KR102066388B1 - Functional food composition for enhancing myofunction and athletic function comprising chamomile extract and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a functional food composition for strengthening muscle function and athletic performance, and a method for manufacturing the same. According to the present invention, it is possible to provide a composition capable of improving energy metabolism and muscle production-related factors, thereby improving athletic performance and muscle function. Can be. Specifically, by increasing the synthesis of mitochondria and muscle cells, and by inhibiting the muscle atrophy can improve the efficiency of muscle exercise and can exhibit the effect of feeling less fatigue. Moreover, such a composition can be applied to various fields, such as food, as a health functional composition.

Description

Functional food composition for enhancing myofunction and athletic function comprising chamomile extract and manufacturing method

The present invention relates to a muscle function and exercise-enhancing functional food composition comprising a chamomile extract and a method of manufacturing the same.

Recently, due to the development of science and modern medicine, the average life expectancy of human beings is gradually increasing, and accordingly, interest for health is increasing to improve the quality of life. This trend is increasing regardless of generation and age. In addition, with the increasing interest in medicines, health supplements, and cosmetics containing natural products, the research and development thereof is increasing. In particular, the efficacy of natural medicines and functions of health supplements and cosmetics containing natural products have been extended to the basic medical range, such as antioxidant, cholesterol suppression, obesity prevention effect, immunity and disease prevention and anti-aging.

Among the natural plants used in natural products, chamomile has the scientific names such as Chamaemelum nobile and Matricaria chamomilla, and the name Matricaria is the Latin word 'matrix', which is derived from medicinal diseases. It is known to be mainly used in car, bathing, beauty, poultice, etc., and is effective for insect repellent, soothing, dysmenorrhea, pain relief, sweating, promoting digestion, and fatigue recovery. In addition, oriental medicine refers to the effects of cardiovascular disease, cold, diarrhea, eczema, hemorrhagic cystitis, hemorrhoids, infant colic, soothing, vaginitis, wound healing.

The present inventors have completed the present invention by checking the effect of the composition containing the chamomile extract to improve muscle function and motor activity while studying the material originating from natural products.

An object of the present invention is to provide a functional food composition for strengthening muscle function and exercise performance.

It is another object of the present invention to provide a method for producing a functional food composition for strengthening muscle function and exercise performance.

In order to solve the above problems, the present invention

Provided with a chamomile extract, muscle functional and athletic enhancing functional food composition.

According to one embodiment, the present invention may comprise 1 to 100% by weight of chamomile extract based on the total weight of the composition.

According to one embodiment, as a composition for the prevention or amelioration of muscle-related diseases, atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia, myotonia, It can be used for the prevention or amelioration of one or more selected from the group consisting of amyotrophic lateral sclerosis, cachexia, sarcopenia.

In addition, according to another embodiment of the present invention, comprising extracting water, an organic solvent having 1 to 6 carbon atoms or a mixture thereof as a solvent with respect to chamomile, muscle function and athletic function functional food composition manufacturing method Can be provided.

Further, according to one embodiment, the method may include obtaining chamomile extract from chamomile by subcritical extraction, supercritical fluid extraction, or ultrahigh pressure extraction.

Other specific details of embodiments of the present invention are included in the following detailed description.

According to the present invention, the muscle function and exercise performance-enhancing functional food composition and a method for producing the same can provide a composition capable of improving energy metabolism and muscle production-related factors, thereby improving athletic performance and muscle function. Furthermore, it can be applied to various fields such as food as a health functional composition.

1 is a graph showing the activity of mTOR according to the embodiment.
2 is a graph showing the activity of PPARδ according to the embodiment.
3 is a graph showing the activity of PGC-1α according to the embodiment.
Figure 4 is an electrophoresis picture showing the muscle protein degradation inhibitory activity according to the embodiment.
Figure 5 is an electrophoresis picture showing mitochondrial biosynthesis and muscle differentiation activity according to the embodiment.

As the present invention allows for various changes and numerous embodiments, particular embodiments will be illustrated in the drawings and described in detail in the written description. However, this is not intended to limit the present invention to specific embodiments, it should be understood to include all transformations, equivalents, and substitutes included in the spirit and scope of the present invention. In the following description of the present invention, if it is determined that the detailed description of the related known technology may obscure the gist of the present invention, the detailed description thereof will be omitted.

Hereinafter, the muscle function and exercise performance-enhancing functional food composition according to the embodiment of the present invention and a manufacturing method thereof will be described in more detail.

As used herein, the term muscle function and motor function enhancement 'means improving the activity of mTOR, PPARδ and PGC-1α, inhibiting the degradation of muscle protein, promoting the biosynthesis of mitochondria, and promoting muscle differentiation. . Specifically, inhibition of muscle protein degradation may be confirmed by the reduction of mRNA expression of atrogin-1 and MuRF-1. In addition, mitochondrial biosynthesis promotion and muscle differentiation promoting activity can be confirmed that the mRNA expression of myogenin, NRF1, Tfam is increased.

According to one embodiment, by strengthening muscle function and exercise ability, it is possible to prevent, treat and improve muscle-related diseases. Prevention, treatment and improvement of muscle related diseases can be confirmed by expression of muscle and exercise related factors such as mTOR, PPARδ and PGC-1α.

Muscle-related diseases may include diseases due to decreased muscle function, muscle wasting, muscle degeneration and the like. Specifically, atony, muscular atrophy, muscular dystrophy, muscle degeneration, myasthenia, myotonia, amyotrophic lateral sclerosis, cachexia Can prevent, treat and ameliorate diseases such as sarcopenia.

The muscle function and exercise enhancing functional food composition according to the present invention includes chamomile extract.

According to one embodiment, the present invention may comprise 1 to 100% by weight, for example 20 to 70% by weight of chamomile extract based on the total weight of the composition.

According to one embodiment, the muscle function and motor function-enhancing functional food composition may be prepared by a method comprising obtaining chamomile extract with water, an organic solvent having 1 to 6 carbon atoms, or a mixture thereof as a solvent. have. For example, the organic solvent having 1 to 6 carbon atoms may include methanol, ethanol, ethyl acetate, hexane, and the like.

According to one embodiment, the composition according to the invention may be prepared by a method comprising the step of extracting alcohol or hot water extracted for chamomile.

According to one embodiment, 10-95%, for example 20-80%, for example 50% alcohol may be used in the alcohol extraction step. In addition, it may include the step of extracting the alcohol by adding 2 to 20 times the volume of the extraction target. It may also be extracted for example for 0.5 to 10 hours, for example 2 to 5 hours, for example at 4 to 80 ° C, for example 20 to 70 ° C, for example 40 to 60 ° C. Can be.

According to one embodiment, the hydrothermal extraction step may include extracting by adding 2 to 20 times the volume of the extraction target volume of water. It may also be extracted for example for 0.5 to 12 hours, for example 2 to 6 hours, for example at 4 to 121 ° C, for example 50 to 121 ° C, for example 80 to 121 ° C. Can be.

According to one embodiment, the muscle function and exercise enhancing functional food composition may be prepared by a method comprising obtaining chamomile extract from chamomile using subcritical extraction, supercritical fluid extraction or ultrahigh pressure extraction.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily practice the present invention. As those skilled in the art would realize, the described embodiments may be modified in various different ways, all without departing from the spirit or scope of the present invention.

Example 1: Hot Water Extraction

After adding 1,500 ml of distilled water to 100 g of chamomile, using a heating mantle equipped with a reflux cooling system (heating mantle, Extraction Rota-Mantle / KBT), the hot water was extracted at 95 ° C. for 4 hours, filtered, concentrated, and lyophilized. An extract was obtained.

Example 2: Alcohol Extract

After adding 1,500 ml of 10%, 30%, 50%, 70%, 90% spirits to 100 g of chamomile, 70 ° C using a heating mantle (Extraction Rota-Mantle / KBT) equipped with a reflux cooling device. Extraction of alcohol for 4 hours at, filtered and concentrated to remove the extraction solvent and lyophilized to obtain a chamomile extract.

Example 3: Methanol Extraction

Chamomile extract was obtained in the same manner as in Example 2 except that methanol was used.

Example 4: Ethyl Acetate Extraction

Chamomile extract was obtained in the same manner as in Example 2, except that ethyl acetate was used.

Example 5: Hexane Extraction

Except for using hexane was carried out in the same manner as in Example 2, whereby the chamomile extract was obtained.

Example 6 Ultra High Pressure Extraction

Chamomile 18%, 76 mL of ethanol was put in a polyethylene pack and sealed and extracted using an ultrahigh pressure extractor (Frescal MFP-7000; Mitsubishi Heavy Industries, Tokyo, Japan). The extraction pressure was set to 320MPa and the extraction time was 5 minutes as the ultrahigh pressure extraction condition. The extracted sample was filtered and concentrated to remove the extraction solvent, and lyophilized to obtain a chamomile extract.

Example 7 Super Grain Boundary Extraction

Chamomile was charged to a sample cartridge and extracted using a supercritical extractor (SFX 3560, Isco Inc., Lincoln, NE, USA). Supercritical fluid extraction conditions were set to extraction pressure 20MPa, extraction temperature 60 ℃, flow rate of supercritical carbon dioxide 60 mL / min, extraction time 60 minutes. When the supercritical fluid extraction was completed, the pressure of the extractor was lowered to release the supercritical fluid state to obtain a chamomile extract.

Experimental Example 1: mTOR Activity

MTOR activity for each extract according to Example 1 was evaluated. mTOR is a protein that plays a role in regulating the mechanism of muscle protein synthesis, and can confirm the improvement of muscle mass by confirming the activity of mTOR. First, a muscle cell master L6 cells (ATCC) 110% fetal bovine serum (FBS; Hyclone, Logan, UT, USA) containing a Dulbecco's modified Eagle's media (DMEM ; Hyclone) 2 Х 10 5 cell with the 6-well plates After inoculation to incubation / mL. When the cell density reached about 80-85%, the medium in the wells was removed and exchanged for DMEM (Hyclone) containing 2% horse serum (HS; Hyclone) to differentiate L6 cells into myotubes. Once every two days, the cells were replaced with fresh medium for a total of six days of differentiation. After differentiation, the extract prepared in Example 1 was dissolved in DMEM medium containing 50 ng / mL of TNF-α at a concentration of 100 or 200 ug / mL, and then treated to cells. After 6 hours, total RNA was isolated using TRIzol reagent (Takara, Osaka, Japan). Total RNA isolated was quantified using NanoDrop 1000; Thermo Fisher Scientific Inc., Waltham, Mass., USA. The results are shown in Table 1.

In addition, the results of treatment of cells with a substance according to Example 1 at a concentration of 100 or 300 ug / mL are shown in FIG. 1.

As shown in Table 1 and Figure 1, it can be seen that the growth of muscle proteins can be induced by the material according to an embodiment of the present invention.

Experimental Example 2: PPARδ Activity

In order to evaluate PPARδ activity for the extract according to Example 1, COS7 cells (ATCC) were placed in DMEM medium containing 10% FBS (Gibco) in a 24-well plate at 1Х10 ⁵ cells / mL. When the cell density reached about 80-85%, Lipofector EXT (Aptabio) was used to induce PPARδ transformation to confirm anti-inflammatory effects. After transformation, each extract was added to the cells together at a concentration of 10 ㎍ / mL. 24 hours treatment. And luciferase activity was evaluated using luciferase assay kit (Promega).

PPARδ is a protein that may be associated with cell differentiation in adipose tissue and sugar production in liver tissue. As a result, PPARδ may be confirmed to improve motor performance by confirming the activity of PPARδ. The results are shown in Table 2.

In addition, the results of treating the cells at 25 or 50 ug / mL according to Example 1 are shown in FIG. 2.

As shown in Table 2 and Figure 2, it can be seen that the athletic performance is improved due to the material according to Example 1 of the present invention.

Experimental Example 3: PGC-1α Activity

In order to evaluate PGC-1α activity on the extract according to Example 1, COS7 cells (ATCC) were placed in DMEM medium containing 10% FBS (Gibco) to 1Х10 ⁵ cells / mL in 24-well plates. When the cell density reached about 80-85%, peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) transformation was induced to identify anti-inflammatory effects using Lipofector EXT (Aptabio). Each extract was treated with cells at a concentration of 10 μg / mL for 24 hours. And luciferase activity was evaluated using luciferase assay kit (Promega).

PGC-1α is a protein related to glycemic regulation and mitochondrial function replication, which can confirm the improvement of exercise ability by confirming the activity of PGC-1α. The results are shown in Table 3.

In addition, the results of treating the cells at 25 or 50 ug / mL according to Example 1 are shown in FIG. 3.

As shown in Table 3 and Figure 3, it can be seen that the athletic performance is improved due to the material according to Example 1 of the present invention.

Experimental Example 4: Muscle protein degradation inhibitory activity

In order to confirm the muscle protein degradation inhibitory activity of Example 1, the expression of atrogin-1, MuRF1 and β-actin was confirmed. atrogin-1 is a muscle breakdown protein that is activated when muscle is lost, MuRF1 is a muscle lyase protein, and β-actin is a protein that makes up muscles, along with myosin, causing muscle contraction and relaxation.

First, a muscle cell master L6 cells (ATCC) 110% fetal bovine serum (FBS; Hyclone, Logan, UT, USA) containing a Dulbecco's modified Eagle's media (DMEM ; Hyclone) with 2Х10 ⁵ cell / mL in a 6-well plate After inoculation to incubate. When the cell density is about 80-85%, the medium in the wells is removed and exchanged for DMEM (Hyclone) containing 2% horse serum (HS; Hyclone) to convert L6 cells into myotubes. Differentiated. Once every two days, the cells were replaced with fresh medium for a total of six days of differentiation. After differentiation, the chamomile extract prepared in Example 1 was dissolved in DMEM medium containing 50 ng / mL of TNF-α at a concentration of 100 or 200 ug / mL, and then treated to cells. After 6 hours, total RNA was isolated using TRIzol reagent (Takara, Osaka, Japan). Total RNA isolated was quantified using NanoDrop 1000; Thermo Fisher Scientific Inc., Waltham, Mass., USA. 16 uL RNA was quantified and mixed with Reverse Transcriptase Premix (ELPIS-Biotech) using a PCR machine (Gene Amp PCR System 2700; Applied Biosystems, Foster City, CA, USA) at 42 ° C, 55 minutes, CDNA was synthesized at 70 ° C. for 15 minutes. PCR samples were prepared by mixing 4 uL of the cDNA, specific primer pairs (Bioneer, Deajeon, Korea), and PCR premix (ELPIS-Biotech) below in the synthesized cDNA, and then, at 95 ° C for 30 seconds and at 60 ° C. PCR was performed by repeating 30 minutes for 1 minute at 72 ° C.

The cDNA amplified by PCR was electrophoresed with 1.5% agarose gel, and cDNA bands were identified using a G; BOX EF imaging system (Syngene, Cambridge, UK). The used sequences are shown in Table 4, and the results are shown in FIG.

division Forward primer Reverse primer Atrogin-1 5'-CCCTGAGTGGCATCGCCCAA-3 ' 5'-AGGTCCCGCCCATCGCTCA-3 ' MuRF-1 5'-GAAATGCTATGCAGAACCTG-3 ' 5'-ATTCCTGCTTGTAGATGTCG-3 ' β-Actin 5'-AGCCATGTACGTAGCCATCC-3 ' 5'-CTCTCAGCTGTGGTGCTGAA-3 '

As shown in Figure 4, it can be seen that the mRNA expression of atrogin-1 and MuRF-1 is reduced in L6 muscle cells as the chamomile extract is treated. This means that the chamomile extract of the present invention has an excellent ability to inhibit muscle protein degradation in muscle cells.

Experimental Example 5: Mitochondrial biosynthesis promoting and muscle differentiation promoting activity

In order to confirm the muscle mitochondrial biosynthesis promoting and muscle differentiation promoting activity of Example 1, the expression of myogenin, NRF1 and Tfam was confirmed. Myogenin controls differentiation and maturation into myotubes, NRF1 is involved in mitochondrial protein production, and Tfam is involved in mitochondrial DNA transcription.

First, a muscle cell master L6 cells (ATCC) 110% fetal bovine serum (FBS; Hyclone, Logan, UT, USA) containing a Dulbecco's modified Eagle's media (DMEM ; Hyclone) 2 Х 10 5 cell with the 6-well plates After inoculation to incubation / mL. When the cell density reached about 80 to 85%, the medium in the well was removed and the chamomile extract prepared in Example 1 was added to DMEM (Hyclone) containing 2% HS of 100 or 200 ug / mL. After dissolving at concentration, the cells were treated to induce differentiation into root canal cells. At this time, the group treated with 0.01% DMSO instead of the sample was used as a control. This process was repeated three times for two days, followed by differentiation for a total of six days, and total RNA was isolated using TRIzol reagent (Takara, Osaka, Japan). Total RNA isolated was quantified using NanoDrop 1000; Thermo Fisher Scientific Inc., Waltham, Mass., USA. 16 uL RNA was quantified and mixed with Reverse Transcriptase Premix (ELPIS-Biotech) using a PCR machine (Gene Amp PCR System 2700; Applied Biosystems, Foster City, CA, USA) at 42 ° C, 55 minutes, CDNA was synthesized at 70 ° C. for 15 minutes. PCR samples were prepared by mixing 4 uL of cDNA, the following specific primer pairs (Bioneer, Deajeon, Korea), and PCR premix (ELPIS-Biotech) in the synthesized cDNA. PCR was performed by repeating 30 minutes for 1 minute at 72 ° C.

The cDNA amplified by PCR was electrophoresed with 1.5% agarose gel, and cDNA bands were identified using a G; BOX EF imaging system (Syngene, Cambridge, UK). The used sequences are shown in Table 5, and the results are shown in FIG.

division Forward primer Reverse primer Myogenin 5'-TGGGCTGCCACAAGCCAGAC-3 ' 5'-CAGCCCAGCCACTGGCATCA-3 ' NRF1 5'-TGGACCCAAGCATTACGGAC-3 ' 5'-GGTCATTTCACCGCCCTGTA-3 ' Tfam 5'-GCTTCCAGGAGGCTAAGGAT-3 ' 5'-CCCAATCCCAATGACAACTC-3 ' β-Actin 5'-CTGTGTGGATTGGTGGCTCTAT-3 ' 5'-GTGTAAAACGCAGCTCAGTAACA-3 '

As shown in Figure 5, it can be seen that the mRNA expression of myogenin, NRF1, Tfam in L6 muscle cells increases as the chamomile extract is treated. This means that the chamomile extract of the present invention is excellent in promoting muscle differentiation and mitochondrial biosynthesis in muscle cells.

As described above, the results according to Experimental Examples 1 to 5 are summarized in Tables 6 and 7.

In addition, the results of Experimental Examples 4 to 8 are summarized in Table 7.

As can be seen from the above results, according to the present invention, it is possible to increase the synthesis of mitochondria and muscle cells and to suppress the muscle atrophy, thereby improving the efficiency of muscle movement and reducing the fatigue. You can check it.

The above description is merely illustrative of the technical idea of the present invention, and those skilled in the art may make various modifications and changes without departing from the essential characteristics of the present invention. In addition, the embodiments disclosed in the present invention are not intended to limit the technical spirit of the present invention but to describe the present invention, and the scope of the technical idea of the present invention is not limited by these embodiments. The protection scope of the present invention should be interpreted by the following claims, and all technical ideas within the equivalent scope should be interpreted as being included in the scope of the present invention.

Claims (5)

Containing chamomile extract,
Suppression of muscle atrophy after exercise or decreased muscle fatigue
Functional food composition for the prevention or improvement of muscular dystrophy, muscular dystrophy or muscular dystrophy. The functional food composition of claim 1, comprising 1 to 100% by weight of chamomile extract, based on the total weight of the composition. delete A method for preparing a functional food composition according to claim 1, comprising the step of obtaining chamomile extract with water, organic solvent having 1 to 6 carbon atoms or mixtures thereof as a solvent. A method for producing a functional food composition according to claim 1, comprising the step of obtaining chamomile extract from chamomile by subcritical extraction, supercritical fluid extraction or ultrahigh pressure extraction.

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