WO2021172726A1 - Health functional food composition containing extract of sanguisorba officinalis l. - Google Patents

Health functional food composition containing extract of sanguisorba officinalis l. Download PDF

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WO2021172726A1
WO2021172726A1 PCT/KR2020/018805 KR2020018805W WO2021172726A1 WO 2021172726 A1 WO2021172726 A1 WO 2021172726A1 KR 2020018805 W KR2020018805 W KR 2020018805W WO 2021172726 A1 WO2021172726 A1 WO 2021172726A1
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extract
functional food
health functional
cucumber
food composition
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PCT/KR2020/018805
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French (fr)
Korean (ko)
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하기태
한정호
배성진
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부산대학교 산학협력단
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/316—Foods, ingredients or supplements having a functional effect on health having an effect on regeneration or building of ligaments or muscles

Definitions

  • the present invention relates to a health functional food composition for improving exercise function comprising a cucumber extract, and a method for improving exercise function, comprising treating an individual with a cucumber extract.
  • Exercise performance refers to the ability to quickly, strongly, long, and skillfully perform physical movements performed in daily life or sports, and includes recovery from muscle fatigue and endurance.
  • Muscle fatigue refers to a state in which physical activity performance is temporarily decreased after intense exercise or due to long-term exercise.
  • the glycolysis system is a process in which glycogen is broken down into lactate in glycogenolysis or glycolysis. Due to the excessive production of lactic acid from glycolysis and glycogenolysis, muscle fatigue increases in the body. Endurance is defined as 'resistance to fatigue'. During high-intensity exercise, if the oxygen supply does not meet the oxygen consumption of the muscle, the concentration of lactic acid in the muscle tissue increases. Excessive accumulation of lactic acid due to exercise causes acidification in the body and suppresses the supply of kinetic energy.
  • Mitochondria are abundantly distributed in muscles and make energy sources such as ATP (adenosine triphosphate). It is known that aerobic exercise increases the number and activity of mitochondria, thereby increasing ATP production and improving endurance.
  • ATP adenosine triphosphate
  • the number and activity of mitochondria is regulated by peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1 ⁇ ), and thus, the corresponding factor is measured and used as an index for exercise performance.
  • POC-1 ⁇ peroxisome proliferator-activated receptor gamma coactivator 1-alpha
  • AMPK AMP activated protein kinase
  • Cucumber ( Sanguisorba) officinalis L.) is distributed in Korea, Japan, Europe, China, Manchuria, and Siberia, and is a perennial plant belonging to the Rosaceae family.
  • the root of the plant is called terrestrial oil and is used as a herbal medicine, and the leaves and above-ground parts are used as food.
  • Oil is effective in reducing fever and astringent hemostasis, so it is effective in stool bleeding, hemorrhoidal bleeding, dysentery abdominal pain and uterine bleeding. Because of this, it is being used as a herbal medicine. Recently, there have been studies showing anti-obesity effects through the regulation of transcription factors related to lipogenesis and lipolysis.
  • cucumber extract has a fatigue recovery function and improves exercise functions such as muscle endurance
  • the present invention was completed.
  • a health functional food composition for improving exercise function comprising a cucumber extract.
  • the present invention is cucumber grass ( Sanguisorba officinalis L ) Provides a health functional food composition for improving exercise function comprising an extract.
  • the present invention is cucumber grass ( Sanguisorba officinalis L ) treating the extract to the subject; It provides a method of improving the motor function of an object comprising a.
  • the present invention is cucumber grass ( Sanguisorba officinalis L ) provides the use of extracts to improve motor function.
  • the present invention is cucumber grass ( Sanguisorba officinalis L ) provides a use for the preparation of a motor function enhancing composition of the extract.
  • the health functional food composition for improving exercise function comprising the cucumber grass extract
  • it is possible to improve the exercise function of the individual without side effects, and it is possible to effectively recover from fatigue. Therefore, it can be widely used in the pharmaceutical industry or health functional food field.
  • 1 is a diagram showing the amount of change in body weight of a mouse according to the administration of a cucumber extract.
  • Figure 2 is a diagram showing by scoring the exercise ability of the mouse according to the administration of cucumber extract.
  • Figure 3 is a diagram showing AST, ALT changes according to the administration of cucumber extract.
  • Figure 4 is a diagram measuring the levels of lactic acid, lactate dehydrogenase and glycogen in the liver according to the administration of cucumber extract.
  • FIG. 5 is a diagram showing the measurement of PGC-1 ⁇ and AMPK levels in muscle cells according to the administration of a cucumber extract.
  • the present invention is cucumber grass ( Sanguisorba officinalis L ) Provides a health functional food composition for improving exercise function comprising an extract.
  • the cucumber grass ( Sanguisorba officinalis L ) is a perennial herb that grows in mountainous areas in Korea.
  • the growing environment is grassy forest with good drainage in semi-shade or sunny places.
  • the height is 30 ⁇ 150cm
  • the leaves are 2.5 ⁇ 5cm long and 1 ⁇ 2.5cm wide, have triangular saw teeth and have an oval shape.
  • Flowers are red, 1-2.5cm long and 0.6-0.8cm in diameter, standing upright, and there are many things without a peduncle around one long flower stalk.
  • Fruits are produced around October-November and are square.
  • the cucumber grass may be at least one selected from the group consisting of leaves, stems, fruits and roots of cucumber grass, and preferably may be roots of cucumber grass.
  • the cucumber extract is water, methanol, ethanol, propanol, isopropanol, C 1 -C 4 alcohols such as butanol, methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl Extracts obtained by leaching using acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof, carbon dioxide, pentane, etc.
  • C 1 -C 4 alcohols such as butanol, methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl Extracts obtained by leaching using acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed
  • the extraction method is cold immersion, reflux, heating, ultrasonic radiation, supercritical extraction, etc. Any method may be applied.
  • the fractionated extract a fraction obtained by suspending the extract in a specific solvent and mixing and standing still with a solvent having a different polarity, and adsorbing the extract to a column filled with silica gel, etc. It is meant to include the fraction obtained as a mobile phase.
  • the meaning of the extract includes a concentrated liquid extract or solid extract from which the extraction solvent is removed by freeze drying, vacuum drying, hot air drying, spray drying, or the like.
  • the cucumber extract may not induce liver toxicity when administered to an individual, and preferably, a significant change in blood aspartate aminotransferase (AST) or ALT (alanine aminotransferase) levels is not observed by administration of the cucumber extract. it may not be
  • the cucumber extract may improve or improve motor function, and preferably, the motor function may be at least one selected from the group consisting of reflexes, muscle strength, agility and muscular endurance, and more preferably muscle It can be endurance.
  • the cucumber extract may be to reduce the level of lactate in the blood or the lactate dehydrogenase activity in the tissue, and may have a fatigue recovery effect based on this effect.
  • the tissue may be an epithelial tissue, a connective tissue, a muscle tissue, or a nervous tissue, and may be a tissue of an organ such as a liver tissue, a kidney tissue, a spleen tissue, or a tissue of a digestive system.
  • the cucumber extract may be to increase the expression level of PGC1-alpha or AMPK in muscle tissue.
  • the dosage of the cucumber extract of the present invention will vary depending on the age, sex, weight and specific disease or pathological condition of the subject to be administered, the route of administration and the judgment of the user. Dosage determination based on these factors is within the level of the skilled artisan, and dosages generally range from 0.01 mg/kg/day to approximately 2000 mg/kg/day. A more preferred dosage may be from 50 mg/kg/day to 1000 mg/kg/day. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way.
  • the health functional food composition for improving exercise function in addition to the active ingredient, adds any compound or natural extract that has already been verified for safety and is known to have an effect of improving exercise function for the purpose of increasing and reinforcing the effect of improving exercise function.
  • any compound or natural extract that has already been verified for safety and is known to have an effect of improving exercise function for the purpose of increasing and reinforcing the effect of improving exercise function.
  • the health functional food of the present invention includes ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients and seasonings.
  • a flavoring agent or natural carbohydrate may be included as an additional ingredient in addition to the cucumber extract as an active ingredient.
  • natural carbohydrates include monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, etc.), oligosaccharides, polysaccharides (eg, dextrin, cyclodextrin, etc.), and sugar alcohols (eg, xylitol, sorbitol, erythritol, etc.).
  • natural flavoring agents eg, taumarin, stevia extract, etc.
  • synthetic flavoring agents eg, saccharin, aspartame, etc.
  • the health functional food of the present invention includes the form of tablets, capsules, pills, or liquids, and the food to which the extract of the present invention can be added includes, for example, various drinks, meat, sausage, bread, candy, Snacks, noodles, ice cream, dairy products, soups, ionized beverages, beverages, alcoholic beverages, gum, tea, and vitamin complexes.
  • the present invention is cucumber grass ( Sanguisorba officinalis L ) treating the extract to the subject; It provides a method of improving the motor function of an object comprising a.
  • the subject may be an individual in need of improvement, enhancement, or improvement in motor function.
  • the cucumber extract will vary depending on the age, sex, weight and specific disease or pathology of the individual, the route of administration and the judgment of the user. Dosage determination based on these factors is within the level of the skilled artisan, and dosages generally range from 0.01 mg/kg/day to approximately 2000 mg/kg/day. A more preferred dosage may be from 50 mg/kg/day to 1000 mg/kg/day. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way.
  • Cucumber extract is not limited thereto, but may be provided to the subject in the form of a health functional food, exercise supplement, or food supplement.
  • the present invention is cucumber grass ( Sanguisorba officinalis L ) extract to improve motor function
  • cucumber grass ( Sanguisorba) officinalis L ) provides a use for the preparation of a motor function enhancing composition of the extract.
  • the detailed description of the exercise function improvement method, use, and use for manufacturing may be equally applicable to the contents described in the health functional food composition for improving exercise function, and will be omitted in order to avoid the complexity of the description.
  • Experimental animals were purchased from Hana Bio (Seongnam, Gyeonggi-do, Korea) as a C57BL/6, 8-week-old model (male), and quarantined and acclimatized for a week at Dongnam Medical Research Institute Animal History (Animal Facility Registration Certificate: No. 412). After breeding, the experiment was carried out. The lighting time during breeding was set to 12 hours (07:00 ⁇ ), and food and water were freely ingested. For the feed, solid feed for experimental animals (Hana Bio, Seongnam, Gyeonggi-do, Korea) was provided, and the change in body weight was measured twice a week.
  • a total of 5 groups were set: a normal group, a negative control group, a positive control group, a low dose group of cucumber extracts, and a group with a high dose of cucumber extracts, and 5 mice were set in each group.
  • Saline was administered to the normal group and the negative control group
  • taurine (5 mg/kg) was administered to the positive control group
  • the cucumber extract low dose group was administered 10 mg/kg cucumber extract
  • the cucumber grass extract high dose group was administered with cucumber grass extract. 50 mg/kg was administered.
  • the drug was orally administered at 10 am every day for 4 weeks.
  • the normal group did not carry out the treadmill exercise experiment of Example 3, and the treadmill exercise experiment of Example 3 was performed for the remaining four experimental groups.
  • a treadmill exercise experiment was conducted to evaluate the effect of cucumber extract administration on muscle endurance.
  • the treadmill exercise experiment was conducted after having an adaptive exercise time once a week for 2 weeks, and the drug was administered to the experimental group of Example 1 3 times a week for 2 weeks of this experiment, and the evaluation was performed after fasting for 2 hours. Starting at 10 m/min, the speed was increased by 3 m/min every 3 min until a final velocity of 25 m/min was reached. If 10 seconds passed after not following the treadmill speed any longer, it was considered exhaustion and a score was given according to the speed.
  • the exercise performance score was higher in the experimental group administered with the cucumber extract, and it was confirmed that the cucumber extract had the effect of increasing exercise capacity, such as muscular endurance.
  • mice of the experimental group were fasted for 16 hours , anesthetized with CO 2 and laparotomy, and then blood was collected from the abdominal vena cava using a 1 mL syringe.
  • the hepatotoxicity indicator enzymes, AST (aspartate aminotransferase) and ALT (alanine aminotransferase) were measured from the blood serum collected using a biochemical analyzer Cobas-c111. The measurement results are shown in FIG. 3 .
  • mice in the experimental group that had finished the treadmill exercise experiment in Example 3 were fasted for 16 hours , anesthetized with CO 2 , and opened, and then blood was collected from the abdominal vena cava using a 1 mL syringe, and tissues of liver, muscle, kidney and spleen. was collected. From the serum of the blood collected, lactate and lactate dehydrogenase were measured.
  • Lactic acid concentration was measured using a commercially available ELISA kit (MyBioSource, MBS756195) using a competitive enzyme immunoassay technique using a conjugation of a polyclonal anti-LAC antibody and LAC-HRP (horseradish peroxidase, chromogenic enzyme). The light intensity was measured.
  • lactate dehydrogenase The activity of lactate dehydrogenase was measured with a commercially available kit using a biochemical analyzer Cobas-c111.
  • a homogenization buffer (12.5 mM sodium phosphate buffer pH 7.0, 400 mM sodium chloride) 10 times the weight of the liver tissue was added to the liver tissue for homogenization, and the supernatant was used as an enzyme source after centrifugation.
  • MyBioSource MBS733396
  • the levels of lactic acid and lactate dehydrogenase in the blood increased and the amount of glycogen in the liver decreased in the negative control group subjected to the treadmill exercise experiment compared to the normal group without exercise.
  • the levels of lactic acid and lactate dehydrogenase in the blood decreased compared to the negative control, and the level of glycogen in the liver was increased compared to the negative control. Therefore, it was confirmed that the cucumber extract showed the effect of decreasing lactic acid and lactate dehydrogenase, which are fatigue substances, and increasing glycogen in the liver.
  • taurine used as a positive control it was confirmed that the Cucumber extract had a similar effect even at a significantly lower concentration.
  • the number and activity of mitochondria is regulated by peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1 ⁇ ), and AMP-activated protein kinase (AMPK) is an enzyme that maintains energy homeostasis in cells.
  • PGC-1 ⁇ peroxisome proliferator-activated receptor gamma coactivator 1-alpha
  • AMPK AMP-activated protein kinase
  • an experiment was conducted to measure the levels of PGC-1 ⁇ and AMPK in muscle tissue. Specifically, 300 ⁇ L of RIPA buffer was added to 50 mg of muscle tissue collected in Example 5 and centrifuged at 4° C. at 12,000 rpm for 20 minutes.
  • Octacosanol powder 15% by weight, lactose hydrolyzate powder 15% by weight, soy protein isolate 15% by weight, chitooligosaccharide 15% by weight yeast extract powder 10% by weight, vitamin and mineral mixture 10% by weight, magnesium stearate 4.6% by weight, titanium dioxide 0.2% by weight, and 0.2% by weight of glycerin fatty acid ester, and 20% by weight of the cucumber extract of the present invention are blended to prepare a tablet-type health functional food in a conventional manner.
  • Honey 5% by weight, fructose 3% by weight, sodium riboflavin hydrochloride 0.0001% by weight, pyridoxine hydrochloride 0.0001% by weight, water 86.9998% by weight and 5% by weight of the cucumber extract of the present invention are blended to prepare a health drink by a conventional method.

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Abstract

The present invention relates to a health functional food composition for improving motor skills, containing an extract of Sanguisorba officinalis L. A health functional food composition for improving motor skills, containing an extract of Sanguisorba officinalis L., according to the present invention, has few side effects, can improve motor skills and enables effective recovery from fatigue. Therefore, the present invention can be widely used in the pharmaceutical industry or the health functional food field.

Description

오이풀 추출물을 포함하는 건강기능식품 조성물Health functional food composition comprising cucumber extract
본 발명은 오이풀 추출물을 포함하는 운동기능 개선용 건강기능식품 조성물 및 오이풀 추출물을 개체에 처리하는 단계를 포함하는 운동기능 개선 방법에 관한 것이다.The present invention relates to a health functional food composition for improving exercise function comprising a cucumber extract, and a method for improving exercise function, comprising treating an individual with a cucumber extract.
최근 서구화된 식습관과 생활 패턴으로 인한 운동 부족 및 스트레스로 인한 비만, 고혈압, 성인병이 증가하고 있는 추세이다. 이를 예방하기 위하여 규칙적인 운동과 효율적인 운동수행능력의 증진에 대한 관심도 증가하고 있다.Recently, obesity, high blood pressure, and geriatric diseases due to lack of exercise and stress due to westernized eating habits and lifestyle are increasing. In order to prevent this, there is an increasing interest in regular exercise and improvement of efficient exercise performance.
운동 수행 능력이란 일상생활이나 스포츠에서 수행되는 신체동작을 빠르게, 강하게, 오래, 능숙하게 할 수 있는 능력을 의미하며 근피로 회복, 지구력 등이 있다. 근피로(muscle fatigue)는 강도가 강한 운동 후 또는 장기간의 운동으로 인해 신체활동 수행능력이 일시적으로 감소된 상태를 의미한다.Exercise performance refers to the ability to quickly, strongly, long, and skillfully perform physical movements performed in daily life or sports, and includes recovery from muscle fatigue and endurance. Muscle fatigue refers to a state in which physical activity performance is temporarily decreased after intense exercise or due to long-term exercise.
해당과정 시스템은 글리코겐 분해(glycogenolysis) 또는 해당과정(glycolysis)에서 글리코겐(glycogen)이 젖산(lactate)으로 분해되는 과정이다. 해당과정 및 글리코겐분해로부터 과도하게 생산된 젖산에 의해 체내에 근피로가 증가하게 된다. 지구력(endurance)은 '피로에 대한 저항력'으로 정의된다. 고강도 운동 시 산소공급량이 근육의 산소소모량에 미치지 못하는 경우 근육조직의 젖산 농도가 증가하게 되고 이때 생성된 젖산은 혈액으로 확산되어 간에서 처리 된다. 운동으로 인해 젖산이 과도하게 축적되면 체내 산성화가 초래되어 운동에너지의 공급이 억제된다. The glycolysis system is a process in which glycogen is broken down into lactate in glycogenolysis or glycolysis. Due to the excessive production of lactic acid from glycolysis and glycogenolysis, muscle fatigue increases in the body. Endurance is defined as 'resistance to fatigue'. During high-intensity exercise, if the oxygen supply does not meet the oxygen consumption of the muscle, the concentration of lactic acid in the muscle tissue increases. Excessive accumulation of lactic acid due to exercise causes acidification in the body and suppresses the supply of kinetic energy.
최근 운동수행능력을 향상시키기 위한 방안으로 미토콘드리아의 양과 활성을 증가시키는 연구가 활발히 진행되고 있다. 근육에는 미토콘드리아가 다량 분포하고 있고 ATP(adenosine triphosphate)와 같은 에너지원을 만든다. 유산소 운동을 하면 미토콘드리아의 수와 활성이 증가하며, 이에 따라 ATP의 생산이 높아져 지구력이 향상된다고 알려져 있다.Recently, research to increase the amount and activity of mitochondria as a way to improve exercise performance is being actively conducted. Mitochondria are abundantly distributed in muscles and make energy sources such as ATP (adenosine triphosphate). It is known that aerobic exercise increases the number and activity of mitochondria, thereby increasing ATP production and improving endurance.
미토콘드리아의 수와 활성은 PGC-1α(peroxisome proliferator-activated receptor gamma coactivator 1-alpha)에 의해 조절되며, 따라서 해당 인자를 측정하여 운동수행능력에 대한 지표로 삼기도 한다.The number and activity of mitochondria is regulated by peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), and thus, the corresponding factor is measured and used as an index for exercise performance.
AMPK(AMP activated protein kinase)는 세포 내의 에너지 항상성 유지 역할을 하는 효소로서, 운동이나 스트레스에 의해 ATP가 고갈되어 AMP/ATP 비율이 증가하는 경우 활성이 증가하게 된다. 이는 PGC1-alpha의 인산화를 증가시켜 미토콘드리아생성 반응에 관여하는 것으로 알려져 있다.AMPK (AMP activated protein kinase) is an enzyme that maintains energy homeostasis in cells, and its activity increases when the AMP/ATP ratio increases due to the depletion of ATP by exercise or stress. It is known to be involved in mitochondrial production by increasing phosphorylation of PGC1-alpha.
오이풀( Sanguisorba officinalis L.)은 한국, 일본, 유럽, 중국 만주, 시베리아 등지에 분포하고 있으며 장미과에 속하는 여러해살이풀이다. 해당 식물의 뿌리를 지유(地楡)라 하며 한약재로 쓰고, 잎과 지상부는 식품으로 사용한다.Cucumber ( Sanguisorba) officinalis L.) is distributed in Korea, Japan, Europe, China, Manchuria, and Siberia, and is a perennial plant belonging to the Rosaceae family. The root of the plant is called terrestrial oil and is used as a herbal medicine, and the leaves and above-ground parts are used as food.
지유는 열을 내리고 수렴지혈(收斂止血)의 효능이 있어서 대변 출혈, 치질 출혈, 이질 복통 출혈 및 자궁 출혈에 효과를 보이며 외과적으로는 화상에 가루를 내어 바르고 치질 환부에 증기를 쐬는 등의 기능이 있어, 한약으로 활용되고 있다. 최근 지방 생성, 지방 분해와 관련된 전사 인자의 조절을 통해 항비만 효과를 나타낸다는 연구 결과가 있었으나, 아직까지 지유의 새로운 기능에 대한 연구는 많이 이루어지지 않았다. Oil is effective in reducing fever and astringent hemostasis, so it is effective in stool bleeding, hemorrhoidal bleeding, dysentery abdominal pain and uterine bleeding. Because of this, it is being used as a herbal medicine. Recently, there have been studies showing anti-obesity effects through the regulation of transcription factors related to lipogenesis and lipolysis.
이에, 본 발명자들은 천연 물질 중에서 운동 수행 능력을 개선시키거나, 근 피로를 해소할 수 있는 물질에 대하여 연구하던 중, 오이풀 추출물이 피로 회복 기능이 있으며, 근 지구력과 같은 운동 기능을 개선시키는 것을 확인하여, 본 발명을 완성하였다.Accordingly, the present inventors confirmed that, while researching on substances that can improve exercise performance or relieve muscle fatigue among natural substances, cucumber extract has a fatigue recovery function and improves exercise functions such as muscle endurance Thus, the present invention was completed.
따라서, 본 발명의 목적은 오이풀 추출물을 포함하는 운동 기능 향상용 건강기능식품 조성물을 제공하는 것이다.Accordingly, it is an object of the present invention to provide a health functional food composition for improving exercise function comprising a cucumber extract.
상기 목적을 달성하기 위하여, 본 발명은 오이풀 ( Sanguisorba officinalis L) 추출물을 포함하는 운동 기능 향상용 건강기능식품 조성물을 제공한다.In order to achieve the above object, the present invention is cucumber grass ( Sanguisorba officinalis L ) Provides a health functional food composition for improving exercise function comprising an extract.
또한 본 발명은 오이풀 ( Sanguisorba officinalis L) 추출물을 개체에 처리하는 단계; 를 포함하는 개체의 운동 기능 향상 방법을 제공한다. In addition, the present invention is cucumber grass ( Sanguisorba officinalis L ) treating the extract to the subject; It provides a method of improving the motor function of an object comprising a.
또한 본 발명은 오이풀 ( Sanguisorba officinalis L) 추출물의 운동기능 향상 용도를 제공한다. In addition, the present invention is cucumber grass ( Sanguisorba officinalis L ) provides the use of extracts to improve motor function.
또한 본 발명은 오이풀 ( Sanguisorba officinalis L) 추출물의 운동기능 향상 조성물의 제조를 위한 용도를 제공한다. In addition, the present invention is cucumber grass ( Sanguisorba officinalis L ) provides a use for the preparation of a motor function enhancing composition of the extract.
본 발명에 따른, 오이풀 추출물을 포함하는 운동 기능 향상용 건강기능식품 조성물을 이용하면, 부작용이 없이 개체의 운동 기능을 개선시킬 수 있으며, 효과적으로 피로 회복이 가능하다. 따라서, 의약 업계 또는 건강기능식품 분야에서 널리 사용될 수 있다.According to the present invention, using the health functional food composition for improving exercise function comprising the cucumber grass extract, it is possible to improve the exercise function of the individual without side effects, and it is possible to effectively recover from fatigue. Therefore, it can be widely used in the pharmaceutical industry or health functional food field.
도 1은 오이풀 추출물 투여에 따른 마우스의 체중 변화량을 나타낸 도이다.1 is a diagram showing the amount of change in body weight of a mouse according to the administration of a cucumber extract.
도 2는 오이풀 추출물 투여에 따른, 마우스의 운동능력을 점수화하여 나타낸 도이다.Figure 2 is a diagram showing by scoring the exercise ability of the mouse according to the administration of cucumber extract.
도 3은 오이풀 추출물 투여에 따른 AST, ALT 변화를 나타낸 도이다.Figure 3 is a diagram showing AST, ALT changes according to the administration of cucumber extract.
도 4는 오이풀 추출물 투여에 따른 혈중 젖산, 젖산 탈수소효소 및 간 내 글리코겐의 수준을 측정한 도이다.Figure 4 is a diagram measuring the levels of lactic acid, lactate dehydrogenase and glycogen in the liver according to the administration of cucumber extract.
도 5는 오이풀 추출물 투여에 따른 근육 세포 내 PGC-1α 및 AMPK 수준을 측정하여 나타낸 도이다.5 is a diagram showing the measurement of PGC-1α and AMPK levels in muscle cells according to the administration of a cucumber extract.
본 발명은 오이풀 ( Sanguisorba officinalis L) 추출물을 포함하는 운동 기능 향상용 건강기능식품 조성물을 제공한다.The present invention is cucumber grass ( Sanguisorba officinalis L ) Provides a health functional food composition for improving exercise function comprising an extract.
본 발명에 있어서, 상기 오이풀( Sanguisorba officinalis L)은 우리나라 각처의 산지에서 자라는 다년생 초본이다. 생육환경은 반그늘 혹은 양지의 물 빠짐이 좋은 풀숲이다. 키는 30~150cm이고, 잎은 길이 2.5~5cm, 폭 1~2.5cm로 삼각형의 톱니가 있고 타원형이다. 꽃은 적색이고 길이 1~2.5cm, 지름 0.6~0.8cm로 곧게 서며 한 개의 긴 꽃대 주위로 꽃자루가 없는 것들이 많이 달린다. 열매는 10~11월경에 맺고 사각형이다.In the present invention, the cucumber grass ( Sanguisorba officinalis L ) is a perennial herb that grows in mountainous areas in Korea. The growing environment is grassy forest with good drainage in semi-shade or sunny places. The height is 30~150cm, the leaves are 2.5~5cm long and 1~2.5cm wide, have triangular saw teeth and have an oval shape. Flowers are red, 1-2.5cm long and 0.6-0.8cm in diameter, standing upright, and there are many things without a peduncle around one long flower stalk. Fruits are produced around October-November and are square.
본 발명에 있어서, 상기 오이풀은 오이풀의 잎, 줄기, 열매 및 뿌리로 이루어진 군에서 선택된 1종 이상일 수 있으며, 바람직하게는 오이풀의 뿌리일 수 있다.In the present invention, the cucumber grass may be at least one selected from the group consisting of leaves, stems, fruits and roots of cucumber grass, and preferably may be roots of cucumber grass.
본 발명에 있어서, 상기 오이풀 추출물은 상기 추출 대상을 물, 메탄올(methanol), 에탄올, 프로판올, 이소프로판올, 부탄올 등의 C 1-C 4 알코올, 메틸렌클로라이드, 에틸렌, 아세톤, 헥산, 에테르, 클로로포름, 에틸아세테이트, 부틸아세테이트, N,N-디메틸포름아미드(DMF), 디메틸설폭사이드(DMSO), 1,3-부틸렌글리콜, 프로필렌글리콜 또는 이들의 혼합 용매를 사용하여 침출하여 얻어진 추출물, 이산화탄소, 펜탄 등 초임계 추출용매를 사용하여 얻어진 추출물 또는 그 추출물을 분획하여 얻어진 분획물을 의미하며, 추출 방법은 활성물질의 극성, 추출 정도, 보존 정도를 고려하여 냉침, 환류, 가온, 초음파 방사, 초임계 추출 등 임의의 방식이 적용될 수 있다. 분획된 추출물의 경우 상기 추출물을 특정 용매에 현탁시킨 후 극성이 다른 용매와 혼합·정치시켜 얻은 분획물, 상기 추출물을 실리카겔 등이 충진된 칼럼에 흡착시킨 후 소수성 용매, 친수성 용매 또는 이들의 혼합 용매를 이동상으로 하여 얻은 분획물을 포함하는 의미이다. 또한 상기 추출물의 의미에는 동결건조, 진공건조, 열풍건조, 분무건조 등의 방식으로 추출 용매가 제거된 농축된 액상의 추출물 또는 고형상의 추출물이 포함된다. In the present invention, the cucumber extract is water, methanol, ethanol, propanol, isopropanol, C 1 -C 4 alcohols such as butanol, methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl Extracts obtained by leaching using acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof, carbon dioxide, pentane, etc. It means an extract obtained by using a supercritical extraction solvent or a fraction obtained by fractionating the extract, and the extraction method is cold immersion, reflux, heating, ultrasonic radiation, supercritical extraction, etc. Any method may be applied. In the case of the fractionated extract, a fraction obtained by suspending the extract in a specific solvent and mixing and standing still with a solvent having a different polarity, and adsorbing the extract to a column filled with silica gel, etc. It is meant to include the fraction obtained as a mobile phase. In addition, the meaning of the extract includes a concentrated liquid extract or solid extract from which the extraction solvent is removed by freeze drying, vacuum drying, hot air drying, spray drying, or the like.
본 발명에 있어서, 상기 오이풀 추출물은 개체에 투여 시 간 독성을 유발하지 않는 것일 수 있으며, 바람직하게는 오이풀 추출물 투여에 의해 혈중 AST(aspartate aminotransferase) 또는 ALT(alanine aminotransferase) 수준에 유의한 변화가 나타나지 않는 것일 수 있다.In the present invention, the cucumber extract may not induce liver toxicity when administered to an individual, and preferably, a significant change in blood aspartate aminotransferase (AST) or ALT (alanine aminotransferase) levels is not observed by administration of the cucumber extract. it may not be
본 발명에 있어서, 상기 오이풀 추출물은 운동 기능을 개선 또는 향상시키는 것 일 수 있으며, 바람직하게 상기 운동 기능은 순발력, 근력, 민첩성 및 근 지구력으로 이루어진 군에서 선택된 하나 이상일 수 있으며, 더 바람직하게는 근 지구력 일 수 있다.In the present invention, the cucumber extract may improve or improve motor function, and preferably, the motor function may be at least one selected from the group consisting of reflexes, muscle strength, agility and muscular endurance, and more preferably muscle It can be endurance.
본 발명에 있어서, 상기 오이풀 추출물은 혈중 젖산 수준 또는 조직 내 젖산 탈수소효소 활성을 감소시키는 것일 수 있으며, 이러한 효과를 바탕으로 피로회복 효과가 있는 것 일 수 있다. 상기 조직은 상피 조직, 결합조직, 근육조직, 신경조직일 수 있으며, 간조직, 신장조직, 비장조직, 소화기관의 조직과 같은 기관(organ)의 조직일 수 있다.In the present invention, the cucumber extract may be to reduce the level of lactate in the blood or the lactate dehydrogenase activity in the tissue, and may have a fatigue recovery effect based on this effect. The tissue may be an epithelial tissue, a connective tissue, a muscle tissue, or a nervous tissue, and may be a tissue of an organ such as a liver tissue, a kidney tissue, a spleen tissue, or a tissue of a digestive system.
본 발명에 있어서, 상기 오이풀 추출물은 근 조직 내 PGC1-alpha 또는 AMPK 발현 수준을 증가시키는 것일 수 있다.In the present invention, the cucumber extract may be to increase the expression level of PGC1-alpha or AMPK in muscle tissue.
본 발명의 오이풀 추출물의 투여량은 투여 대상의 연령, 성별, 체중과 특정 질환 또는 병리 상태, 투여경로 및 사용자의 판단에 따라 달라질 것이다. 이러한 인자에 기초한 투여량 결정은 당업자의 수준 내에 있으며, 일반적으로 투여량은 0.01㎎/㎏/일 내지 대략 2000㎎/㎏/일의 범위이다. 더 바람직한 투여량은 50㎎/㎏/일 내지 1000㎎/㎏/일 일 수 있다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다. The dosage of the cucumber extract of the present invention will vary depending on the age, sex, weight and specific disease or pathological condition of the subject to be administered, the route of administration and the judgment of the user. Dosage determination based on these factors is within the level of the skilled artisan, and dosages generally range from 0.01 mg/kg/day to approximately 2000 mg/kg/day. A more preferred dosage may be from 50 mg/kg/day to 1000 mg/kg/day. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way.
본 발명에 있어서, 운동기능 향상용 건강기능식품 조성물은 유효성분 이외에, 운동기능 향상 효과의 상승·보강을 위하여 이미 안전성이 검증되고 운동기능 향상 효과를 갖는 것으로 공지된 임의의 화합물이나 천연 추출물을 추가로 포함할 수 있다.In the present invention, the health functional food composition for improving exercise function, in addition to the active ingredient, adds any compound or natural extract that has already been verified for safety and is known to have an effect of improving exercise function for the purpose of increasing and reinforcing the effect of improving exercise function. can be included as
본 발명의 건강기능식품은 식품 제조 시 통상적으로 첨가되는 성분을 포함하며, 예를 들어, 단백질, 탄수화물, 지방, 영양소 및 조미제를 포함한다. 예컨대, 드링크제로 제조되는 경우에는 유효성분으로서 오이풀 추출물 이외에 향미제 또는 천연 탄수화물을 추가 성분으로 포함시킬 수 있다. 예를 들어, 천연 탄수화물은 모노사카라이드(예컨대, 글루코오스, 프럭토오스 등), 디사카라이드(예컨대, 말토스, 수크로오스 등), 올리고당, 폴리사카라이드(예컨대, 덱스트린, 시클로덱스트린 등),및 당알코올(예컨대, 자일리톨, 소르비톨, 에리쓰리톨 등)을 포함한다. 향미제로서 천연 향미제(예컨대, 타우마린, 스테비아 추출물 등) 및 합성 향미제(예컨대, 사카린, 아스파르탐 등)를 이용할 수 있다.The health functional food of the present invention includes ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients and seasonings. For example, when it is prepared as a drink, a flavoring agent or natural carbohydrate may be included as an additional ingredient in addition to the cucumber extract as an active ingredient. For example, natural carbohydrates include monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, etc.), oligosaccharides, polysaccharides (eg, dextrin, cyclodextrin, etc.), and sugar alcohols (eg, xylitol, sorbitol, erythritol, etc.). As flavoring agents, natural flavoring agents (eg, taumarin, stevia extract, etc.) and synthetic flavoring agents (eg, saccharin, aspartame, etc.) can be used.
본 발명의 건강기능식품은 정제, 캡슐제, 환제 또는 액제 등의 형태를 포함하며, 본 발명의 추출물을 첨가할 수 있는 식품으로는, 예를 들어, 각종 드링크제, 육류, 소세지, 빵, 캔디류, 스넥류, 면류, 아이스크림, 유제품, 스프, 이온음료, 음료수, 알코올 음료, 껌, 차 및 비타민 복합제 등이 있다.The health functional food of the present invention includes the form of tablets, capsules, pills, or liquids, and the food to which the extract of the present invention can be added includes, for example, various drinks, meat, sausage, bread, candy, Snacks, noodles, ice cream, dairy products, soups, ionized beverages, beverages, alcoholic beverages, gum, tea, and vitamin complexes.
또한 본 발명은 오이풀 ( Sanguisorba officinalis L) 추출물을 개체에 처리하는 단계; 를 포함하는 개체의 운동 기능 향상 방법을 제공한다. In addition, the present invention is cucumber grass ( Sanguisorba officinalis L ) treating the extract to the subject; It provides a method of improving the motor function of an object comprising a.
상기 개체는 운동 기능 향상, 강화, 또는 개선이 필요한 개체일 수 있다. The subject may be an individual in need of improvement, enhancement, or improvement in motor function.
본 발명에 있어, 오이풀 추출물은 개체의 연령, 성별, 체중과 특정 질환 또는 병리 상태, 투여경로 및 사용자의 판단에 따라 달라질 것이다. 이러한 인자에 기초한 투여량 결정은 당업자의 수준 내에 있으며, 일반적으로 투여량은 0.01㎎/㎏/일 내지 대략 2000㎎/㎏/일의 범위이다. 더 바람직한 투여량은 50㎎/㎏/일 내지 1000㎎/㎏/일 일 수 있다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다. In the present invention, the cucumber extract will vary depending on the age, sex, weight and specific disease or pathology of the individual, the route of administration and the judgment of the user. Dosage determination based on these factors is within the level of the skilled artisan, and dosages generally range from 0.01 mg/kg/day to approximately 2000 mg/kg/day. A more preferred dosage may be from 50 mg/kg/day to 1000 mg/kg/day. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way.
오이풀 추출물은 이에 제한되는 것은 아니나, 건강기능식품, 운동 보조제, 식품 보조제의 형태로 개체에 제공될 수 있다. Cucumber extract is not limited thereto, but may be provided to the subject in the form of a health functional food, exercise supplement, or food supplement.
또한 본 발명은 오이풀 ( Sanguisorba officinalis L) 추출물의 운동기능 향상 용도, 오이풀 ( Sanguisorba officinalis L) 추출물의 운동기능 향상 조성물의 제조를 위한 용도를 제공한다. In addition, the present invention is cucumber grass ( Sanguisorba officinalis L ) extract to improve motor function, cucumber grass ( Sanguisorba) officinalis L ) provides a use for the preparation of a motor function enhancing composition of the extract.
상기 운동기능 향상 방법, 용도, 제조를 위한 용도에 대한 상세한 설명은 운동 기능 향상용 건강기능식품 조성물에 기재된 내용을 동일하게 적용할 수 있으며, 명세서 기재의 복잡성을 피하기 위해 생략한다. The detailed description of the exercise function improvement method, use, and use for manufacturing may be equally applicable to the contents described in the health functional food composition for improving exercise function, and will be omitted in order to avoid the complexity of the description.
본 명세서에서 달리 정의되지 않은 용어들은 본 발명이 속하는 기술분야에서 통상적으로 사용되는 의미를 갖는 것이다.Terms not otherwise defined herein have meanings commonly used in the art to which the present invention pertains.
실험동물 준비laboratory animal preparation
실험 동물은 하나바이오 (경기도 성남, 한국)로부터 C57BL/6, 8주령 모델 (수컷)을 분양 받아 (주)동남의화학연구원 동물사 (동물시설등록증: 제 412호)에서 일주일간 검역과 순화·사육을 거친 후 실험하였다. 사육 시 조명 시간은 12 시간 (07:00∼)으로 설정했고, 식이와 급수는 자유 섭취시켰다. 사료는 실험 동물용 고형 사료 (하나바이오, 경기도 성남, 한국)를 제공하고, 체중 변화는 주 2 회 측정하였다.Experimental animals were purchased from Hana Bio (Seongnam, Gyeonggi-do, Korea) as a C57BL/6, 8-week-old model (male), and quarantined and acclimatized for a week at Dongnam Medical Research Institute Animal History (Animal Facility Registration Certificate: No. 412). After breeding, the experiment was carried out. The lighting time during breeding was set to 12 hours (07:00~), and food and water were freely ingested. For the feed, solid feed for experimental animals (Hana Bio, Seongnam, Gyeonggi-do, Korea) was provided, and the change in body weight was measured twice a week.
오이풀 추출물 제조방법Cucumber extract preparation method
3차 증류수 1리터에 한약재 오이풀 100g을 넣어 100℃에서 1시간 끓인 후, 필터로 여과한 용액을 감압농축기로 농축하였다. 그 후 농축액을 동결 건조하여 수분을 없애고 분말을 얻었다.100 g of herbal medicine cucumber grass was added to 1 liter of tertiary distilled water, boiled at 100° C. for 1 hour, and the filtered solution was concentrated with a vacuum concentrator. Thereafter, the concentrate was freeze-dried to remove moisture to obtain a powder.
통계 분석statistical analysis
실험 결과는 평균치와 표준편차로 나타내었으며, 각 군의 유의성 검정은 StatView program에서 Anova t-test를 이용하여 통계 처리하였다.Experimental results were expressed as mean and standard deviation, and the significance test of each group was statistically processed using Anova t-test in StatView program.
실시예Example 1. One. 실험군experimental group 설정 Settings
실험군은 정상군, 음성대조군, 양성대조군, 오이풀 추출물 저용량 투여군, 오이풀 추출물 고용량 투여군으로 총 5 군을 설정하였으며 각각의 군에는 마우스를 5 마리씩 설정하였다. 정상군과 음성대조군에는 식염수(saline)를 투여하였고, 양성 대조군에는 타우린(5 mg/kg)을 투여하였으며, 오이풀 추출물 저용량 투여군은 오이풀 추출물 10 mg/kg을 투여하였고, 오이풀 추출물 고용량 투여군은 오이풀 추출물 50 mg/kg을 투여하였다. 약물은 4 주간 매일 오전 10 시에 경구투여 하였다. 정상군은 실시예 3의 트레드밀 운동실험을 실시하지 않았으며, 나머지 4개 실험군에 대해서 실시예 3의 트레드밀 운동실험이 진행되었다.For the experimental group, a total of 5 groups were set: a normal group, a negative control group, a positive control group, a low dose group of cucumber extracts, and a group with a high dose of cucumber extracts, and 5 mice were set in each group. Saline was administered to the normal group and the negative control group, taurine (5 mg/kg) was administered to the positive control group, the cucumber extract low dose group was administered 10 mg/kg cucumber extract, and the cucumber grass extract high dose group was administered with cucumber grass extract. 50 mg/kg was administered. The drug was orally administered at 10 am every day for 4 weeks. The normal group did not carry out the treadmill exercise experiment of Example 3, and the treadmill exercise experiment of Example 3 was performed for the remaining four experimental groups.
실시예Example 2. 오이풀 추출물 투여에 따른 체중 변화 2. Weight change according to Cucumber extract administration
실시예 1의 실험군을 대상으로 4주간 체중 변화를 확인하였으며, 그 결과를 도 1에 나타내었다. Changes in body weight for 4 weeks were confirmed for the experimental group of Example 1, and the results are shown in FIG. 1 .
도 1에 나타낸 바와 같이, 대조군에 비하여 오이풀 추출물 투여군에서 체중 증가량이 감소하는 경향을 확인하였으나, 통계적인 유의성은 나타나지 않았다.As shown in FIG. 1 , it was confirmed that the weight gain decreased in the group receiving the cucumber extract compared to the control group, but no statistical significance was observed.
실시예Example 3. 오이풀 추출물 투여에 의한 근 지구력 변화 측정 시험 3. Test for measuring changes in muscle endurance by administration of cucumber extract
오이풀 추출물 투여가 근 지구력에 미치는 영향을 평가하기 위해 트레드밀 운동 실험을 실시하였다. 트레드밀 운동 실험은 2 주간 주 1 회 적응 운동 시간을 가진 후 진행하였으며, 본 실험 2 주 동안 실시예 1의 실험군에 대하여, 해당 약물을 주 3 회씩 투여 하고, 2 시간 금식 후 평가를 실시하였다. 10 m/min 속도로 시작하여 최종 속도 25 m/min 에 도달할 때까지 매 3분 마다 3 m/min 씩 속도를 증가시켰다. 더 이상 트레드밀 속도를 따라가지 못한 채 10 초가 지난 경우, 탈진 상태로 간주하며 속도에 따라 점수를 부여하였다. 탈진 시기에 따라 10 m/min, 13 m/min, 16 m/min, 19 m/min, 22 m/min, 25 m/min 에 각각 0, 1, 2, 3, 4, 5 점을 부여하였고 탈진하지 않으면 6 점으로 평가하였다. 평가 결과를 도 2에 나타내었다.A treadmill exercise experiment was conducted to evaluate the effect of cucumber extract administration on muscle endurance. The treadmill exercise experiment was conducted after having an adaptive exercise time once a week for 2 weeks, and the drug was administered to the experimental group of Example 1 3 times a week for 2 weeks of this experiment, and the evaluation was performed after fasting for 2 hours. Starting at 10 m/min, the speed was increased by 3 m/min every 3 min until a final velocity of 25 m/min was reached. If 10 seconds passed after not following the treadmill speed any longer, it was considered exhaustion and a score was given according to the speed. 0, 1, 2, 3, 4, and 5 points were assigned to 10 m/min, 13 m/min, 16 m/min, 19 m/min, 22 m/min, and 25 m/min according to the time of exhaustion, respectively. If there was no exhaustion, it was rated as 6 points. The evaluation results are shown in FIG. 2 .
도 2에 나타낸 바와 같이, 음성 대조군에 비하여 오이풀 추출물을 투여한 실험군에서 운동 수행 능력 점수가 높게 측정되었는 바, 오이풀 추출물이 근 지구력과 같은 운동 능력을 증가시키는 효과가 있는 것을 확인하였다.As shown in FIG. 2 , as compared to the negative control group, the exercise performance score was higher in the experimental group administered with the cucumber extract, and it was confirmed that the cucumber extract had the effect of increasing exercise capacity, such as muscular endurance.
실시예Example 4. 오이풀 추출물 투여에 의한 4. By administration of cucumber extract 간독성hepatotoxicity 시험 exam
오이풀 추출물에 의하여 간손상이 나타나는지 여부를 확인하기 위한 실험을 실시하였다. 실시예 3에서 트레드밀 운동실험을 종료한 실험군 마우스를 16 시간 금식 시킨 후, CO 2 로 마취시켜 개복한 후 1 mL 주사기를 이용하여 복부대정맥에서 채혈하였다. 채혈한 혈액의 혈청으로부터 간독성 지표 효소인 AST(aspartate aminotransferase), ALT(alanine aminotransferase)를 생화학분석기 Cobas-c111를 이용하여 측정하였다. 측정 결과를 도 3에 나타내었다.An experiment was conducted to determine whether liver damage was caused by the cucumber extract. After completing the treadmill exercise experiment in Example 3, the mice of the experimental group were fasted for 16 hours , anesthetized with CO 2 and laparotomy, and then blood was collected from the abdominal vena cava using a 1 mL syringe. The hepatotoxicity indicator enzymes, AST (aspartate aminotransferase) and ALT (alanine aminotransferase), were measured from the blood serum collected using a biochemical analyzer Cobas-c111. The measurement results are shown in FIG. 3 .
도 3에 나타낸 바와 같이, AST와 ALT 수치가 모든 실험군에서 통계적으로 유의미한 차이가 나타나지 않았는 바, 오이풀 추출물에 의해 간손상이 나타나지 않는 것을 확인하였다.As shown in FIG. 3 , there was no statistically significant difference in AST and ALT levels in all experimental groups, and it was confirmed that liver damage was not caused by the cucumber extract.
실시예Example 5. 오이풀 추출물 투여에 따른 피로 관련 물질의 변화 측정 5. Measurement of changes in fatigue-related substances according to the administration of cucumber extract
오이풀 추출물투여에 따라 혈중 피로 관련 물질 및 간 내 글리코겐 수준이 변화하는지 여부를 확인하였다. 구체적으로, 실시예 3에서 트레드밀 운동실험을 종료한 실험군 마우스를 16 시간 금식 시킨 후, CO 2 로 마취시켜 개복한 후 1 mL 주사기를 이용하여 복부대정맥에서 채혈하였으며, 간, 근육, 신장 및 비장 조직을 채취하였다. 채혈한 혈액의 혈청으로부터, 젖산(Lactate), 젖산 탈수소효소(lactate dehydrogenase)를 측정하였다. It was checked whether blood fatigue-related substances and liver glycogen levels were changed according to the administration of cucumber extract. Specifically, the mice in the experimental group that had finished the treadmill exercise experiment in Example 3 were fasted for 16 hours , anesthetized with CO 2 , and opened, and then blood was collected from the abdominal vena cava using a 1 mL syringe, and tissues of liver, muscle, kidney and spleen. was collected. From the serum of the blood collected, lactate and lactate dehydrogenase were measured.
젖산 농도는 상용화된 ELISA 키트(MyBioSource, MBS756195)를 사용하여 polyclonal anti LAC antibody와 LAC-HRP(Horseradish peroxidase, 발색효소)의 접합을 이용한 경쟁 효소 면역 분석 기술로 측정하였으며, 마이크로 플레이트 리더기로 450nm에서 분광 광도를 측정하였다. Lactic acid concentration was measured using a commercially available ELISA kit (MyBioSource, MBS756195) using a competitive enzyme immunoassay technique using a conjugation of a polyclonal anti-LAC antibody and LAC-HRP (horseradish peroxidase, chromogenic enzyme). The light intensity was measured.
젖산탈수소효소의 활성은 생화학분석기 Cobas-c111를 이용하여 상용화된 키트로 측정하였다. The activity of lactate dehydrogenase was measured with a commercially available kit using a biochemical analyzer Cobas-c111.
글리코겐의 측정을 위해, 간 조직 무게 10 배의 균질화 완충액 (12.5 mM sodium phosphate buffer pH 7.0, 400 mM sodium chloride)을 간 조직에 넣어 균질화하고, 원심분리 후 상층액을 효소원으로 사용하였다. 상용화된 ELISA 키트(MyBioSource, MBS733396)를 사용하여, polyclonal anti-GLY 항체와 GLY-HRP(Horseradish peroxidase, 발색효소)의 접합을 이용한 경쟁 효소 면역 분석 기술로 측정하였다.For the measurement of glycogen, a homogenization buffer (12.5 mM sodium phosphate buffer pH 7.0, 400 mM sodium chloride) 10 times the weight of the liver tissue was added to the liver tissue for homogenization, and the supernatant was used as an enzyme source after centrifugation. Using a commercially available ELISA kit (MyBioSource, MBS733396), it was measured by a competitive enzyme immunoassay technique using the conjugation of a polyclonal anti-GLY antibody and GLY-HRP (horseradish peroxidase, chromogenic enzyme).
상기 젖산, 젖산 탈수소효소, 글리코겐 측정 결과를 도 4에 나타내었다.The measurement results of lactic acid, lactate dehydrogenase, and glycogen are shown in FIG. 4 .
도 4에 나타낸 바와 같이, 운동을 시키지 않은 정상군에 비하여 트레드밀 운동 실험을 실시한 음성대조군에서는 혈중 젖산 및 젖산탈수소효소의 수준이 증가하고, 간 내 글리코겐의 양은 감소하였다. 이에 반하여, 오이풀 추출물 투여 시, 혈중 젖산 및 젖산탈수소효소의 수준이 음성 대조군에 비하여 감소한 것을 확인하였으며, 간 내 글리코겐 수준은 음성 대조군에 비하여 증가한 것을 확인하였다. 따라서, 오이풀 추출물이 피로 물질인 젖산과 젖산 탈수소효소는 낮추며, 간 내 글리코겐은 증가시키는 효과를 나타내는 것을 확인하였다. 또한, 양성대조군으로 사용한 타우린과 비교하여, 오이풀 추출물이 현저히 적은 농도에서도 유사한 효과가 나타나는 것을 확인하였다. As shown in FIG. 4 , the levels of lactic acid and lactate dehydrogenase in the blood increased and the amount of glycogen in the liver decreased in the negative control group subjected to the treadmill exercise experiment compared to the normal group without exercise. On the other hand, when Cucumber extract was administered, it was confirmed that the levels of lactic acid and lactate dehydrogenase in the blood decreased compared to the negative control, and the level of glycogen in the liver was increased compared to the negative control. Therefore, it was confirmed that the cucumber extract showed the effect of decreasing lactic acid and lactate dehydrogenase, which are fatigue substances, and increasing glycogen in the liver. In addition, compared to taurine used as a positive control, it was confirmed that the Cucumber extract had a similar effect even at a significantly lower concentration.
실시예Example 6. 오이풀 추출물 투여에 따른 근육 내 세포 에너지 대사의 변화 6. Changes in intramuscular cellular energy metabolism according to the administration of cucumber extract
미토콘드리아의 수와 활성은 PGC-1α(peroxisome proliferator-activated receptor gamma coactivator 1-alpha)에 의해서 조절되며, AMPK(AMP activated protein kinase)는 세포 내 에너지 항상성 유지 역할을 하는 효소로써 운동이나 스트레스에 의해 ATP가 고갈되어 AMP/ATP 비율이 증가하는 경우 활성도가 증가하게 된다. 오이풀 추출물이 근육 내 세포 에너지 대사에 미치는 영향을 확인하기 위해, 근 조직 내 PGC-1α 및 AMPK 수준을 측정하는 실험을 실시하였다. 구체적으로, 실시예 5에서 채취한 근 조직 50mg 에 RIPA 완충액 300 μL를 넣고 4 ℃에서 12,000 rpm 으로 20 분 간 원심 분리하였다. Bradford 어세이 방법으로 단백질을 정량한 후, 단백질의 SDS-PAGE (Polyacrylamide Gel Electrophoresis)를 이용하여 크기 별로 분리하였다. Semidry transfer system (Bio-Rad, USA)를 이용하여 PVDF 막에 이동시킨 후, 5% 탈지 우유(skim milk)를 함유한 차단 완충액(0.5% 탈지 우유(skim milk), 1×TBS/T 완충액)을 1시간 동안 처리하였다. 1×TBS/T 완충액으로 10 분, 3회 세척 후, 일차 항체인 PGC1α, p-AMPK, AMPK, GAPDH를 1:1000의 비율로 처리한 후, 4 ℃에서 밤새 반응시켜, 1×TBS/T 완충액으로 10분, 3회 세척하였다. 웨스턴 블롯 검출 키트 (Biorad, hercules, USA)을 이용하여 막과 반응시킨 후 Chemi-Doc 장비를 이용하여 발현 정도를 관찰하였으며, 그 결과를 도 5에 나타내었다.The number and activity of mitochondria is regulated by peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), and AMP-activated protein kinase (AMPK) is an enzyme that maintains energy homeostasis in cells. When the AMP/ATP ratio increases due to depletion, the activity increases. In order to confirm the effect of cucumber extract on cellular energy metabolism in muscle, an experiment was conducted to measure the levels of PGC-1α and AMPK in muscle tissue. Specifically, 300 μL of RIPA buffer was added to 50 mg of muscle tissue collected in Example 5 and centrifuged at 4° C. at 12,000 rpm for 20 minutes. After quantification of proteins by the Bradford assay method, proteins were separated by size using SDS-PAGE (Polyacrylamide Gel Electrophoresis). After transfer to the PVDF membrane using a semidry transfer system (Bio-Rad, USA), a blocking buffer containing 5% skim milk (0.5% skim milk, 1×TBS/T buffer) was treated for 1 hour. After washing 3 times for 10 minutes with 1×TBS/T buffer, primary antibodies PGC1α, p-AMPK, AMPK, and GAPDH were treated at a ratio of 1:1000, and then reacted at 4°C overnight, 1×TBS/T Washed 3 times for 10 minutes with buffer. After reacting with the membrane using a Western blot detection kit (Biorad, hercules, USA), the expression level was observed using Chemi-Doc equipment, and the results are shown in FIG. 5 .
도 5에 나타낸 바와 같이, 근 조직 내 PGC1-alpha와 AMPK의 단백질 발현을 확인한 결과, 음성 대조군(C) 대비 양성 대조군(P) 및 오이풀 추출물 고용량 투여군(H)군에서 PGC1-alpha와 AMPK의 발현이 유의한 수준으로 증가한 것을 확인하였다. 상기 결과를 통해 오이풀 추출물 50 mg/kg 투여가 근육 내 세포의 에너지 대사에 도움을 주며, 이러한 기전을 바탕으로 운동기능 개선 효과를 나타내는 것을 확인하였다.As shown in FIG. 5 , as a result of confirming the protein expression of PGC1-alpha and AMPK in muscle tissue, the expression of PGC1-alpha and AMPK in the positive control group (P) and the cucumber extract high-dose group (H) compared to the negative control group (C) It was confirmed that this increased to a significant level. From the above results, it was confirmed that 50 mg/kg of cucumber extract was administered to help the energy metabolism of cells in the muscle, and based on this mechanism, it was confirmed that the effect of improving the motor function was shown.
이상, 본 발명내용의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적인 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의해 정의된다고 할 것이다.Above, specific parts of the present invention have been described in detail, for those of ordinary skill in the art, it is clear that these specific descriptions are only preferred embodiments, and the scope of the present invention is not limited thereby. something to do. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.
이하, 본 발명의 오이풀 추출물을 포함하는 운동기능 향상용 건강기능식품 조성물의 제제예를 설명하나, 본 발명을 한정하고자 함이 아니고 단지 이를 구체적으로 설명하고자 함이다.Hereinafter, a formulation example of a health functional food composition for improving exercise function comprising the cucumber grass extract of the present invention will be described, but it is not intended to limit the present invention, but merely to describe it in detail.
제제예Formulation example 1. 정제형 건강기능식품 1. Tablet health functional food
옥타코사놀분말 15중량%, 유당가수분해물분말 15중량%, 분리대두단백분말 15중량%, 키토올리고당 15중량% 효모추출물분말 10중량%, 비타민미네랄혼합제재 10중량%, 스테아린산 마그네슘 4.6중량%, 이산화티타늄 0.2 중량%, 및 글리세린지방산에스테르 0.2중량%와 본 발명의 오이풀 추출물 20중량%를 배합하여 통상의 방법으로 정제형 건강기능식품을 제조한다.Octacosanol powder 15% by weight, lactose hydrolyzate powder 15% by weight, soy protein isolate 15% by weight, chitooligosaccharide 15% by weight yeast extract powder 10% by weight, vitamin and mineral mixture 10% by weight, magnesium stearate 4.6% by weight, titanium dioxide 0.2% by weight, and 0.2% by weight of glycerin fatty acid ester, and 20% by weight of the cucumber extract of the present invention are blended to prepare a tablet-type health functional food in a conventional manner.
제제예Formulation example 2. 건강 음료 2. Healthy Drinks
꿀 5중량%, 과당 3중량%, 염산리보플라빈나트륨 0.0001중량%, 염산피리독신 0.0001중량%, 물 86.9998중량% 및 본 발명의 오이풀 추출물 5중량%를 배합하여 통상의 방법으로 건강 음료를 제조한다. Honey 5% by weight, fructose 3% by weight, sodium riboflavin hydrochloride 0.0001% by weight, pyridoxine hydrochloride 0.0001% by weight, water 86.9998% by weight and 5% by weight of the cucumber extract of the present invention are blended to prepare a health drink by a conventional method.

Claims (8)

  1. 오이풀 ( Sanguisorba officinalis L) 추출물을 포함하는 운동 기능 향상용 건강기능식품 조성물.Cucumber grass ( Sanguisorba officinalis L ) A health functional food composition for improving exercise function comprising an extract.
  2. 제1항에 있어서, 상기 오이풀 추출물은 50mg/kg 내지 1000mg/kg으로 투여되는 것인, 운동 기능 향상용 건강기능식품 조성물.The health functional food composition for improving exercise function according to claim 1, wherein the cucumber extract is administered in an amount of 50 mg/kg to 1000 mg/kg.
  3. 제1항에 있어서, 상기 운동 기능은 근 지구력인, 운동 기능 향상용 건강기능식품 조성물.According to claim 1, wherein the exercise function is muscular endurance, health functional food composition for improving exercise function.
  4. 제1항에 있어서, 상기 오이풀 추출물은 혈중 젖산 수준 또는 조직 내 젖산 탈수소효소 활성을 감소시키는 것인, 운동 기능 향상용 건강기능식품 조성물.The health functional food composition for improving exercise function according to claim 1, wherein the cucumber extract reduces blood lactate levels or lactate dehydrogenase activity in tissues.
  5. 제4항에 있어서, 상기 조직은 간 조직 또는 근육 조직인, 운동 기능 향상용 건강기능식품 조성물.According to claim 4, wherein the tissue is liver tissue or muscle tissue, health functional food composition for improving exercise function.
  6. 제1항에 있어서, 상기 오이풀 추출물은 근 조직 내 PGC1-alpha 또는 AMPK 발현 수준을 증가시키는 것인, 운동 기능 향상용 건강기능식품 조성물.According to claim 1, wherein the cucumber extract is to increase the expression level of PGC1-alpha or AMPK in muscle tissue, health functional food composition for improving exercise function.
  7. 오이풀 ( Sanguisorba officinalis L) 추출물을 개체에 처리하는 단계를 포함하는 개체의 운동 기능 향상 방법. Cucumber grass ( Sanguisorba officinalis L ) A method for improving motor function in a subject comprising the step of treating the extract to the subject.
  8. 제7항에 있어서, 상기 운동 기능은 근 지구력인, 운동 기능 향상 방법. The method of claim 7 , wherein the motor function is muscular endurance.
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