

Among them, in lung-soluble muscle atrophy due to gypsum fixation, an increase in TBA (thiobarbituric acid) reactive positive substance (TBARS), an increase in oxidized glutathione (GSSG), and a ratio of GSSG to total glutathione in atrophic muscles Has been confirmed, suggesting an increase in lipid peroxidation and an increase in oxidative stress [Kondo, H. et al., Acta. Physiol. Scand., 142, 527-528 (1991). The increase in oxidative stress in atrophy muscles is also suggested by the fact that the activity of antioxidant enzymes in atrophy muscles is increased. Kondo, H. et al., Am. J. Physiol., 265, E839-E844 (1993). In addition, in pulmonary soluble muscle atrophy by gypsum fixation, an increase in TBARS is observed with an increase in iron in the microsomal fraction of the atrophic muscle, which increases the production of hydroxy radicals, thereby releasing lipid peroxide. Increasing is suggested [Kondo, H. et al., Acta. Physiol. Scand., 142, 527-528 (1991).

Therefore, oxidative stress is thought to affect the progression of pulmonary soluble muscle atrophy, and the effect of oxidative stress on the progression of pulmonary soluble muscular atrophy has been reviewed in a trial in which vitamin E is administered as an antioxidant [Kondo, H. et al., Acta. Physiol. Scand., 142, 527-528 (1991)], suggests that vitamin E reduces oxidative stress of atrophy muscles, thereby inhibiting the progression of muscle atrophy. In addition, it has been reported that pulmonary soluble muscular atrophy due to gypsum fixation is suppressed by intraperitoneal administration of vitamin E by Appel et al. [Appell, H.-J. et al., Int. J. Sports Med., 18, 157-160 (1997).

In addition, there is a report reviewing whether iron increase in atrophic muscles during lung-soluble muscle atrophy is involved in the progression of muscle atrophy by administration of deferoxamine (DFX) as an iron chelating agent [Kondo, H]. et al., Pflugers Arch., 421, 295-297 (1992)], suggests that iron plays an important role in the oxidative stress of lung soluble muscular dystrophy.

On the other hand, it is well known that polyphenols have antioxidant capacity by radical scavenging action. Flavonoids, a type of polyphenols, have been reported to possess not only radical scavenger action but also chelating action of transition metals. Afanasev, I. B. et al., Biochem. Pharmacol., 38, 1763-1769 (1989).

In addition, there has been a positive correlation between muscle mass and osteomyelitis density to date [Nichols, D. L., et al., Med. Sci. Sports Exerc., 27, 178-182 (1995)], has been reported to decrease bone mass and bone salt as well as decrease muscle mass during immobilization (Grobus, RK, et al., Enfocrinology, 114, 2264). -2270 (1984); Weinreb, M., et al., Bone, 10, 187-194 (1989).

As a manufacturing method of a polyphenol, the method of extracting and extracting from the immature fruit of the Rosaceae fruit (refer Japanese Unexamined-Japanese-Patent No. 7-285876) or grape fruit Method of producing proanthocyanidins in which water or leek seeds of extracts at 70 ° C. or higher are removed by contacting with water at a temperature below 70 ° C. as a pretreatment to remove water-soluble substances (see Japanese Patent No. 2694748). This is known.

It is an object of the present invention to provide a muscle atrophy inhibitor, in particular a muscle atrophy inhibitor which can have an inhibitory effect on lung-soluble muscle atrophy by gypsum fixation.

The inventors of the present invention, in the course of intensive studies to achieve the above object, the substance having a variety of antioxidant action in view of the effective intake of a substance having an antioxidant action in the suppression of lung soluble muscle atrophy suggesting the involvement of oxidative stress Repeated review. As a result, it was found that certain polyphenols had an inhibitory effect on lung-soluble muscle atrophy, and also suppressed a decrease in dry weight and ash weight of bone and a decrease in bone calcium content due to lung-soluble muscle atrophy. It has been found that the present invention has been completed thereby.

According to the present invention, there is provided a muscle atrophy inhibiting composition, which comprises fruit polyphenol as an active ingredient.

In particular, according to the present invention, there is provided a muscle atrophy inhibiting composition which is a lung-soluble muscle atrophy inhibiting composition.

Further, according to the present invention, there is provided a composition for inhibiting bone weight and bone salt lowering in pulmonary soluble muscle atrophy, which comprises fruit polyphenol as an active ingredient.

Best Mode for Carrying Out the Invention

The fruit polyphenol used as an active ingredient in the composition of the present invention will be described in detail. As the fruit polyphenol, the polyphenol fraction purified from the juice or extract of the immature fruits of the Rosaceae, in particular of the immature fruits of apples, pears and peaches Or proanthocyanidins extracted from grape juice or seeds of grape fruit. Purification of such a polyphenol fraction is carried out by treating the juice and extract with an adsorbent, and the polyphenol is contained in a fraction adsorbed to the adsorbent (hereinafter referred to as an adsorption fraction). The polyphenol fraction is purified by eluting the adsorption fraction with a hydrous alcohol (ethanol or the like).

This polyphenol fraction can then be concentrated to yield a liquid formulation. In addition, powder formulations can be obtained by spray drying or freeze drying the concentrate of the polyphenol fraction.

As the raw material, fruits belonging to the family Rosaceae, for example, apples, pears, peaches and the like can be mentioned, and apples are particularly preferable. In addition, although both mature and immature fruits can be used as fruits, immature fruits are particularly preferable because they contain more polyphenol compounds and a large amount of various active ingredients having a wide range of physiological functions.

As a juice method, a juice is obtained by crushing and crushing raw material as it is or by adding sulfurous acid, Preferably, pectin degrading enzyme is added. Subsequently, the method of obtaining a blue light juice by means, such as centrifugation or filtration, is mentioned. In addition, as an extraction method, the washed raw materials are mixed with an alcohol (for example, ethanol or methanol), crushed and extracted while being immersed, pressed or heated under reflux, followed by distillation of the alcohol under reduced pressure, followed by centrifugation or filtration, or organic A method of dividing and filtration with a solvent (for example, hexane or chloroform, etc.) to obtain a blue extract is mentioned.

As the purification method, an adsorbent capable of selectively adsorbing and eluting polyphenols, for example, styrene-divinylbenzene-based synthetic adsorption resin, anion exchange resin, and octadecyl group chemically bonded silica gel (ODS) The polyphenol fraction is adsorbed by passing the above-mentioned blue juice or blue water extract through a column filled with the lamp. The polyphenol fraction can then be recovered by eluting the column with distilled water and then eluting the polyphenol fraction by passing a 20-100% alcohol (eg ethanol) solution, preferably about 50% alcohol solution, through the column. By distilling off the obtained polyphenol solution under reduced pressure, alcohol can be distilled off and the fruit polyphenol liquid formulation (preferably addition of organic acid, such as malic acid) can be obtained. The liquid polyphenol powder formulation can also be obtained by spray drying or lyophilizing the liquid formulation as it is or by adding powder preparation such as dextrin.

The composition of the fruit polyphenol used in the present invention is a simple polyphenol compound as a caffeic acid derivative, p-coumaric acid derivative, flavan-3-ols (catechins), flavonols (quercetin glucosides), or dihydrochalcones. (Fluoretine glucoside) and the like, and also condensed tannins as the polymeric polyphenol compound.

The muscle atrophy inhibiting composition of the present invention contains fruit polyphenols as an active ingredient, and is prepared in general pharmaceutical and food forms to be administered or supplied to mammals including humans.

When the composition of the present invention is prepared in the form of a pharmaceutical, the obtained pharmaceutical form is prepared by a general formulation means using an active ingredient and a conventional preparation carrier. Here, as the preparation carrier, diluents or excipients such as fillers, extenders, binders, humectants, disintegrating agents, surfactants and suspending agents which are usually used may be exemplified depending on the form of use of the preparation, and these are the dosage units of the resulting preparation. It is appropriately selected and used depending on the form.

As the dosage unit form of the pharmaceutical preparation, various forms may be selected according to the therapeutic purpose, and representative examples thereof include tablets, pills, powders, solutions, suspensions, emulsions, granules, capsules, and injections (for example, solutions and suspensions). Etc.) can be mentioned.

When molded into the form of tablets, as the preparation carrier, for example, excipients such as lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid and potassium phosphate; Binders such as water, ethanol, propanol, single syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, methylcellulose and polyvinylpyrrolidone; Sodium carboxymethylcellulose, calcium carboxymethylcellulose, low-substituted hydroxypropylcellulose, sodium croscarmellose, dry starch, sodium alginate, agar powder, laminaran powder, sodium bicarbonate, calcium carbonate, sodium carboxymethyl starch, partial Disintegrants such as alpha starch; Surfactants such as polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate and monoglyceride stearate; Disintegration inhibitors such as sucrose, stearin, cacao butter and hydrogenated oil; Absorption accelerators such as quaternary ammonium base and sodium lauryl sulfate; Humectants, such as glycerin and starch; Adsorbents such as starch, lactose, kaolin, bentonite and colloidal silicic acid; And lubricants such as purified talc, stearate, boric acid powder, and polyethylene glycol can be used.

In addition, tablets can be peeled into tablets which have been conventionally coated as needed, for example, dragee tablets, gelatin coated tablets, enteric coated tablets, film coated tablets, double tablets and multilayer tablets. .

In the case of molding in the form of pills, as a preparation carrier, for example, excipients such as glucose, lactose, starch, cacao butter, hardened vegetable oil, kaolin and talc; Binders such as gum arabic powder, tragacanth powder, gelatin and ethanol; And disintegrants such as laminaran and agar.

Capsules may be prepared by mixing the active ingredient with the various carriers exemplified above and then filling the hard gelatin capsules or soft gelatin capsules.

In addition, solutions (suspensions), granules and the like can also be easily prepared in accordance with conventional methods using the above-mentioned various carriers.

Moreover, a pharmaceutical agent can contain a coloring agent, a preservative, a fragrance | flavor, a flavoring agent, a sweetener, etc., and other pharmaceutical products as needed.

The amount of the fruit polyphenol which is the active ingredient to be contained in the above-mentioned pharmaceutical formulation is not particularly limited and is appropriately selected in a wide range, but is preferably contained in about 1 to 70% by weight in the conventional pharmaceutical formulation.

The dosage of the above-described pharmaceutical preparation is appropriately selected according to its usage, patient's age, sex and other conditions, and the severity of the disease, but it is generally preferred that the active ingredient is used in an effective amount capable of exhibiting its original action. . The amount is appropriately determined according to the active ingredient used and is not particularly limited, but is generally about 0.5 to 500 mg per 1 kg of body weight per adult per day, the preparation is once a day Or divided into two to four times.

In addition, the composition of the present invention can be actually used as a food form. Here, the food form includes all of the usual food forms such as beverages, tablets, chewing tablets, confectionary, block forms, cookies and cakes.

The composition of the present invention in food form contains various excipients commonly known in active ingredients, such as sugars (except oligosaccharides), sugar alcohols, sweeteners and other excipients, binders, disintegrants, lubricants, thickeners, and the like. , Surfactant, electrolyte, flavoring agent, colorant, pH adjuster, fluidity improving agent, vitamins and the like can be appropriately added and blended to prepare. As said additive, For example, starches, such as wheat starch, potato starch, corn starch, and dextrin; Sugars such as sucrose, glucose, fructose, maltose, xylose and lactose; Sugar alcohols such as sorbitol, mannitol, maltitol and xylitol; Excipients such as calcium phosphate and calcium sulfate; Binders or thickeners such as starch, sugars, gelatin, gum arabic, dextrin, methylcellulose, polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropylcellulose, xanthan gum, pectin, tragacanth gum, casein and alginic acid; Lubricants such as leucine, isoleucine, L-valine, sugar esters, hydrogenated oils, stearic acid, magnesium stearate, talc, macrogol; Disintegrants such as CMC, CMC-Na, and CMC-Ca; Surfactants such as polysorbate and lecithin; Dipeptides such as aspartame and alitam; Fluidity improving agents such as silicon dioxide; Sweeteners such as stevia and saccharin and the like can be exemplified, and these can be appropriately selected and used. Preparation of the compositions of the present invention in these food forms may be in accordance with conventional techniques.

The food of the present invention thus obtained is taken orally. Its intake (dosage) amount can generally be aimed at the amount of several tablets prepared in tablets, for example, about 0.5 to 6 g per tablet.

In order to demonstrate this invention below, the preparation example of the composition of this invention is given to an Example, and the test example performed about the composition of this invention is then illustrated. In each example,% is based on weight.

Examples 1-6

Each raw material compound was mixed and dissolved in water so that the composition of the following Table 1 may be prepared, and the composition of this invention was prepared. In the composition of each Example, fragrance | flavor and / or vitamins are mix | blended suitably again. Water was added to each formulation to make the total amount 1000 ml.                 

Example number One 2 3 4 5 6 Cation (mEq / l) Na ⁺ 21 15 21 15 8 27 K ⁺ 5 5 5 5 4 5 Ca ²⁺ One One One 2 One One Mg ²⁺ 0.5 0.5 0.5 0.5 0.5 0.5 Sum 27.5 21.5 27.5 22.5 13.5 33.5 Anion (mEq / ℓ) Cl ^- 16.5 10.5 16.5 10.5 6.5 17.5 Citrate Ion ( ^2- ) 10 10 8 10 4 11 Lactic acid ion ( ^- ) One One One 2 One One Tartaric acid ion ( ^2- ) 0 0 One 0 One 2 Maleic acid ion ( ^2- ) 0 0 One 0 One 2 Sum 27.5 21.5 27.5 22.5 13.5 33.5 Rebaudioside A (mg / L) 80 75 83 73 70 85 Sugar Fructose (g / ℓ) Glucose (g / ℓ) 20 2 18 1 17 2 16 3 15 2 22 1 Apple Phenon 50 (g / ℓ) 100 50 10 60 30 200

Example 7

The following components (total amount 5 g) are mixed and tableted directly to prepare (tablet), or each component is weighed and mixed to distribute (powder), or each component is weighed and mixed to dry granules. Then, it was distributed (granulated) to prepare a composition of the present invention in the form of each formulation.

Apple Phenon 50 34%

L-ascorbic acid 21%

20% L-tartaric acid

Sweetener

Sodium bicarbonate 21%                 

Sodium Chloride

Potassium Carbonate 0.5%

Very small amount of flavoring and coloring

Total 10%

In addition, said "applephenone 50" is fruit polyphenol powder (concentration 50%, hereafter called "AP") made from Nikka Whiskey Distilling Co., Ltd.

Examples 8-10

In the same manner as in Example 7, the composition of the present invention in the form of tablets, powders, and granules containing the following components were prepared.

<Example 8 Prescription>

40% AP

L-ascorbic acid 10%

L-tartaric acid 23%

Sweetener

Sodium bicarbonate 22%

Sodium citrate

Potassium Carbonate 0.4%

Very small amount of flavoring and coloring

100% in total (total amount: 5g)

Example 9 Prescription

40% AP

L-ascorbic acid 11%

L-tartaric acid 23%

Sweetener

Sodium bicarbonate 22%

Ammonium Citrate O.8%

Cyanocobalamin Trace

Sodium Citrate Trace

Potassium Carbonate 0.4%

Very small amount of flavoring and coloring

100% in total (total amount: 4.6g)

<Example 10 Prescription>

40% AP

L-tartaric acid 29%

Sweetener

Sodium bicarbonate 24%                 

Ammonium Ferric Citrate 3.6%

Cyanocobalamin Trace

Potassium Carbonate 0.5%

Very small amount of flavoring and coloring

100% in total (4 g total)

Examples 11-18

In the same manner as in Examples 7 to 10, a composition of the present invention in the form of a foam formulation having a composition shown in Table 2 below was prepared.

Ingredient (%) Example number 11 12 13 14 15 16 17 18 AP 40 30 40 60 50 35 45 35 L-ascorbic acid 11 16 10 8 10 10 10 13 L-tartaric acid 23 23 23 13 19 20 20 25 sweetener Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity Sodium bicarbonate 22 22 23 15 15 20 20 23 Ammonium Ferric Citrate 0.8 0.8 1.0 0.8 0.8 - - 0.7 Sodium ferrous citrate - - - - - 1.2 - - Ferric citrate - - - - - - 0.8 - Cyanocobalamin a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Sodium citrate Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity Spices & Colors Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity Gross weight (g) 4.6 4.6 4.7 4.6 4.6 4.7 4.7 5.4

Examples 19-25

Each component (mg) shown in Table 3 below was mixed and the composition of the present invention in the form of a chewing tablet was prepared by direct compression.

Ingredient (mg) Example number 19 20 21 22 23 24 25 AP 1800 3600 4620 700 3330 600 350 Sugar ester 40 80 841 20 80 15 7 Polydextrose 150 400 300 100 300 60 60 Sucrose 100 200 200 20 200 80 20 Vitamin c 150 320 200 20 200 - - Orange juicer powder 80 150 - - 100 20 5 Lemon juicer powder - - 84 40 - 10 5 Tartaric acid - - - - 460 95 50 NaHCO ₃ - - - - 500 100 50 K ₂ CO ₃ 30 40 40 40 30 8 8 Spices & Sweeteners Quantity Quantity Quantity Quantity Quantity Quantity Quantity Total amount (mg) 2400 4950 5580 1000 5200 1000 5000

Examples 26-34

The components of the present invention in the form of healthy beverages were prepared by mixing the components shown in Table 4 and adding water to make the total amount 100 ml.

Ingredients (in 100 ml) Example number 26 27 28 29 30 31 32 33 34 β-carotene (mg) 3 5 10 15 One 2 30 5 3 Polydextrose (g) 5 3 5 7 4 2 10 20 20 Emulsifier (mg) 6 10 20 30 5 6 20 15 8 Emulsion (mg) 100 90 80 120 50 50 200 70 60 Fructose (g) - 15 10 - 15 15 15 5 10 Citric Acid (mg) 200 400 100 300 50 20 - - - Tartaric acid (mg) - - 50 - 50 10 100 200 - Lactic acid (mg) - - - - 50 10 100 - 200 Ascorbic acid (mg) 300 200 100 50 30 150 200 1000 50 Tocopherol (mg) 10 5 10 20 20 0.5 20 10 5 AP (g) 2 5 10 3 8 7 5 12 10 Spices & Sweeteners Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity

Examples 35-45

Each of the ingredients shown in Table 5 were mixed and water was added to make the total amount 100 ml to prepare a composition of the present invention in the form of a beverage.

In addition, the gas volume value in a table | surface is an index which shows the carbon dioxide content computed by making the case where the gas of carbon dioxide of the same volume as a solution was melt | dissolved as 1, and a numerical value shows that there is more carbon dioxide content.

Ingredients (in 100 ml) Example number 35 36 37 38 39 40 41 42 43 44 45 AP (g) 3 12 9 One 8 2 4 10 15 7 13 Isomerized sugar (g) 8 - - 7 7 8 5 - - 9 - Refined Sucrose (g) - One 8 3 7 5 3 - - - - Fructose (g) 2 6 - One - - 3 - - - 60 Glucose (g) 2 2 - - 2 - 3 - - 2 4 Citric Acid (mg) 3 2 - - 8 - - 2 5 - - Tartaric acid (mg) - 2 - 2 - - - - - 10 - Malic acid (mg) 4 - 8 - 5 - 4.5 - - - - Lactic acid (mg) 8 - - 2 - 20 - - - 10 - Sodium citrate (mg) 20 30 10 - 80 - - 100 55 70 - Sodium Tartrate (mg) 60 - - 60 25 70 - - - 30 20 Sodium Maleate (mg) - 80 150 - - 100 45 - 10 - 50 Calcium lactate (mg) - - - - 15 10 - - - - 5 Sodium chloride (mg) - - 4 - One 1.5 - - 2 - - Potassium chloride (mg) - 3 - 2 - - 5 - One One - Magnesium chloride (mg) 2 - One - - - - - One - - crush(%) 3 - One 0.5 0.1 - - - - 2 0.3 Spices & Sweeteners Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity Gas volume - - - - - 3.0 2.0 2.5 2.3 3.3 1.5 pH 5.0 6.3 5.8 4.9 5.8 5.3 5.5 5.6 6.4 5.6 5.9

Test Example 1

Muscle atrophy inhibition test example

The test was conducted by the following method.

(1) the public animal:

A total of 21 Wister male rats (12 weeks old, microbiological quality: SPF / VAF) were used.

(2) experimental group:

Experimental group: AP administration group (Group A, n = 11)

Control group: (C group, n = 10)

(3) Experimental Method:

Rats were bred under one condition per cage and used a 12-hour light cycle (light cycle 8: 00-20: 00).

Each rat was subjected to gypsum fixation with the ankle joint extended to the right ankle joint under general anesthesia with pentobarbital sodium (50 mg / kg). Gypsum fixation was maintained for 12 days to induce lung soluble muscle atrophy. Immediately after the release of plaster, both soleus muscles and tibias are collected.

(4) Test Formula Administration Method

During the gypsum fixation period, Group A provides a powdered formula (referred to as "AD") with 1% AP added to commercially available rat powder formula AIN-93G (its composition is shown in Table 1). Powderless AIN-93G without AP was provided.

Composition (g / kg diet) AIN-93G AD Corn starch ^{※ 1} 529.49 524.49 Casein ^{※ 1} 200.00 200.00 Sucrose ^{* 2} 100.00 100.00 Soybean oil ^{* 1} 70.00 70.00 Cellulose Powder ^{※ 1} 50.00 45.00 Mineral mixture (type AIN-93) ^{※ 1} 35.00 35.00 Vitamin mixture (type AIN-93) ^{※ 1} 10.00 10.00 L-cysteine ^{* 3} 3.00 3.00 Choline Tartaric Acid ^{※ 3} 2.50 2.50 AP 0.00 10.00 Tert-Butylhydroxyquinone ^{* 3} 0.014 0.014 Total energy (MJ) 15.60 15.60

In Table 6, * 1 is an ingredient made of Oriental Yeast, Co., Ltd., * 2 is a ingredient made by KS Co., Ltd. * 3 is a Wako Pure-Yaku high school kabukishi. It is a component made by Wako Pure Chemical Industries, Ltd.

Meals were provided twice a day by pair feeding, feeding the food while adjusting the amount of feeding so that there was no difference in feeding between groups. Meal time was 8: 00-9: 00 and 20: 00-21: 00. Drinking water was freely consumed during the experiment.

(5) measurement items

ㆍ Weight and Feeding:

Body weight was measured every morning after breakfast (9:00). The feeding amount was measured before and after the meal at 9:00 and 21:00, and the weight of the food box was measured and the difference was used as the feeding amount. Automatic balance was used for all measurements.

Fluke atrophy rate:

The flounder was quickly weighed after harvesting, frozen with liquid nitrogen and cryopreserved (-80 ° C.) until analysis. Muscle atrophy was calculated by the following equation.

Muscle atrophy rate (%) = [(contralateral muscle wet weight-atrophic muscle wet weight) / contralateral muscle wet weight] × 100

Free iron concentration in the soleus muscle:

Free iron in muscles is described by Zhang, D. et al., Biochem. Mol. Biol. Int., 35, 635-641 (1995), which is modified by Ma, Y. et al., Path. Int., 47, 203-208 (1997)]. Approximately 20 mg of muscle tissue was homogenized with 1.8 ml of Hunks' solution, and 500 ml of this homogenized solution was added with 800 mM nitrotriacetic acid (NTA) solution (pH 7.0, 50 µl), followed by stirring for 30 minutes at room temperature. It was left for a while. This solution was centrifugally filtered using an ultrafiltration filter (fraction molecular weight 30,000, ultra-free CL UFC4LTK25, manufactured by Millipore Co.) (4 ° C., 3000 rpm, 60 minutes). The recovered filtrate was ultracentrifuged (4 ° C., 21460 g, for 60 minutes) to recover the supernatant. This supernatant was used as a sample solution for free iron measurement. The iron concentration of the sample solution was measured by atomic absorption spectrometry by graphite atomization.

Dry weight of tibia and ash weight:

The bone sample was placed in a test tube previously weighed to constant weight and dried in a vacuum oven until constant weight (100 ° C., vacuum suction, for 6 days). The weight at this time was weighed to calculate the dry weight of the bone from the difference from the weight of the test tube alone. The dried bone sample was also incinerated with an oven (reference: 600 ° C. for 3 days), cooled to room temperature, and then weighed. The weight of the ash was calculated from the difference between the weight at this time and the weight of the test tube alone.

Calcium content of tibia:

About 2 ml of concentrated nitric acid was added to the sample to which the weight of the ash was measured, and it heated and dried at 130 degreeC by heating and blotting. This operation was repeated twice to dissolve the sample obtained by dissolving with 1% nitric acid and sequentially diluting. To the final diluted sample solution, lanthanum chloride was added as a stabilizer. The sample solution was then measured by atomic absorption spectrometry (compressed air-acetylene gas, measurement wavelength: 422.7 nm, slits: 1.3 mm, photomal voltage: 560 V, lamp current: 7.5 mA) by the flame method. . As for all glass containers, what used the acid treatment with 1% nitric acid was used.

(6) statistical method

All data are expressed as mean and standard deviation. Comparison of gypsum-fixed atrophy-side Fuji and control-side Fuji of the same individual was performed by a paired t test to examine the effects of gypsum fixation. Comparison of group A and group C was performed by unpaired t test. p <0.05 was judged to be a significant difference.

(7) results

The results are shown in Tables 7 and 8 below.                 

(Mean ± standard deviation) A group (group of the present invention) C group (control) Weight before gypsum (g) Weight after gypsum (g) 374.5 ± 21.4 360.2 ± 21.6 372.6 ± 22.5 359.3 ± 26.3 Average Feed (g / d) 13.6 ± 1.3 13.6 ± 1.2 AP intake (mg / kg / d) 379 ± 36 - Fluke muscle weight (mg) Atrophic side Control side muscular atrophy rate 112 ± 10 ^* 193 ± 12 41.8 ± 5.5 ^# 104 ± 9 ^* 200 ± 14 48.1 ± 4.1 Free Iron Concentration (μg / g) Atrophy Control Side Atrophy / Control Side (%) 1.7 ± 0.6 2.0 ± 0.8 91.2 ± 24.5 ^# 2.1 ± 1.1 ^* 1.6 ± 0.7 131.9 ± 35.4   *: p <0.05 vs control (paired t test) #: p <0.05 vs. C group (nonpaired t test)

(Mean ± standard deviation) A group (group of the present invention) C group (control) Tibia dry weight (mg) Atrophy side Control side Atrophy side / Control side (%)  491.1 ± 30.8 497.5 ± 22.6 98.8 ± 6.4 489.8 ± 19.2 ^* 506.2 ± 30.2 96.9 ± 3.7 Ash weight (mg) atrophy side control side atrophy side / control side (%)  297.2 ± 18.5 308.2 ± 14.6 96.5 ± 5.8 294.8 ± 7.5 ^* 312.5 ± 15.7 94.5 ± 2.9 Calcium content (mg) Atrophy side Control side Atrophy side / Control side (%)  95.0 ± 7.2 98.2 ± 4.8 96.8 ± 6.4 91.5 ± 3.3 ^* 96.5 ± 4.9 94.9 ± 2.7   *: p <0.05 vs control (paired t test)

The following facts become clear from Tables 7 and 8.

Body weight and feeding rate:                 

During the gypsum fixation period, weight was reduced in both groups (Group A and Group C) but no difference was found between the groups. The amount of feeding during gypsum fixation was not different between groups. The average AP intake of group A was 379 ± 36 mg / kg / d (mean ± standard deviation).

Muscular dystrophy:

The muscle atrophy rate of group A was 13% significantly lower than that of group C (41.8 ± 5.5% vs. 48.1 ± 4.1%, p <0.01)

Free Iron Concentration in Soleus Muscles:

The free iron concentration in the soleus muscle was significantly higher in the atrophy side than the control side in the C group, but no difference was observed between the atrophic side and the control side in the A group.

No difference was found between group A and C. The ratio of free iron in the muscle on the atrophy side to the control side was significantly lower in group A than in group C.

Dry weight of tibia, ash weight and calcium content:

The dry weight of the tibia showed a significantly lower value in the atrophy side than the control side in group C, but no difference was found between the atrophy side and the control side in group A. No significant difference was found between the A and C groups in the atrophy / control side (%).

Tibial reweight was significantly lower in the atrophic side than in the control group in group C, but no difference was found between the atrophic and control sides in group A. No significant difference was found between the A and C groups in the atrophy / control side (%).

The tibia calcium content was significantly lower in the atrophy side than the control side in group C, but no difference was found between the atrophy side and the control side in group A. No significant difference was found between the A and C groups in the atrophy / control side (%).

From the above, it has been clarified that administration of AP during gypsum fixation is effective in inhibiting atrophy of soleus muscle and also in suppressing the reduction of dry weight of ash, weight of ash and calcium content. Inhibition of atrophy of the soleus muscle by administration of AP is thought to be involved in the inhibition of the increase of free iron in the atrophy muscle.

According to the present invention, there is provided a muscle atrophy inhibiting composition and a bone weight and bone salt lowering inhibitory composition, and such a composition is obtained by ingesting it in the elderly who are believed to be involved in pulmonary trophic atrophy due to gypsum fixation or the like, as well as oxidative stress. It can be expected to suppress the decrease in muscle mass even for sarcopenia and progressive muscle atrophy. In addition, suppressing the decrease in muscle mass leads to suppressing the decrease in muscle strength proportional to the muscle mass. In particular, maintenance of strength in the elderly leads to maintaining the body's ability to balance, leading to lowering the risk of falling or preventing the risk of forcing a fracture or a life that cannot occur due to falling. In addition, since muscle is the maximum energy consuming tissue in vivo, maintaining muscle mass can be expected to prevent carbohydrate metabolism and lipid metabolism from deteriorating, and to prevent lifestyle diseases such as diabetes and atherosclerosis. In addition, it is expected that the decrease in the weight of bone and the density of osteoarthritis, which is reported to decrease with the decrease in muscle mass, can be suppressed.

Claims (12)

delete delete delete delete delete Bone weight and bone salt reduction inhibitory composition at the time of lung-soluble muscle atrophy, which contains fruit polyphenol as an active ingredient. The bone weight and bone salt lowering inhibitory composition according to claim 6, wherein the fruit polyphenol is separated and purified from the fruit belonging to the Rosaceae family. The bone weight and bone salt lowering inhibitory composition according to claim 7, wherein the fruit belonging to the Rosaceae is apple, pear or peach. The bone weight and bone salt lowering inhibitory composition according to claim 6, which is a pharmaceutical composition or a food composition. delete delete delete