KR20240099758A - Composition for preventing or treating age-related sarcopenia containing adult green tea extract as an active ingredient - Google Patents
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Classifications
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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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
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- A—HUMAN NECESSITIES
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- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
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- 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
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- 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
Abstract
본 발명에 의하면 성체 녹차추출물을 유효성분으로 포함하는 근육 질환을 겪는 대상체로부터 근육 질환의 예방 또는 치료를 위한 약제학적 조성물이 제공될 수 있다.According to the present invention, a pharmaceutical composition for preventing or treating muscle disease in a subject suffering from muscle disease containing adult green tea extract as an active ingredient can be provided.
Description
본 발명은 성체 녹차추출물을 유효성분으로 포함하는 노화성 근감소증 예방 또는 치료를 위한 약제학적 조성물에 관한 것으로서, 보다 구체적으로는 MuRF1 억제 및 Akt 신호전달 경로의 활성화를 통하여 MyoD-매개 근원세포(myoblast)의 분화를 촉진하고 근육 재생 능력을 강화하기 위한 성체 녹차추출물을 유효성분을 포함하는 노화성 근감소증 예방 또는 치료를 위한 약제학적 조성물에 관한 것이다.The present invention relates to a pharmaceutical composition for preventing or treating age-related sarcopenia containing adult green tea extract as an active ingredient. More specifically, it relates to a pharmaceutical composition for preventing or treating age-related sarcopenia, and more specifically, to inhibit MyoD-mediated myoblasts by inhibiting MuRF1 and activating the Akt signaling pathway. ) It relates to a pharmaceutical composition for preventing or treating age-related sarcopenia containing adult green tea extract as an active ingredient to promote differentiation and enhance muscle regeneration ability.
근육은 신체를 움직일 수 있게 하는 가장 기본적인 장기이며, 근위축증, 노화에 의한 근감소증, 악액질 등의 이유로 손상되거나 감소될 수 있다. 근육 질환의 치료는 손상된 근육의 복구나 새로운 근육의 생성을 위해 근원세포(myoblast)의 분화를 촉진하는 근육발생(myogenesis) 과정을 유도하는 방법을 통해 이뤄질 수 있다. 이와 같은 근육 질환의 치료를 위해 다양한 연구들이 진행 중이며, 천연물을 통한 의약품 개발도 이에 포함된다. 특히 녹차는 카테킨을 풍부하게 함유하고 있는 식물로, 그 중 에피카테킨에 의한 근원세포 분화 촉진 효과에 대해 보고된 바 있다(SJ Lee et al., PLoS One 12, 2017).Muscles are the most basic organs that allow the body to move, and can be damaged or reduced due to reasons such as muscular dystrophy, sarcopenia due to aging, and cachexia. Treatment of muscle diseases can be achieved by inducing the myogenesis process, which promotes the differentiation of myoblasts to repair damaged muscles or create new muscles. A variety of research is underway to treat these muscle diseases, including the development of medicines using natural products. In particular, green tea is a plant rich in catechin, and the effect of epicatechin on promoting myogenic differentiation has been reported (SJ Lee et al., PLoS One 12, 2017).
이와 관련하여, 근육 손실은 근육량의 점진적 손실, 특히 골격근 또는 수의근 및 심장근육의 약화 및 퇴행을 특징으로 하며, 위축증 및 비대증이 일어나는 과정은 포유동물 종 전체에서 관측되는데, 다양한 연구에 의해 동일한 분자기전, 세포 및 생리학적 과정이 설치류와 인간 모두의 위축증에서 동일함이 입증되었다.In this regard, muscle loss is characterized by the progressive loss of muscle mass, especially the weakening and degeneration of skeletal or voluntary and cardiac muscles, and the processes leading to atrophy and hypertrophy are observed across mammalian species, and various studies have shown that the same molecular mechanisms are involved. , it has been demonstrated that the cellular and physiological processes are the same in atrophy in both rodents and humans.
근육 손실은 다양한 원인에 기인하며, 다양한 병증, 질병 및 질환과 관련된다. 이들로는 유전자 이상에 의해 야기된 근이영양증, 예를 들면, 뒤센(Duchenne)형 근이영양증(Duchenne Muscular Dystrophy (DMD)), 진행성 근이영양증, 베커(Becker)형 근이영양증, 디제린-란도즈(Dejerine-Landouzy) 근이영양증, 에르브(Erb)씨 근이영양증, 척수성 근위축증 및 영아 신경축삭퇴행위축증이 포함되나, 이들로 한정되지는 않는다.Muscle loss is due to a variety of causes and is associated with a variety of conditions, diseases, and disorders. These include muscular dystrophies caused by genetic abnormalities, such as Duchenne Muscular Dystrophy (DMD), progressive muscular dystrophy, Becker muscular dystrophy, and Dejerine-Landouzy. These include, but are not limited to, muscular dystrophy, Erb's muscular dystrophy, spinal muscular atrophy, and infantile neuroaxonal atrophy.
근육 손실은 또한 다양한 만성 질환 및 노화 과정에 의해 야기될 수 있다. 신체가 노화됨에 따라, 새롭게 생성되는 골격근의 일부가 섬유 조직으로 대체된다. 그러므로, 인간의 정상적 노화는 약화 및 퇴화의 원인이 되는, 근육감소증으로 불리는 상태인 골격 근육량 및 강도의 진행성 감소와 관련된다.Muscle loss can also be caused by various chronic diseases and the aging process. As the body ages, some of the newly created skeletal muscle is replaced by fibrous tissue. Therefore, normal aging in humans is associated with a progressive decline in skeletal muscle mass and strength, a condition called sarcopenia, which causes weakness and deterioration.
노화성 근감소증(Sarcopenia)은 노화 과정에서 발생되는 골격근의 손실에 따른 질환으로, 2045년 65세 이상인구가 30%를 넘을 것으로 예상되는 초고령화 사회를 진입으로 노화 및 노화에 따른 근감소증에 대한 사회적 관심이 증대되고 있는 실정이며, 최근 2016년 미국 FDA는 노화성 근감소증을 질병으로 분류한 바 있고(M62.84), 2018년 일본에서도 노화성 근감소증을 질병으로 분류하였으며, 2021년 1월 발표된 한국표준질병사인분류(KCD) 8차 개정안에도 노화성 근감소증에 대해 질병코드가 부여된 바 있으며, 관련 시장은 2017년 22억 달러 규모로 예측되었으며, 2024년에는 30억 달러로 증가할 것으로 예측된 바 있다.Sarcopenia is a disease caused by the loss of skeletal muscle that occurs during the aging process. As we enter a super-aging society in which the population aged 65 or older is expected to exceed 30% in 2045, the need for aging and age-related sarcopenia is increasing. Social interest is increasing, and recently in 2016, the U.S. FDA classified age-related sarcopenia as a disease (M62.84), and in 2018, Japan also classified age-related sarcopenia as a disease, and as of January 2021 In the announced 8th revision of the Korean Standard Classification of Diseases (KCD), a disease code was assigned to age-related sarcopenia, and the related market was predicted to be worth $2.2 billion in 2017 and will increase to $3 billion in 2024. It has been predicted that
근육 손실은 또한 암, 자가면역 질환, 감염 질환, HIV 감염, AIDS, 만성 염증, 관절염, 영양실조, 신장 질환, 만성 폐쇄성 폐질환(COPD), 폐기종, 구루병, 만성 하부 척추 통증, 말초신경 손상, 척수 손상, 화학적 손상, 중추신경계(CNS) 손상과 같은 다른 질환도 근육 손실과 연관되거나 근육 손실을 야기할 수 있다. 최종적으로, 근육 손실을 야기하는 상태는 질병으로 인한 장기 고정과 같은 폐용 상태, 또는 장기간 휠체어 사용, 장기 침상 요양, 골절 또는 외상과 같은 무력함으로부터 비롯될 수 있다. 수술 후 침상 요양은 주당 약 10%의 골격 근육량의 손실을 야기하는 것으로 평가된다. Muscle loss can also be linked to cancer, autoimmune diseases, infectious diseases, HIV infection, AIDS, chronic inflammation, arthritis, malnutrition, kidney disease, chronic obstructive pulmonary disease (COPD), emphysema, rickets, chronic lower spine pain, peripheral nerve damage, Other conditions, such as spinal cord injury, chemical injury, and central nervous system (CNS) injury, may also be associated with or cause muscle loss. Finally, conditions causing muscle loss may result from disabling conditions such as prolonged immobilization due to disease, or from incapacitation such as prolonged wheelchair use, prolonged bed rest, fractures, or trauma. Postoperative bed rest is estimated to cause a loss of approximately 10% of skeletal muscle mass per week.
근육 손실을 유발하는 암으로는 근육에 발생하는 연부조직 육종(Soft-tissue sarcoma)이 있으며, 구체적으로는 평활근육종(Leiomyosarcoma), 횡문근육종(Rhabdomyosarcoma) 등이 있다. 평활근육종(Leiomyosarcoma) 및 횡문근육종(Rhabdomyosarcoma)은 악성 종양으로서 덩어리짐, 통증, 두통, 메스꺼움, 출혈등의 증상을 수반하는 질환이다. 이러한 질환의 치료를 위해서 외과적 수술, 화학요법, 방사선 치료 등이 동원된다. 약물치료를 위해서 할라벤(Halaven, 에리불린 메실레이트)등의 약물이 개발되고 있으나, 피로감, 호중구 감소증, 구역, 탈모, 변비, 복통, 발열 등 다양한 부작용이 보고되고 있다.Cancers that cause muscle loss include soft-tissue sarcoma, which occurs in muscles, and specifically leiomyosarcoma and rhabdomyosarcoma. Leiomyosarcoma and Rhabdomyosarcoma are malignant tumors that are accompanied by symptoms such as lumpiness, pain, headache, nausea, and bleeding. To treat these diseases, surgery, chemotherapy, and radiation therapy are used. Drugs such as Halaven (eribulin mesylate) are being developed for drug treatment, but various side effects such as fatigue, neutropenia, nausea, hair loss, constipation, abdominal pain, and fever are reported.
치료되지 않은 근육 손실 질환은 심각한 건강상의 문제를 초래할 수 있으며, 근육 손실 시에 일어나는 변화는 약화된 신체 상태를 유발하여 무능한 신체 수행력 및 유해한 건강 상태를 야기할 수 있다. 만성 질환에 기인한 근육 손실은 운동성의 조기 손실을 유발하고 질환-관련 사망률의 위험성을 증가시킬 수 있다. 폐용으로 인한 근육 손실은 근육 기능 및 근육량에 있어 연령 관련 결함을 이미 가질 수 있는 노년층에서 특히 심각한 문제로, 영구적 무력함 및 조기 사망뿐 아니라 골절률의 증가를 야기한다. 상기 상태의 임상적 중요성에도 불구하고, 상기 상태를 예방하거나 역전하기 위한 치료법이 거의 존재하지 않아 이에 대한 해결방안이 요구된다.Untreated muscle loss disorders can lead to serious health problems, and the changes that occur during muscle loss can cause a weakened body condition, resulting in impaired physical performance and adverse health conditions. Muscle loss due to chronic disease can cause premature loss of mobility and increase the risk of disease-related mortality. Muscle loss due to lung use is a particularly serious problem in older adults who may already have age-related deficits in muscle function and mass, leading to increased fracture rates as well as permanent disability and premature death. Despite the clinical importance of this condition, few treatments exist to prevent or reverse the condition, so solutions are needed.
근육 활성은 신체의 다양한 장기 기능과 밀접하게 연관되어 있으며, 근감소증은 당뇨, 비알콜성 지방간, 고혈압이나 동맥경화와 같은 심혈관질환 유병 및 증세를 악화시키는 것으로 알려져 있다. 따라서 근육 재생을 위한 조성물, 근육 질환에 적용하기 위한 조성물을 제공하는 것, 노화성 근감소증을 예방 또는 치료하기 위한 약제학적 조성물 또는 노화성 근감소증을 예방 또는 개선하기 위한 식품 조성물을 제공하는 것은 향후 제약시장 및 건강기능식품 조성물 분야에서의 사업화에 큰 이점이 있다.Muscle activity is closely related to the function of various organs in the body, and sarcopenia is known to worsen the prevalence and symptoms of cardiovascular diseases such as diabetes, non-alcoholic fatty liver disease, high blood pressure, and arteriosclerosis. Therefore, providing a composition for muscle regeneration, a composition for application to muscle disease, a pharmaceutical composition for preventing or treating age-related sarcopenia, or a food composition for preventing or improving age-related sarcopenia will not be possible in the future. There are great advantages to commercialization in the pharmaceutical market and health functional food composition fields.
종래의 건강보조제는 단백질 및 아미노산과 같은 근육형성을 위한 성분을 포함하면서, 항산화 및 항염증 물질을 포함하는 것을 통해 근육 성분을 보충하는 역할을 수행하는 것에 그치나, 근육 자체의 기능 및 근육량 자체의 향상에는 직접적인 영향을 미치지 못한다는 문제점이 있었다. Conventional health supplements contain ingredients for muscle building, such as proteins and amino acids, and only serve to supplement muscle ingredients by containing antioxidant and anti-inflammatory substances, but improve the function of the muscle itself and muscle mass itself. There was a problem in that it had no direct influence.
나아가, 노화에 의한 근감소증으로 인해 노년층에서의 근기능 유지에 대한 수요가 확산되고 있지만, 노화성 근감소증에 대한 미국 FDA의 허가 또는 시장에 출시된 의약품은 세계적으로 전무한 실정이며, 기존의 단백질 보충제나 개발 중인 근감소 억제 기전의 치료제는 효과가 불분명하거나 부작용이 심각하다는 문제점이 있다. 이러한 배경 하에, 근육 줄기세포의 분화조절에 의한 근본적 치료제 개발의 필요성이 커지고 있다.Furthermore, although the demand for maintaining muscle function in the elderly is increasing due to sarcopenia caused by aging, there are no drugs approved by the US FDA or released on the market for aging sarcopenia worldwide, and existing protein supplements or Treatments that are being developed with mechanisms to suppress muscle loss have problems such as unclear effectiveness or serious side effects. Under this background, the need to develop fundamental treatments by controlling the differentiation of muscle stem cells is increasing.
성체 녹차추출물(Green Tea Extract, GTE)은 에피갈로카테킨 갈레이트(EGCG, Epigallocatechin Gallate), 에피카테킨 갈레이트(ECG, Epicatechin Gallate), 갈로카테킨 갈레이트(GCG, Gallocatechine Gallate), 에피갈로카테킨(EGC, Epigallocatechin), 에피카테킨(EC, Epicatechin), 갈로카테킨(GC, Gallocatechine) 등을 포함하는 녹차추출물일 수 있다. 음용으로 사용되는 녹차에는 어린잎이 주로 사용되는 것과 대조적으로, 다 자란 녹차잎은 음용으로 사용되지 않아 경제적이지 않은 것으로 취급되었으나, 근육량을 증대시키고, 근력을 강화하고, 근육 재생 능력을 강화할 수 있는 유효성분은 오히려 다 자란 녹차잎에 훨씬 풍부한 것으로 알려져 있다.Adult green tea extract (GTE) contains epigallocatechin gallate (EGCG), epicatechin gallate (ECG), gallocatechin gallate (GCG), and epigallocatechin ( It may be a green tea extract containing Epigallocatechin (EGC), Epicatechin (EC), Gallocatechin (GC), etc. In contrast to the fact that young leaves are mainly used in green tea for drinking, mature green tea leaves are not used for drinking and are considered uneconomical. However, they can increase muscle mass, strengthen muscle strength, and enhance muscle regeneration ability. It is known that the active ingredients are much more abundant in fully grown green tea leaves.
성체 녹차추출물에 포함된 유효성분 중, 에피카테킨은 녹차의 총 건조중량의 약 2~5%을 구성하는 성분으로, 심혈관질환 및 심근손상의 발병위험을 줄이는 것으로 알려져 있고(Buijsse, Weikert, Drogan, Bergmann, & Boeing, 2010; Ramirez-Sanchez, Maya, Ceballos, & Villarreal, 2010), 골형성증식, 분화 및 미네랄화를 현저하게 촉진하며(Zeng et al.,2014). 근육성장 및 강화에 이로운 효과((Gutierrez-Salmean et al., 2014)를 갖는 것으로 알려져 있다.Among the active ingredients contained in adult green tea extract, epicatechin constitutes approximately 2-5% of the total dry weight of green tea and is known to reduce the risk of cardiovascular disease and myocardial damage (Buijsse, Weikert, Drogan, Bergmann , & Boeing, 2010; Ramirez-Sanchez, Maya, Ceballos, & Villarreal, 2010) and significantly promote osteogenic proliferation, differentiation, and mineralization (Zeng et al., 2014). It is known to have beneficial effects on muscle growth and strengthening (Gutierrez-Salmean et al., 2014).
성체 녹차추출물은 전술한 바와 같은 에피카테킨의 유용한 효과에 더하여 골근육세포의 에스트로겐 발현을 억제하고, ERK 신호전달을 억제하며, unloading-mediated 근육 위축증을 예방할 수 있는 것으로 알려져 있다. 그러나, 전술한 바와 같은 성체 녹차추출물의 유용한 효과에 관한 연구에도 불구하고, 근육성장에 대한 성체 녹차추출물의 양성적 효과에 대한 구체적인 메커니즘은 아직 밝혀지지 않은 실정이다.In addition to the beneficial effects of epicatechin as described above, adult green tea extract is known to inhibit estrogen expression in osteomuscular cells, inhibit ERK signaling, and prevent unloading-mediated muscular dystrophy. However, despite the research on the beneficial effects of adult green tea extract as described above, the specific mechanism for the positive effect of adult green tea extract on muscle growth has not yet been revealed.
이에 본 발명자는 근육 질환을 겪는 대상체로부터 근육 질환의 예방 또는 치료를 위한 약제학적 조성물, 근육 질환을 겪는 대상체로부터 근육을 재생하기 위한 약제학적 조성물, 노화성 근감소증 예방 또는 치료를 위한 약제학적 조성물로서, 보다 구체적으로는 성체 녹차추출물이 MuRF1 억제 및 Akt의 신호전달 경로의 활성화를 통해 MyoD-매개 근원세포의 분화를 촉진하는 현상을 규명하여 본 발명을 완성하였다.Accordingly, the present inventors provide a pharmaceutical composition for preventing or treating muscle disease in a subject suffering from a muscle disease, a pharmaceutical composition for regenerating muscle in a subject suffering from a muscle disease, and a pharmaceutical composition for preventing or treating age-related sarcopenia. , More specifically, the present invention was completed by identifying the phenomenon in which adult green tea extract promotes MyoD-mediated differentiation of myogenic cells through MuRF1 inhibition and activation of the Akt signaling pathway.
또한, 본 발명자는 근육 질환을 겪는 대상체로부터 근육 질환의 예방 또는 치료를 위한 약제학적 조성물, 근육 질환을 겪는 대상체로부터 근육을 재생하기 위한 약제학적 조성물, 노화성 근감소증의 예방 또는 치료를 위한 약제학적 조성물로서, 상기 조성물이 근육량을 증대시키고, 근력을 강화하고, 근육 재생 능력을 강화할 수 있는 현상을 규명하여 본 발명을 완성하였다.In addition, the present inventors provide pharmaceutical compositions for preventing or treating muscle diseases in subjects suffering from muscle diseases, pharmaceutical compositions for regenerating muscles in subjects suffering from muscle diseases, and pharmaceutical compositions for preventing or treating age-related sarcopenia. As a composition, the present invention was completed by identifying the phenomenon in which the composition can increase muscle mass, strengthen muscle strength, and enhance muscle regeneration ability.
본 발명이 해결하고자 하는 과제들은 이상에서 언급된 과제로 제한되는 것은 아니며, 언급되지 않은 본 발명이 해결하고자 하는 과제들은 아래의 기재로부터 본 발명이 속하는 기술분야에서 보통의 지식을 가진 자("통상의 기술자")에게 명확하게 이해될 수 있을 것이다.The problems to be solved by the present invention are not limited to the problems mentioned above, and the problems to be solved by the present invention that are not mentioned above can be understood from the description below by anyone with ordinary knowledge in the technical field to which the present invention pertains. It will be clearly understandable to "technologists").
본 발명의 목적은 근육 질환의 예방 또는 치료를 위한 조성물을 제공하고자 하는 것으로, 보다 구체적으로는 근육 질환을 치료, 예방 또는 개선하기 위한, 성체 녹차추출물을 유효성분으로 포함하는 약제학적 조성물, 건강 식품용 조성물, 동물 사료용 조성물을 제공하고자 하는 것이다.The purpose of the present invention is to provide a composition for preventing or treating muscle diseases, and more specifically, a pharmaceutical composition and health food containing adult green tea extract as an active ingredient for treating, preventing or improving muscle diseases. The aim is to provide a composition for use and a composition for animal feed.
본 발명의 또 다른 목적은 성체 녹차추출물을 유효성분으로 포함하는 근육 질환을 치료, 예방 또는 개선하기 위한, 성체 녹차추출물을 유효성분을 포함하는 약제학적 조성물 및 건강 식품용 조성물을 제공하고자 하는 것이다.Another object of the present invention is to provide a pharmaceutical composition and a health food composition containing adult green tea extract as an active ingredient for treating, preventing or improving muscle diseases, including adult green tea extract as an active ingredient.
본 발명의 또 다른 목적은 성체 녹차추출물을 유효성분으로 포함하는 노화성 근감소증을 치료, 예방 또는 개선하기 위한, 성체 녹차추출물을 유효성분을 포함하는 약제학적 조성물 및 건강 식품용 조성물을 제공하고자 하는 것이다.Another object of the present invention is to provide a pharmaceutical composition and a health food composition containing adult green tea extract as an active ingredient for treating, preventing or improving age-related sarcopenia, which contains adult green tea extract as an active ingredient. will be.
본 발명의 또 다른 목적은 근육을 재생하기 위한 조성물을 제공하고자 하는 것이다.Another object of the present invention is to provide a composition for regenerating muscles.
본 발명의 또 다른 목적은 성체 녹차추출물의 근육 재생 용도에 관한 것이며, 근육 재생을 필요로 하는 대상체에게 치료학적 유효량의 성체 녹차추출물을 투여하는 단계를 포함하는, 근육 재생을 촉진하는 방법을 제공하고자 한다.Another object of the present invention relates to the use of adult green tea extract for muscle regeneration, and to provide a method for promoting muscle regeneration comprising administering a therapeutically effective amount of adult green tea extract to a subject in need of muscle regeneration. do.
전술한 기술적 과제를 달성하기 위하여, 본 발명은 성체 녹차추출물을 유효성분으로 포함하는 근육재생용 조성물을 제공한다.In order to achieve the above-described technical problem, the present invention provides a composition for muscle regeneration containing adult green tea extract as an active ingredient.
상기 조성물은 근육전사 인자인 MyoD의 활성을 조절하여 근원세포의 근관세포로의 분화유도 또는 섬유아세포를 근원세포로 분화유도할 수 있다.The composition can regulate the activity of MyoD, a muscle transcription factor, to induce differentiation of myogenic cells into myotube cells or fibroblasts into myogenic cells.
상기 조성물은 근육위축증을 예방 또는 치료하기 위한 약제학적 조성물일 수 있다.The composition may be a pharmaceutical composition for preventing or treating muscular dystrophy.
상기 조성물은 근육위축증을 개선 또는 완화하기 위한 건강 식품용 조성물일 수 있다.The composition may be a composition for health food to improve or alleviate muscular dystrophy.
상기 근육위축증은 노화성 근육감소증, 퇴행성 근육감소증, 당뇨병성 근육감소증, 지대형근위축증, 베커근위축증(Becker Muscular Dystrophy,BMD), 뒤세엔트근위축증(Duchenne Muscular Dystrophy, DMD), 근긴장성근위축증(Myotonic Muscular Dystrophy), 에머리-드레이푸스근위축증(Emery-Dreifuss Muscular Dystrophy, EDMD), 원위 근위축증(Distal Muscular Dystrophy, DD), 안면겹갑상완근우위축증(Facioscapulohumeral Muscular Dystrophy, FSHD) 또는 선천성 근위축증(Congenital Muscular Dystrophy, CMD)을 포함할 수 있다. The muscular dystrophy includes aging sarcopenia, degenerative sarcopenia, diabetic sarcopenia, girdle muscular atrophy, Becker Muscular Dystrophy (BMD), Duchenne Muscular Dystrophy (DMD), and Myotonic Muscular Dystrophy. ), Emery-Dreifuss Muscular Dystrophy (EDMD), Distal Muscular Dystrophy (DD), Facioscapulohumeral Muscular Dystrophy (FSHD), or Congenital Muscular Dystrophy (CMD) It can be included.
상기 성체 녹차추출물(Green Tea Extract, GTE)은 에피갈로카테킨 갈레이트(EGCG, Epigallocatechin Gallate), 에피카테킨 갈레이트(ECG, Epicatechin Gallate), 갈로카테킨 갈레이트(GCG, Gallocatechine Gallate), 에피갈로카테킨(EGC, Epigallocatechin), 에피카테킨(EC, Epicatechin), 갈로카테킨(GC, Gallocatechine) 등을 포함하는 녹차추출물일 수 있다.The adult green tea extract (GTE) contains epigallocatechin gallate (EGCG), epicatechin gallate (ECG), gallocatechin gallate (GCG), and epigallocatechin. It may be a green tea extract containing (EGC, Epigallocatechin), epicatechin (EC, Epicatechin), gallocatechin (GC, Gallocatechin), etc.
또한, 전술한 기술적 과제를 달성하기 위하여, 본 발명은 성체 녹차추출물을 유효성분으로 포함하는 노화성 근감소증의 치료 또는 예방을 위한 약제학적 조성물을 제공한다.In addition, in order to achieve the above-mentioned technical problem, the present invention provides a pharmaceutical composition for the treatment or prevention of age-related sarcopenia containing adult green tea extract as an active ingredient.
상기 노화성 근감소증의 예방 또는 치료를 위한 약제학적 조성물은 근육전사인자인 MyoD의 활성을 조절하여 근원세포의 근관세포로의 분화를 유도하는 것일 수 있다.The pharmaceutical composition for preventing or treating age-related sarcopenia may induce differentiation of myogenic cells into myotube cells by regulating the activity of MyoD, a muscle transcription factor.
상기 노화성 근감소증의 예방 또는 치료를 위한 약제학적 조성물은 근육전사인자인 MyoD의 활성을 조절하여 섬유아세포의 근원세포로의 분화를 유도하는 것일 수 있다.The pharmaceutical composition for preventing or treating age-related sarcopenia may induce differentiation of fibroblasts into myogenic cells by regulating the activity of MyoD, a muscle transcription factor.
상기 노화성 근감소증은 근육에서 발생하는 것일 수 있다.The age-related sarcopenia may occur in muscles.
또한, 전술한 기술적 과제를 달성하기 위하여, 본 발명은 성체 녹차추출물을 포함하는, 근육 질환을 겪는 대상체로부터 근육 질환의 예방 또는 치료를 위한 약제학적 조성물을 제공한다.In addition, in order to achieve the above-mentioned technical problem, the present invention provides a pharmaceutical composition for preventing or treating muscle disease in a subject suffering from muscle disease, comprising an adult green tea extract.
또한, 전술한 기술적 과제를 달성하기 위하여, 본 발명은 성체 녹차추출물을 포함하는, 근육 질환을 겪는 대상체로부터 근육을 재생하기 위한 약제학적 조성물을 제공한다.In addition, in order to achieve the above-described technical problem, the present invention provides a pharmaceutical composition for regenerating muscle from a subject suffering from muscle disease, comprising an adult green tea extract.
또한, 전술한 기술적 과제를 달성하기 위하여, 본 발명은 성체 녹차추출물을 포함하는, 근육 재생용 조성물을 제공한다.In addition, in order to achieve the above-mentioned technical problem, the present invention provides a composition for muscle regeneration comprising adult green tea extract.
또한, 전술한 기술적 과제를 달성하기 위하여, 본 발명은 성체 녹차추출물을 포함하는, 근육 질환의 예방 또는 개선을 위한 조성물을 포함하는, 식품을 제공한다.In addition, in order to achieve the above-mentioned technical problem, the present invention provides a food containing a composition for preventing or improving muscle disease, including adult green tea extract.
또한, 전술한 기술적 과제를 달성하기 위하여, 본 발명은 성체 녹차추출물을 포함하는, 근육 질환의 예방 또는 개선을 위한 조성물을 포함하는, 건강기능식품을 제공한다.In addition, in order to achieve the above-described technical problem, the present invention provides a health functional food containing a composition for preventing or improving muscle disease, including adult green tea extract.
또한, 전술한 기술적 과제를 달성하기 위하여, 본 발명은 성체 녹차추출물을 포함하는, 근육 질환의 예방 또는 개선을 위한 조성물을 포함하는, 동물 사료를 제공한다.In addition, in order to achieve the above-described technical problem, the present invention provides animal feed containing a composition for preventing or improving muscle disease, including adult green tea extract.
상기 근육 질환은 암, 노화, 근육 기능 저하, 근육 퇴화, 근육 손상을 포함하는 군으로부터 선택되는 것일 수 있고, 암 또는 노화로 인하여 유발되는 것일 수 있다.The muscle disease may be selected from the group including cancer, aging, decreased muscle function, muscle degeneration, and muscle damage, and may be caused by cancer or aging.
상기 근육 질환은 근위축증(muscular atrophy), 악액질(cachexia), 근육소실증 및 근육감소증(sarcopenia)을 포함하는 군으로부터 선택되는 것일 수 있고, 노화성 근감소증일 수 있다.The muscle disease may be selected from the group including muscular atrophy, cachexia, muscle loss, and sarcopenia, and may be age-related sarcopenia.
상기 약제학적 조성물은 근육전사인자인 MyoD의 활성을 조절하여 근원세포의 근관세포로의 분화를 유도하는 것을 특징으로 하는, 약제학적 조성물일 수 있다.The pharmaceutical composition may be a pharmaceutical composition characterized in that it induces differentiation of myogenic cells into myotube cells by regulating the activity of MyoD, a muscle transcription factor.
상기 약제학적 조성물은 PGC-1α활성 유도를 통한 근육 내 미토콘드리아 기능 활성을 증진시키는 것을 특징으로 하는, 약제학적 조성물일 수 있다.The pharmaceutical composition may be a pharmaceutical composition characterized by enhancing mitochondrial functional activity in muscle through inducing PGC-1α activity.
본 발명에 의하면 성체 녹차추출물을 유효성분으로 포함하는 노화성 근감소증의 예방 또는 치료를 위한 약제학적 조성물이 제공될 수 있고, 보다 구체적으로는 MuRF1 억제 및 Akt 신호전달 경로의 활성화를 통하여 근육전사인자인 MyoD의 활성을 조절하여 근원세포의 근관세포로의 분화를 유도하거나, 섬유아세포의 근원세포로의 재분화를 유도하는, 성체 녹차추출물을 유효성분으로 포함하는 노화성 근감소증의 예방 또는 치료를 위한 약제학적 조성물이 제공될 수 있다.According to the present invention, a pharmaceutical composition for the prevention or treatment of age-related sarcopenia containing adult green tea extract as an active ingredient can be provided, and more specifically, the muscle transcription factor is activated through MuRF1 inhibition and activation of the Akt signaling pathway. For the prevention or treatment of age-related sarcopenia containing adult green tea extract as an active ingredient, which regulates the activity of MyoD to induce differentiation of myogenic cells into myotube cells or redifferentiation of fibroblasts into myogenic cells. Pharmaceutical compositions may be provided.
또한, 본 발명에 의하면 성체 녹차추출물을 유효성분으로 포함하는 노화성 근감소증의 예방 또는 개선을 위한 식품 조성물이 제공될 수 있고, 보다 구체적으로는 MuRF1 억제 및 Akt 신호전달 경로의 활성화를 통하여 근육전사인자인 MyoD의 활성을 조절하여 근원세포의 근관세포로의 분화를 유도하거나, 섬유아세포의 근원세포로의 재분화를 유도하는, 성체 녹차추출물을 유효성분으로 포함하는 노화성 근감소증의 예방 또는 개선을 위한 식품 조성물이 제공될 수 있다.In addition, according to the present invention, a food composition for preventing or improving age-related sarcopenia containing adult green tea extract as an active ingredient can be provided, and more specifically, muscle transcription can be achieved through MuRF1 inhibition and activation of the Akt signaling pathway. Prevention or improvement of age-related sarcopenia containing adult green tea extract as an active ingredient, which regulates the activity of the factor MyoD to induce differentiation of myogenic cells into myotube cells or redifferentiation of fibroblasts into myogenic cells. A food composition may be provided for.
본 발명에 따른 우수한 및/또는 유용한 효과는 상기한 본 발명의 효과로 제한되는 것이 아니고, 통상의 기술자는 본 명세서의 개시를 바탕으로 본 명세서에 명시적으로 개시되지 않은 본 발명의 우수한 및/또는 유용한 효과를 또한 자명하게 인식할 수 있을 것이며, 이는 본 명세서가 의도적으로 개시하는 것으로서, 본 개시의 범위에 자명하게 포함됨이 이해되어야 한다.The excellent and/or useful effects according to the present invention are not limited to the above-described effects of the present invention, and those skilled in the art can understand the excellent and/or useful effects of the present invention not explicitly disclosed herein based on the disclosure of the present specification. Useful effects will also be readily apparent, and it should be understood that this is intentionally disclosed by this specification and is clearly included within the scope of the present disclosure.
도 1은 성체 GTE로 처리된 C2C12 근원세포에서 MHC, 미오게닌, HSP90 수준을 측정한 결과를 나타낸 도이다.
도 2 및 3은 각각 성체 GTE로 처리된 C2C12 근원세포에서 미오게닌 및 미오신 중쇄1, 2, 3, 4, 및 7 수준을 측정한 결과를 나타낸 도이다.
도 4는 3일 동안 성체 GTE 처리된 C2C12 근원세포를 MHC immunostaining에 의해 정량분석한 결과를 나타낸 도이다
도 5는 3일 동안 성체 GTE 처리된 C2C12 근원세포의 미오튜브의 지름은 측정한 결과를 나타낸 도이다.
도 6은 분화한 근원세포에 근위축(atrophy)을 유도한 후 성체 GTE 처리된 후 MuRF1 및 pAkt의 수준 측정한 결과를 나타낸 도이다.
도 7은 성체 GTE 처리된 LLC 또는 Dexa 유도된 근위축에서 확보한 C2C12 근원세포를 MHC immunostaining에 의해 정량분석한 결과를 나타낸 도이다.
도 8 내지 도 10는 성체 GTE 처리된 C2C12 근원세포에서 JC-1 다이머 및 JC-1 모노머를 사용하여 미토콘드리아의 형성의 결과 및 SIRT1 수치를 측정한 결과를 나타낸 도이다.
도 11 내지 도 16는 68세의 사람에게서 유래한 근원세포에 성체 GTE 처리한 결과를 나타낸 도이다.
도 17 및 도 18은 성체 GTE를 2주 간 투여한 마우스의 전경골근에 근육의 괴사를 cardiotoxin를 통해 유도한 후 근육 재생의 결과를 나타낸 도이다.
도 19는 근육 성능의 향상에 있어 성체 GTE의 효과를 평가한 결과를 나타낸 도이다.Figure 1 shows the results of measuring MHC, myogenin, and HSP90 levels in C2C12 myogenic cells treated with adult GTE.
Figures 2 and 3 show the results of measuring the levels of myogenin and myosin heavy chain 1, 2, 3, 4, and 7 in C2C12 myoblasts treated with adult GTE, respectively.
Figure 4 shows the results of quantitative analysis of C2C12 myogenic cells treated with adult GTE for 3 days by MHC immunostaining.
Figure 5 is a diagram showing the results of measuring the diameter of myotubes of C2C12 myoblasts treated with adult GTE for 3 days.
Figure 6 shows the results of measuring the levels of MuRF1 and pAkt after inducing muscle atrophy in differentiated myogenic cells and then treating them with adult GTE.
Figure 7 shows the results of quantitative analysis of C2C12 myoblasts obtained from adult GTE-treated LLC or Dexa-induced muscular atrophy by MHC immunostaining.
Figures 8 to 10 show the results of mitochondrial formation and SIRT1 levels measured using JC-1 dimer and JC-1 monomer in adult GTE-treated C2C12 myogenic cells.
Figures 11 to 16 show the results of adult GTE treatment on myogenic cells derived from a 68-year-old person.
Figures 17 and 18 show the results of muscle regeneration after muscle necrosis was induced using cardiotoxin in the tibialis anterior muscle of mice administered adult GTE for 2 weeks.
Figure 19 is a diagram showing the results of evaluating the effect of adult GTE in improving muscle performance.
이하, 본 발명에 대하여 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 명세서 및 특허청구범위에 사용된 용어 또는 단어는 통용되는 사전적 의미로 한정하여 해석되도록 의도된 것이 아니며, 본 발명이 속하는 기술분야에서 보통의 지식을 가진 자는 상기 용어 또는 단어를 본 명세서 및 특허청구범위가 자명하게 전달하고자 하는 관념을 표현하는 범위 내에서 본 발명이 의미하는 바를 전달하기 위한 범위의 의미로 사용된 것으로 명확하게 이해할 수 있을 것이다.The terms or words used in this specification and patent claims are not intended to be construed as limited to their commonly used dictionary meanings, and those of ordinary skill in the technical field to which the present invention pertains should not use the terms or words in this specification and patent claims. It can be clearly understood that the claims are used in the sense of a scope for conveying the meaning of the present invention within the scope of expressing the idea that is clearly intended to be conveyed.
또한, 본 명세서에 기재된 실시예 및 상기 실시예에 기재된 구성들은 통상의 기술자에게 본 발명이 이해되고 또 이를 재현할 수 있도록 하기 위해 단지 예시로 제시된 바람직한 실시예에 불과할 뿐이고, 본 발명을 이에 한정시키고자 하는 의도가 아님을 통상의 기술자는 명확하게 이해할 수 있을 것이다.In addition, the embodiments described in this specification and the configurations described in the above embodiments are merely preferred embodiments presented as examples to enable those skilled in the art to understand and reproduce the present invention, and the present invention is limited thereto. A person skilled in the art will be able to clearly understand that this is not the intention.
또한, 본 명세서에서 기술되는 각각의 구성에 관한 설명 및 구체적인 실시 양태는 각각의 다른 설명 및 실시 양태에도 자명하게 적용될 수 있다. 즉, 본 명세서에서 개시되는 다양한 구성과 구체적인 실시 양태의 모든 조합은 본 발명이 개시하는 범위에 속하는 것으로 통상의 기술자는 명확하게 이해할 수 있을 것이다.In addition, the description and specific implementation aspects of each configuration described in this specification can be obviously applied to each other description and implementation aspect. That is, those skilled in the art will clearly understand that all combinations of various configurations and specific embodiments disclosed in this specification fall within the scope disclosed by the present invention.
본 명세서에서 사용되는 용어 '및/또는'은, 언급된 항목들의 각각 및 하나 이상의 모든 조합을 포함하는 용어이다. 또한, 단수형 용어는 별도로 언급되지 않는 한 복수형도 포함한다.The term 'and/or' used in this specification is a term that includes each and all combinations of one or more of the mentioned items. Additionally, singular terms also include plural terms unless otherwise noted.
본 명세서에서 사용하는 '포함한다(comprising)' 및/또는 '포함하는(comprising)'의 용어는 언급된 항목들 외의 다른 항목의 존재 또는 추가를 배제하지 않는 용어이다.The terms 'comprising' and/or 'comprising' used in this specification are terms that do not exclude the presence or addition of items other than the mentioned items.
본 명세서에서 용어 '내지'를 사용하여 나타낸 수치의 범위는, 상기 용어의 앞과 뒤에 기재된 값을 각각 하한 값과 상한 값으로써 포함하는 수치의 범위를 나타낸다. 임의의 수치 범위의 상한과 하한으로서의 수치 값이 각각 복수 개로 개시된 경우, 본 명세서에서 개시하는 수치의 범위는 복수의 하한 값 중 임의의 하나의 값 및 복수의 상한 값 중 임의의 하나의 값을 각각 하한 값 및 상한 값으로 하는 임의의 수치의 범위로 이해될 수 있다.The range of values expressed using the term 'to' in this specification refers to the range of values that include the values described before and after the term as the lower limit and upper limit, respectively. When a plurality of numerical values as the upper and lower limits of an arbitrary numerical range are disclosed, the numerical range disclosed in this specification includes any one value among the plurality of lower limit values and any one value among the plurality of upper limit values, respectively. It can be understood as a range of arbitrary values with a lower limit and an upper limit.
본 명세서에서 사용되는 용어 '약' 또는 '대략'은, 사용되는 경우, 상기 용어의 뒤에 기재된 값 또는 수치 범위의 10% 이내의 값 또는 수치 범위를 의미한다.As used herein, the term 'about' or 'approximately', when used, means a value or numerical range within 10% of the value or numerical range stated after the term.
본 명세서 및 특허청구범위에 사용된 용어 또는 단어는 통상적인 사전적인 의미로 한정해서 해석되어서는 안 되며, 발명자는 그 자신의 발명을 가장 최선의 방법으로 설명하기 위해 용어의 개념을 적절하게 정의할 수 있다는 원칙에 입각하여 본 발명의 기술적 사상에 부합하는 의미와 개념으로 해석되어야 한다. 따라서, 본 명세서에 기재된 실시예에 기재된 구성은 본 발명의 가장 바람직한 일 실시예에 불과할 뿐이고, 본 발명의 기술적 사상을 모두 대변하는 것은 아니므로, 본 출원시점에 있어서 이들을 대체할 수 있는 다양한 균등물과 변형예들이 있을 수 있음을 이해하여야 한다. Terms or words used in this specification and claims should not be construed as limited to their ordinary dictionary meanings, and the inventor may appropriately define the concept of terms in order to explain his or her invention in the best way. It should be interpreted with meaning and concept consistent with the technical idea of the present invention based on the principle that it can be done. Therefore, the configuration described in the embodiments described in this specification is only one of the most preferred embodiments of the present invention, and does not represent the entire technical idea of the present invention, so various equivalents can be substituted for them at the time of filing the present application. It should be understood that there may be variations.
한편, 본 명세서에서 개시된 각각의 설명 및 실시형태는 각각의 다른 설명 및 실시 양태에도 적용될 수 있다. 즉, 본 명세서에서 개시된 다양한 요소들의 모든 조합이 본 발명의 범주에 속하고, 일 구현예에서 생략된 기재는 다른 구현예에서 기재된 바와 같은 방식으로 해석될 수 있다. 또한, 하기 기술된 구체적인 서술에 의하여 본 발명의 범주가 제한된다고 볼 수 없다.Meanwhile, each description and embodiment disclosed in this specification may also be applied to each other description and embodiment. That is, all combinations of various elements disclosed in this specification fall within the scope of the present invention, and descriptions omitted in one embodiment may be interpreted in the same way as described in other embodiments. Additionally, the scope of the present invention cannot be considered to be limited by the specific description described below.
본 발명의 일 구현예에 따르면, 성체 녹차추출물을 유효성분을 포함하는 근육재생용 조성물이 제공될 수 있고, 보다 구체적으로는 MuRF1 억제 및 Akt 신호전달 경로의 활성화를 통하여 근육전사인자인 MyoD의 활성을 조절하여 근원세포의 근관세포로의 분화를 유도하거나, 섬유아세포의 근원세포로의 재분화를 유도하는, 성체 녹차추출물을 유효성분으로 포함하는 노화성 근감소증의 예방 또는 치료를 위한 약제학적 조성물이 제공될 수 있다.According to one embodiment of the present invention, a composition for muscle regeneration containing adult green tea extract as an active ingredient can be provided, and more specifically, activation of MyoD, a muscle transcription factor, through inhibition of MuRF1 and activation of the Akt signaling pathway. A pharmaceutical composition for the prevention or treatment of age-related sarcopenia containing adult green tea extract as an active ingredient, which regulates the differentiation of myogenic cells into myotube cells or induces the re-differentiation of fibroblasts into myogenic cells. can be provided.
상기 근육재생은 손상된 근육세포의 재생일 수 있고, 상기 손상은 수술, 암, 염증, 감염 등 유전적 인자와 환경적 인자에 의한 것일 수 있다. 근육 재생은 근육세포 분화를 통해 이루어질 수 있고, 상기 근육세포 분화는 근육세포로 분화 가능한 근원세포 또는 섬유아세포 등의 근원세포 및 근육세포로의 분화일 수 있다.The muscle regeneration may be the regeneration of damaged muscle cells, and the damage may be caused by genetic and environmental factors such as surgery, cancer, inflammation, and infection. Muscle regeneration can be achieved through muscle cell differentiation, and the muscle cell differentiation can be differentiation into myogenic cells such as myogenic cells or fibroblasts capable of differentiating into muscle cells, and muscle cells.
상기 조성물은 근육전사 인자인 MyoD의 활성을 조절하여 근원세포의 근관세포로의 분화유도 또는 섬유아세포를 근원세포로 분화유도할 수 있다.The composition can regulate the activity of MyoD, a muscle transcription factor, to induce differentiation of myogenic cells into myotube cells or fibroblasts into myogenic cells.
본 발명의 일 구현예에 따르면, 성체 녹차추출물을 유효성분으로 포함하는 노화성 근감소증의 예방 또는 치료를 위한 약제학적 조성물이 제공될 수 있고, 보다 구체적으로는 MuRF1 억제 및 Akt 신호전달 경로의 활성화를 통하여 근육전사인자인 MyoD의 활성을 조절하여 근원세포의 근관세포로의 분화를 유도하거나, 섬유아세포의 근원세포로의 재분화를 유도하는, 성체 녹차추출물을 유효성분으로 포함하는 노화성 근감소증의 예방 또는 치료를 위한 약제학적 조성물이 제공될 수 있다.According to one embodiment of the present invention, a pharmaceutical composition for the prevention or treatment of age-related sarcopenia containing adult green tea extract as an active ingredient can be provided, and more specifically, inhibition of MuRF1 and activation of the Akt signaling pathway. It regulates the activity of MyoD, a muscle transcription factor, to induce differentiation of myogenic cells into myotube cells or redifferentiation of fibroblasts into myogenic cells, a treatment for age-related sarcopenia containing adult green tea extract as an active ingredient. Pharmaceutical compositions for prevention or treatment may be provided.
상기 성체 녹차추출물(Green Tea Extract, GTE)은 에피갈로카테킨 갈레이트(EGCG, Epigallocatechin Gallate), 에피카테킨 갈레이트(ECG, Epicatechin Gallate), 갈로카테킨 갈레이트(GCG, Gallocatechine Gallate), 에피갈로카테킨(EGC, Epigallocatechin), 에피카테킨(EC, Epicatechin), 갈로카테킨(GC, Gallocatechine) 등을 포함하는 녹차추출물일 수 있다.The adult green tea extract (GTE) contains epigallocatechin gallate (EGCG), epicatechin gallate (ECG), gallocatechin gallate (GCG), and epigallocatechin. It may be a green tea extract containing (EGC, Epigallocatechin), epicatechin (EC, Epicatechin), gallocatechin (GC, Gallocatechin), etc.
노화성 근감소증의 예방 또는 치료에 사용하기 위해 본 발명에 따른 성체 녹차추출물 자체를 투여하는 것이 가능하나, 약제학적 조성물의 활성 성분으로서 성체 녹차추출물이 포함되는 것이 바람직하다.Although it is possible to administer the adult green tea extract according to the present invention itself for use in the prevention or treatment of age-related sarcopenia, it is preferable that the adult green tea extract is included as an active ingredient in the pharmaceutical composition.
본 발명에 따른 약제학적 조성물은 성체 녹차추출물을 유효성분으로 함유할 수 있고, 약제학적으로 허용 가능한 담체를 포함할 수 있다. 상기 약제학적으로 허용 가능한 담체는 제제시에 통상적으로 이용되는 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 사이클로덱스트린, 덱스트로즈 용액, 말토덱스트린 용액, 글리세롤, 에탄올, 리포좀 등을 포함하지만 이에 한정되지 않으며, 필요에 따라 항산화제, 완충액 등 다른 통상의 첨가제를 추가로 포함할 수 있다.The pharmaceutical composition according to the present invention may contain adult green tea extract as an active ingredient and may include a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is commonly used in preparation and includes, but is limited to, saline solution, sterile water, Ringer's solution, buffered saline solution, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposome, etc. If necessary, other common additives such as antioxidants and buffers may be additionally included.
본 발명에 따른 약제학적 조성물은 성체 녹차추출물을 유효성분으로 함유할 수 있고, 희석제, 분산제, 계면활성제, 결합제, 윤활제 등을 추가로 포함할 수 있고, 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립 또는 정제로 제제화될 수 있다.The pharmaceutical composition according to the present invention may contain adult green tea extract as an active ingredient, and may further include diluents, dispersants, surfactants, binders, lubricants, etc., and may be formulated as injectable formulations such as aqueous solutions, suspensions, emulsions, etc. , may be formulated as pills, capsules, granules or tablets.
본 발명에 따른 약제학적 조성물에 적합한 약제학적으로 허용되는 담체 및 제제화는 레밍턴의 문헌 (Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA)에 개시되어 있는 방법을 이용하여 약제학적 조성물에 포함되는 성분의 특성에 따라 바람직하게 제제화될 수 있다. Pharmaceutically acceptable carriers and formulations suitable for the pharmaceutical composition according to the present invention are obtained by determining the characteristics of the ingredients included in the pharmaceutical composition using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA. It can be preferably formulated according to.
본 발명에 따른 약제학적 조성물의 제형은 특별히 제한되지는 않으나, 예를 들면 주사제, 흡입제, 피부 외용제 등으로 제제화될 수 있다.The formulation of the pharmaceutical composition according to the present invention is not particularly limited, but may be formulated as, for example, injections, inhalants, topical skin preparations, etc.
본 발명에 따른 약제학적 조성물의 투여방법은 특별히 제한되지는 않으나, 본 발명에 따른 약제학적 조성물은 정맥 내, 피하, 복강 내, 흡입, 피부도포 또는 국소적용과 같이 비경구 투여되거나 경구 투여될 수 있다.The method of administration of the pharmaceutical composition according to the present invention is not particularly limited, but the pharmaceutical composition according to the present invention may be administered parenterally or orally, such as intravenously, subcutaneously, intraperitoneally, inhaled, applied to the skin, or topically applied. there is.
본 발명에 따른 약제학적 조성물의 투여량은 특별히 제한되지는 않으나, 환자의 체중, 연령, 성별, 건강상태, 식이, 투여시간, 투여방법, 배설율 및 질환의 중증도 등에 따라 그 범위가 다양할 수 있다.The dosage of the pharmaceutical composition according to the present invention is not particularly limited, but the range may vary depending on the patient's weight, age, gender, health condition, diet, administration time, administration method, excretion rate, and severity of disease. there is.
일일 투여량은 치료를 필요로 하는 개체에 투여됨으로써 경감된 질병 상태에 대한 치료에 충분한 본 발명의 치료용 물질의 양을 의미한다. 치료용 물질의 효과적인 양은 특정 화합물, 질병 상태 및 그의 심각도, 치료를 필요로 하는 개체에 따라 달라지며, 이는 통상의 기술자에 의해 적절히 결정될 수 있다. Daily dosage refers to the amount of therapeutic agent of the invention sufficient to treat the disease condition alleviated by administration to an individual in need of treatment. The effective amount of a therapeutic agent depends on the particular compound, the disease state and its severity, and the individual in need of treatment, and can be determined appropriately by a person skilled in the art.
비제한적인 예로서, 본 발명에 따른 조성물의 인체에 대한 투여량은 환자의 나이, 몸무게, 성별, 투여 형태, 건강 상태 및 질환 정도에 따라 달라질 수 있으며, 몸무게가 70 ㎏인 성인 환자를 기준으로 할 때, 일반적으로는 0.01∼㎎/일, 바람직하게는 1∼㎎/일이며, 일정시간 간격으로 1일 1회 내지 수회에 걸쳐 분할 투여될 수 있다.As a non-limiting example, the dosage for the human body of the composition according to the present invention may vary depending on the patient's age, weight, gender, administration form, health condition, and disease degree, and is based on an adult patient weighing 70 kg. When administered, it is generally 0.01 to ㎎/day, preferably 1 to 1 mg/day, and may be administered in divided doses from once to several times a day at regular time intervals.
본 발명의 일 구현예에 따르면, 성체 녹차추출물을 유효성분으로 포함하는 노화성 근감소증의 예방 또는 개선을 위한 식품 조성물이 제공될 수 있고, 보다 구체적으로는 MuRF1 억제 및 Akt 신호전달 경로의 활성화를 통하여 근육전사인자인 MyoD의 활성을 조절하여 근원세포의 근관세포로의 분화를 유도하거나, 섬유아세포의 근원세포로의 재분화를 유도하는, 성체 녹차추출물을 유효성분으로 포함하는 노화성 근감소증의 예방 또는 개선을 위한 식품 조성물이 제공될 수 있다.According to one embodiment of the present invention, a food composition for preventing or improving age-related sarcopenia containing adult green tea extract as an active ingredient can be provided, and more specifically, inhibiting MuRF1 and activating the Akt signaling pathway. Prevention of age-related sarcopenia using adult green tea extract as an active ingredient, which regulates the activity of MyoD, a muscle transcription factor, to induce differentiation of myogenic cells into myotube cells or redifferentiation of fibroblasts into myogenic cells. Alternatively, a food composition for improvement may be provided.
본 발명의 조성물이 식품 조성물로 제조되는 경우, 유효성분으로서 성체 녹차추출물을 함유할 뿐만 아니라, 식품 조성물의 제조시에 통상적으로 첨가되는 성분을 포함할 수 있고, 예를 들어, 단백질, 탄수화물, 지방, 영양소, 조미제 및 향미제를 포함할 수 있다. When the composition of the present invention is manufactured as a food composition, it may not only contain adult green tea extract as an active ingredient, but may also contain ingredients commonly added during the production of food compositions, such as proteins, carbohydrates, and fats. , may include nutrients, seasonings, and flavoring agents.
비제한적인 예로서, 본 발명의 식품 조성물이 드링크제로 제조되는 경우에는 본 발명에 따른 성체 녹차추출물 이외에 구연산, 액상과당, 설탕, 포도당, 초산, 사과산, 과즙 등이 추가로 포함될 수 있다.As a non-limiting example, when the food composition of the present invention is manufactured as a drink, citric acid, high fructose corn syrup, sugar, glucose, acetic acid, malic acid, fruit juice, etc. may be additionally included in addition to the adult green tea extract according to the present invention.
이하, 본 발명을 다음 실시예 및 실험예를 사용하여 더욱 상세히 설명한다. 하기 실시예 및 실험예에 명시되지 않은 공정 조건 및 준비 단계는, 본 발명이 속하는 기술분야에서 자명한 공정 조건 또는 준비 단계일 수 있고, 통상의 기술자는 본 개시를 바탕으로 이를 별다른 어려움 없이 선정하여 본 발명의 과제해결원리를 재현할 수 있을 것이다.Hereinafter, the present invention will be described in more detail using the following examples and experimental examples. Process conditions and preparation steps not specified in the following examples and experimental examples may be process conditions or preparation steps that are obvious in the technical field to which the present invention pertains, and those skilled in the art can select them without much difficulty based on the present disclosure. It will be possible to reproduce the problem-solving principle of the present invention.
실시예 1: 준비Example 1: Preparation
연중 7월 내지 9월 수확한 녹차로부터 추출한 성체 녹차추출물(Green Tea Extract, GTE)을 전체 성체 녹차추출물 조성물 100질량%를 기준으로 57.12질량%의 에피갈로카테킨 갈레이트, 15.74질량%의 에피카테킨 갈레이트, 4.25질량%의 갈로카테킨 갈레이트, 7.70질량%의 에피갈로카테킨, 4.80질량%의 에피카테킨, 및 0.06질량%의 갈로카테킨을 포함하는 성체 녹차추출물로서 준비하였다.Green tea extract (GTE) extracted from green tea harvested from July to September of the year contains 57.12% by mass of epigallocatechin gallate and 15.74% by mass of epicatechin gall based on 100% by mass of the total adult green tea extract composition. It was prepared as an adult green tea extract containing 4.25% by mass of gallocatechin gallate, 7.70% by mass of epigallocatechin, 4.80% by mass of epicatechin, and 0.06% by mass of gallocatechin.
실시예 2: 세포 배양Example 2: Cell Culture
근원세포 C2C12, 배아 섬유아세포 10T1/2 세포주 및 배아 신장 293T 세포를 배양하였다(Bae et al., 2009; Bae et al., 2010). C2C12 근원세포 (myoblast)의 분화를 유도하기 위해, 융합점 근처의 세포주를 37, CO2 인큐베이터에서 15% FBS (growth medium, GM)를 포함하는 DMEM에서 2% HS (differentiation medium, DM)를 함유하는 DMEM으로 이주하였고, 약 48 내지 72시간 동안 근관세포(myotube) 형성을 관찰하였다. 근관세포(myotube) 형성의 효율성은 임시 분화 분석(transient differentiation assay)를 이용하여 정량화되었다(Bae et al.,2010). MuRF1 및 Akt 억제제가 사용된 본 발명의 실험결과에서, 새로운 배양배지에서 30분간 C2C12 세포는 1μM의 LY294002으로 선배양된 후 1ng/ml 성체 GTE로 처리되었다. Myoblast C2C12, embryonic fibroblast 10T1/2 cell line, and embryonic kidney 293T cells were cultured (Bae et al., 2009; Bae et al., 2010). To induce differentiation of C2C12 myoblasts, cell lines near the point of confluence were cultured in DMEM containing 15% FBS (growth medium, GM) containing 2% HS (differentiation medium, DM) in a CO2 incubator. They were transferred to DMEM, and myotube formation was observed for about 48 to 72 hours. The efficiency of myotube formation was quantified using a transient differentiation assay (Bae et al., 2010). In the experimental results of the present invention using MuRF1 and Akt inhibitors, C2C12 cells were pre-incubated with 1 μM LY294002 for 30 minutes in fresh culture medium and then treated with 1 ng/ml adult GTE.
Cdo+/+ 마우스에서 분리된 마우스 배아 섬유아세포(Mouse embryonic fibroblasts, MEFs)가 배양되었고(Cole & Krauss, 2003), MEFs는 20% FBS 및 염기 섬유아세포 성장인자(bFGF; 100ng/mL)를 포함하는 F10 배지에서 배양되었다. 트랜스(전환)-분화 연구를 위해, 100mM 배양 접시에 10T1/2 세포주와 MEFs 세포들을 10% FBS가 함유된 DMEM에서 배양하여, 5㎍ MyoD 또는 대조군 벡터(pBp)로 형질전환하였다. 24시간 내지 48시간 동안 배양한 후, 10T1/2 세포주 및 MEFs 세포는 2일 동안 2% HS에서 1ng/ml 성체 GTE로 처리되었다.Mouse embryonic fibroblasts (MEFs) isolated from Cdo +/+ mice were cultured (Cole & Krauss, 2003), and MEFs were supplemented with 20% FBS and basic fibroblast growth factor (bFGF; 100 ng/mL). were cultured in F10 medium. For trans-differentiation studies, 10T1/2 cell lines and MEFs cells were cultured in DMEM containing 10% FBS in 100mM culture dishes and transfected with 5 μg MyoD or control vector (pBp). After culturing for 24 to 48 hours, 10T1/2 cell lines and MEFs cells were treated with 1 ng/ml adult GTE in 2% HS for 2 days.
실시예 3: 웨스턴블럿 및 면역침전(Western blotting analysis and immunoprecipitation analysis)Example 3: Western blotting analysis and immunoprecipitation analysis
세포주를 온전한 proteinase inhibitor cocktail (Roche)를 포함하는 추출 버퍼(10mM Tris-Hcl, [pH7.2], 150mM NaCl, 1mM EDTA, 1% Triton X-100)에서 분쇄(lysis)시키고 SDS PAGE를 수행하였다. 일차 항체로는 항-Cdo, 항-MuRF1, 항-Akt, 항-p-Akt (Akt의 인산화된 활성형), 항-MHC, 항-미오게닌 및 항-MyoD를 사용하였다. 면역침전분석을 위해서, C2C12 또는 293T 세포가 36시간 동안 MyoD로 형질전환되었다. 24시간 동안 성체 GTE로 처리한 후, 세포 추출액 전체는 항-E2A 및 단백질 G 아가로스 비드(Roche Diagnostics)로 4℃오버나이트 배양하였다. 비드는 추출 버퍼로 3회 세척하였고, 추출 버퍼안에서 재현탁하였다. 그리고 샘플을 웨스턴 블롯으로 분석하였다.Cell lines were lysed in extraction buffer (10mM Tris-Hcl, [pH7.2], 150mM NaCl, 1mM EDTA, 1% Triton X-100) containing intact proteinase inhibitor cocktail (Roche), and SDS PAGE was performed. . Primary antibodies used were anti-Cdo, anti-MuRF1, anti-Akt, anti-p-Akt (phosphorylated active form of Akt), anti-MHC, anti-myogenin, and anti-MyoD. For immunoprecipitation analysis, C2C12 or 293T cells were transfected with MyoD for 36 h. After treatment with adult GTE for 24 hours, the entire cell extract was incubated overnight at 4°C with anti-E2A and protein G agarose beads (Roche Diagnostics). The beads were washed three times with extraction buffer and resuspended in extraction buffer. The samples were then analyzed by Western blot.
실시예 4: 면역 세포 화학 및 현미경 검사(Immunocytochemistry and microscopy)Example 4: Immunocytochemistry and microscopy
C2C12 세포주를 4 % 파라 포름알데하이드(paraformaldehyde)로 20분간 고정시키고, 인산완충식염수(phosphate buffered saline)의 0.2 % Triton X-100로 투과시킨 후 밀봉하여 항-MHC로 염색하였다. 이어서 Alexa Fluor 568-접합 2차 항체(Molecular Probes)로 처리하였다. 이미지는 Nikon ECLIPSE TE-2000U microscope, NISElements F software (Nikon), Photoshop CS5 (Adobe)로 촬영 및 처리하였다.The C2C12 cell line was fixed with 4% paraformaldehyde for 20 minutes, permeabilized with 0.2% Triton X-100 in phosphate buffered saline, sealed, and stained with anti-MHC. It was then treated with Alexa Fluor 568-conjugated secondary antibody (Molecular Probes). Images were captured and processed using a Nikon ECLIPSE TE-2000U microscope, NISElements F software (Nikon), and Photoshop CS5 (Adobe).
실험예 1: 근원세포 분화를 촉진하는 성체 GTEExperimental Example 1: Adult GTE promoting myoblast differentiation
녹차추출물을 근원세포에 처리한 채로 근원세포의 분화를 유도하여, 녹차추출물을 처리하지 않았을 때에 비해 분화가 더 활발히 이뤄짐을 확인하였다. 근원세포 분화에 있어서 성체 GTE의 효과를 실험하기 위해, vehicle DMSO 또는 다양한 성체 GTE의 농도 조건에서, C2C12 근원세포를 2일 동안 분화유도하였고, 근육-특이적 단백질을 사용하는 면역블롯(immunoblotting)을 이용하여 근원세포의 분화에 미치는 영향을 평가하였다. 성체 GTE으로 처리된 C2C12 근원세포는 MHC 및 미오게닌의 발현이 0.1 내지 10ng/ml 성체 GTE에서 가장 우수하였고, HSP90 수준은 변동이 없었다(도 1). 미오게닌은 특히 성체 GTE 처리된 경우 mRNA 발현이 우수하였음이 확인되었고(도 2), 미오신 중쇄1, 2, 3, 4, 및 7은 특히 성체 GTE 처리된 경우 mRNA 발현이 우수하였음이 확인되었다(도 3). 3일 동안 성체 GTE 처리된 C2C12 근원세포는 더 많은 핵을 갖고, 보다 길어진 MHC-양성 근관세포를 형성하였다. 이는 MHC immunostaining에 의해 정량분석하고, 이를 도식화하였다(도 4). 미오튜브의 지름은 성체 GTE 처리된 경우 상대적으로 증가되었음이 확인되었다(도 5). 성체 GTE 처리는 복수의 핵을 가진 근관세포의 비중을 증가시키고, 단핵세포의 비중을 급격히 감소시켰다. 이러한 결과들은 생화학적 측면에서나 형태학적 측면에서 성체 GTE가 C2C12 근원세포의 분화를 촉진시키고 있음을 의미한다.Differentiation of myogenic cells was induced by treating them with green tea extract, and it was confirmed that differentiation occurred more actively compared to when green tea extract was not treated. To test the effect of adult GTE on myoblast differentiation, C2C12 myoblasts were induced to differentiate for 2 days in vehicle DMSO or various concentrations of adult GTE, and immunoblotting using muscle-specific proteins was performed. The effect on the differentiation of myogenic cells was evaluated. C2C12 myoblasts treated with adult GTE had the best expression of MHC and myogenin at 0.1 to 10 ng/ml adult GTE, and HSP90 levels were unchanged (Figure 1). It was confirmed that myogenin had excellent mRNA expression, especially when treated with adult GTE (Figure 2), and myosin heavy chains 1, 2, 3, 4, and 7 showed excellent mRNA expression, especially when treated with adult GTE. (Figure 3). C2C12 myoblasts treated with adult GTE for 3 days formed longer MHC-positive myotubes with more nuclei. This was quantitatively analyzed by MHC immunostaining and schematized (Figure 4). It was confirmed that the diameter of myotubes was relatively increased when adults were treated with GTE (Figure 5). Adult GTE treatment increased the proportion of myotube cells with multiple nuclei and sharply decreased the proportion of mononuclear cells. These results indicate that adult GTE promotes differentiation of C2C12 myogenic cells from a biochemical and morphological perspective.
실험예 2: 근위축을 완화하는 성체 GTEExperimental Example 2: Adult GTE alleviating muscle atrophy
분화한 근원세포에 근위축(atrophy)을 유도한 후 녹차추출물을 처리할 시에 근위축이 완화됨을 확인하였다. LLC 또는 Dexa 유도된 근위축에서 확보한 C2C12 근원세포를 1ng/ml의 성체 GTE 처리하고 면역블롯(immunoblotting)에서 MHC, HSP90, MuRF1, 및 Akt(pAkt)에 대한 항체를 사용하여 MHC, HSP90, MuRF1 및 Akt의 활성화 상태를 분석하였다. 성체 GTE 처리는 MuRF1의 수준을 억제하고 pAkt의 수준을 상당히 향상시킨 반면, 전체 단백질의 발현수준은 상대적으로 일정하였다(도 6). 성체 GTE 처리된 LLC 또는 Dexa 유도된 근위축에서 확보한 C2C12 근원세포를 MHC immunostaining에 의해 정량분석하고, 이를 도식화하였다(도 7). 그 결과 미오튜브의 지름은 LLC 유도된 근위축 및 Dexa 유도된 근위축 모두에서 증대되었음이 확인되어, 성체 GTE가 근위축 환경에서 MuRF1의 수준을 억제하고 Akt(pAkt)의 수준을 향상시켜 근위축을 완화함을 확인하였다.After inducing muscle atrophy in differentiated myogenic cells, it was confirmed that muscle atrophy was alleviated when treated with green tea extract. C2C12 myoblasts obtained from LLC- or Dexa-induced muscular atrophy were treated with 1 ng/ml adult GTE and immunoblotted for MHC, HSP90, MuRF1, and MuRF1 using antibodies against MHC, HSP90, MuRF1, and Akt (pAkt). and the activation status of Akt was analyzed. Adult GTE treatment suppressed the level of MuRF1 and significantly enhanced the level of pAkt, while the expression level of the total protein was relatively constant (Figure 6). C2C12 myoblasts obtained from adult GTE-treated LLC or Dexa-induced muscular atrophy were quantitatively analyzed by MHC immunostaining, and the results were schematized (Figure 7). As a result, it was confirmed that myotube diameter was increased in both LLC-induced muscle atrophy and Dexa-induced muscle atrophy, suggesting that adult GTE inhibits the level of MuRF1 and improves the level of Akt (pAkt) in the muscle atrophy environment, thereby promoting muscle atrophy. It was confirmed that it was alleviated.
실험예 3: 미토콘드리아 발생 및 기능을 촉진하는 성체 GTEExperimental Example 3: Adult GTE that promotes mitochondrial development and function
미토콘드리아 발생 및 기능을 촉진함에 있어서 성체 GTE의 효과를 실험하기 위해, 성체 GTE 1ng/ml를 처리한 C2C12 근원세포를 3일 동안 분화유도하였고, JC-1 다이머 및 JC-1 모노머를 사용하여 형광표지하고 이를 확인한 결과 성체 GTE 처리된 C2C12 근원세포에서 미토콘드리아의 형성이 상대적으로 증가되었음이 확인되었다(도 8 및 도 9). 또한, 항-시르투인-1 항체를 이용하여 면역블롯한 결과, 성체 GTE 처리된 C2C12 근원세포에서 SIRT1 수치가 유의미하게 증가되었음이 확인되었다(도 10).To test the effect of adult GTE in promoting mitochondrial development and function, C2C12 myogenic cells treated with 1 ng/ml of adult GTE were differentiated for 3 days and fluorescently labeled using JC-1 dimer and JC-1 monomer. As a result, it was confirmed that the formation of mitochondria was relatively increased in adult GTE-treated C2C12 myogenic cells (FIGS. 8 and 9). Additionally, as a result of immunoblotting using an anti-sirtuin-1 antibody, it was confirmed that SIRT1 levels were significantly increased in adult GTE-treated C2C12 myoblasts (FIG. 10).
실험예 4: 노화근육의 개선을 유도하는 성체 GTEExperimental Example 4: Adult GTE that induces improvement in aging muscles
68세의 사람에게서 유래한 근원세포에 성체 GTE 처리된 경우 근원세포의 분화가 촉진되고 근위축이 완화되었음을 확인하였다. 노화근육의 개선에 있어서 성체 GTE의 효과를 실험하기 위해, skMDC Skeleton Muscle-Derived Cell(68 Male Caucasian Rectus Abdominis, Hypertension, Smoke, BMI 25 (old))을 입수하여 성체 GTE 1ng/ml 처리하고 D1, D2, D3에서 각각 관측하였다. 그 결과 근세포의 연결이 증대되었음이 확인되었다(도 11). 또한, 미오게닌은 특히 성체 GTE 처리된 경우 D3에서 mRNA 발현이 현저히 우수하였음이 확인되었고(도 12), 미오신 중쇄3 및 7은 특히 성체 GTE 처리된 경우 D3에서 mRNA 발현이 현저히 우수하였음이 확인되었다(도 13). 또한, PGC-1a는 특히 성체 GTE 처리된 경우 D4에서 mRNA 발현이 현저히 우수하였음이 확인되어(도 14), 근육 구성이 유산소적으로 변화되었음이 확인되었다. 종합적으로, 노화근육에서 전사인자가 증대됨에 따라 근육구성 단백질이 증가하고, 근육줄기세포가 증가함이 확인되었다. 이는 곧 노화에 의한 근감소증이 녹차추출물에 의해 완화될 수 있음을 시사한다.It was confirmed that when myoblasts derived from a 68-year-old person were treated with adult GTE, differentiation of myoblasts was promoted and muscle atrophy was alleviated. To test the effect of adult GTE in improving aging muscles, skMDC Skeleton Muscle-Derived Cells (68 Male Caucasian Rectus Abdominis, Hypertension, Smoke, BMI 25 (old)) were obtained, treated with 1ng/ml of adult GTE, and cultured at D1, Observed at D2 and D3, respectively. As a result, it was confirmed that the connection between muscle cells was increased (FIG. 11). In addition, it was confirmed that the mRNA expression of myogenin was significantly superior in D3, especially when treated with adult GTE (Figure 12), and that myosin heavy chains 3 and 7 had significantly superior mRNA expression in D3, especially when treated with adult GTE. (Figure 13). In addition, PGC-1a was confirmed to have significantly superior mRNA expression at D4, especially when treated with adult GTE (FIG. 14), confirming that muscle composition was aerobically changed. Overall, it was confirmed that as transcription factors increase in aging muscles, muscle building proteins increase and muscle stem cells increase. This suggests that sarcopenia due to aging can be alleviated by green tea extract.
7일 동안 성체 GTE 처리된 C2C12 근원세포는 더 많은 핵을 갖고, 보다 길어진 MHC-양성 근관세포를 형성하였다. 이는 MHC immunostaining에 의해 정량분석하고, 이를 도식화하였다(도 15). 다핵성 MHC+ 튜브의 비율을 확인한 결과, 성체 GTE 처리는 복수의 핵을 가진 근관세포의 비중을 증가시키고, 단핵세포의 비중을 급격히 감소시켰다. 이러한 결과들은 생화학적 측면에서나 형태학적 측면에서 성체 GTE가 C2C12 근원세포의 분화를 촉진시키고 있음을 의미한다(도 16). C2C12 myoblasts treated with adult GTE for 7 days formed longer MHC-positive myotubes with more nuclei. This was quantitatively analyzed by MHC immunostaining and schematized (Figure 15). As a result of confirming the ratio of multinucleated MHC+ tubes, adult GTE treatment increased the proportion of myotube cells with multiple nuclei and sharply decreased the proportion of mononuclear cells. These results mean that adult GTE promotes differentiation of C2C12 myogenic cells from a biochemical and morphological perspective (FIG. 16).
실험예 5: 손상근육의 줄기세포에 의한 재생을 유도하는 성체 GTEExperimental Example 5: Adult GTE that induces regeneration of damaged muscle by stem cells
손상근육의 재생에 있어 성체 GTE의 효과를 실험하기 위해, 성체 GTE를 2주 간 투여한 마우스의 전경골근에 근육의 괴사를 cardiotoxin를 통해 유도한 후 근육의 재생을 살펴보았다(도 17). D데이로부터 14일 전부터 마우스에 25㎍/day 성체 GTE 처리하고 D데이에 CTX로 데미지를 가한 후, PID0, PID3, 및 PID7에 TA/BW(%)를 측정하였다. 그 결과 PID0에는 성체 GTE 처리된 경우의 효과가 미미하였으나, PID7에는 성체 GTE 처리된 경우 TA/BW 비율이 대조군에 비해 현저히 상승되었음이 확인되어(도 18), 초기 근육 손상에 있어 성체 GTE 처리가 탁월한 효과가 있음이 확인되었다. 그 결과 녹차추출물을 투여한 군에서 근육 재생 신호가 더 활발하게 이뤄졌고, 실제로 근육이 더 많이 재생되었음을 확인하였다.To test the effect of adult GTE on the regeneration of damaged muscle, muscle necrosis was induced using cardiotoxin in the tibialis anterior muscle of mice administered adult GTE for 2 weeks, and then muscle regeneration was examined (FIG. 17). Mice were treated with 25 ㎍/day adult GTE starting 14 days before day D, and damaged with CTX on day D, and TA/BW (%) was measured at PID0, PID3, and PID7. As a result, the effect of adult GTE treatment was minimal at PID0, but at PID7, the TA/BW ratio was confirmed to be significantly increased compared to the control group when treated with adult GTE (Figure 18), suggesting that adult GTE treatment is effective in early muscle damage. It was confirmed that it had excellent effects. As a result, it was confirmed that muscle regeneration signals were more active in the group administered green tea extract, and more muscles were actually regenerated.
실험예 6: 근육 성능의 향상을 유도하는 성체 GTEExperimental Example 6: Adult GTE induces improvement in muscle performance
근육 성능의 향상에 있어 성체 GTE의 효과를 실험하기 위해, D데이로부터 마우스에 25㎍/day 성체 GTE 처리하고 4주 후 트레드밀 평가하였다. 그 결과 성체 GTE 처리된 경우 러닝타임이 현저히 증대된 것으로 확인되어, 근육 성능의 향상에 성체 GTE 처리가 탁월한 효과가 있음이 확인되었다(도 19).To test the effect of adult GTE in improving muscle performance, mice were treated with 25 μg/day adult GTE from day D and evaluated on a treadmill 4 weeks later. As a result, it was confirmed that the running time was significantly increased when treated with adult GTE, confirming that adult GTE treatment has an excellent effect on improving muscle performance (FIG. 19).
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