KR20190082054A - Composition for improving muscular quality comprising scytosiphon lomentaria extract or its fraction - Google Patents

Abstract

The present invention relates to a composition for improving muscularity containing a Scytosiphon lomentaria extract or a fraction thereof as an active component. The Scytosiphon lomentaria hot-water extract binds with myostatin in a concentration-dependent manner without cytotoxicity to hinder combination of myostatin and a receptor, thereby exhibiting an anti-myostatin effect. In addition, by confirming the improvement in a muscle density and muscle strength of a mouse animal model in which the Scytosiphon lomentaria hot-water extract is orally administered, the composition containing the Scytosiphon lomentaria extract can be provided as the composition for improving muscularity and the composition for preventing or alleviating myostatin-related diseases.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for improving muscular quality comprising an extract of Koriachia or a fraction thereof as an active ingredient,

The present invention relates to a composition for improving muscle strength, which comprises a cyclic extract or a fraction thereof as an active ingredient.

The muscle of a person decreases by 1% or more every year since the age of 40 years. At 80 years old, 50% of the maximum amount of muscle is decreased, and the decrease of the muscle of the old age is recognized as the most important factor that lowers the overall body function.

As the aging progresses, the body shape changes such as muscle and fat content and skeletal distortion are recognized. The prevalence of obesity due to the decrease of old age muscle is continuously increasing at more than 30% level worldwide. In addition, when insulin secretion abnormality can not supply the energy to the cell properly, it can cause muscle development disorder, and the diabetic patients have increased myopenia. Such myopenia is also known to be closely related to chronic diseases such as osteoporosis, insulin resistance and arthritis.

In addition, muscle atrophy, another disease caused by muscle decline, is caused by a gradual decrease in muscle mass, which occurs when protein degradation occurs more than synthesis due to inactivity, oxidative stress, chronic inflammation, . Increased mitochondrial production, suppression of muscle protein degradation, and anti-inflammatory drugs have been proposed as treatments for muscle weakness, but there are no clear remedies.

Recently, many people have chosen protein supplements as an alternative, but protein supplements are likely to cause side effects because they cause excessive protein intake, and even in the presence of kidney disease, high protein diets are not available and kidney function is reduced by aging , A new alternative is needed to prevent and improve myopenia.

Myostatin is one of the TGF-β (Transforming Growth Factor-β) superfamily, a protein that inhibits muscle growth and differentiation. These myostatin binds to specific receptors with serine / threonine kinase activity on the cell surface similar to the TGF mechanism, activating the SMAD signal transduction system and interfering with the further synthesis of muscle in the nucleus The excessive release of myostatin inhibits the cell division of the nucleus and thus inhibits the production of muscle.

It is an algae of decayed chiffon necklace, which is a light brown-colored, thin-walled shape with tapering upward and downward. It has a constricted part in every place and it is gathered from the base of the button shape. Have not yet been reported.

Korean Patent Publication No. 10-2018-0048416 (disclosed on May 10, 2018)

The present invention aims to provide a composition for improving muscle strength, which exhibits a safe and superior muscular quality improvement effect to a living body.

The present invention provides a composition for improving muscular quality, comprising a cyclic extract or a fraction thereof as an active ingredient.

The present invention provides a feed additive for muscular improvement comprising an aliskiren extract or a fraction thereof as an active ingredient.

The present invention provides a health food for preventing or ameliorating a myostatin-related disease, which comprises an extract of Krismae or a fraction thereof as an active ingredient.

The present invention provides a pharmaceutical composition for preventing or treating a myostatin-related disease, which comprises an antagonist extract or a fraction thereof as an active ingredient.

The present invention also provides a veterinary composition for the prevention or treatment of myostatin-related diseases, which contains an extract of Kishimaki or a fraction thereof as an active ingredient.

According to the present invention, the hydrolyzate of Corynebacterium exhibits an antimyostatin effect by inhibiting the binding of myostatin to the receptor by binding to myostatin in a concentration-dependent manner without cytotoxicity, As the muscle density and muscle strength of the animal model have been confirmed, the composition containing the horsetail extract as an active ingredient can be provided as a composition for improving muscle strength and a composition for preventing or ameliorating myostatin-related diseases.

FIG. 1 is a schematic view showing a process for producing a cyclic hydrothermal extract.
FIG. 2 shows the results of confirming anti-myostatin (MSTN), anti-actin A, anti-DDF11 activity, cytotoxicity and anti-oxidative activity of the cyclic hydrothermal extract (SLWE) Fig. 2 (A) shows the result of confirming the recombinant myostatin inhibitory activity, Fig. 2 (B) shows the result of confirming the recombinant activin A inhibitory activity, Fig. 2 (C) D) is the result of confirming DPPH radical scavenging ability.
Figure 3 shows that in HepG2 cells Western blot analysis confirmed the effect of SLWE as an MSTN antagonist as a result of confirming the myostatin signal-blocking effect of the horseradish hydrothermal extract (SLWE). Line 1 was untreated, line 2 was treated with 10 nM MSTN, 3 was treated with 10 nM MSTN and 10 μM SB431542, line 4 was treated with 10 nM MSTN and SLWE (IC100), and line 5 was treated with 10 nM MSTN and SLWE (IC10).
FIG. 4 shows the result of the oral administration of a cyclic hydrothermal extract (SLWE) at a dose of 690 μg / 200 μl once a day for 14 days in a mouse model, FIG. 4 (B) shows the results of grip strength, and FIG. 4 (C) shows swimming endurance at 0, 7 and 14 days. and (n = 12), the black bars are SLWE treatment group (n = 10), each error bars are the standard errors are shown (* p <0.1, ** p <0.01, *** p <0.005 and * *** p <0.001 compared with the control).
FIG. 5 shows the result of confirming the muscle development level of a mouse model in which oral ginseng hydrothermal extract (SLWE) was orally administered at a dose of 690 μg / 200 μl for 14 days once a day. FIG. 5 (A) shows the mass of the forelimb and hindlimb muscles FIG. 5 (B) shows the result of checking the volume of the forelimb and hindlimb muscles, and FIG. 5 (C) shows the results of checking the density of the forelimb and hindlimb muscles.
FIG. 6 is a graph showing the relationship between total cholesterol (TC), triglyceride (TC) and total cholesterol (TC) in serum isolated from the blood collected from the abdominal vein of a mouse model, which was orally administered once a day for 6 days at 690 μg / Glucose (GLU), lactate dehydrogenase (LDH) activity, glutathione peroxydase (GPx) activity, HDL and LDL / VLDL, free fatty acid (FFA), triglyceride .
7 (A) is a schematic view showing the preparation process of the organic solvent fraction of the cyclic hydrothermal extract (SLWE), FIG. 7 (B) is a view showing the antimyostatin effect of the organic solvent fraction of the cyclic hydrothermal extract to be.

Hereinafter, the present invention will be described in more detail.

The present invention relates to a composition for improving muscular function, and a protein supplement is alternatively selected as a method for improving muscular weakness. However, protein supplement is highly likely to cause side effects due to excessive protein intake, In the case of decreased renal function, there is a difficulty in high protein diet. Accordingly, the inventors of the present invention have completed the present invention based on the finding that a natural extract, Glycyrrhiza uralensis extract exhibits excellent effects on muscle density and muscle strength while studying new alternatives for prevention and improvement of myopenia.

The present invention can provide a composition for improving muscular quality, which contains a horseradish extract or a fraction thereof as an active ingredient.

The horsetail extract may be extracted with water, C1 to C4 alcohol or a mixed solvent thereof, more preferably, it may be a hot-water extract, but is not limited thereto.

In addition, the extract may be subjected to an additional fractionation process. More preferably, it may be a butanol fraction obtained by fractionating and extracting a water fraction obtained by fractionating a cyclic hydrothermal extract with ethyl acetate and a water mixture solution using a mixed solution of butanol and water.

The cyclic extract or its fractions may exhibit antimyostatin activity.

According to an embodiment of the present invention, the degree of phosphorylation of the Smad2 transcription factor was examined by western blot analysis in order to examine the effect of the cyclic hydrothermal extract on the myostatin signaling pathway. As a result, In the experimental group treated with myostatin and SLWE (IC100), the expression of phosphatidylated SAMd2 was increased by the signal of myostatin, whereas in the experimental group treated with myostatin and SLWE And inhibited the phosphorylation of Samd2. In the experimental group treated with low concentration of SLWE, the phosphorylation of Samd2 was confirmed because it did not completely suppress the myostatin signal.

From the above results, it was confirmed that SLWE binds to myostatin in a concentration-dependent manner, and myostatin inhibits receptor binding, thereby inhibiting Samd2 signaling.

In addition, the horseradish extract or its fraction may be one that increases muscle density to improve muscle strength.

According to another embodiment of the present invention, 4-week-old male ICR mice were orally administered once a day for 14 days, and weight and strength tests were performed on days 0, 7 and 14, and as a result, Body weight was not significantly different from the control group. Also, referring to FIG. 3 (B), the muscle strength was significantly increased by 15.7% at 7 days after oral administration of the extract of the horseradish hydrothermal extract, and was significantly increased by about 8.8% at 14 days after the oral administration. As a result of confirming the swimming endurance, it was confirmed that there was no significant difference in the treated group treated with the hot water extract of the coli compared with the control group, as shown in FIG. 3 (C) It was confirmed that it was significantly increased.

The present invention can provide a feed additive for improving muscular quality containing an aliskiren extract or a fraction thereof as an active ingredient.

The feed additive may be used for livestock to improve the meat quality of livestock. Examples of suitable animals include poultry such as laying hen, duck, turkey and goose, ruminants such as cattle, horse, goat and sheep, Of rodents and fishes.

The substance contained in the feed additive of the present invention is not particularly limited as long as it is a substance used in ordinary feed additives for livestock in addition to the extract of Coleoptera.

In addition, the present invention can provide a health food for preventing or ameliorating myostatin-related diseases, which comprises the extract of Coleoptera or a fraction thereof as an active ingredient.

The myostatin-related diseases may be selected from the group consisting of muscle wasting diseases, metabolic diseases, degenerative bone diseases, hypogonadism and cachexia.

The muscular dyspepsia may be selected from the group consisting of muscular dystrophy, rigid spine syndrome, muscle-eye-brain disease, amyotrophic lateral sclerosis, A chronic inflammatory neuropathy, and distal myopathy. &Lt; Desc / Clms Page number 2 &gt;

The metabolic disease may be selected from the group consisting of type 2 diabetes, noninsulin-dependent diabetes mellitus, hyperglycemia, obesity and diabetic complications.

The degenerative bone disease may be osteoporosis.

The health food may be used in combination with other food or food additives other than the above-mentioned coliform extract or its fraction, and may be appropriately used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its use purpose, for example, prevention, health or therapeutic treatment.

The effective dose of the compound contained in the above-mentioned health food may be used in accordance with the effective dose of the therapeutic agent, but may be less than the above range for health and hygiene purposes or for long-term intake for health control purposes, It is clear that the component can be used in an amount of more than the above range since there is no problem in terms of safety.

There is no particular limitation on the type of the health food, and examples thereof include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, Drinks, alcoholic beverages and vitamin complexes.

In addition, the present invention can provide a pharmaceutical composition for preventing or treating myostatin-related diseases, which comprises a cyclic extract or a fraction thereof as an active ingredient.

The present invention can provide a veterinary composition for the prevention or treatment of myostatin-related diseases, which comprises the extract of Coleoptera or a fraction thereof as an active ingredient.

In one embodiment of the present invention, the pharmaceutical composition for preventing or treating myostatin-related diseases containing the cyclic extract or its fractions as an active ingredient may be administered orally or parenterally in the form of injections, granules, powders, tablets, , Suppositories, gels, suspensions, emulsions, drops, or liquid preparations can be used.

In another embodiment of the present invention, the pharmaceutical composition for prevention or treatment of myostatin-related diseases containing the horseradish extract or the fraction thereof as an active ingredient may be formulated with a suitable carrier, excipient, disintegrant, sweetener A lubricant, a lubricant, a flavoring agent, an antioxidant, a buffer, a bacteriostatic agent, a diluent, a dispersant, a surfactant, a binder and a lubricant.

Specific examples of carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. Solid formulations for oral administration may be in the form of tablets, pills, powders, granules, capsules These solid preparations can be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., into the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, syrups and the like, and various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin which are commonly used simple diluents. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As the suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

According to one embodiment of the present invention, the pharmaceutical composition may be administered orally, intraarterally, intraperitoneally, intramuscularly, intraarterally, intraperitoneally, intrasternally, transdermally, nasally, inhaled, topically, rectally, &Lt; / RTI &gt; can be administered to the subject in a conventional manner.

The desired dosage of the cyclic extract or its fraction may vary depending on the condition and body weight of the subject, the kind and degree of disease, the drug form, the administration route and the period, and may be appropriately selected by those skilled in the art. According to one embodiment of the present invention, the daily dose may be 0.01 to 200 mg / kg, specifically 0.1 to 200 mg / kg, more specifically 0.1 to 100 mg / kg, though it is not limited thereto. The administration may be performed once a day or divided into several times, and thus the scope of the present invention is not limited thereto.

In the present invention, the 'subject' may be a mammal including a human, but is not limited thereto.

The present invention relates to a method for inhibiting myostatin activity, which comprises the step of extracting Koriachis or a fraction thereof as an active ingredient, wherein the Koriachma extract or a fraction thereof inhibits myostatin activity in cells in vitro The reagent composition can be provided.

In addition, the present invention can provide a method for inhibiting myostatin activity, comprising the step of treating the cells with a horsetail extract or a fraction thereof in vitro.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

< Example  1> Ring  Preparation of extract

The horsetail extract was prepared in the same manner as in Fig.

20 mg of the dried ring was added to 1 ml of distilled water, and the mixture was subjected to hot water extraction at 90 ° C for 20 minutes. The above procedure was repeated three times.

The supernatant was recovered by centrifugation at 4000 rpm for 20 min.

After the particles were completely removed from the supernatant using a pore size 0.2 μM filter, the concentration of the sample was measured using a centrifugal rotary concentrator.

As a result, it was confirmed that the hydrolyzate extract of Corynebacterium was obtained at a yield of about 16%.

< Example  2> Ring  Of the extract Myostatin  And term Activin  A activity inhibition

Luciferase assay was performed to confirm antimyostatin activity at the cellular level.

HEK293 cells were plated at 2.0 × 10 ⁴ cells per well in a 96-well plate and cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin / streptomycin, 1% gentamicin, CO ₂ incubator.

After 24 hours of incubation, the medium was replaced with DMEM containing no FBS. 1 nM recombinant myostatin (R & D system, USA) and cyclic hydrothermal extract (SLWE) were treated with 1, 10 and 100 μg / ₂ incubator for 24 hours.

After incubation, the medium was removed and 6 μl of DMEM medium and 6 μl of reagent (Bright-Glo luciferase assay system, USA) were treated and the luminescence was measured in a microplate luminometer.

The measured values were expressed as the antimyostatin activity (%) using the values of the positive control and the negative control and the following formula 1.

[Equation 1]

(%) = [(Firefly RLU of 1 nM Myostatin (positive control) - Firefly RLU of the group treated with the extracts] × 100) / (Firefly RLU of 1 nM Myostatin treatment group (negative group)]

In addition, in order to confirm the inhibitory activity of the isolate extract against actin A belonging to the superfamily such as myostatin, the activity of inhibiting actin A was confirmed by the Luciferase assay.

As for the concentration of actin A, inhibitory activity was confirmed by treating the cells with 1 nM concentration as in the case of myostatin in the previous experiment, and the antactivin A activity (%) was calculated using the following equation 2.

[Equation 2]

(%) = [(Firefly RLU of 1 nM ActivinA (positive group) - Firefly RLU of the group treated with the extracts x 100] / (Firefly RLU of 1 nM ActivinA treatment (negative group)]

As a result, as shown in FIG. 1 (A), the IC ₅₀ value of the hydrolyzate extract (SLWE) against 1 nM myostatin was 11.9 μg / ml, and SLWE at 1, 10 and 100 μg / No inhibitory activity against actin A (1 nM) belonging to the superfamily such as myostatin was observed in the experimental group.

From the above results, it was confirmed that a specific myostatin inhibitory activity is exhibited at an SLWE concentration of 10 μg / ml or less without inhibitingactin A activity.

< Example  3> Ring  Identification of cytotoxicity and antioxidant activity of extract

HSK293 cells were plated at a density of 1 × 10 ⁴ cells per well in a 96-well plate and cultured in a 5% CO ₂ incubator for 24 hours. To examine the cytotoxicity of the horseradish hydrothermal extract (SLWE), 10% fetal bovine serum ) And DMEM medium containing 1% penicillin / streptomycin were used.

After 24 hours of incubation, the extracts were treated with 0.1, 1, 10, and 100 μg / ml of the extracts of P. pertussis for 12 hours in a 5% CO ₂ incubator, and 10 μl of WST reagent (DaeilLab, Korea) And the absorbance at 415 nm was measured. The cytotoxicity was shown by the following equation 3.

Positive control cells were treated with untreated cells and H ₂ O ₂ treated cells were negative control cells.

As a result, no cytotoxicity was observed at all concentrations of the cyclic hydrothermal extract as shown in Fig. 1 (C).

[Equation 3]

Cell viability (%) = [(absorbance value of positive control-absorbance value of the group treated with extracts) × 100 / (absorbance value of negative control)].

On the other hand, DPPH analysis was performed to confirm the antioxidative activity of the hydrolyzate extract (SLWE).

100 μl of the DPPH reagent and 100 μl of the hot-water extract of the horseradish peroxidase were treated at 1, 10 and 100 μg / ml for 30 min at room temperature, and the absorbance was measured at 562 nm. As a control group and a positive control group, DPPH analysis was performed by treating the material with 10 μg / ml of vitamin C.

As a result, as shown in Fig. 1 (D), it was confirmed that the antioxidant effect of about 70% was exhibited at a concentration of 100 μg / ml of the hydrothermal extract (SLWE).

From the above results, it was confirmed that the extract of Corydalis hydrothermal extract is a safe substance which does not show cytotoxicity, and antioxidant activity and antioxidative effect are exhibited.

< Example  4> Ring Hot water extract  ( SLWE )On by Myostatin  Confirm signaling inhibition

Western blot analysis of the degree of phosphorylation of the Smad2 transcription factor was performed to confirm the effect of the cyclic hydrothermal extract on the myostatin signaling pathway.

First, HepG2 cells were plated on a 6-well plate at 2.0 × 10 ⁵ cells per well and cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin in a 5% CO ₂ incubator.

After 24 hours of culture, the cells were replaced with DMEM containing no FBS. After 4 hours, the cells were treated with 10 nM recombinant myostatin and cyclic hydrothermal extract (SLWE) IC100 and IC10 for 30 minutes, respectively. (The IC100 value of SLWE is 1500 μg / ml and the IC10 value is 15 μg / ml)

After 30 minutes of treatment, the cells were washed twice with PBS and incubated with RIPA buffer [20 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1 mM Na ₂ EDTA, 1 mM EGTA, 1% NP- 40, 1% sodium deoxychloate, 1 mM β-glycerophosphate, 1 mM NA ₃ VO ₄ , and 1 μg / ml leupeptin (cell signaling, USA)].

The collected cells were disrupted using an ultrasonic sonicator and centrifuged at 12,000 rpm for 20 minutes at 4 ° C to obtain supernatant. The supernatant was quantitated by BCA analysis and electrophoresed on a 10% polyacrylamide gel.

In order to confirm expression of p-Smad2, p-Smad2 primary antibody (monoclonal antibody, Cell signaling, USA) was reacted at room temperature for 3 hours. Smad2 (monoclonal antibody, Cell signaling, USA) and β-actin (monoclonal antibody, Cell signaling, USA) were reacted for 12 hours at 4 ℃.

Then, the secondary antibody anti-rabbit IgG (Samd2, p-Smad2), anti-mouse IgG (b-actin) (Cell signaling, USA) was washed with TBS-T buffer at room temperature for 3 minutes, And reacted for 3 hours.

After the reaction, the membrane was washed with TBS-T buffer three times for 10 minutes at room temperature and sensitized to X-ray film using SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Scientific, USA).

As a result, as shown in FIG. 2, it was confirmed that the expression of phosphorylated Samd2 was increased by the signal of myostatin in line 2 treated with only myostatin. In addition, line 3 did not show an increase in the expression level of phosphorylated Samd2 as a positive control which simultaneously treated small molecule SB431542, which inhibits phosphorylation of receptors bound to myostatin and myostatin.

In line 4, it was confirmed that the phosphorylation of Samd2 was inhibited by suppressing the myostatin signal by SLWE, as in the positive control treated with SB431542, while the line 5 was the same group treated with myostatin and SLWE (IC100) In the experimental group treated with myostatin and a low concentration of SLWE (IC10), it was confirmed that low concentration of SLWE did not completely inhibit the myostatin signal, indicating the phosphorylation of Samd2.

From the above results, it was confirmed that SLWE binds to myostatin in a concentration-dependent manner and interferes with the binding of myostatin to the receptor, thereby inhibiting Samd2 signaling.

< Example  5> Using the mouse model Ring Hot water extract  ( SLWE )

1. Check for changes in weight and strength

To determine the effects of the cyclic hydrothermal extract on the muscles at the iv vivo level, changes in body weight and muscle strength were observed by oral administration to ICR mice.

4 weeks old male ICR mice were orally administered once a day for 14 days and weight and strength tests were performed at 0, 7, and 14 days.

As a result, as shown in Fig. 3 (A), the body weight of the mice was not significantly different from that of the control group. Also, referring to FIG. 3 (B), the muscle strength was significantly increased by 15.7% at 7 days after oral administration of the extract of the horseradish hydrothermal extract, and was significantly increased by about 8.8% at 14 days after the oral administration. As a result of checking the swimming endurance, no significant difference was observed in the experimental group treated with the cyclic hydrothermal extract as compared with the control group as shown in FIG. 3 (C).

From the above results, it was confirmed that the cyclic hydrothermal extract significantly increases the muscle strength without changing the body weight.

2. Identification of muscle weight and volume changes

The weight and volume of the forelimb and hindlimb muscles were measured and the muscle density was determined for 14 days after oral administration of cyclic hydrothermal extract (690 μg / 200 μl).

As a result, as shown in Fig. 4 (A), there was no significant difference in muscle mass compared to the control group.

As shown in FIG. 4 (B), the volume of the forelimb muscle was 10.8% lower than that of the control group (p value = 0.025). The volume of the hindlimb muscle was not significantly different from that of the control group.

As shown in FIG. 4 (C), the density of the forelimb muscle was significantly increased by 14.6% as compared with that of the control, and the density of the hindlimb muscle was not significantly different from that of the control group.

From the above results, it was confirmed that the muscle density was significantly increased as the volume reduction rate was increased compared with the muscle weight increase / decrease rate.

3. Blood analysis

Blood was collected from the abdominal vein of mice administered orally for 14 days with cyclic hydrothermal extract (690 μg / 200 μl) in the same manner as in the previous experiment and total cholesterol (TC), triglyceride (TG ), Glucose (GLU), lactate dehydrogenase (LDH) activity, glutathione peroxydase (GPx) activity, HDL and LDL / VLDL, and free fatty acid .

Glucose (GL), lactate dehydrogenase (LDH) activity, glutathione peroxidase (GPx) activity, HDL and LDL / VLDL , And free fatty acid (FFA) were not significantly different from those of the control group, but triglyceride (TG) was significantly decreased by 39.9% (p value = 0.034).

From the above results, it was confirmed that the content of triglyceride in the blood was significantly reduced by the hot water extract of Corynebacterium glutamicum.

Group Total cholesterol
(mg / dl) Triglyceride
(mg / dl) Glucose
(md / dl) LDH activity ^†
(mU / ml) GPx activity ^†
(mU / dl) HDL ^†
(μg / μl) LDL / VLDL ^†
(μg / μl) FFA
(mM) Control 115.2 ± 5.26 127.0 ± 17.04 354.3 ± 20.07 501.8 ± 82.5 6990.7 ± 1567.4 0.979 + 0.029 0.577 + - 0.052 0.779 + 0.107 SLWE 123.3 ± 6.78 76.3 ± 11.67 ^* 320.0 ± 10.95 647.7 ± 162.1 5476.6 ± 1192.9 1.087 + 0.074 0.571 + 0.040 0.679 + 0.067 p value 0.364 0.034 0.173 0.453 0.471 0.222 0.931 0.446

Values are means ± SE (n = 6, †: n = 4). * P <0.1, ** P <0.01, *** P <0.005 and **** P <0.001 compared with the control. Statistical

< Example  6> Ring Hot water extract  ( SLWE ) Organic solvent  Fraction and its activity confirmation

One. Ring Of hot-water extract Organic solvent  Fraction

Antimyostatin was isolated from the extracts of Corynebacterium glutamicum using an organic solvent.

The dried coconut powder was hydrolyzed at a concentration of 20 mg / ml at 90 ° C for 20 minutes. The solvent was removed from the hot water extract using a rotary evaporator and the mixture was partitioned in a separatory funnel by adding a mixture of ethyl acetate and water (1: 1, v: v).

The solvent was removed from the ethyl acetate fraction layer and the water fraction layer using a rotary evaporator.

To the ethyl acetate fraction layer from which the solvent had been removed, 90% methanol and hexane mixture (1: 1, v: v) was added and fractionated in a separating funnel. The water fraction layer was washed with a mixture of butanol and water ) Was added and fractionated in a separating funnel.

The solvent was removed from the 90% methanol, hexane, butane and water fraction layers fractionated using a rotary evaporator.

2. Ring Hot water extract  The term of the fraction Myostatin  Verify Active

The concentrations of the 90% methanol, hexane, butane and water fractions prepared in the above procedure were measured using a centrifugal rotary evaporator, and the antimyostatin activity was confirmed using Luciferase analysis at a concentration of 100 μg / ml of each fraction layer Respectively.

As a result, as shown in FIG. 6, about 97% anti-myostatin activity was confirmed in the butanol fraction layer, and thus it was confirmed that a substance exhibiting anti-myostatin activity was present in the butanol fraction layer of the extract of Kishima.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (13)

A composition for improving muscular tone comprising an extract of Koriachia or a fraction thereof as an active ingredient. [Claim 2] The composition for improving muscle strength according to claim 1, wherein the horsetail extract is extracted with water, C1 to C4 alcohols or a mixed solvent thereof. The composition of claim 1, wherein the fraction is a butanol fraction. [Claim 2] The composition for improving muscle strength according to claim 1, wherein the extract or the fraction thereof exhibits an antioxidant activity. [Claim 2] The composition for improving muscle strength according to claim 1, wherein the horseradish extract or the fraction thereof increases muscle strength to improve muscular strength. A feed additive for muscular improvement comprising an extract of Koriachia or a fraction thereof as an active ingredient. A health food for preventing or ameliorating a myostatin-related disease containing an extract or a fraction thereof as an active ingredient. [8] The method according to claim 7, wherein the myostatin-related disease is selected from the group consisting of muscle wasting disease, metabolic disease, degenerative bone disease, hypogonadism and cachexia. Health food for preventing or improving related diseases. 9. The method of claim 8, wherein the muscular dystrophy is selected from the group consisting of muscular dystrophy, rigid spine syndrome, muscle-eye-brain disease, amyotrophic lateral sclerosis ), Chalcot-Marie-Tooth disease, chronic inflammatory neuropathy, and distal myopathy. The present invention relates to a method for preventing or ameliorating a myostatin-related disease, Health food for. 9. The method of claim 8, wherein the metabolic disease is selected from the group consisting of type 2 diabetes, noninsulin-dependent diabetes mellitus, hyperglycemia, obesity and diabetic complications Health food for preventing or improving myostatin-related diseases. [Claim 8] The health food for preventing or ameliorating myostatin-related diseases according to claim 8, wherein the degenerative bone disease is osteoporosis. A pharmaceutical composition for preventing or treating a myostatin-related disease, which comprises an extract of Koriachia or a fraction thereof as an active ingredient. A veterinary composition for the prevention or treatment of myostatin-related diseases, which comprises an extract of Koriachia or a fraction thereof as an active ingredient.

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