KR102133954B1 - Composition for improving muscle disease comprising dark tea extract - Google Patents
Composition for improving muscle disease comprising dark tea extract Download PDFInfo
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- KR102133954B1 KR102133954B1 KR1020190157363A KR20190157363A KR102133954B1 KR 102133954 B1 KR102133954 B1 KR 102133954B1 KR 1020190157363 A KR1020190157363 A KR 1020190157363A KR 20190157363 A KR20190157363 A KR 20190157363A KR 102133954 B1 KR102133954 B1 KR 102133954B1
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- muscle
- dark tea
- tea
- extract
- dark
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Abstract
Description
본 발명은 다크티(dark tea) 추출물을 포함하는 근육 질환 개선용 조성물에 관한 것으로서, 더욱 상세하게는 차잎을 금화균(Eurotium cristatum strains)을 이용하여 후발효시킨 다크티 추출물 및 이로 인한 근육 질환 개선 효과에 관한 것이다.The present invention relates to a composition for improving muscle disease comprising a dark tea extract, more specifically, a dark tea extract obtained by post-fermenting tea leaves using Eurotium cristatum strains , and thereby improving muscle disease It's about effectiveness.
우리나라는 출산율 감소와 평균수명 연장으로 인하여 고령화 인구가 급격히 증가하고 있다. 국내 통계청 자료에 의하면 2011년 65세 이상 노인인구가 전체 11.3%, 2020년에는 15.7%, 2030년에는 24.3%, 2060년에는 40%에 이를 것으로 전망하고 있으며, 이러한 변화는 OECD 국가 중에서도 가장 빠른 속도로 진행되고 있다.In Korea, the aging population is rapidly increasing due to a decrease in fertility rates and an extended life expectancy. According to data from the National Statistical Office, the elderly population aged 65 and over in 2011 is expected to reach 11.3% in total, 15.7% in 2020, 24.3% in 2030, and 40% in 2060, and this change is the fastest among OECD countries. Is going on.
근감소증(sarcopenia)은 그리스어에서 기원한 근육(muscle)을 뜻하는 "sarco"와 감소되어 있다는 뜻의 "penia"가 합성된 단어로 골격근의 감소(loss of skeletal muscle mass)를 의미하며 노화에 의하여 수반되는 증상이다. 우리나라 국민건강 영양조사에서 근감소증의 유병률을 조사한 결과 6.6%로 나타났고, 미국의 경우 근감소증의 유병률은 70세 미만에서는 15 내지 25%, 80세 이후에는 남자에서 50%, 여자에서 40% 이상으로 나타났다. 근감소증은 골격근의 양(mass), 질(quality), 강도(strength)의 전반적인 손실을 수반하는 퇴행성 반응으로서 노화의 대표적 증상이다. 근육손실은 근육 단백질의 생성과 퇴행의 불균형, 근세포 사멸 증가 및 근육 재생력 감소의 통합적 결과이다.Sarcopenia is a combination of Greek word "sarco," meaning muscle, and "penia," meaning reduced, which means loss of skeletal muscle mass. It is a concomitant symptom. In the National Health and Nutrition Survey of Korea, the prevalence of muscular dystrophy was found to be 6.6%, and in the United States, the prevalence of muscular dystrophy was 15-25% under 70 years old, 50% in men after 80 years, and 40% over women. Appeared. Myopathy is a degenerative reaction involving overall loss of skeletal muscle mass, quality, and strength, and is a representative symptom of aging. Muscle loss is an integrated result of imbalances in muscle protein production and degeneration, increased myocyte death, and decreased muscle regeneration.
근감소증은 한 번 발병하면 빠르게 나빠지는 경향이 있으며, 국내 65세 이상 성인 560명을 6년 동안 추적 관찰한 결과 근육량과 근력이 부족한 근감소증 환자의 사망 위험이 4.1배 높게 나타났고, 특히 암 환자에게 근감소증이 있을 경우 생존기간이 더 짧고 재발 등의 예후가 더 나쁘다는 보고도 있다. 뿐만 아니라 근감소증은 골다공증, 인슐린저항성 및 관절염과 같은 노인성 만성질환과도 밀접한 관계가 있는 것으로 알려져, 근감소증의 예방 또는 개선을 통해서 노화로 인한 신체 활동력의 감소를 억제할 수 있다.Myopathy tends to deteriorate rapidly with one onset, and as a result of following six years of 560 adults over 65 years of age in Korea, the risk of dying of muscular dystrophy patients with muscle mass and muscle strength was 4.1 times higher, especially for cancer patients. There are reports of shorter survival and a worse prognosis, such as relapse, in patients with muscular dystrophy. In addition, muscular dystrophy is known to be closely related to senile chronic diseases such as osteoporosis, insulin resistance, and arthritis, and it is possible to suppress a decrease in physical activity due to aging through prevention or improvement of muscular dystrophy.
국내외적으로 근감소증 개선을 위한 소재 특히, 천연소재를 이용한 근육 노화 제어 원천기술 개발 및 원인 타겟 중심 치료 약물 개발이 진행 중이나, 현재까지 뚜렷한 효과를 보이거나 허가 받은 소재는 없는 실정이다.Materials for improving muscular dystrophy at home and abroad, in particular, are developing source technology for controlling muscle aging using natural materials and developing drug for target-based treatment, but there are no materials that show a clear effect or have been approved so far.
한편, 다크티(dark tea, dark brick tea; 흑차)는 항산화 작용뿐만 아니라 고혈압과 동맥경화 억제 작용, 수은이나 카드뮴 등 유독성 중금속의 해독 작용 등 다양한 생리활성 효능이 있는 것으로 알려져 있다.On the other hand, dark tea (dark tea, dark brick tea; black tea) is known to have various bioactive effects, such as anti-oxidative action, anti-hypertension and atherosclerosis inhibitory action, and detoxification of toxic heavy metals such as mercury or cadmium.
이에 본 발명자들은 근감소 마우스 모델에 다크티(dark tea, dark brick tea; 흑차) 추출물을 공급하였을 경우 근감소 개선과 운동수행능력의 개선 효과가 월등히 우수한 것을 확인하였다.Accordingly, the present inventors confirmed that when the dark tea (dark tea, dark brick tea; black tea) extract was supplied to the muscle-reducing mouse model, the effect of improving muscle loss and improving exercise performance was significantly superior.
이에, 본 발명의 목적은 다크티 추출물을 포함하는 근육 질환 개선용 건강기능식품 조성물을 제공하는 것이다.Accordingly, an object of the present invention is to provide a health functional food composition for improving muscle disease comprising a dark tea extract.
본 발명의 또 다른 목적은 다크티 추출물을 포함하는 근육 질환 예방 및 치료용 약학 조성물을 제공하는 것이다.Another object of the present invention is to provide a pharmaceutical composition for the prevention and treatment of muscle diseases comprising a dark tea extract.
본 발명의 또 다른 목적은 다크티 추출물을 포함하는 근육량 증대용 건강기능식품 조성물을 제공하는 것이다.Another object of the present invention is to provide a health functional food composition for increasing muscle mass comprising a dark tea extract.
본 발명의 또 다른 목적은 다크티 추출물을 포함하는 운동수행능력 개선용 건강기능식품 조성물을 제공하는 것이다.Another object of the present invention is to provide a health functional food composition for improving exercise performance, including a dark tea extract.
본 발명의 또 다른 목적은 다크티 추출물을 포함하는 염증 개선용 건강기능식품 조성물을 제공하는 것이다.Another object of the present invention is to provide a health functional food composition for improving inflammation comprising a dark tea extract.
본 발명의 또 다른 목적은 다크티 추출물을 포함하는 근육 질환 예방 및 치료용 약학 조성물을 제공하는 것이다.Another object of the present invention is to provide a pharmaceutical composition for the prevention and treatment of muscle diseases comprising a dark tea extract.
본 발명의 또 다른 목적은 다크티 추출물의 근육 질환 개선 용도에 관한 것이다.Another object of the present invention relates to the use of dark tea extract to improve muscle disease.
본 발명은 다크티(dark tea) 추출물을 포함하는 염증 예방 또는 치료용 약제학적 조성물에 관한 것으로, 근육 질환에 대하여 개선 효과를 나타낸다.The present invention relates to a pharmaceutical composition for preventing or treating inflammation including a dark tea extract, and exhibits an improvement effect on muscle diseases.
이에 본 발명자들은 근감소 마우스 모델에 다크티 추출물을 공급하였을 경우 근감소 개선과 운동수행능력의 개선 효과가 월등히 우수함을 확인하였다.Accordingly, the present inventors confirmed that when the dark tea extract was supplied to the muscle-reducing mouse model, the effect of improving muscle loss and improving exercise performance was significantly superior.
이하 본 발명을 더욱 자세히 설명하고자 한다.Hereinafter, the present invention will be described in more detail.
본 발명의 일 양태는 다크티 추출물을 포함하는 근육 질환 개선용 건강기능식품 조성물이다.One aspect of the present invention is a health functional food composition for improving muscle disease comprising a dark tea extract.
본 명세서상의 용어 "다크티"는 차잎을 금화균(Eurotium cristatum strains)을 이용하여 후발효시킨 것을 의미한다.The term "dark tea" in the present specification means that the tea leaves are post-fermented using a gold germ ( Eurotium cristatum strains ).
구체적으로, 차는 차나무(Camella Sinensis) 잎을 가공한 것으로, 차의 분류는 통상 발효정도에 따라 구분되는 것이 기본이다. 제다과정 중 차 잎의 산화효소 및 가수분해효소 등의 작용에 의하여 차 잎의 성분이 생화학적으로 변화하는데, 차 잎 중의 내생효소의 작용정도에 따라 불발효차, 반발효차 및 발효차로 나뉜다.Specifically, the tea is processed from the leaves of the tea tree (Camella Sinensis), and the classification of the tea is usually classified according to the degree of fermentation. During the tea making process, the composition of tea leaves is biochemically changed by the action of oxidase and hydrolase of tea leaves, which are divided into unfermented tea, semi-fermented tea and fermented tea according to the degree of action of endogenous enzyme in tea leaves.
녹차와 같이 제다공정의 첫 단계에서 차 잎을 가열하여 효소의 활성을 실활시켜 녹색이 그대로 유지되도록 만든 차를 "불발효차"라 하며, 우롱차와 같이 수확한 차 잎을 위조와 교반을 하여 내생효소를 작용시켜 찻잎의 폴리페놀 성분의 10~65% 정도가 변환되도록 만든 차를 "반발효차"라 한다. 또 홍차와 같이 위조와 발효 공정을 거쳐 내생효소의 작용에 의해 폴리페놀 성분의 85%이상이 변환되도록 하여 만든 차를 "발효차"라고 한다. 즉 이 분류 방법은 미생물에 의한 발효 정도가 기준이 아니라 찻잎 중에 존재하는 내생효소의 작용 정도가 기준이 되고 있다. 그래서 실제로 찻잎 자체의 내생효소가 아니라 미생물이 작용되어 발효 과정을 거친 차를 "미생물발효차"또는 "후발효차"로 분류하기도 한다.A tea made by heating tea leaves in the first stage of the Jeddah process, such as green tea, to deactivate the activity of enzymes to maintain green, is called "fermented tea". Forged and stirred tea leaves harvested like oolong tea A tea made by converting 10 to 65% of the polyphenol content of a tea leaf by acting an endogenous enzyme is called "reverse fermented tea." In addition, a tea made by fermentation and fermentation processes such as black tea, such that 85% or more of the polyphenol component is converted by the action of endogenous enzymes, is called "fermented tea. That is, in this classification method, the degree of fermentation by microorganisms is not the standard, but the degree of the action of endogenous enzymes present in the tea leaves. So, in fact, not the endogenous enzyme of the tea leaves themselves, but the microorganisms are applied and the fermented tea is classified as "microbial fermented tea" or "post fermented tea".
다크티(dark tea, dark brick tea; 흑차)는 가공공정에서 녹차에 비해 후발효 과정이 추가된다. 다크티의 원료가 비교적 거칠기 때문에 퇴적시켜 긴 시간 동안 발효시키는 과정을 통해서 차잎을 부드럽게 하고 떫은맛을 없애고 차의 맛을 맑고 부드럽게 해준다. 이러한 과정을 통해 만들어진 차잎은 길쭉한 모양에 검고 반지르르하며 흑갈색으로 변해서 다크티라고 부른다. 육보차, 복전차, 천량차, 운남보이차가 유명하다.Dark tea (dark tea, dark brick tea; black tea) is added to the post-fermentation process compared to green tea in the processing process. Since the raw material of dark tea is relatively rough, it is deposited and fermented for a long time to soften the tea leaves, remove the astringent taste, and make the taste of tea clear and soft. The tea leaves made through this process are elongated in shape, black and round, and turn dark brown to be called dark tea. Yukbo Tea, Futian Tea, Chun Liang Tea, and Yunnan Boy Tea are famous.
중국의 후난성(Hunan Province)에서 주로 생산되는 다크티는 후발효차의 하나로, 모차(잎차)를 일차적으로 가공한 후 다시 퇴적(일차 가공한 찻잎을 쌓아 두는 방법)이나 악퇴(일차 가공한 찻잎에 물을 뿌리고 쌓아 두어 찻잎 자체의 온도와 습도를 높이는 방법)의 과정을 거쳐 2차 가공을 하거나 저장하면서 차에 미생물이 발생하도록 한다.Dark tea, mainly produced in Hunan Province in China, is one of the later fermented teas, which is primarily processed after processing macha (leaf tea) and deposited again (how to stack the primary processed tea leaves) or bad decay (primarily processed tea leaves). After sprinkling water and stacking it, a method of increasing the temperature and humidity of the tea leaves itself) is followed by secondary processing or storage to generate microorganisms in the car.
일반적으로 생엽을 채집(채엽)하여 시들게 하고(위조), 덖은 후(살청), 비벼서(유념), 건조(쇄청)시키는 과정을 거쳐 모차를 제조한다. 이후 상기 모차에 증기를 쐬어 부드럽게 하고, 80℃에서 밤새 퇴적하여 벽돌(brick) 모양으로 압축시킨 후, 15 내지 17일 동안 온도 20 내지 30℃ 습도 70 내지 85%에서 금화균(Eurotium cristatum strains; Golden flora; 관돌산냥균)으로 후발효 과정을 거친다. 발효 시 찻잎의 함수량이 13 내지 14%를 넘지 않고 공기의 상대습도가 75% 전후인 비교적 건조한 조건에서 금화균의 생장이 특히 양호하다. 이후 찻잎을 건조시켜 6개월 정도 숙성시켜 제조된다.In general, the raw leaves are collected (pickled) and then withered (forged), smashed (salted), rubbed (remembered), and dried (crushed) to produce a mocha. Thereafter, the steam was softened by steaming the vehicle, deposited overnight at 80° C., compressed into a brick shape, and then aged 15 to 17 days at a temperature of 20 to 30° C. at a humidity of 70 to 85%. Eurotium cristatum strains; Golden flora; gwandol sankyyun). During fermentation, the growth of gold germs is particularly good in relatively dry conditions where the water content of the tea leaves does not exceed 13 to 14% and the relative humidity of the air is around 75%. After that, the tea leaves are dried and aged for about 6 months.
금화균이란 다크티의 하나인 복전차 등으로부터 자연 발생적으로 나타나는 유익한 노란색의 곰팡이균들 중에서, 황금색 꽃(golden flora)이 핀 것 같은 미생물 균총을 의미하고, 유로티움 속(Eurotium)이 여기에 속한다. 다크티의 제조에 있어서는 찻잎의 발효를 위해 조건을 조성해줌으로써 인위적으로 금화균의 생장을 촉진시키고 번식시키는 공정이 수행되며, 이에 따라 발효되는 찻잎은 노란색을 띄는 금화균의 포자에 의하여 표면에 금가루가 뿌려진 것처럼 보이는 특징을 갖게 된다.Among the beneficial yellow fungi that naturally occur from a dark tea, such as an electric car, gold-fired bacteria means a microbial microflora like a golden flora, and the genus Eurotium belongs to this. . In the manufacture of dark tea, artificial fertilization is promoted and propagated by creating conditions for fermentation of tea leaves, and the fermented tea leaves are gold powdered on the surface by spores of yellowish gold bacteria. You will have a characteristic that appears to be scattered.
상기 근육 질환은 근위축증(muscular atrophy), 근육감소증(Sacopenia), 근육퇴행위축증, 심위축증, 긴장감퇴증(atony), 근이영양증(muscular dystrophy), 근육 퇴화증 및 근무력증으로 이루어진 군으로부터 선택되는 것일 수 있고, 예를 들어, 근위축증일 수 있으나, 이에 한정되는 것은 아니다.The muscle disease may be selected from the group consisting of muscle atrophy, sarcopenia, muscular atrophy, cardiac atrophy, atony, muscular dystrophy, muscle degeneration and myasthenia gravis. , For example, may be muscular dystrophy, but is not limited thereto.
상기 근위축증은 노화성 근위축증 또는 비만성 근위축증인 것일 수 있다.The muscular dystrophy may be aging muscular dystrophy or obese muscular dystrophy.
상기 '추출물'은 용매 조추출물, 특정 용매 가용 추출물(용매 분획물) 및 용매 조추출물의 용매 분획물을 포함하며, 상기 다크티 추출물은 용액, 농축물 또는 분말 상태일 수 있다.The'extract' includes a solvent crude extract, a specific solvent soluble extract (solvent fraction) and a solvent fraction of the solvent crude extract, and the dark tea extract may be in solution, concentrate or powder form.
상기 다크티 추출물은 물, 탄소수 1 내지 6의 유기용매, 아임계 유체 및 초임계 유체로 이루어진 군으로부터 선택되는 하나 이상의 용매로 추출된 조추출물일 수 있고, 예를 들어, 물을 용매로 하여 추출된 것일 수 있으나, 이에 한정되는 것은 아니다.The dark tea extract may be a crude extract extracted with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, a subcritical fluid, and a supercritical fluid, for example, water as a solvent It may be, but is not limited to this.
다크티의 조추출물 제조에 사용되는 한 용매에 물과 알코올의 혼합물을 사용하는 경우에는 10%이상 내지 100%(v/v)미만, 20%이상 내지 100%(v/v)미만, 30%이상 내지 100%(v/v)미만, 40%이상 내지 100%(v/v)미만, 50%이상 내지 100%(v/v)미만, 60%이상 내지 100%(v/v)미만, 70%이상 내지 100%(v/v)미만, 10%이상 내지 90%(v/v)미만, 20%이상 내지 90%(v/v)미만, 30%이상 내지 90%(v/v)미만, 40%이상 내지 90%(v/v)미만, 50%이상 내지 90%(v/v)미만, 60%이상 내지 90%(v/v)미만 또는 70%이상 내지 90%(v/v)의 탄소수 1 내지 4개의 직쇄 또는 분지형 알코올 수용액, 예를 들어, 80%(v/v)의 탄소수 1 내지 4개의 직쇄 또는 분지형 알코올 수용액일 수 있다.When using a mixture of water and alcohol in one solvent used to prepare the dark tea crude extract, 10% or more to 100% (v/v) or less, 20% or more to 100% (v/v) or less, 30% Above to 100% (v/v), above 40% to below 100% (v/v), above 50% to below 100% (v/v), above 60% to below 100% (v/v), 70% to 100% (v/v), 10% to 90% (v/v), 20% to 90% (v/v), 30% to 90% (v/v) Less than, 40% to 90% (v/v), 50% to 90% (v/v), 60% to 90% (v/v) or 70% to 90% (v/v) v) 1 to 4 carbon atoms straight or branched alcohol aqueous solution, for example, 80% (v/v) 1 to 4 carbon atoms straight chain or branched alcohol aqueous solution.
본 발명에 따른 다크티 추출물은 용매 조추출물을 추가의 용매로 분획한 용매 분획물일 수 있으며, 예를 들면 상기 용매 조추출물에 에틸에테르, 아세트산에틸, 및 부탄올로 이루어지는 군에서 선택된 1종 이상의 용매를 사용한 용매 분획물일 수 있다.The dark tea extract according to the present invention may be a solvent fraction obtained by fractionating the solvent crude extract with an additional solvent, for example, one or more solvents selected from the group consisting of ethyl ether, ethyl acetate, and butanol in the solvent crude extract. It may be the solvent fraction used.
예를 들면, 상기 다크티를 물 및 탄소수 1 내지 4개의 직쇄 또는 분지형 알코올로 이루어지는 군에서 선택된 1종 이상의 용매로 추출한 용매 조추출물을 에틸에테르, 아세트산에틸, 및 부탄올로 이루어지는 군에서 선택된 1종 이상의 용매를 사용한 용매 분획물일 수 있다.For example, the solvent extract extracted from the dark tea with one or more solvents selected from the group consisting of water and straight or branched alcohol having 1 to 4 carbon atoms is selected from the group consisting of ethyl ether, ethyl acetate, and butanol. It may be a solvent fraction using the above solvent.
본 발명의 일 양태는 다크티 추출물을 제조하는 방법에 관한 것이다.One aspect of the present invention relates to a method for preparing a dark tea extract.
상기 다크티 추출물은 다크티의 용매 조추출물, 용매 분획물을 포함하며 상술한 바와 같다.The dark tea extract includes a crude solvent extract and a solvent fraction of dark tea and is as described above.
상기 다크티 추출물은 다크티의 잎, 줄기 및 뿌리로 이루어지는 군에서 선택된 1종 이상을, 물 및 탄소수 1 내지 4개의 직쇄 또는 분지형 알코올로 이루어진 군에서 선택된 1종 이상의 용매로 추출하여 얻어진 조추출물일 수 있다.The dark tea extract is a crude extract obtained by extracting one or more kinds selected from the group consisting of dark tea leaves, stems and roots, and one or more solvents selected from the group consisting of water and straight or branched alcohol having 1 to 4 carbon atoms. Can be
상기 용매는 10%이상 내지 100%(v/v)미만, 20%이상 내지 100%(v/v) 미만, 30%이상 내지 100%(v/v)미만, 40%이상 내지 100%(v/v)미만, 50%이상 내지 100%(v/v)미만, 60%이상 내지 100%(v/v)미만, 70%이상 내지 100%(v/v)미만, 10%이상 내지 90%(v/v)미만, 20%이상 내지 90%(v/v)미만, 30%이상 내지 90%(v/v)미만, 40%이상 내지 90%(v/v)미만, 50%이상 내지 90%(v/v)미만, 60%이상 내지 90%(v/v)미만 또는 70%이상 내지 90%(v/v)의 탄소수 1 내지 4개의 직쇄 또는 분지형 알코올 수용액, 예를 들어, 80%(v/v)의 탄소수 1 내지 4개의 직쇄 또는 분지형 알코올 수용액일 수 있다.The solvent is 10% or more to less than 100% (v/v), 20% or more to less than 100% (v/v), 30% or more to less than 100% (v/v), 40% or more to 100% (v /v) Less than, 50% to 100% (v/v) less, 60% to 100% (v/v) less, 70% to 100% (v/v) less, 10% to 90% (v/v) or less, 20% or more to 90% (v/v) or less, 30% or more to 90% (v/v) or less, 40% or more to 90% (v/v) or less, 50% or more to 90% (v/v) or less, 60% or more to 90% (v/v) or 70% to 90% (v/v) of a straight chain or branched alcohol aqueous solution having 1 to 4 carbon atoms, for example 80% (v/v) of a straight chain or branched alcohol solution having 1 to 4 carbon atoms.
본 발명에 따른 다크티 추출물의 제조 과정을 보다 상세하게 설명하면 다음과 같다: 다크티의 중량에 대하여 약 10 내지 50배 중량의 추출용매로 상온 추출한다. 추출 후 여과하여 여과액을 모은다. 추출 온도는 특별한 제한은 없지만 15 내지 110, 바람직하게는 20 내지 90인 것이 좋다.The manufacturing process of the dark tea extract according to the present invention will be described in more detail as follows: Extraction at room temperature with an extraction solvent of about 10 to 50 times the weight of the dark tea. After extraction, the filtrate is collected by filtration. The extraction temperature is not particularly limited, but is preferably 15 to 110, preferably 20 to 90.
추출공정은 1회 또는 수회 반복할 수 있으며, 본 발명의 한 바람직한 예에서는 1차 추출 후 다시 재추출하는 방법을 채택할 수 있는데, 이는 생약추출물을 대량 생산하는 경우 효과적으로 여과를 한다 하더라도 생약 자체의 수분 함량이 높기 때문에 손실이 발생하게 되어 1차 추출만으로는 추출효율이 떨어지므로 이를 방지하기 위함이다. 또한, 각 단계별 추출효율을 검증한 결과 2차 추출에 의해 전체 추출량의 80 내지 90% 정도가 추출되는 것으로 밝혀졌다.The extraction process may be repeated once or several times, and in one preferred example of the present invention, a method of re-extraction after the primary extraction may be adopted, which, even when effectively filtering large quantities of herbal extracts, is performed even if the herbal extracts are filtered effectively. This is to prevent the loss due to the high moisture content, and the extraction efficiency is reduced only by the first extraction. In addition, as a result of verifying the extraction efficiency at each step, it was found that about 80 to 90% of the total extraction amount is extracted by secondary extraction.
본 발명의 일 예에서, 추출공정을 2회 반복하는 경우, 상기 얻어진 잔사에 다시 추출용매, 약 5 내지 15 부피배, 바람직하게는 8 내지 12 부피배로 환류 추출한다. 추출 후 여과하고 이전에 얻어진 여과액과 합쳐서 감압농축을 하여 다크티 추출물을 제조한다. 이와 같이 2차에 걸친 추출 및 각각의 추출 후 얻어진 여과액을 혼합함으로써 추출 효율을 높일 수 있으나, 본 발명의 추출물이 추출 회수에 한정되는 것은 아니다.In one example of the present invention, when the extraction process is repeated twice, the obtained residue is refluxed again with an extraction solvent, about 5 to 15 volume times, preferably 8 to 12 volume times. After extraction, it is filtered and combined with the filtrate obtained previously, and concentrated under reduced pressure to prepare a dark tea extract. Thus, the extraction efficiency can be increased by mixing the extract obtained over the second time and the filtrate obtained after each extraction, but the extract of the present invention is not limited to the number of extractions.
상기 다크티 추출물 제조 시에 사용되는 용매의 양이 너무 적으면 교반이 어렵게 되고 추출물의 용해도가 낮아져 추출효율이 떨어지게 되고, 지나치게 많은 경우는 다음의 정제단계에서 사용되는 용매의 사용량이 많아져 경제적이지 못하여 취급상 문제가 발생할 수 있으므로, 용매의 사용량은 상기 범위로 하는 것이 좋다.When the amount of the solvent used in the preparation of the dark tea extract is too small, stirring becomes difficult and the solubility of the extract is lowered to lower the extraction efficiency, and in the case of too much, the amount of the solvent used in the next purification step is increased, which is economical. Since it may not cause handling problems, the amount of the solvent used is preferably within the above range.
이와 같이 얻어진 여과된 추출물은 의약품 원료로 사용하기에 적합하도록 잔존하는 저급 알코올의 함량을 조절하기 위하여 농축물 총량의 약 10 내지 30 중량배, 바람직하게는 15 내지 25 중량배, 보다 바람직하게는 약 20 중량배의 물로 1 내지 5회, 바람직하게는 2 내지 3회 공비 농축하고 재차 동량의 물을 가하여 균질하게 현탁시킨 후 동결건조하여 분말상태의 다크티 추출물로서 제조될 수 있다.The filtered extract obtained in this way is about 10 to 30 times the total amount of concentrate, preferably 15 to 25 times, more preferably about to adjust the content of the lower alcohol remaining to be suitable for use as a pharmaceutical raw material. After azeotropic concentration of 1 to 5 times, preferably 2 to 3 times, with 20 times the weight of water, and then homogeneously suspended by adding the same amount of water, lyophilization may be prepared as a powdery dark tea extract.
본 발명에 사용된 추출 방법은 통상적으로 사용되는 모든 방법일 수 있으며, 예컨대, 냉침, 열수추출, 초음파 추출, 또는 환류 냉각 추출법일 수 있으나, 이에 한정되는 것은 아니다.The extraction method used in the present invention may be any method commonly used, for example, cold immersion, hot water extraction, ultrasonic extraction, or reflux cooling extraction, but is not limited thereto.
본 발명의 건강기능식품 조성물을 식품 첨가물로 사용할 경우, 상기 건강기능식품 조성물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용할 수 있고, 통상적인 방법에 따라 적절하게 사용할 수 있다. 일반적으로, 식품 또는 음료의 제조 시에 본 발명의 건강기능식품 조성물은 원료에 대하여 15 중량% 이하, 바람직하게는 10 중량% 이하의 양으로 첨가될 수 있다.When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as it is or used together with other food or food ingredients, and may be suitably used according to a conventional method. In general, the health functional food composition of the present invention in the manufacture of a food or beverage may be added in an amount of 15% by weight or less, preferably 10% by weight or less based on the raw material.
상기 식품의 종류에는 특별한 제한은 없다. 상기 물질을 첨가할 수 있는 식품의 예로는 육류, 소세지, 빵, 초콜릿, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 식품을 모두 포함한다.There are no particular restrictions on the type of food. Examples of foods to which the above substances can be added are meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, dairy products including gums, ice creams, various soups, beverages, teas, drinks, Alcoholic beverages, vitamin complexes, and the like, and include all foods in the ordinary sense.
상기 음료는 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 당업자의 선택에 의해 적절하게 결정될 수 있다.The beverage may contain various flavoring agents or natural carbohydrates as an additional component. The natural carbohydrates described above may include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharin and aspartame. . The proportion of the natural carbohydrate can be appropriately determined by the choice of those skilled in the art.
상기 외에 본 발명의 건강기능식품 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 건강기능식품 조성물은 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율 또한 당업자에 의해 적절히 선택될 수 있다.In addition to the above, the health functional food composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin , Carbonic acid used in alcoholic beverages, carbonated beverages, and the like. In addition, the health functional food composition of the present invention may contain flesh for the preparation of natural fruit juice, fruit juice beverage and vegetable beverage. These ingredients can be used independently or in combination. The proportions of these additives can also be appropriately selected by those skilled in the art.
본 발명의 다른 양태는 다크티 추출물을 포함하는 근육 질환 예방 및 치료용 약학 조성물이다.Another aspect of the present invention is a pharmaceutical composition for preventing and treating muscle disease comprising a dark tea extract.
상기 근육 질환은 근위축증, 근육감소증, 근육퇴행위축증, 심위축증, 긴장감퇴증, 근이영양증, 근육 퇴화증 및 근무력증으로 이루어진 군으로부터 선택되는 것일 수 있고, 예를 들어, 근위축증일 수 있으나, 이에 한정되는 것은 아니다.The muscle disease may be selected from the group consisting of muscular dystrophy, muscular dystrophy, muscular dystrophy, cardiac atrophy, tension reduction, muscular dystrophy, muscular dystrophy and myasthenia, for example, muscular dystrophy, but is not limited thereto. no.
상기 근위축증은 노화성 근위축증 또는 비만성 근위축증인 것일 수 있다.The muscular dystrophy may be aging muscular dystrophy or obese muscular dystrophy.
상기 추출물은 물, 탄소수 1 내지 6의 유기용매, 아임계 유체 및 초임계 유체로 이루어진 군으로부터 선택되는 하나 이상의 용매로 추출된 것일 수 있고, 예를 들어, 물을 용매로 하여 추출된 것일 수 있으나, 이에 한정되는 것은 아니다.The extract may be extracted with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, a subcritical fluid, and a supercritical fluid, for example, water may be extracted using a solvent. , But is not limited thereto.
상기 다크티 추출물은 50 내지 300 mg/kg BW, 80 내지 250 mg/kg BW, 100 내지 200 mg/kg BW, 예를 들어, 150 내지 200 mg/kg BW의 농도로 활용되는 것일 수 있다.The dark tea extract may be utilized at a concentration of 50 to 300 mg/kg BW, 80 to 250 mg/kg BW, 100 to 200 mg/kg BW, for example, 150 to 200 mg/kg BW.
본 발명의 약학 조성물은 다크티 추출물의 약제학적 유효량 및/또는 약제학적으로 허용되는 담체를 포함하는 약제학적 조성물로 이용될 수 있다.The pharmaceutical composition of the present invention may be used as a pharmaceutical composition comprising a pharmaceutically effective amount of dark tea extract and/or a pharmaceutically acceptable carrier.
본 명세서에서 용어 "약제학적 유효량"은 상술한 다크티 추출물의 효능 또는 활성을 달성하는 데 충분한 양을 의미한다.The term "pharmaceutically effective amount" as used herein means an amount sufficient to achieve the efficacy or activity of the aforementioned dark tea extract.
본 발명의 약학 조성물에 포함되는 약제학적으로 허용되는 담체는 제제시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 약제학적 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다.The pharmaceutically acceptable carrier included in the pharmaceutical composition of the present invention is commonly used in formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, silicic acid Calcium, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, but is not limited thereto. It is not. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components.
본 발명에 따른 약학 조성물은 인간을 포함하는 포유동물에 다양한 경로로 투여될 수 있다. 투여 방식은 통상적으로 사용되는 모든 방식일 수 있으며, 예컨대, 경구, 피부, 정맥, 근육, 피하 등의 경로로 투여될 수 있으며, 바람직하게는 경구로 투여될 수 있다.The pharmaceutical composition according to the present invention can be administered to various mammals, including humans, by various routes. The mode of administration may be any of the commonly used modes, for example, oral, skin, intravenous, intramuscular, subcutaneous, and the like, and may be administered, preferably, orally.
본 발명의 약학 조성물의 적합한 투여량은 제제화 방법, 투여방식, 환자의 연령, 체중, 성별, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하며, 보통으로 숙련된 의사는 소망하는 치료 또는 예방에 효과적인 투여량을 용이하게 결정 및 처방할 수 있다. 본 발명의 바람직한 구현예에 따르면, 본 발명의 약학 조성물의 1일 투여량은 100 내지 400 mg/kg이다.Suitable dosages of the pharmaceutical compositions of the invention vary by factors such as formulation method, mode of administration, patient's age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion, and response sensitivity, usually As a result, a skilled doctor can easily determine and prescribe a dose effective for the desired treatment or prevention. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 100 to 400 mg/kg.
본 발명의 약학 조성물은 당해 발명이 속하는 기술 분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약제학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기내에 내입시켜 제조될 수 있다. 이때 제형은 오일 또는 수성 매질중의 용액, 현탁액 또는 유화액 형태이거나 엑스제, 분말제, 과립제, 정제, 캅셀제 또는 젤(예컨대, 하이드로젤) 형태일 수도 있으며, 분산제 또는 안정화제를 추가적으로 포함할 수 있다.The pharmaceutical composition of the present invention is prepared in a unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person skilled in the art to which the present invention pertains. Or it can be manufactured by incorporating into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of ex-agent, powder, granule, tablet, capsule or gel (eg, hydrogel), and may further include a dispersant or stabilizer. .
본 발명의 또 다른 양태는 다크티 추출물을 포함하는 근육량 증대용 건강기능식품 조성물이다.Another aspect of the present invention is a health functional food composition for increasing muscle mass comprising a dark tea extract.
본 발명의 또 다른 양태는 다크티 추출물을 포함하는 운동수행능력 개선용 건강기능식품 조성물이다.Another aspect of the present invention is a health functional food composition for improving exercise performance, comprising a dark tea extract.
본 발명의 또 다른 양태는 다크티 추출물을 포함하는 염증 개선용 건강기능식품 조성물이다.Another aspect of the present invention is a health functional food composition for improving inflammation comprising a dark tea extract.
본 발명의 또 다른 양태는 다크티 추출물을 포함하는 염증 예방 또는 치료용 약제학적 조성물이다.Another aspect of the present invention is a pharmaceutical composition for preventing or treating inflammation comprising a dark tea extract.
본 발명은 다크티(dark tea) 추출물을 포함하는 근육 질환 개선용 조성물에 관한 것으로서, 근감소 마우스 모델에 다크티(dark tea) 추출물을 공급하였을 경우 근감소 개선과 운동수행능력의 개선 효과가 월등히 우수하였으므로, 이를 효과적으로 근육 기능 증강에 이용할 수 있다.The present invention relates to a composition for improving muscle disease including a dark tea extract, and when a dark tea extract is supplied to a muscle-reduced mouse model, the effect of improving muscle loss and improving exercise performance is significantly improved. Since it was excellent, it can be effectively used to enhance muscle function.
도 1a는 다크티(dark tea) 추출물이 전경골근(tibialis anterior; TA)의 근육량에 미치는 영향을 보여주는 그래프이다.
도 1b는 다크티 추출물이 장지신근(extensor digitorum long; EDL)의 근육량에 미치는 영향을 보여주는 그래프이다.
도 1c는 다크티 추출물이 비복근(Gastroc nemius)의 근육량에 미치는 영향을 보여주는 그래프이다.
도 1d는 다크티 추출물이 비장근(Soleus)의 근육량에 미치는 영향을 보여주는 그래프이다.
도 1e는 다크티 추출물이 대퇴사두근(Quadricept)의 근육량에 미치는 영향을 보여주는 그래프이다.
도 1f는 다크티 추출물이 삼두박근(Tricept)의 근육량에 미치는 영향을 보여주는 그래프이다.
도 2는 골격근의 크기에 미치는 다크티의 영향을 나타낸 그래프이다.
도 3은 다크티 추출물의 그립강도 개선 효과를 나타낸 그래프이다.
도 4a는 다크티 추출물의 운동수행거리 증진 효과를 나타낸 그래프이다.
도 4b는 다크티 추출물의 운동시간 증진 효과를 나타낸 그래프이다.
도 5a는 다크티 추출물의 염증성 사이토카인 TNFα 감소 효과를 나타낸 그래프이다.
도 5b는 다크티 추출물의 염증성 사이토카인 IL-1β 감소 효과를 나타낸 그래프이다.
도 5c는 다크티 추출물의 염증성 사이토카인 MCP1 감소 효과를 나타낸 그래프이다.
도 5d는 다크티 추출물의 염증성 사이토카인 IL-6 감소 효과를 나타낸 그래프이다.Figure 1a is a graph showing the effect of dark tea (dark tea) extract on the muscle mass of the tibialis anterior (TA).
1B is a graph showing the effect of dark tea extract on muscle mass of extensor digitorum long (EDL).
Figure 1c is a graph showing the effect of dark tea extract on the muscle mass of gastrocnemius (Gastroc nemius).
Figure 1d is a graph showing the effect of dark tea extract on the muscle mass of the spleen muscle (Soleus).
Figure 1e is a graph showing the effect of dark tea extract on the muscle mass of the quadriceps (Quadricept).
Figure 1f is a graph showing the effect of dark tea extract on the muscle mass of the triceps muscle (Tricept).
2 is a graph showing the effect of dark tea on the size of skeletal muscle.
3 is a graph showing the effect of improving the grip strength of the dark tea extract.
Figure 4a is a graph showing the effect of increasing the exercise distance of dark tea extract.
Figure 4b is a graph showing the effect of increasing the exercise time of the dark tea extract.
5A is a graph showing the effect of reducing the inflammatory cytokine TNFα of the dark tea extract.
Figure 5b is a graph showing the effect of reducing the inflammatory cytokine IL-1β of the dark tea extract.
Figure 5c is a graph showing the effect of reducing the inflammatory cytokine MCP1 of the dark tea extract.
Figure 5d is a graph showing the effect of reducing the inflammatory cytokine IL-6 of the dark tea extract.
이하, 본 발명을 하기의 실시예에 의하여 더욱 상세히 설명한다. 그러나 이들 실시예는 본 발명을 예시하기 위한 것일 뿐이며, 본 발명의 범위가 이들 실시예에 의하여 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail by the following examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.
본 명세서 전체에 걸쳐, 특정 물질의 농도를 나타내기 위하여 사용되는 "%"는 별도의 언급이 없는 경우, 고체/고체는 (중량/중량)%, 고체/액체는 (중량/부피)%, 그리고 액체/액체는 (부피/부피)%이다.Throughout this specification, "%" used to indicate the concentration of a specific substance, unless otherwise specified, solids/solids (weight/weight)%, solids/liquids (weight/volume)%, and The liquid/liquid is (volume/volume)%.
실시예 1: 다크티 추출물 및 시험동물의 준비Example 1: Preparation of dark tea extract and test animals
본 실험에 사용된 시료인 다크티는 중국에서 생산된 제품을 각각 사용하였다(HUNAN TEA CO.,LTD.). 또한 추출물의 제조는 증류수 500 mL에 시료 50 g을 넣어(10%, v/w) 80℃에서 10분간 환류냉각 추출한 후 여과지(filter paper, Whatman No. 2. UK)로 여과 및 동결건조하여 시료로 사용하였다.Dark tea, a sample used in this experiment, was used for products manufactured in China ( HUNAN TEA CO.,LTD. ). In addition, the extract was prepared by putting 50 g of sample in 500 mL of distilled water (10%, v/w), extracting by reflux cooling at 80° C. for 10 minutes, filtering and lyophilizing the sample with filter paper (Whatman No. 2. UK) It was used as.
동물은 4주령의 C57BL/6J 마우스를 1주일간 환경에 적응시킨 뒤 근감소를 유도하기 위하여 고지방식이(research diet 60% kcal, high fat diet, HFD)로 9주간 사육한 뒤, 고지방식이 대조군, 고지방식이 다크티 추출물 저농도군, 고지방식이 다크티 추출물 고농도군의 3그룹으로 나누었고 각군당 10마리로 무작위 배치하였다.Animals were raised for 4 weeks with a high-fat diet (
고지방식이 대조군에는 고지방식이와 식염수를 경구 투여하였고 다크티 추출물군은 고지방식이와 동시에 다크티 추출물을 100 mg/kg BW(DT), 200 mg/kg BW(DT)를, 정상식이군(Chow)의 경우 다크티 추출물 없이 정상식이를 각각 경구 투여하였으며 11주 동안 사육하였다. 물과 실험식이는 자유롭게 섭취하도록 하였다. 사육이 끝난 동물은 마취 하에 해부하여 혈액 채취 및 근육의 전 조직을 구분하여 채취하고 무게를 소수점 4자리까지 취하여 측정하였다.High-fat diet and saline were administered orally to the high-fat diet control group, and the dark tea extract group received 100 mg/kg BW(DT), 200 mg/kg BW(DT), and the dark diet extract at the same time as the high-fat diet. Chow), each of the normal diet was administered orally without dark tea extract and kept for 11 weeks. Water and experimental diet were allowed to be consumed freely. Animals after breeding were dissected under anesthesia, and blood was collected and all tissues of the muscles were collected separately.
실시예 2: 다크티의 근감소 개선 효과 확인Example 2: Confirm the effect of dark tea on improving muscle loss
다크티 추출물에 의한 근감소 개선 및 근육량 증대 효과를 확인하기 위해 시험동물의 근육조직(Gastrocnemius)을 확인하였다.The muscle tissue ( Gastrocnemius ) of the test animal was confirmed to confirm the effect of improving muscle loss and increasing muscle mass by the dark tea extract.
구체적으로, 실시예 1에서 준비된 고지방식이 대조군(HFD), 고지방식이 다크티 추출물 저농도군(100 mg/kg BW), 고지방식이 다크티 추출물 고농도군(200 mg/kg BW) 및 대조군(정상식이; Chow)에 해당하는 시험동물의 근육조직을 적출하여 4% 포름알데하이드 용액에 24시간 동안 고정하고 물로 충분히 수세하였다. 이후 78%, 80%, 90% 및 100% 에탄올로 이용하여 단계적으로 탈수시킨 다음 파라핀(paraffin) 투과과정을 거쳐 포매하였다. 포매된 조직은 박절편기로 4 um의 두께로 박절하여 헤마톡실린-에오신(hematoxylin-eosin, H&E)으로 염색하고, 염색된 근육조직 샘플은 현미경(digital imaging software 부착)을 이용하여 관찰하였으며, CSA(cross sectional area, um2)측정하여 계산하였다.Specifically, the high-fat diet prepared in Example 1 (HFD), the high-fat diet dark tea extract low concentration group (100 mg/kg BW), the high-fat diet dark tea extract high concentration group (200 mg/kg BW) and the control group ( The muscle tissue of the test animal corresponding to the normal diet (Chow) was extracted, fixed in a 4% formaldehyde solution for 24 hours, and washed sufficiently with water. Subsequently, it was dehydrated step by step using 78%, 80%, 90% and 100% ethanol, and then embedded through a paraffin permeation process. The embedded tissue was sliced to a thickness of 4 um with a thin slicer, stained with hematoxylin-eosin (H&E), and the stained muscle tissue sample was observed using a microscope (digital imaging software attachment) and CSA (cross sectional area, um 2 ) was measured and calculated.
도 1 및 표 1에서 확인할 수 있듯이, 비만으로 인한 근감소 마우스 모델에서 다크티를 100 mg/kg, 200 mg/kg를 11주 동안 경구 투여한 결과, 정상식이군(Chow)에 비하여 고지방식이 대조군에서는 측정한 모든 부위의 근육이 유의하게 감소한 반면 고지방식이와 함께 다크티 추출물을 첨가한 군에서는 대조군에 비하여 근육량이 증가하였고, 특히 전경골근(tibialis anterior; TA), 비복근(Gastroc nemius), 대퇴사두근(Quadricept), 삼두박근(Tricept)에서는 용량의존적으로 근육량이 증가하였다.As can be seen in Figure 1 and Table 1, as a result of oral administration of
실시예 3: 다크티의 골격근비대 유도 효과 확인Example 3: Confirming the effect of dark tea on skeletal muscle hypertrophy
다크티 추출물에 의한 근감소 개선 및 근육량 증대 효과를 확인하기 위해 실시예 1에서 준비된 고지방식이 대조군(HFD), 고지방식이 다크티 추출물 저농도군(100 mg/kg BW), 고지방식이 다크티 추출물 고농도군(200 mg/kg BW) 및 대조군(Chow)에 해당하는 시험동물의 근섬유 크기를 확인하였다.High-fat diet control (HFD) prepared in Example 1, high-fat diet dark tea extract low concentration group (100 mg/kg BW), high-fat diet dark tea to improve muscle loss and increase muscle mass by dark tea extract The muscle fiber size of the test animals corresponding to the extract high concentration group (200 mg/kg BW) and the control group (Chow) was confirmed.
도 2 및 표 2에서 확인할 수 있듯이, 마우스 근육조직(gastronemius)을 염색하여 근섬유(muscle fiber)의 크기를 분석한 결과 정상식이군에 비하여 고지방식이군에서는 감소하였으나, 다크티 추출물군은 고지방식이 대조군에 비하여 근섬유의 크기가 증가하는 것을 확인하였다.As can be seen in Figure 2 and Table 2, as a result of analyzing the size of the muscle fibers by staining the mouse muscle tissue (gastronemius), compared to the normal diet group, the high-fat diet group decreased, but the dark tea extract group was a high-fat diet control group. Compared to, it was confirmed that the size of the muscle fibers increased.
실시예 4: 다크티의 근육기능 개선 효과의 확인Example 4: Confirmation of dark tea muscle function improvement effect
다크티의 근육기능 및 운동수행능력 개선 효능을 확인하기 위하여 실시예 1에서 준비된 고지방식이 대조군(HFD), 고지방식이 다크티 추출물 저농도군(100 mg/kg BW), 고지방식이 다크티 추출물 고농도군(200 mg/kg BW) 및 대조군(Chow)에 해당하는 시험동물에 대하여 그립강도 및 트레드밀(treadmill)을 이용한 운동부하검사를 실시하였다.High-fat diet control (HFD) prepared in Example 1, high-fat diet dark tea extract low-concentration group (100 mg/kg BW), high-fat diet dark tea extract to confirm the improvement of muscle function and exercise performance ability of dark tea The test loads corresponding to the high concentration group (200 mg/kg BW) and the control group (Chow) were tested for exercise load using grip strength and treadmill.
시험동물의 근육기능 분석을 위하여 앞다리 그립 강도 테스트(Forelimb grip strength test)를 진행하였다. 상기 테스트는 시험동물의 목덜미와 꼬리를 잡고 시험동물이 바(bar)를 양손으로 잘 잡을 수 있도록 자세를 유도하고, 꼬리를 잡아 쥐가 앞다리로 잡았던 막대를 놓치게 되는 순간의 최대 악력을 측정하는 방법이다. 각각의 시험동물에게 연속적으로 그립 테스트를 5회씩 실시하였으며, 5번의 시도 중 최고값과 최저값을 제외한 평균값을 데이터로 사용하였다. 데이터상의 수치는 측정된 힘의 값을 체중(g)으로 표준화하여 나타내었다.Forelimb grip strength test was performed to analyze muscle function of test animals. The test induces a posture so that the test animal can grasp the bar with both hands by holding the neck and tail of the test animal, and the method of measuring the maximum grip force at the moment when the rod catches the rod held by the forelimb by holding the tail. to be. Each test animal was subjected to a
도 3 및 표 3에서 확인할 수 있듯이, 고지방식이 대조군은 정상군에 비하여 그립강도가 유의하게 감소하였으나 다크티 추출물군은 대조군에 비하여 농도의존적으로 증가하였다.As can be seen in Figure 3 and Table 3, the high-fat diet control group significantly decreased grip strength compared to the normal group, but the dark tea extract group increased concentration-dependently compared to the control group.
최대 운동 수행능력은 15˚ 경사의 트레드밀에서 마우스가 탈진할 때까지 이동한 거리(running distance, m)로 측정하였다. 트레드밀 운동 훈련을 위해 첫째 날에는 5 m/min의 속도로 10분 동안, 10 m/min으로 10분 동안, 둘째 날에는 5 m/min으로 5분 동안, 10 m/min으로 15분 동안 훈련시켰다. 시험 당일인 셋째 날에는 10 m/min으로 20분 동안 운동 후, 2분마다 2 m/min으로 속도를 증가시켰다. 최대 운동 수행능력은 운동을 멈춘 후 그리드(grid)에서 10초 이상 머무를 때까지의 달린 거리로 환산하였다.도 4 및 표 4에서 확인할 수 있듯이, 트레드밀에 의한 운동수행능력을 분석한 결과 고지방식이 대조군은 정상식이군에 비하여 유의하게 감소되었으나 다크티 추출물군은 대조군에 비하여 유의하게 증가하여 다크티 추출물군이 근육기능 개선에 효과적임을 확인하였다.The maximum exercise performance was measured by a running distance (m) until the mouse exhausted in a treadmill with a slope of 15 degrees. For treadmill exercise training, trained for 10 minutes at a speed of 5 m/min on the first day, 10 minutes at 10 m/min, for 5 minutes at 5 m/min on the second day, and 15 minutes at 10 m/min. . On the third day of the test day, after exercising for 20 minutes at 10 m/min, the speed was increased to 2 m/min every 2 minutes. The maximum exercise performance was converted to the running distance from the grid until it stopped for more than 10 seconds after stopping the exercise. As can be seen from FIG. 4 and Table 4, as a result of analyzing the exercise performance by the treadmill, the notification method was The control group was significantly reduced compared to the normal diet group, but the dark tea extract group increased significantly compared to the control group, confirming that the dark tea extract group was effective in improving muscle function.
실시예 5: 다크티의 염증 개선 효과Example 5: Dark tea's inflammation improvement effect
실시예 1에서 준비된 고지방식이 대조군(HFD), 고지방식이 다크티 추출물 저농도군(100 mg/kg BW), 고지방식이 다크티 추출물 고농도군(200 mg/kg BW) 및 대조군(정상식이; Chow)에 해당하는 시험동물에 대한 혈액 중 염증성 사이토카인을 키트(ELISA kit)를 이용하여 분석하였다.High-fat diet prepared in Example 1 (HFD), high-fat diet dark tea extract low-concentration group (100 mg/kg BW), high-fat diet dark tea extract high-concentration group (200 mg/kg BW) and control (normal diet; The inflammatory cytokines in blood for test animals corresponding to Chow) were analyzed using a kit (ELISA kit).
도 5 및 표 5에서 확인할 수 있듯이, 다크티가 근육감소 원인의 하나로 알려져 있는 염증에 미치는 영향을 분석한 결과, 다크티 추출물은 고지방식이에 의하여 증가된 혈액 중 염증성 사이토카인인 TNFα, IL-1β, MCP1, IL-6의 수준을 유의하게 감소시켜, 염증 개선 효과가 있음을 확인하였다.As can be seen in Figure 5 and Table 5, as a result of analyzing the effect of dark tea on inflammation, which is known as one of the causes of muscle loss, the dark tea extract is TNFα, IL-, which are inflammatory cytokines in the blood increased by high-fat diet. It was confirmed that the levels of 1β, MCP1, and IL-6 were significantly reduced, thereby improving inflammation.
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