KR20080110212A - Lentinus edodes composition as a bone growth stimulating effector - Google Patents
Lentinus edodes composition as a bone growth stimulating effector Download PDFInfo
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- KR20080110212A KR20080110212A KR1020070058684A KR20070058684A KR20080110212A KR 20080110212 A KR20080110212 A KR 20080110212A KR 1020070058684 A KR1020070058684 A KR 1020070058684A KR 20070058684 A KR20070058684 A KR 20070058684A KR 20080110212 A KR20080110212 A KR 20080110212A
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- growth
- shiitake
- hot water
- bone
- water extract
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Abstract
Description
도1은 표고버섯의 열수 추출물이 조골세포 배양에서 IGF-1과 IGFBP-3의 발현을 촉진하는 결과를 나타낸 그래프이다.1 is a graph showing the results of promoting the expression of IGF-1 and IGFBP-3 in the osteoblastic culture of shiitake mushrooms.
(A)는 표고버섯의 열수 추출물이 조골세포 배양에서 IGF-1의 발현을 촉진하는 결과를 나타내고 (B)는 표고버섯의 열수 추출물의 IGFBP-3의 발현을 촉진하는 결과를 나타낸다.(A) shows that the hydrothermal extract of shiitake mushroom promotes the expression of IGF-1 in osteoblast culture and (B) shows the result of promoting the expression of IGFBP-3 of the hydrothermal extract of shiitake mushroom.
도2는 표고버섯의 열수 추출물을 쥐에 경구 투여하였을 때 성장판 길이를 증진시킨 결과를 나타낸 도면이다.Figure 2 is a view showing the result of increasing the growth plate length when orally administered hot water extract of shiitake mushrooms to mice.
(A)는 증류수를 투여한 대조군 쥐 중 대표적인 개체의 경골 성장판 부위를 확대한 성장판 사진이며 (B)는 표고버섯의 열수 추출물을 투여한 쥐 중 대표적인 개체의 경골 성장판 부위를 확대한 성장판 사진이다. (C)는 대조군 및 표고버섯 열수 추출물 투여군의 성장판 길이를 측정한 결과를 나타낸 그래프이다.(A) is an enlarged plate image showing the tibial growth plate area of the representative individual of the control rats administered distilled water, (B) is an enlarged plate image of the tibial growth plate area of the representative individual among the mice administered the hydrothermal extract of shiitake mushroom. (C) is a graph showing the results of measuring the growth plate length of the control group and shiitake hot water extract administration group.
도3은 표고버섯의 열수 추출물을 쥐에 경구 투여하였을 때 하루 동안 골길이 성장 을 증진시킨 결과를 나타낸 도면이다.Figure 3 is a view showing the result of the bone length growth during the day when oral administration of the hydrothermal extract of shiitake mushrooms to mice.
(A)는 증류수를 투여한 대조군 쥐 중 대표적인 개체의 경골 성장판 부위를 형광현미경으로 촬영하여 확대한 테트라사이클린 밴드 사진이며 (B)는 표고버섯의 열수 추출물을 투여한 쥐 중 대표적인 개체의 경골 성장판 부위를 형광 현미경으로 촬영하여 확대한 테트라사이클린 밴드 사진이다. (C)는 대조군 및 표고버섯 열수 추출물 투여군의 골길이 성장을 측정한 결과를 나타낸 그래프이다.(A) is a tetracycline band photograph of the tibial growth plate region of a representative individual of the control rats administered with distilled water by fluorescence microscopy. (B) is a tibial growth plate region of a representative individual among the rats treated with hydrothermal extract of shiitake mushroom. Is a tetracycline band photograph taken by fluorescence microscope. (C) is a graph showing the results of measuring the bone length growth of the control group and shiitake hot water extract administration group.
도4는 표고버섯의 열수 추출물을 쥐에 경구 투여하였을 때 장골길이를 성장시킨 결과를 나타낸 그래프이다.Figure 4 is a graph showing the growth of iliac bone when orally administered hot water extract of shiitake mushrooms to mice.
도5는 표고버섯의 열수 추출물을 쥐에 경구 투여하였을 때 장골무게를 증진시킨 결과를 나타낸 그래프이다.5 is a graph showing the results of enhancing iliac bone when oral administration of hydrothermal extract of shiitake mushrooms to rats.
도6은 표고버섯의 열수 추출물을 쥐에 경구투여 하였을 때 성장호르몬의 수치를 증가시킨 결과를 나타낸 그래프이다.Figure 6 is a graph showing the result of increasing the level of growth hormone when orally administered hot water extract of shiitake mushrooms to mice.
본 발명은 표고버섯에 증류수를 첨가하여 추출한 표고버섯 열수 추출물을 유효성분 으로 함유하는 골길이 성장에 효능이 있는 소재에 관한 것으로서, 보다 상세하게는 골길이 성장에 관여하는 성장호르몬 및/또는 IGF-1의 발현을 촉진하여 성장판 길이와 골길이 성장을 촉진하는 것을 특징으로 하는 표고버섯 열수 추출물을 유효성분으로 하는 성장기 성장 촉진 및 성장 장애 예방용 조성물, 발효유, 음료 및 건강기능식품에 관한 것이다.The present invention relates to a material having an effect on bone length growth containing shiitake mushroom hydrothermal extract extracted by adding distilled water to shiitake mushroom as an active ingredient, more specifically, growth hormone and / or IGF- involved in bone length growth. The present invention relates to a growth promoting growth and growth disorders preventive composition, fermented milk, beverages and health functional foods using the shiitake mushroom hydrothermal extract, characterized by promoting growth of plate length and bone length growth by promoting the expression of 1.
성장이란 세포의 크기와 수가 증가하는 것으로 성장기의 영양 상태는 일생에 걸쳐 영향을 줄 뿐만 아니라 영양소 간의 상호 작용은 유전적 요인의 발현에도 영향을 준다. 특히 청소년기의 성장은 상당부분 식이 섭취에 의해 결정되며 이 시기의 식이 섭취 패턴은 중년 이후의 골격 건강에까지 영향을 끼치므로 가장 중요하다고 해도 과언이 아니다. 이 중 키가 성장하는 것은 일정한 시기가 있어 사춘기가 시작되고 2~3년이 되면 성장이 정지되기 때문에 적절한 시기를 놓치지 않도록 하는 것 또한 매우 중요하다. Growth is the increase in the size and number of cells. The growth status of nutrients affects not only lifelong but also the interactions between nutrients affects the expression of genetic factors. In particular, growth in adolescence is largely determined by dietary intake, and it is no exaggeration to say that the dietary intake pattern of this period is most important since it affects skeletal health after middle age. It is also very important that the height of growth is a certain period of time so that puberty begins and growth stops in two to three years.
일반적으로 키가 크는 것은 유전적 요인이 작용한다고 생각하기 쉬우나 지금까지 알려진 바로는 키가 자라는 것은 선천적인 요인보다 후천적인 요인에 의해 영향을 더 받는다고 보고되었다. 이 중 영양 상태와 운동이 중요한 인자들로 알려져 있다. In general, it is easy to think that height is a genetic factor, but so far it has been reported that height growth is influenced by acquired factors rather than congenital factors. Among them, nutritional status and exercise are known as important factors.
최근 경제성장에 따른 생활수준의 향상과 영양 개선, 유년기 생활환경 및 생활습관 등의 요인에 의해 소아의 평균 신장, 체중 등이 점차 증가되고 있으며 이 때문에 부모나 어린이들은 키에 대한 관심과 요구가 증가되고 있다.In recent years, the average height and weight of children are gradually increasing due to factors such as improvement of living standards, nutrition, and childhood living conditions and lifestyles, and this is why parents and children have increased interest and demand for height. It is becoming.
인위적으로 키를 키우기 위한 방법으로 성장호르몬 요법과 일리자로프 수술 등이 있다. 성장호르몬 요법은 성장 호르몬 결핍증, 터너증후군 환자들을 중심으로 시술되고 있으나 1년에 걸쳐 거의 매일 주사를 맞아야 하는 번거로움뿐만 아니라 그 비용도 만만치 않고 간혹 부작용도 보고되고 있다. 일리자로프 수술은 정강이 뼈에 금을 낸 다음 뼈를 늘리는 수술로서 어쩔 수 없는 장애나 질병이 아니면 일반적으로 적용하기는 어려운 방법이다. Growth hormone therapy and ilizarov surgery are some of the ways to artificially increase your height. Growth hormone therapy has been performed mainly for patients with growth hormone deficiency and Turner syndrome, but it is not only hassle that has to be injected almost every day over a year, but also its cost is sometimes reported. Ilizarov surgery is a surgery that cracks the shin bone and then stretches the bone, which is usually difficult to apply unless it is inevitable.
이러한 문제점 때문에 최근에는 성장을 자극하는 운동요법, 운동 보조기구 등과 건강기능식품이 키를 키우기 위한 보다 간편하고 부작용이 없는 대안으로 제시되고 있다. 예를 들어, 대한민국 특허등록 제404455호에는 성장촉진추출물 및 이를 함유하는 약학적 제제에 관하여, 대한민국 특허등록제463263호에는 골길이 성장 촉진 효과를 가지는 가시오가피 추출물 및 이를 함유하는 약학적 제제에 관하여, 대한민국 특허등록 제561286호에는 건조효모, 천연물 추출분말 및 영양성분 혼합분말을 함유하여 성장발육에 도움을 주는 건강기능성 조성물에 관하여, 대한민국 특허등록 제597563호에는 골성장 촉진활성을 갖는 엘루테로사이드 Ｅ를 함유하는 건강기능식품이 소개되어 있다. 이러한 성장에 효과가 있다고 알려진 천연물제들을 이용한 기능성 식품들은 천연물의 특유한 성질로 인해 소비자의 맛이나 기호도를 만족시키지 못하는 형태로 섭취되고 있을 뿐만 아니라 성장의 효과 또한 미비한 단점이 있다.Because of these problems, recently, exercise therapy, exercise aids, and other health functional foods to stimulate growth have been proposed as a simpler and no side effect for raising height. For example, Korean Patent Registration No. 404455 relates to growth promoters and pharmaceutical preparations containing the same, and Korean Patent Registration No. 463263 relates to prickly pear extracts and bone medicinal preparations having bone growth growth effects, Patent registration No. 561286 relates to a health functional composition that helps the growth and development by containing dry yeast, natural product extraction powder and nutrient mixture powder, Republic of Korea Patent Registration No. 575763 Eluteroside E having bone growth promoting activity Contains dietary supplements. Functional foods using natural ingredients known to be effective for such growth are consumed in a form that does not satisfy the taste or taste of consumers due to the unique properties of the natural products, as well as the effect of growth is inadequate.
한편, 표고버섯은 식용버섯으로 사용되며 특히 동북아시아에서는 약용으로 사용되어 왔다. 표고버섯은 영양학적으로 우수할 뿐만 아니라 약리활성이 있는 물질들이 보고되었다. 표고버섯의 (1-3)-beta-D-glucan인 렌티난(lentinan)은 항종양 및 면역활성을 증가시키는 것으로 알려졌다. 일본에서 렌티난은 면역증강제 및 항암제로 도 사용되고 있다. 표고버섯에는 단백질과 지방질, 당질이 많이 포함되어 있다. 비타민A와 C는 함유하고 있지 않지만 비타민B1, B2의 함유량은 야채의 두배 이상을 함유하고 있다. 또한 표고버섯은 산소를 운반하는 혈액 중의 헤모글로빈을 생성하는 철분도 다량 포함하고 있으며 식이섬유가 많이 들어 있어 변비 예방에 도움을 준다. 뿐만 아니라, 표고버섯에는 비타민D의 효과를 지닌 에르고스테롤이 많고, 에리다데닌이 함유되어 있어 혈액 중의 콜레스테롤을 제거, 고혈압 예방에 효과가 있다. Shiitake mushrooms, on the other hand, are used as edible mushrooms, especially in Northeast Asia. Shiitake mushrooms have been reported to be not only nutritionally superior but also pharmacologically active. Lentinan (1-3) -beta-D-glucan of shiitake mushrooms has been known to increase antitumor and immune activity. In Japan, lentinan is also used as an adjuvant and anticancer agent. Shiitake mushrooms are high in protein, fat and sugar. It does not contain vitamins A and C, but vitamins B1 and B2 contain more than twice the amount of vegetables. Shiitake mushrooms also contain a large amount of iron that produces hemoglobin in the blood that carries oxygen and contains a lot of dietary fiber to help prevent constipation. In addition, shiitake mushrooms contain a lot of ergosterol with the effect of vitamin D, and contains eridadenin, which is effective in removing cholesterol from the blood and preventing hypertension.
그러나, 상기와 같은 표고버섯의 다양한 약리작용은 잘 알려져 있었으나 골길이 성장과 관련된 연구는 충분히 이루어지지 않았다.However, although various pharmacological effects of shiitake mushrooms are well known, studies related to bone length growth have not been sufficiently conducted.
이에 본 발명은 앞서 설명한 바와 같이 기존의 종래 기술의 문제점을 더욱 효율적으로 해결하기 위하여 안출된 것으로써, 본 발명의 목적은 성장호르몬 및/또는 IGF-1의 발현을 촉진하여 성장판 길이 및 골길이 성장을 촉진하는 표고버섯으로부터 분리된 열수 추출물을 유효성분으로 포함하는 골길이 성장 촉진 및 성장장애 예방용 조성물, 발효유, 음료 및 건강기능식품을 제공하는 데 있다.Therefore, the present invention has been made in order to solve the problems of the prior art more efficiently as described above, the object of the present invention is to promote the growth hormone and / or IGF-1 expression plate growth length and bone length growth It is to provide a composition for promoting bone length growth and growth disorder prevention, fermented milk, beverages and health functional foods containing hot water extract isolated from shiitake mushrooms to promote.
상기와 같은 목적을 달성하기 위하여, 본 발명은 성장 호르몬 및/또는 IGF-1의 발현을 증가시켜 골길이 성장을 촉진하는 효능을 갖는 표고버섯 열수 추출물을 유효 성분으로 하여 성장기 골길이 성장 도움 및 성장장애 예방용 조성물, 발효유, 음료 및 건강기능식품을 제공하는 것을 특징으로 한다.In order to achieve the above object, the present invention is to increase the expression of growth hormone and / or IGF-1 by using the shiitake mushroom hydrothermal extract having the effect of promoting bone length growth as an active ingredient growth and bone length growth help It is characterized by providing a disorder preventing composition, fermented milk, beverages and health functional food.
이하 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
표고버섯으로부터 성장판 길이 및 골길이 성장을 촉진하는 것을 특징으로 하는 표고버섯의 열수추출물을 효율적으로 분리하는 과정을 상세히 살펴보면 다음과 같다.Looking at the process of efficiently separating the hydrothermal extract of shiitake mushrooms, characterized in that to promote growth plate length and bone length growth from shiitake mushrooms.
경북 문경의 농협에서 구매한 표고버섯 슬라이스를 증류수에 넣고 100~120℃에서 4시간 동안 환류 냉각하면서 2~3회 추출한다. 이때 수용액의 사용량은 표고버섯 부피의 10~20배 정도가 적당하다. 상기에서 분리된 표고버섯의 열수추출액을 750mmHg 감압 조건과 35~55℃의 조건하에서 증발 농축한 후 -20℃에서 1차 동결하고 -80℃에서 2차 동결한 다음 동결건조기를 사용해 48시간 동안 건조하거나 감압건조기를 사용하여 10시간 동안 건조하여 추출액을 분말상태로 만들거나 기타 통상적인 방법으로 분말상태로 제조할 수 있다. 그리고 상기 표고버섯 열수 추출분말 10mg을 물 1ml에 녹여 골길이 성장 촉진 및 성장장애 예방 물질로 사용한다.Sliced shiitake mushrooms purchased from Nonghyup in Mungyeong, Gyeongbuk, are placed in distilled water and extracted 2-3 times with reflux cooling at 100-120 ° C for 4 hours. At this time, the amount of the aqueous solution is suitable 10 ~ 20 times the volume of shiitake mushrooms. The hot water extract of shiitake mushroom was separated by evaporation under 750mmHg decompression and 35 ~ 55 ℃, and then freeze at -20 ℃ for the first time, and freeze at -80 ℃ for 2nd time, and then dried for 48 hours using freeze dryer. Alternatively, the extract may be dried in a powder state by drying with a reduced pressure dryer for 10 hours or prepared in powder form by other conventional methods. And 10mg of the shiitake mushroom hot water extract powder is dissolved in 1ml of water and used as a substance for promoting bone length growth and preventing growth disorder.
본 발명에 따른 표고버섯의 열수 추출물을 유효성분으로 하는 골길이 성장 촉진 및 성장장애 예방 조성물은 경구, 경피, 피하, 정맥 또는 근육을 포함한 여러 경로를 통해 투여될 수 있다. 또, 본 발명의 표고버섯 추출물을 유효성분으로 하는 골길이 성장 촉진 및 상장 장애 예방 조성물은 여러 가지 제형으로 제제화 할 수 있는데 제제화 할 경우에는 통상적으로 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제 및 계면활성제 등의 희석제 또는 부형제를 사용하여 제제화 할 수 있다. 경구투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제, 및 캡슐제 등이 포함되며, 이러한 고형 제제는 표고버섯의 열수추출물에 적어도 하나 이상의 부형제(예를 들면, 전분, 수크로스, 락토오스 및 젤라틴) 등이 섞여 조제된다. 또한 단순한 부형제 이외에 윤활제들도 사용될 수 있다. 경구투여를 위한 액상제제로는 현탁제, 내용액제, 유제 및 시럽제 등을 들 수 있는데, 흔히 사용되는 단순 희석제인 물, 액체 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제 및 보존제 등이 포함될 수 있다. 비 경구 투여를 위한 제제에는 멸균된 수용액, 비수용성 용제, 현탁제, 유제, 동결건조 제제 및 좌제가 포함된다. 비수용성용제와 현탁용제로는 프로필렌글리콜, 폴리에틸렌글리콜, 올리브 오일과 같은 식물성기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 글리세롤 및 젤라틴 등이 사용될 수 있다.Bone length growth promotion and growth disorder prevention composition comprising the hydrothermal extract of shiitake mushrooms according to the present invention as an active ingredient can be administered through various routes including oral, transdermal, subcutaneous, intravenous or intramuscular. In addition, bone length growth promotion and seizure prevention composition comprising the shiitake mushroom extract of the present invention as an active ingredient can be formulated into a variety of formulations, when formulated, commonly used fillers, extenders, binders, wetting agents, disintegrant and It can be formulated using diluents or excipients such as surfactants. Solid form preparations for oral administration include tablets, pills, powders, granules, and capsules, and the like form at least one excipient (e.g., starch, sucrose, lactose and gelatin) in the hydrothermal extract of shiitake mushrooms. ) Is mixed and prepared. Lubricants may also be used in addition to simple excipients. Liquid preparations for oral administration include suspensions, solvents, emulsions, and syrups. In addition to the commonly used simple diluents, water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives, etc. May be included. Formulations for non-oral administration include sterile aqueous solutions, water-insoluble solvents, suspensions, emulsions, lyophilized formulations and suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. Glycerol and gelatin may be used as the base of the suppository.
사람의 경우 통상적인 1일 투여량은 1~30 mg/kg 체중의 범위일 수 있고, 1회 또는 수회로 나누어 투여할 수 있다. 그러나 실제 투여량은 투여경로, 연령, 성병 및 체중, 건강 상태 등의 여러 관련 인자에 비추어 결정되어야 한다.For humans, a typical daily dose may range from 1 to 30 mg / kg body weight and may be administered once or in divided doses. However, the actual dosage should be determined in light of several relevant factors such as route of administration, age, sexually transmitted disease and weight, and health status.
본 발명의 열수 추출물은 식품으로 사용되는 표고버섯을 원료로 하여 독성 및 부작용이 거의 없으므로 골길이 성장 촉진 및 성장장애 예방 목적으로 장기간 복용 시에도 안심하고 사용할 수 있는 제제이다.Since the hot water extract of the present invention is made from shiitake mushrooms used as food, there is almost no toxicity and side effects, and thus it is a preparation that can be used safely even for long-term use for promoting bone length growth and preventing growth disorders.
한편, 본 발명은 상기 추출 방법에 의하여 얻어진 표고버섯의 열수 추출물을 유효 성분으로 함유하는 골길이 성장 증진 및 성장장애 예방용 건강식품, 유제품 및 음료로 사용가능하다.On the other hand, the present invention can be used as a health food, dairy products and beverages for bone length growth enhancement and growth disorder prevention containing hot water extract of shiitake mushroom obtained by the extraction method as an active ingredient.
상기 골길이 성장 증진 및 성장장애 예방용 발효유는 탈지분유로 무지유고형분 함량을 조정한 원료유에 락토바실러스 카제이 HY2782 {Lactobacillus casei HY2782; 기탁기관: 한국 종균 협회, 기탁번호 KFCC-10813, 기탁일자 1993년 12월 7일, 특허등록 제0125219호(발명의 명칭:프로파아지가 제거된 락토바실러스카제이 HY2782 및 이를 이용한 제품)}를 접종한 배양액에 과즙 농축액, 식이섬유, 포도당, 올리고당, 칼슘, 비타민, 표고버섯의 열수 추출물 등을 녹여 제조된 시럽을 일정 비율로 혼합, 교반하여 균질화한 뒤 용기에 포장하여 발효유를 제조한다.The fermented milk for bone length growth and growth disorder prevention is lactobacillus casei HY2782 { Lactobacillus casei HY2782 in raw milk adjusted to the non-fat solid content of skim milk powder; Deposited Organization: Korean spawn association, Accession No. KFCC-10813, Date of Deposit, December 7, 1993, Patent Registration No. 0125219 (Name of invention: Lactobacillus skazy HY2782 and product using it) Dissolve juice concentrates, dietary fiber, glucose, oligosaccharides, calcium, vitamins, hot water extracts of shiitake mushrooms, etc. in a culture solution mixed with a certain ratio, stirred and homogenized, and then packaged in a container to prepare fermented milk.
또한 상기의 골길이 성장 증진 및 성장장애 예방용 음료는 표고버섯 추출물이 유효성분으로 포함되는 것 이외에 칼슘, 가시오가피 농축액, 액상과당, 정제수를 첨가 혼합하여 드링크용 병에 충진하여 살균한 후 실온으로 냉각하여 음료를 제조한다.In addition, the above bone length growth and growth disorders preventive beverages include shiitake mushroom extract as an active ingredient, calcium, thorny oak concentrate, liquid fructose, and purified water are added and mixed in the drink bottle and sterilized and then cooled to room temperature. To prepare a beverage.
이하 실시예를 통하여 본 발명을 보다 상세하게 설명한다. 그러나 다음의 실시 예는 본 발명의 범위를 한정하는 것은 아니며, 본 발명의 기술적 사상의 범위 내에서 당업자에 의한 통상적인 변화가 가능하다.Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are not intended to limit the scope of the present invention, it is possible for the ordinary change by those skilled in the art within the scope of the technical idea of the present invention.
<실시예 1><Example 1>
표고버섯 열수 추출물의 분리 정제Separation and Purification of Shiitake Hot Water Extract
표고버섯을 구입하여 수세, 풍건한 후 길이 1 cm 간격으로 분쇄하였다. 분쇄된 시 료 1kg을 추출병에 넣은 후 증류수 10L를 첨가하였다. 그런 다음 상기 증류수를 첨가한 시료를 110℃에서 4시간 동안 추출한 후 60 mesh로 1차 여과하고 여과지(Whatman No. 41)로 2차 여과하였다. 상기 여과지에 의한 여과액을 진공회전농축기(Eyela NE, Japan)를 사용하여 감압 농축한 후 동결건조하여 표고버섯의 열수 추출물을 제조하였다. 상기의 건조된 시료 10mg을 정제수에 녹여 골성장 활성 소재로 사용하였다.Shiitake mushrooms were washed with water, air dried, and then ground at 1 cm intervals. 1 kg of the ground sample was placed in an extraction bottle, and 10 L of distilled water was added thereto. Then, the sample to which the distilled water was added was extracted for 4 hours at 110 ° C., followed by primary filtration with 60 mesh and secondary filtration with filter paper (Whatman No. 41). The filtrate by the filter paper was concentrated under reduced pressure using a vacuum rotary concentrator (Eyela NE, Japan) and then lyophilized to prepare a hot water extract of shiitake mushroom. 10 mg of the dried sample was dissolved in purified water and used as an active material for bone growth.
<시험예 1><Test Example 1>
표고버섯 열수 추출물의 조골세포에서 IGF-1의 발현을 증가시키는 효능의 측정Measurement of Efficacy of Increasing IGF-1 Expression in Osteoblasts of Shiitake Mushroom Water Extract
조골세포에서 인슐린 유사 성장 인자 (insulin-like growth factor type 1 ;IGF-1)의 발현 증가는 ELISA 방법을 이용하였으며, Diagnostic Systems Laboratories사의 IGF-1 ELISA kit를 사용하였다. 측정방법은 다음과 같다.The expression of insulin-like growth factor type 1 (IGF-1) in osteoblasts was increased by ELISA and IGF-1 ELISA kit by Diagnostic Systems Laboratories. The measurement method is as follows.
사람에서 유래한 조골세포주인 MG63 조골세포주를 24 well plate에 8ⅹ104/well로 접종하여 5% FBS가 포함된 DMEM 배지에서 37℃, 5% CO2 조건에서 24시간 배양하였다. 12시간 후 2% FBS가 포함된 DMEM 배지를 첨가하고 표고버섯 열수 추출물의 최종 농도가 각각 0.25㎎/㎖, 0.5㎎/㎖, 1㎎/㎖이 되도록 첨가하였다. 24시간 후 50μl 배양액의 IGF-1을 200μl 추출 완충액 (87.5% 에탄올, 12.5% 2N HCl)으로 30분간 교반하면서 반응하여 추출하였다. 100μl 추출액에 100μl 중화 완충액 (0.855 M Tris Base)을 첨가하여 중화하였다. 이 중화액을 적절한 농도로 carbonate- bicarbonate 완충액에 희석하고 96 well plate에 4℃에서 12시간 정치하였다. TBST(Tris-buffered saline, 0.02% Tween 20) 완충액으로 3회 세척하고 1% BSA가 함유된 TBST로 1시간 동안 반응하였다. 1:10,000으로 희석한 anti-IGF-1 항체(Santa Cruz Biotech, USA)로 1시간 동안 실온에서 반응하고 TBST로 3회 세척한 다음 1:1,000으로 희석한 HRP-conjugated anti-goat IgG (Santa Cruz Biotech, USA)를 실온에서 1시간 반응하였다. 다시 TBST로 3회 세척한 후 OPD peroxidase substrate (Sigma, USA)로 15분간 반응한 후 2N H2SO4를 첨가하여 반응을 종료시키고 493nm에서 흡광도를 측정하여 IGF-1 농도를 결정하였다. 데이터는 평균값 ±SD 값으로 표시하였으며 IGF-1의 차이는 T Test를 이용하여 통계처리 하였다. 모든 데이터는 P<0.05 이하일 경우 통계적으로 유의적이라고 결정하였다.MG63 osteoblasts, human-derived osteoblasts, were inoculated at 8ⅹ10 4 / well in 24 well plates and incubated in DMEM medium containing 5% FBS for 24 hours at 37 ° C and 5% CO 2 . After 12 hours, DMEM medium containing 2% FBS was added, and the final concentration of shiitake hot water extract was added to 0.25 mg / ml, 0.5 mg / ml, and 1 mg / ml, respectively. After 24 hours, IGF-1 of 50 µl culture was extracted by reaction with 200 µl extraction buffer (87.5% ethanol, 12.5% 2N HCl) for 30 minutes with stirring. To 100 μl extract was neutralized by addition of 100 μl neutralization buffer (0.855 M Tris Base). The neutralized solution was diluted in carbonate-bicarbonate buffer at an appropriate concentration and allowed to stand on a 96 well plate at 4 ° C for 12 hours. Washed three times with TBST (Tris-buffered saline, 0.02% Tween 20) buffer and reacted with TBST containing 1% BSA for 1 hour. React with an anti-IGF-1 antibody diluted at 1: 10,000 (Santa Cruz Biotech, USA) for 1 hour at room temperature, wash three times with TBST and then diluted HRP-conjugated anti-goat IgG (Santa Cruz) at 1: 1,000 Biotech, USA) was reacted at room temperature for 1 hour. After washing 3 times with TBST again, the reaction was performed with OPD peroxidase substrate (Sigma, USA) for 15 minutes, and then 2N H 2 SO 4 was added to terminate the reaction. The absorbance was measured at 493 nm to determine the IGF-1 concentration. Data were expressed as mean ± SD value and the difference of IGF-1 was statistically processed using T Test. All data were determined to be statistically significant when P <0.05 or less.
표고버섯 열수 추출물의 조골세포주에서 IGF-1의 발현을 증가시키는 효능을 측정한 결과는 표1과 도1의 (A)에 도시하였다.The results of measuring the effect of increasing the expression of IGF-1 in osteoblast cell line of Shiitake hot water extract are shown in Table 1 and FIG.
상기 표1 및 도1(A)에서 확인할 수 있는 바와 같이, 표고버섯의 열수 추출물을 첨가하지 않은 대조군에서 IGF-1이 100%를 기준으로 할 때 유의적으로 실험군1은 10%, 실험군2는 6%, 실험군3은 10%로 각각 IGF-1의 발현을 촉진함을 알 수 있어 표고버섯 열수 추출물이 골성장에 효과가 있음을 알 수 있었다.As can be seen in Table 1 and FIG. 1 (A), when IGF-1 is 100% in the control group without adding hydrothermal extract of shiitake mushroom,
<시험예 2><Test Example 2>
표고버섯 열수 추출물의 조골세포에서 IGFBP-3의 발현을 증가시키는 효능의 측정Determination of Efficacy of Increasing Expression of IGFBP-3 in Osteoblasts of Shiitake Hot Water Extract
조골세포에서 인슐린 유사 성장인자 결합단백질 3(Insulin-like Growth Factor Binding Protein-3; IGFBP-3)의 발현 증가는 ELISA 방법을 이용하였으며, Diagnostic Systems Laboratories사의 IGFBP-3 ELISA kit를 사용하였다. 측정방법은 다음과 같다.The expression of insulin-like Growth Factor Binding Protein-3 (IGFBP-3) in osteoblasts was increased by ELISA and IGFBP-3 ELISA kit by Diagnostic Systems Laboratories. The measurement method is as follows.
사람에서 유래한 조골세포주인 MG63 조골세포주를 24 well plate에 8ⅹ104/well로 접종하여 5% FBS가 포함된 DMEM 배지에서 37℃, 5% CO2 조건에서 24시간 배양하였다. 12시간 후 2% FBS가 포함된 DMEM 배지를 첨가하고 표고버섯 의 열수 추출물의 최종 농도가 각각 0.25㎎/㎖, 0.5㎎/㎖, 1㎎/㎖이 되도록 첨가하였다. 24시간 후 배양액 20μl와 Zero Standard Solution A 100μl를 혼합하였다. 여기서 25μl를 취하여 Assay Buffer 50μl와 혼합하고 실온에서 2시간 동안 흔들어 주면서 반응하였다. TBST로 5회 세척하고 물기를 제거하였다. Antibody-Enzyme Conjugate Solution 100μl를 각 well에 투입하고 실온에서 1시간 동안 흔들어 주면서 반응하였다. TBST로 5회 세척한 다음 물기를 제거하고 TMB Chromogen Solution 100μl를 각 well에 첨가하고 실온에서 10분 동안 흔들어 주면서 반응하였다. Stopping Solution 100μl를 각 well에 첨가하고 450 nm에서 흡광도를 측정하여 IGFBP-3의 함량을 결정하였다. 데이터는 평균값 ±SD 값으로 표시 하였으며 IGFBP-3의 차이는 T Test를 이용하여 통계처리 하였다. 모든 데이터는 P<0.05 이하일 경우 통계적으로 유의적이라고 결정하였다.MG63 osteoblasts, human-derived osteoblasts, were inoculated at 8ⅹ10 4 / well in 24 well plates and incubated in DMEM medium containing 5% FBS for 24 hours at 37 ° C and 5% CO 2 . After 12 hours, DMEM medium containing 2% FBS was added, and the final concentration of the hydrothermal extract of shiitake mushrooms was added to 0.25 mg / ml, 0.5 mg / ml, and 1 mg / ml, respectively. After 24 hours, 20 μl of the culture solution and 100 μl of Zero Standard Solution A were mixed. Here 25μl was taken and mixed with 50μl Assay Buffer and reacted by shaking for 2 hours at room temperature. Wash 5 times with TBST and drain. 100μl of Antibody-Enzyme Conjugate Solution was added to each well and reacted by shaking for 1 hour at room temperature. After washing 5 times with TBST, the water was removed, and 100 μl of TMB Chromogen Solution was added to each well and reacted by shaking at room temperature for 10 minutes. The content of IGFBP-3 was determined by adding 100 μl of Stopping Solution to each well and measuring the absorbance at 450 nm. Data were expressed as mean value ± SD value and the difference of IGFBP-3 was statistically processed using T Test. All data were determined to be statistically significant when P <0.05 or less.
표고버섯 열수 추출물의 조골세포주에서 IGFBP-3의 발현을 증가시키는 효능을 측정한 결과는 표2와 도1의 (B)에 도시하였다.The results of measuring the effect of increasing the expression of IGFBP-3 in osteoblast cell line of Shiitake hot water extract are shown in Table 2 and FIG.
상기 표2 및 도1(B)에서 확인할 수 있는 바와 같이, 표고버섯의 열수 추출물을 첨가하지 않은 대조군에서 IGFBP-3이 100%를 기준으로 할 때 유의적으로 실험군1은 17%, 실험군2는 27%, 실험군3은 66%로 각각 IGF-1의 성장촉진에 따라 IGF-1에 결합하는 IGFBP-3이 증가함을 알 수 있어 표고버섯 열수 추출물이 골성장에 효과가 있음을 알 수 있었다.As can be seen in Table 2 and FIG. 1 (B), when IGFBP-3 is 100% in the control group without adding the hydrothermal extract of shiitake mushroom,
<시험예 3><Test Example 3>
표고버섯 열수 추출물이 쥐에서 경골의 성장판 및 골길이 성장을 촉진하는 효과의 측정Measurement of the Effect of Shiitake Mushroom Water Extract on the Growth Plate and Bone Length Growth of Tibia in Rats
표고버섯 열수 추출물을 증류수에 용해하여 350㎎/kg의 양으로 4주령의 쥐에 5일 연속으로 경구투여 하였다. 대조군에는 증류수를 투여하였고 실험군당 8마리의 쥐를 사용하였다. 시료 투여 4일째와 5일째에 테트라싸이클린을 증류수에 용해하여 10㎎을 투여하였다. 투여 후 6일째에 쥐를 희생하여 경골의 성장판 부위를 적출하였다. 분리된 조직을 10% 포르말린 용액에서 24시간 동안 고정한 다음 시료를 탈회하고 파라핀에 고정하였다. 마이크로톰을 사용하여 수직으로 1μm의 두께로 절단하여 각각의 절편을 얻었다. 슬라이드 글라스에 고정하고 H&E 염색을 실시하고 마이크로 현미경을 사용해 성장판 부위의 사진을 촬영하였다. 길이 측정 프로그램을 사용해 성장판 길이를 측정하였다. 1일 골성장 길이는 형광 현미경을 사용해 형광 사진을 얻은 후 동일한 프로그램을 사용해 테트라싸이클린 밴드사이의 길이를 측정하여 결정하였다. 데이터는 평균값 ±SD 값으로 표시 하였으며 데이터간의 차이는 T Test를 이용하여 통계처리 하였다. 모든 데이터는 P<0.05 이하일 경우 통계적으로 유의적이라고 결정하였다.Shiitake mushroom hot water extract was dissolved in distilled water and orally administered to 4 week old rats in an amount of 350 mg / kg for 5 consecutive days. Distilled water was administered to the control group and 8 rats were used per experimental group. On the 4th and 5th day of sample administration, tetracycline was dissolved in distilled water and 10 mg was administered. On day 6 after administration, the rats were sacrificed and the growth plate area of the tibia was removed. The detached tissue was fixed in 10% formalin solution for 24 hours, then the sample was demineralized and fixed in paraffin. Each section was obtained by vertically cutting to a thickness of 1 μm using a microtome. Fixed to slide glass, subjected to H & E staining and photographed the growth plate area using a micro microscope. The growth plate length was measured using a length measuring program. The daily bone growth length was determined by fluorescence microscopy, followed by measuring the length between tetracycline bands using the same program. The data were expressed as mean value ± SD value and the difference between the data was statistically processed using T Test. All data were determined to be statistically significant when P <0.05 or less.
표고버섯 열수 추출물을 투여한 쥐에서 경골의 성장판 길이 및 골길이 성장을 촉진하는 효능을 측정한 결과를 표3과 도2 및 도3에 도시하였다.Table 3 and Figures 2 and 3 show the results of measuring the effect of promoting the growth plate length and bone length growth of the tibia in rats administered Shiitake hot water extract.
상기 표3과 도2 및 도3에서 확인할 수 있는 바와 같이, 표고버섯의 열수 추출물을 쥐에게 투여하지 않은 대조군과 투여한 실험군의 성장판 길이의 차이는 71μm 이고, 골길이 성장의 차이는 36μm로서 생체 내에서도 표고버섯 열수 추출물이 골성장에 효과가 있음을 알 수 있었다.As can be seen in Table 3 and Figures 2 and 3, the difference in the growth plate length of the control group and the control group not administered the hydrothermal extract of shiitake mushrooms to the rat is 71μm, the difference in bone length growth is 36μm in vivo Shiitake mushroom hot water extract was found to be effective in bone growth.
<시험예 4><Test Example 4>
표고버섯 열수 추출물이 쥐의 장골길이 성장 및 성장호르몬에 미치는 영향Effects of Shiitake Hot Water Extract on Growth Length and Growth Hormone of Rats
5주령인 수컷 SD 쥐를 사용하였으며 대조군에는 증류수를, 표고버섯 열수 추출물 투여군은 350㎎/kg을 존대로 4주간 매일 경구로 투여하였으며 실험군당 15마리를 사용하였다. 사료는 상용화된 사료를 급여하였고 사료와 물은 자유로이 급식하였다. 실험방은 매일 12시간 등을 켜두고 12시간 점등하였다. 모든 쥐들은 1주일에 한번씩 총 4주간 체중을 측정하였다. 사료섭취량은 각 군당 총 사료섭취량을 측정한 후 쥐 개체수로 나눈 평균 사료섭취량으로 결정하였다. 4주간 투약시험이 종료된 후 모든 쥐들을 안락사하고 근육과 인대 등을 모두 분리하고 왼쪽 대퇴골을 분리해 내었다. 대퇴골의 길이는 버어니어 켈리퍼스로 측정하였으며 대퇴골의 무게는 일반 저울로 측정하였다. 혈청 성장호르몬 농도는 상용화되어 있는 Rat Growth Hormone Radioimmunoassay (RIA) kit (LINCO Research, USA)를 이용하여 측정하였다. 데이터는 평균값 ±SEM 값으로 표시 하였으며 성장호르몬 차이는 Wilcoxon signed-rank test를 이용하여 통계처리 하였다. 모든 데이터는 SPSS version 12.0 통계프로그램을 이용하여 분석하였고, P<0.05 이하일 경우 통계적으로 유의하다고 판단하였다.Male SD rats, 5 weeks old, were used, and distilled water was used as a control group and 350 mg / kg of shiitake mushroom hot water extracts were orally administered daily for 4 weeks, and 15 rats were used per experimental group. Feed was fed commercially available feed, and feed and water were freely fed. The test room was turned on for 12 hours with a 12-hour light on every day. All mice were weighed for a total of four weeks once a week. Feed intake was determined by measuring total feed intake per group and then average feed intake divided by rat population. After four weeks of dosing, all rats were euthanized, their muscles and ligaments were separated, and their left femur was removed. The length of the femur was measured with a vernier caliper and the weight of the femur was measured with a general balance. Serum growth hormone concentration was measured using a commercially available Rat Growth Hormone Radioimmunoassay (RIA) kit (LINCO Research, USA). Data were expressed as mean ± SEM and growth hormone differences were statistically analyzed using Wilcoxon signed-rank test. All data were analyzed using the SPSS version 12.0 statistical program and judged to be statistically significant when P <0.05 or less.
표고버섯 열수 추출물을 투여한 쥐의 장골길이 성장에 미치는 영향을 표4에 표시하였으며, 또한 사료섭취량 대비 장골길이의 비율을 도4에 나타내었고 사료섭취량 대비 장골무게의 비율을 도5에 나타내었다. The effect on the growth of iliac bone length of rats treated with Shiitake hot water extract is shown in Table 4, and the ratio of iliac length to feed intake is shown in FIG. 4, and the ratio of iliac weight to feed intake is shown in FIG.
표고버섯 열수 추출물을 투여한 쥐의 혈중 성장호르몬 수치에 대한 영향은 표5 및 도6에 나타내었다.The effects on the blood growth hormone levels of the rats treated with Shiitake mushroom hot water extract are shown in Table 5 and FIG. 6.
상기 표4와 도4 및 도5에서 확인할 수 있는 바와 같이, 표고버섯의 열수 추출물을 쥐에게 투여하지 않은 대조군과 투여한 실험군의 사료섭취량 대비 장골길이의 차이는 0.008mm/g이고, 사료섭취량 대비 장골무게의 차이는 0.154㎎/g로서 생체내에서 표고버섯 열수 추출물이 장골길이의 성장 및 장골무게의 증가에 효과가 있음을 알 수 있었다.As can be seen in Table 4 and Figures 4 and 5, the difference between the feed intake of the control group and the experimental group that did not administer the hydrothermal extract of shiitake mushrooms to the rat was 0.008mm / g, compared to the feed intake The difference in iliac bone weight was 0.154 mg / g, and it was found that shiitake mushroom hot water extract was effective in the growth of iliac bone length and iliac bone weight in vivo.
상기 표5 및 도6에서 확인할 수 있는 바와 같이, 4주 동안의 기간 동안 표고버섯의 열수 추출물을 쥐에게 투여하지 않은 대조군과 투여한 실험군의 혈중 성장호르몬 수치의 차이는 각각 67.9ng/㎗, 262.6ng/㎗로서 혈중 성장호르몬의 수치가 표고버섯 열수 추출물을 투여한 실험군에서 월등히 증가함을 알 수 있었다.As can be seen in Table 5 and Figure 6, the difference in blood growth hormone levels of the control group and the experimental group that did not receive the hydrothermal extract of shiitake mushrooms in the mice for 4 weeks was 67.9ng / ㎗, 262.6 As ng / ㎗, blood growth hormone levels were significantly increased in the experimental group administered with shiitake hot water extract.
이상의 상기 시험예 1, 2, 3 및 4의 결과를 종합해 보면, 표고버섯 열수 추출물은 조골세포에서 IGF-1 수치를 대조군에 비하여 증가시켰으며 6일간 쥐에 350 ㎎/kg을 투여한 경우, 경골의 성장판 길이 및 골길이 성장을 유의적으로 증진시켰고, 또한 4주간 쥐에 350 ㎎/kg의 표고버섯 열수 추출물을 투여한 경우, 장골의 길이 및 장골의 무게를 유의적으로 증진시켰으며 혈중 성장호르몬 수치 또한 대조군에 비해서 증가되어 있는 것을 확인할 수 있었다. 이러한 결과들은 표고버섯 열수 추출물이 성장기 골길이 성장을 촉진하는 효과가 있음을 보여준다고 할 수 있다.In summary, the results of Test Examples 1, 2, 3 and 4 above, the shiitake mushroom hot water extract increased the IGF-1 level in osteoblasts compared to the control group, and the rats were treated with 350 mg / kg for 6 days. Tibial plate length and bone length growth were significantly increased. Also, when 350 mg / kg of shiitake hot water extract was administered to rats for 4 weeks, the length of the long bone and the weight of the long bone were significantly increased. Hormone levels were also increased compared to the control. These results show that shiitake mushroom hot water extract has an effect of promoting growth of bone length.
<실시예 2><Example 2>
표고버섯 열수 추출물을 이용한 약학적 조성물 제조Preparation of pharmaceutical composition using shiitake mushroom hot water extract
본 발명의 표고버섯 열수추출물을 유효 성분으로 포함하는 약학 조성물의 제제예를 설명하나, 본 발명은 이를 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다.The preparation example of the pharmaceutical composition comprising the shiitake mushroom hot water extract of the present invention as an active ingredient will be described, but the present invention is not intended to be limited thereto but only to be described in detail.
캡슐제의 제조Preparation of Capsules
표고버섯 열수 추출물 시료 100㎎, 옥수수 전분 100㎎, 유당 100㎎, 스테아린산 마그네슘 2㎎을 완전히 혼합한 후 통상의 캡슐제의 제조 방법에 따라서 경질 젤라틴 캡슐에 충전하여 캡슐제를 제조하였다.100 mg of shiitake hot water extract sample, 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate were thoroughly mixed and then filled into hard gelatin capsules according to a conventional method for preparing capsules to prepare capsules.
산제의 제조Manufacture of powder
표고버섯 열수 추출물 시료 2g, 유당 1g을 완전히 혼합한 후 기밀포에 충진하여 통상의 산제의 제조 방법에 따라 산제를 제조하였다.2 g of shiitake mushroom hot water extract samples and 1 g of lactose were completely mixed, and then filled in an airtight cloth to prepare a powder according to a conventional powder preparation method.
<실시예 3><Example 3>
표고버섯 열수 추출물을 이용한 발효유 제조Preparation of Fermented Milk Using Hot Water Extract of Shiitake Mushroom
탈지분유로 무지유고형분 함량을 8~20중량%로 조정한 원료유를 72~75℃에서 15초간 살균하였다. 살균된 원료유를 37℃로 냉각시킨 후 락토바실러스 카제이 HY 2782를 106cfu/㎖의 농도로 접종하여 배양액이 pH 4~5가 될 때까지 37℃에서 배양하고 냉각시켰다. 시럽은 과즙 농축액 0.1~50중량%, 식이섬유 0.1~20중량%, 포도당 0.5~30중량%, 올리고당 0.1~15중량%, 칼슘 0.001~10 중량%, 비타민 0.0001~5중량% 및 표고버섯 열수 추출물 0.01~5중량%를 녹여 제조하고 살균한 뒤 냉각하였다. 이렇게 제조된 시럽과 상기 배양액을 1:3의 부피비율로 혼합, 교반하여 균질화한 뒤 용기에 포장하여 발효유를 제조하였다. 제조된 발효유는 관능검사 결과 풍미, 물성, 전체적인 맛에서 양호한 결과를 보였다.Crude milk was sterilized for 15 seconds at 72 ~ 75 ℃ to adjust the nonfat milk solid content to 8 to 20% by weight. The sterilized crude oil was cooled to 37 ° C., and then inoculated with Lactobacillus casei HY 2782 at a concentration of 10 6 cfu / ml, and then cultured and cooled at 37 ° C. until the culture solution had a pH of 4-5. Syrup is 0.1-50% by weight of juice concentrate, 0.1-20% by weight of dietary fiber, 0.5-30% by weight of glucose, 0.1-15% by weight of oligosaccharide, 0.001-10% by weight of calcium, 0.0001-5% by weight of shiitake mushroom It was prepared by dissolving 0.01-5% by weight, sterilizing and cooling. The syrup and the culture solution thus prepared were mixed and stirred at a volume ratio of 1: 3, homogenized, and then packaged in a container to prepare fermented milk. The produced fermented milk showed good results in flavor, physical properties and overall taste.
<실시예 4><Example 4>
표고버섯 열수 추출물을 이용한 음료 제조Beverage Preparation Using Shiitake Mushroom Hot Water Extract
표고버섯 열수 추출물 14중량%, 대추엑기스 1.5중량% 및 당귀농축액 7중량%에 액상과당 7.4중량%를 혼합하고 여기에 정제수를 첨가, 혼합하여 드링크용 병에 충진하여 90~95℃에서 살균한 후 실온으로 냉각하여 음료를 제조하였다.14% by weight of Shiitake hot water extract, jujube extract 1.5%, and 7% by weight of Angelica Concentrate were mixed with 7.4% by weight of liquid fructose, and purified water was added and mixed into the drink bottle and sterilized at 90-95 ° C. The beverage was prepared by cooling to room temperature.
<실시예 5>Example 5
표고버섯 열수 추출물을 이용한 건강보조식품 제조Preparation of Health Supplements Using Shiitake Mushroom Hot Water Extract
표고버섯 열수 추출물 0.01~30중량%에 영양보조성분(비타민 B1, B2, B5, B6, E 및 초산에스테르, 니코틴산 아미드), 올리고당, 50% 에탄올을 상기의 표고버섯 열수 추출물 100중량%에 대하여 5~10중량%가 되도록 첨가하여 고속회전 혼합기에서 혼합하였다. 상기 혼합물에 멸균 정제수 10중량%를 첨가, 혼합하고 직경 1~2 mm의 과립상으로 성형하였다. 상기 성형된 과립은 40℃~50℃의 진공건조기에서 건조시킨 후, 12~14 메쉬(mesh)를 통과시켜 균일하게 과립을 제조하였다. 상기와 같이 제조된 과립은 적당량씩 압출 성형되어 정제 또는 분말로 되거나 경질캡슐에 충전되어 경질캡슐제품으로 제조될 수 있다.0.01 ~ 30% by weight of shiitake hot water extract, nutritional supplements (vitamins B1, B2, B5, B6, E and acetate esters, nicotinic acid amide), oligosaccharides, 50% ethanol 5 to 100% by weight of shiitake hot water extract It was added to ˜10% by weight and mixed in a high speed rotary mixer. 10% by weight of sterile purified water was added to the mixture, mixed, and molded into granules having a diameter of 1 to 2 mm. The molded granules were dried in a vacuum dryer at 40 ° C.-50 ° C., and then uniformly prepared granules were passed through 12 to 14 mesh. The granules prepared as described above may be extruded by appropriate amounts into tablets or powders or filled into hard capsules to produce hard capsule products.
이상에서 살펴 본 바와 같이, 본 발명의 표고버섯 열수 추출물은 성장 호르몬 및/또는 IGF-1의 발현을 증가시킴으로서 성장기 골길이 성장을 촉진하는 작용이 탁월하여 성장기 골길이 성장 도움 및 골길이 성장장애의 치료 및 예방을 목적으로 유제품, 건강식품 및 음료로 이용될 수 있다.As described above, the shiitake mushroom hydrothermal extract of the present invention has an excellent effect of promoting growth of bone length by increasing the expression of growth hormone and / or IGF-1, thus helping to grow bone length and bone length growth disorder. It can be used as dairy products, health foods and beverages for the purpose of treatment and prevention.
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