KR102041852B1 - Composition for preventing, improving or treating fatty liver disease comprising Oxya chinensis extract as effective component - Google Patents
Composition for preventing, improving or treating fatty liver disease comprising Oxya chinensis extract as effective component Download PDFInfo
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- KR102041852B1 KR102041852B1 KR1020170181811A KR20170181811A KR102041852B1 KR 102041852 B1 KR102041852 B1 KR 102041852B1 KR 1020170181811 A KR1020170181811 A KR 1020170181811A KR 20170181811 A KR20170181811 A KR 20170181811A KR 102041852 B1 KR102041852 B1 KR 102041852B1
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- fatty liver
- liver disease
- rice locust
- prevention
- rice
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- A—HUMAN NECESSITIES
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- 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
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K35/56—Materials from animals other than mammals
- A61K35/63—Arthropods
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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
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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
- A23V2250/00—Food ingredients
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Abstract
본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 지방간 질환의 예방, 개선 또는 치료용 조성물에 관한 것으로, 본 발명의 유효성분인 벼메뚜기 추출물이 간암 세포주(HepG2)에서 중성지방의 축적을 효과적으로 감소시키는 효과가 있으며, 벼메뚜기 추출물의 투여에 따른 동물 모델의 체중변화; 간, 피하지방, 신장지방 조직 및 소장 지방조직의 무게 변화; 지질생화학적 지표들의 변화가 유의하게 나타나는 것을 확인하였다. 따라서 본 발명의 조성물은 지방간 질환의 예방, 개선 또는 치료를 위한 건강기능식품, 의약품 또는 사료첨가제로 유용하게 사용될 수 있다.The present invention relates to a composition for the prevention, improvement or treatment of fatty liver disease comprising rice locust extract as an active ingredient, the effective effect of the rice locust extract of the present invention to effectively reduce the accumulation of triglycerides in liver cancer cell line (HepG 2 ) There is, the weight change of the animal model according to the administration of rice locust extract; Weight changes in liver, subcutaneous fat, kidney fat tissue and small intestine adipose tissue; It was confirmed that the change of the lipid biochemical markers appeared significantly. Therefore, the composition of the present invention can be usefully used as a dietary supplement, medicine or feed additive for the prevention, improvement or treatment of fatty liver disease.
Description
본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 지방간 질환의 예방, 개선 또는 치료용 조성물에 관한 것이다.The present invention relates to a composition for the prevention, improvement or treatment of fatty liver disease comprising rice locust extract as an active ingredient.
국내 간질환 사망률은 인구 십만명 당 23.5명으로 매우 높으며, 40대 사망원인 1위(41.1명/10만 명), 50대 사망원인 2위(72.4명/10만 명), 30대 사망원인 3위(10명/10만 명)를 차지하는 등 간질환은 한국 중년층 인구의 주요 사망원인이다. The mortality rate of liver disease in Korea is very high, with 23.5 deaths per 100,000 population, the number one cause of death in the 40s (41.1 / 100,000), the second cause of death in the 50s (72.4 / 10 million), and the third cause of death in the 30s. Liver disease is the leading cause of death among the Korean middle-aged population.
상기 간질환 중에서 지방간은 정상세포 내에는 존재하지 않는 중성지방이 간 세포 내에 비정상적으로 침착되어 보이는 현상이 나타난 것을 말한다. 정상 간은 약 5%가 지방조직으로 구성되어 있으며 중성지방, 지방산, 인지질, 콜레스테롤 및 콜레스테롤 에스터가 지방의 주요 성분이나, 일단 지방간이 발생하면 대부분의 성분이 중성지방으로 대체되며 중성지방의 양이 간 중량의 5% 이상이면 지방간으로 진단된다. 지방간이 악화되어 간세포 속의 지방 덩어리가 커지면 핵을 포함한 세포의 중요한 구성성분이 한쪽으로 밀려 간세포의 기능이 저하되며, 세포 내에 축적된 지방으로 인하여 팽창된 간세포들이 간세포 사이에 있는 미세 혈관과 임파선을 압박하여 간 내의 혈액과 임파액의 순환에 장애가 생기게 된다. 이렇게 되면 간세포는 산소와 영양공급을 적절히 받을 수 없어 간기능이 저하되는 것이다. Among the liver diseases, fatty liver refers to a phenomenon in which neutral fat, which does not exist in normal cells, appears abnormally deposited in liver cells. About 5% of normal liver is composed of fatty tissue, and triglycerides, fatty acids, phospholipids, cholesterol, and cholesterol esters are the major components of fat, but once fatty liver occurs, most of the components are replaced with triglycerides. If more than 5% of the weight of the liver is diagnosed as fatty liver. Fatty liver deteriorates and the mass of fat in the hepatocytes is pushed to important components of the cells, including the nucleus, to one side, reducing the function of the hepatocytes, and due to the accumulation of fat in the cells, the expanded hepatocytes squeeze the microvascular and lymph glands between the hepatocytes. This leads to impaired circulation of blood and lymph in the liver. If this happens, the liver cells can not receive adequate oxygen and nutrition, liver function is reduced.
비알콜성 지방간 질환(non-alcoholic fatty liver disease, NAFLD)은 알코올에 의한 간 손상이 아니라, 지방산이 중성지방의 형태로 간의 실질세포 내에 5% 이상 축적된 경우로 정의한다. 병리학적으로는 단순 지방간(simple steatosis)과 염증을 동반한 지방간염(steatohepatitis)으로 분류되는데, 장기간 방치 시, 간염, 간 섬유, 간경변 등의 심각한 간 질환으로 이행될 수 있다. 국내에서도 생활양식의 변화로 인해 비알코올성 간질환 발생빈도가 증가하는 추세이다.Non-alcoholic fatty liver disease (NAFLD) is not defined as alcohol damage to liver, but is defined as fatty acids accumulated in the parenchymal cells of the liver more than 5% in the form of triglycerides. Pathologically, it is classified as simple steatosis and steatohepatitis with inflammation, which can lead to serious liver diseases such as hepatitis, liver fibers, and cirrhosis of the liver for a long time. In Korea, the incidence of nonalcoholic liver disease is increasing due to lifestyle changes.
또한, 알콜성 지방간은 과다한 음주로 인해 발생하는데, 개인별로 유전적인 특징, 성별에 따라 차이가 있지만, 하루 80g 이상의 알코올의 섭취를 하는 경우 알콜성 지방간과 같은 간질환이 발생할 가능성이 크다. 여성의 경우 이보다 적은 양으로도 알코올성 간질환이 발생할 가능성이 커진다. 일반적으로 소주 1잔, 맥주 1잔, 양주 1잔, 막걸리 1홉에 약 10g의 알코올이 포함된 것으로 계산할 수 있다. 증상으로는 가장 경미한 형태인 알코올성 지방간 환자들의 경우 대부분 무증상이지만, 경미한 간비대(간이 정상보다 크게 된 상태)가 나타난 경우 오른쪽 상복부에 가벼운 압통을 호소할 수 있다.In addition, alcoholic fatty liver is caused by excessive drinking, the genetic characteristics of each person, depending on gender, but if you consume more than 80g of alcohol per day, liver disease such as alcoholic fatty liver is likely to occur. For women, even lower amounts are more likely to develop alcoholic liver disease. In general, a glass of soju, a glass of beer, a glass of liquor, 1 hop of makgeolli can be calculated to contain about 10g of alcohol. Symptoms are mostly asymptomatic in the mildest form of alcoholic fatty liver, but minor hepatomegaly (liver larger than normal) may cause mild tenderness in the right upper abdomen.
한편, 벼메뚜기는 몸길이 30~38mm로, 몸빛깔은 황록색이나 머리와 가슴은 황갈색이다. 겹눈은 달걀 모양이고 광택이 있는 회갈색이다. 앞가슴등판에 가느다란 3개의 가로 홈이 있고 양쪽에 갈색의 세로줄이 있다. 날개는 황갈색이고 배 끝보다 길지만 별로 날아다니지 않는다. 논이나 경작지 근처의 풀밭에 서식한다. 연 1회 발생하며, 땅 속에서 알무더기로 월동한다. 알무더기는 아교질의 엷은 막으로 싸여 있다. 다 자란 약충은 날개딱지가 제4배 마디에 이르며 몸길이는 약 30mm이다. 벼의 주요 해충으로 농약을 사용하면서 그 수가 줄어들었으나 차차 늘어나고 있다. 1930년 황해도에서 발생해 알 덩어리를 정부에서 사들인 일도 있다. 한국·일본·중국 등지에 분포한다. On the other hand, rice locusts are 30 ~ 38mm long, with yellowish green color but yellowish brown hair and chest. Compound eyes are oval, glossy greyish brown. There are three narrow grooves on the pronotum and brown vertical lines on both sides. The wings are yellowish brown and longer than the tip of the ship, but do not fly very much. Inhabits grasses near rice fields or farmland. Occurs once a year, wintering in the ground. The pellets are wrapped in a gelatinous membrane. The mature nymphs have the fourth scab, and the body length is about 30mm. The number of pesticides used as a major pest of rice has decreased, but gradually increased. It occurred on the Yellow Sea in 1930 and the egg masses were bought by the government. It is distributed in Korea, Japan, and China.
한국등록특허 제1648812호에 벼메뚜기 성분을 포함하는 다이어트 식품 조성물 제조 방법에 관하여 개시되어 있고, 한국공개특허 제2010-0002661호에 히알루론산을 포함하는 기능성 복합체 및 그 제조방법에 관하여 개시되어 있으나, 본 발명의 벼메뚜기 추출물을 유효성분으로 포함하는 지방간 질환의 예방, 개선 또는 치료용 조성물에 대해서는 개시된 바 없다.Korean Patent No. 1648612 discloses a method for producing a diet food composition comprising a rice locust component, and Korean Patent Publication No. 2010-0002661 discloses a functional complex including hyaluronic acid and a method for preparing the same, There is no disclosure about the composition for the prevention, improvement or treatment of fatty liver disease comprising the rice locust extract of the invention as an active ingredient.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 지방간 질환의 예방, 개선 또는 치료용 조성물을 제공하고, 본 발명의 벼메뚜기 추출물을 처리한 간세포에서 중성지방의 함량이 효과적으로 감소하는 것과 벼메뚜기 추출물의 투여에 따른 동물 모델의 체중변화; 간, 피하지방, 신장지방 조직 및 소장 지방조직의 무게 변화; 지질생화학적 지표들이 유의미하게 변화한다는 것을 확인함으로써, 본 발명을 완성하였다.The present invention is derived from the above requirements, the present invention provides a composition for the prevention, improvement or treatment of fatty liver disease comprising rice locust extract as an active ingredient, triglycerides in liver cells treated with the rice locust extract of the present invention Weight loss of animal model according to the effective decrease of the content of and the administration of rice locust extract; Weight changes in liver, subcutaneous fat, kidney fat tissue and small intestine adipose tissue; The present invention was completed by confirming that the lipid biochemical indicators changed significantly.
상기 목적을 달성하기 위하여, 본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 지방간 질환의 예방 또는 개선용 건강기능식품 조성물을 제공한다.In order to achieve the above object, the present invention provides a dietary supplement for the prevention or improvement of fatty liver disease comprising rice locust extract as an active ingredient.
또한, 본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 지방간 질환의 예방 또는 치료용 약학 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of fatty liver disease comprising rice locust extract as an active ingredient.
또한, 본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 인간을 제외한 동물의 지방간 질환의 예방 또는 치료용 수의학적 조성물을 제공한다.The present invention also provides a veterinary composition for preventing or treating fatty liver disease in animals other than humans, including rice locust extract as an active ingredient.
또한, 본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 인간을 제외한 동물의 지방간 질환의 예방 또는 개선용 사료 첨가제를 제공한다.In addition, the present invention provides a feed additive for the prevention or improvement of fatty liver disease in animals other than humans, including rice locust extract as an active ingredient.
또한, 본 발명은 벼메뚜기 추출물을 인간을 제외한 개체에게 투여하는 단계를 포함하는 지방간 질환의 예방 또는 치료방법을 제공한다.The present invention also provides a method for preventing or treating fatty liver disease, comprising administering rice locust extract to an individual except a human.
본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 지방간 질환의 예방, 개선 또는 치료용 조성물에 관한 것으로, 본 발명의 유효성분인 벼메뚜기 추출물은 식용 가능한 것으로 알려진 벼메뚜기가 원료이므로, 부작용 및 독성이 없을 뿐만 아니라, 간암 세포주(HepG2)에서 중성지방의 축적을 효과적으로 감소시키고, 벼메뚜기 추출물의 투여에 따른 동물 모델의 체중변화; 간, 피하지방, 신장지방 조직 및 소장 지방조직의 무게 변화; 지질생화학적 지표들의 변화가 유의하게 나타났으므로, 본 발명의 조성물은 지방간 질환의 예방, 개선 또는 치료를 위한 건강기능식품, 의약품 또는 사료첨가제로 유용하게 사용될 수 있다.The present invention relates to a composition for the prevention, improvement or treatment of fatty liver disease comprising rice locust extract as an active ingredient, rice locust extract as an active ingredient of the present invention is a raw rice locust known to be edible, as well as no side effects and toxicity , Effectively reduce the accumulation of triglycerides in liver cancer cell line (HepG 2 ), body weight change of the animal model following the administration of rice locust extract; Weight changes in liver, subcutaneous fat, kidney fat tissue and small intestine adipose tissue; Since the changes in the lipid biochemical indicators have been significant, the composition of the present invention can be usefully used as a dietary supplement, medicine or feed additive for the prevention, improvement or treatment of fatty liver disease.
도 1은 간암 세포주(HepG2)에 올레이트 및 팔미테이트가 2:1의 농도비로 혼합된 1.0mM의 유리지방산 및 벼메뚜기 추출물을 처리한 후, 중성지방의 함량을 나일레드(아디포레드) 염색하여 측정한 결과이다. ####은 1.0mM의 유리 지방산을 처리하지 않은 음성대조군(Control)에 비해 1.0mM의 유리 지방산을 처리한 군에서 나일레드 발현(%)이 통계적으로 유의미하게 증가하였다는 것으로, p<0.0001임을 의미하며, ****은 유리지방산 처리군에 비해 유리지방산과 벼메뚜기 추출물을 함께 처리한 군에서, 나일레드 발현이 통계적으로 의미있게 감소하였다는 것을 의미하며, p<0.0001이다.
도 2는 간암 세포주(HepG2)에 올레이트 및 팔미테이트가 2:1의 농도비로 혼합된 1.0mM의 유리지방산 및 벼메뚜기 추출물을 처리한 후, 총 중성지방의 함량을 측정한 결과이다. ####은 1.0mM의 유리 지방산을 처리하지 않은 음성대조군(Control)에 비해 1.0mM의 유리 지방산을 처리한 군에서, 총 중성지방(mg/dl)이 통계적으로 유의미하게 증가하였다는 것으로, p<0.0001임을 의미하며, ***, ****은 유리지방산 처리군에 비해 유리지방산과 벼메뚜기 추출물을 함께 처리한 군에서 총 중성지방(mg/dl)이 통계적으로 의미있게 감소하였다는 것을 의미하며, ***은 p<0.001이고, ****은p<0.0001이다.
도 3은 간암 세포주(HepG2)에 올레이트 및 팔미테이트가 2:1의 농도비로 혼합된 1.0mM의 유리지방산 및 벼메뚜기 추출물을 처리한 후, 지방 흡수율(lipid uptake)을 측정한 결과이다. ####은 1.0mM의 유리 지방산을 처리하지 않은 음성대조군(Control)에 비해 1.0mM의 유리 지방산을 처리한 군에서, 상대적 지방 흡수율이 통계적으로 유의미하게 증가하였다는 것으로, p<0.0001임을 의미하며, ****은 유리지방산 처리군에 비해 유리지방산과 벼메뚜기 추출물을 함께 처리한 군에서 상대적 지방 흡수율이 통계적으로 의미있게 감소하였다는 것을 의미하며, p<0.0001이다.
도 4는 간암 세포주(HepG2)에 에탄올 및 벼메뚜기 추출물을 처리한 후, 세포생존률(%)을 확인한 결과이다. ####은 1.0mM의 유리 지방산을 처리하지 않은 음성대조군(Control)에 비해 1.0mM의 유리 지방산을 처리한 군에서, 세포생존률(%)이 통계적으로 유의미하게 감소하였다는 것으로, p<0.0001임을 의미하며, **, ****은 유리지방산 처리군에 비해 유리지방산과 벼메뚜기 추출물을 함께 처리한 군에서 세포생존률(%)이 통계적으로 의미있게 증가하였다는 것을 의미하며, **은 p<0.01이고, ****은 p<0.0001이다.
도 5는 간암 세포주(HepG2)에 에탄올 및 벼메뚜기 추출물을 처리한 후, Caspase-3의 발현량을 확인한 결과이다. ####은 1.0mM의 유리 지방산을 처리하지 않은 음성대조군(Control)에 비해 1.0mM의 유리 지방산을 처리한 군에서, 상대적 Caspase-3의 발현량이 통계적으로 유의미하게 증가하였다는 것으로, p<0.0001임을 의미하며, ****은 유리지방산 처리군에 비해 유리지방산과 벼메뚜기 추출물을 함께 처리한 군에서 상대적 Caspase-3의 발현량이 통계적으로 의미있게 감소하였다는 것을 의미하며, p<0.0001이다.
도 6은 고지방 식이에 의해 증가한 체중이 벼메뚜기 추출물의 투여에 의해 감소하는 것을 확인한 결과이다.
도 7은 고지방 식이에 의해 증가한 마우스의 간, 피하지방, 신장지방, 소장지방 중량이 벼메뚜기 추출물의 투여에 의해 감소하는 것을 확인한 결과이다. ####은 정상식이군(Nor)에 비해 고지방식이군(HFD)의 간, 피하지방, 신장지방조직 및 소장 지방조직의 중량이 통계적으로 유의미하게 증가하였다는 것으로, p<0.0001임을 의미하며, *, *** 및 ****는 고지방식이군에 비해 고지방방식이 및 벼메뚜기 추출물(MTG)을 모두 섭취한 군이 고지방식이군에 비해 통계적으로 유의미하게 간, 피하지방, 신장지방조직 및 소장지방조직의 중량이 감소하였다는 것으로, *는 p<0.05이고, ***는 p<0.001이며, ****는 p<0.0001임을 의미하고, M100은 양성대조군으로, 밀크씨슬이다.
도 8은 고지방 식이에 의해 증가한 마우스의 혈액내 ALT 및 AST 함량이 벼메뚜기 추출물의 투여에 의해 감소하는 것을 확인한 결과이다. ##, ####은 정상식이군(Nor)에 비해 고지방식이군(HFD)의 GPT(ALT) 및 GOT(AST)가 통계적으로 유의미하게 증가하였다는 것으로, ##은 p<0.01이고, ####은 p<0.0001임을 의미하며, *, *** 및 ****는 고지방식이군에 비해 고지방방식이 및 벼메뚜기 추출물(MTG)을 모두 섭취한 군이 고지방식이군에 비해 통계적으로 유의미하게 GPT(ALT) 및 GOT(AST)의 함량이 감소하였다는 것으로, *는 p<0.05이고, ***는 p<0.001이며, ****는 p<0.0001임을 의미하고, M100은 양성대조군으로, 밀크씨슬이다.
도 9는 고지방 식이에 의해 증가한 마우스의 혈액 내 중성지방, 총콜레스테롤, 저밀도지단백, 유리지방산의 농도가 벼메뚜기 추출물의 투여에 의해 감소하는 것을 확인한 결과이다. ####은 정상식이군(Nor)에 비해 고지방식이군(HFD)의 중성지방, 총콜레스테롤, 저밀도지단백 및 유리지방산의 농도가 통계적으로 유의미하게 증가하였다는 것으로, p<0.0001임을 의미하며, *, ** 및 ***는 고지방식이군에 비해 고지방방식이 및 벼메뚜기 추출물(MTG)을 모두 섭취한 군이 고지방식이군에 비해 통계적으로 유의미하게 중성지방, 총콜레스테롤, 저밀도지단백 및 유리지방산의 농도가 감소하였다는 것으로, *는 p<0.05이며, **는 p<0.01이고, ***는 p<0.001임을 의미하고, M100은 양성대조군으로, 밀크씨슬이다.
도 10은 고지방 식이에 의해 증가한 마우스의 혈당 농도가 벼메뚜기 추출물의 투여에 의해 감소하는 것을 확인한 결과이다. ####은 정상식이군(Nor)에 비해 고지방식이군(HFD)의 혈청 내 혈당 농도가 통계적으로 유의미하게 증가하였다는 것으로, p<0.0001임을 의미하며, * 및 ***는 고지방식이군에 비해 고지방방식이 및 벼메뚜기 추출물(MTG)을 모두 섭취한 군이 고지방식이군에 비해 통계적으로 유의미하게 혈청 내 혈당 농도가 감소하였다는 것으로, *는 p<0.05이고, ***는 p<0.001임을 의미하고, M100은 양성대조군으로, 밀크씨슬이다.1 is treated with 1.0 mM free fatty acid and rice locust extract, in which oleate and palmitate are mixed at a concentration ratio of 2: 1 in liver cancer cell lines (HepG 2 ), and the content of triglycerides is stained with nile (adifored). It is the result measured. #### is a statistically significant increase in Nile expression (%) in the group treated with 1.0 mM free fatty acid compared to the control group without treatment with 1.0 mM free fatty acid, p <0.0001 **** indicates that the NAIL expression was significantly decreased in the group treated with the free fatty acid and the rice locust extract, compared to the free fatty acid treatment group, and p <0.0001.
2 is a result of measuring the total triglyceride content after treatment with 1.0 mM free fatty acid and rice locust extract mixed with oleate and palmitate at a concentration ratio of 2: 1 in liver cancer cell line (HepG 2 ). #### is a statistically significant increase in total triglycerides (mg / dl) in the group treated with 1.0 mM free fatty acid compared to the control group without treatment with 1.0 mM free fatty acid. p <0.0001, and *** and **** indicate that the total triglycerides (mg / dl) were significantly decreased in the group treated with free fatty acid and rice locust extract compared to the free fatty acid treated group. Where *** is p <0.001 and **** is p <0.0001.
Figure 3 is a result of measuring the lipid uptake after treatment of 1.0 mM free fatty acid and rice locust extract mixed with oleate and palmitate at a concentration ratio of 2: 1 in liver cancer cell line (HepG 2 ). #### represents a statistically significant increase in the relative fat absorption rate in the group treated with 1.0 mM free fatty acid compared to the control group without treatment with 1.0 mM free fatty acid, indicating p <0.0001. And **** means that the relative fat absorption rate was statistically decreased in the group treated with the free fatty acid and the rice locust extract, compared to the free fatty acid treatment group, and p <0.0001.
4 is a result of confirming the cell survival rate (%) after treating ethanol and rice locust extract in liver cancer cell line (HepG 2 ). #### is a statistically significant decrease in cell survival rate (%) in the group treated with 1.0 mM free fatty acid compared to the control group without treatment with 1.0 mM free fatty acid, p <0.0001 **, **** means that the cell survival rate (%) increased significantly in the group treated with free fatty acid and rice locust extract compared to the free fatty acid treatment group, ** is p <0.01 and **** are p <0.0001.
5 is a result of confirming the expression level of Caspase-3 after treating ethanol and rice locust extract in liver cancer cell line (HepG 2 ). #### is a statistically significant increase in the relative expression of Caspase-3 in the group treated with 1.0 mM free fatty acid compared to the control group without treatment with 1.0 mM free fatty acid. **** indicates that the expression of Caspase-3 in the group treated with free fatty acid and rice locust extract was significantly decreased compared to the free fatty acid treatment group, and p <0.0001.
6 is a result confirming that the weight increased by the high fat diet is reduced by the administration of rice locust extract.
7 is a result confirming that the weight of the liver, subcutaneous fat, kidney fat, and small intestine fat increased by the administration of rice locust extract increased in the high fat diet. #### means that the weight of liver, subcutaneous fat, renal adipose tissue and small intestine adipose tissue of HFD group was significantly higher than that of normal diet group (Nor), which means p <0.0001. *, *** and **** are significantly higher in liver, subcutaneous fat, renal adipose tissue and small intestine in the group that consumed both high fat diet and rice locust extract (MTG) compared to the high fat diet group. The weight of the adipose tissue decreased, * means p <0.05, *** means p <0.001, **** means p <0.0001, M100 is a positive control, milk thistle.
8 is a result confirming that the ALT and AST content in the blood of the mice increased by the high fat diet decreased by the administration of the rice locust extract. ##, #### is a statistically significant increase in GPT (ALT) and GOT (AST) of the high-fat diet group (HFD) compared to the normal diet group (Nor), where ## is p <0.01, # ### means p <0.0001, and *, ***, and **** are statistically significant in the high fat diet and rice locust extract (MTG) groups compared to the high fat diet group. Therefore, the contents of GPT (ALT) and GOT (AST) were decreased, * means p <0.05, *** means p <0.001, **** means p <0.0001, and M100 means positive control. It is milk thistle.
9 is a result confirming that the concentration of triglycerides, total cholesterol, low density lipoprotein, and free fatty acid in the blood of mice increased by high fat diet decreased by administration of rice locust extract. #### means that the levels of triglyceride, total cholesterol, low density lipoprotein and free fatty acid in HFD were significantly increased compared to normal diet (Nor), which means p <0.0001. , ** and *** are significantly higher in triglycerides, total cholesterol, low-density lipoprotein, and free fatty acids in the high-fat diet and rice locust extract (MTG) than the high-fat diet. Is reduced, * means p <0.05, ** means p <0.01, *** means p <0.001, and M100 is a positive control and milk thistle.
10 is a result confirming that the blood glucose concentration of the mouse increased by the high fat diet is reduced by the administration of rice locust extract. #### means that the blood glucose level in the HFD group was significantly increased compared to the normal diet group (Nor), which means that p <0.0001, and * and *** were higher in the high-fat diet group. Compared to the high fat diet group, the high-fat diet and the rice locust extract (MTG) showed statistically significant decreases in serum glucose levels, where * was p <0.05 and *** was p <0.001. M100 is a positive control, milk thistle.
본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 지방간 질환의 예방 또는 개선용 건강기능식품 조성물에 관한 것이다.The present invention relates to a dietary supplement for the prevention or improvement of fatty liver disease comprising rice locust extract as an active ingredient.
상기 벼메뚜기 추출물은 하기의 단계를 포함하는 방법에 의해 제조할 수 있으나, 이에 한정하지 않는다:The rice locust extract may be prepared by a method comprising the following steps, but is not limited thereto:
(1) 벼메뚜기에 추출용매를 가하여 추출하는 단계;(1) extracting by adding an extraction solvent to the rice grasshopper;
(2) 단계 (1)의 추출물을 여과하는 단계; 및 (2) filtering the extract of step (1); And
(3) 단계 (2)의 여과한 추출물을 감압 농축하고 건조하여 추출물을 제조하는 단계. (3) concentration of the filtered extract of step (2) under reduced pressure and drying to prepare an extract.
상기 단계 (1)에서 추출용매는 물, C1~C4의 저급 알코올 또는 이들의 혼합물 중에서 선택하는 것이 바람직하며, 더 바람직하게는 C1~C4의 저급 알코올이고, 더욱더 바람직하게는 70%(v/v) 에탄올이지만 이에 한정하지 않는다.The extraction solvent in step (1) is preferably selected from water, lower alcohols of C 1 ~ C 4 or mixtures thereof, more preferably lower alcohols of C 1 ~ C 4 , even more preferably 70% (v / v) ethanol but not limited thereto.
상기 제조방법에 있어서, 추출방법은 여과법, 열수 추출, 침지 추출, 환류 냉각 추출 및 초음파 추출 등의 당 업계에 공지된 모든 통상적인 방법을 이용할 수 있다. 상기 추출용매는 건조된 벼메뚜기 중량의 1~20배 첨가하여 추출하는 것이 바람직하며, 더 바람직하게는 5~15배 첨가하는 것이다. 추출온도는 4~50℃인 것이 바람직하나 이에 한정하지 않는다. 또한, 추출시간은 0.5~10시간인 것이 바람직하며, 0.5~5시간이 더욱 바람직하나 이에 한정하지 않는다. 상기 방법에 있어서, 단계 (3)의 감압농축은 진공 감압 농축기 또는 진공회전증발기를 이용하는 것이 바람직하나 이에 한정하지 않는다. 또한, 건조는 감압건조, 진공건조, 비등건조, 분무 건조 또는 동결 건조하는 것이 바람직하나 이에 한정하지 않는다.In the above production method, the extraction method may use all conventional methods known in the art, such as filtration, hot water extraction, immersion extraction, reflux cooling extraction and ultrasonic extraction. The extraction solvent is preferably extracted by adding 1 to 20 times the weight of the dried rice locust, more preferably 5 to 15 times. Extraction temperature is preferably 4 ~ 50 ℃ but is not limited thereto. In addition, the extraction time is preferably 0.5 to 10 hours, more preferably 0.5 to 5 hours, but is not limited thereto. In the above method, the vacuum concentration in step (3) is preferably used as a vacuum vacuum concentrator or a vacuum rotary evaporator, but not always limited thereto. In addition, the drying is preferably reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto.
상기 지방간 질환은 비알콜성 또는 알콜성 단순 지방간; 비알콜성 또는 알콜성 지방간염; 및 비알콜성 또는 알콜성 간경변 중에서 선택된 어느 하나인 것이 바람직하지만 이에 한정하는 것은 아니다. The fatty liver disease is non-alcoholic or alcoholic simple fatty liver; Nonalcoholic or alcoholic fatty hepatitis; And non-alcoholic or alcoholic cirrhosis is preferred, but is not limited thereto.
상기 벼메뚜기 추출물은 간 조직 내 중성지방의 함량을 감소시키는 것이 특징이며, 간 조직 내 총 중성지방의 함량을 감소시키며, 지방 흡수율(lipid uptake)을 감소시키는 것이 특징이다.The rice locust extract is characterized by reducing the content of triglycerides in liver tissue, reducing the content of total triglycerides in liver tissue, characterized by reducing the fat uptake (lipid uptake).
상기 본 발명의 조성물을 지방간 질환의 예방 또는 개선을 목적으로 식품에 첨가하여 건강기능성 식품을 제조할 수 있는데, 상기 건강기능 식품은 특별히 이에 제한되지 않으며, 건강 기능성 식품, 영양 보조제, 영양제, 파머푸드(pharmafood), 건강 식품, 뉴트라슈티칼(nutraceutical), 디자이너 푸드, 식품 첨가제 등의 모든 형태의 식품이 될 수 있는데, 바람직하게는 육류, 소세지, 빵, 쵸코렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 될 수 있다. 이때, 첨가하는 방법은 통상적인 방법에 따라 적절하게 사용되고, 첨가량은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. The composition of the present invention can be added to the food for the purpose of preventing or improving fatty liver disease to produce a health functional food, the health functional food is not particularly limited to this, health functional food, nutritional supplements, nutrients, farmers food (pharmafood), health foods, nutraceuticals, designer foods, food additives, all forms of food, preferably meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen And other dairy products including noodles, gums, ice creams, various soups, beverages, teas, drinks, alcoholic beverages and vitamin complexes. At this time, the method of addition is suitably used according to a conventional method, and the addition amount can be suitably determined according to the purpose of use (prevention, health or therapeutic treatment).
상기 본 발명의 조성물은 유효성분 이외의 다양한 성분을 추가로 포함할 수도 있는데, 이들 다양한 성분은 특별히 이에 제한되지 않으나, 바람직하게는 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등이 될 수 있다. 또한, 기호성 및/또는 기능성을 부가하기 위하여 천연 과일 주스, 과일 주스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있으며, 이들 성분들은 독립적으로 또는 조합하여 사용할 수 있다.The composition of the present invention may further include various components other than the active ingredient, but these various components are not particularly limited thereto, but preferably various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and Salts, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks and the like. It may also contain pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks to add palatability and / or functionality, and these ingredients may be used independently or in combination.
또한, 본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 지방간 질환의 예방 또는 치료용 약학 조성물에 관한 것이다.The present invention also relates to a pharmaceutical composition for the prevention or treatment of fatty liver disease comprising rice locust extract as an active ingredient.
본 발명의 약학 조성물은 약학적으로 허용되는 담체를 포함할 수 있다. 본 발명의 약학 조성물에 포함되는 약학적으로 허용되는 담체는 제제 시에 통상적으로 이용되는 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 덱스트로즈 용액, 말토덱스트린 용액, 글리세롤, 에탄올, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. The pharmaceutical composition of the present invention may comprise a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are conventionally used in the preparation, and include saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, lactose, dex Straw, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methyl cellulose, methylhydroxybenzoate, propyl Hydroxybenzoate, talc, magnesium stearate and mineral oil, and the like.
본 발명의 약학적 조성물은 상기 성분들 이외에 항산화제, 완충액, 정균제, 희석제, 계면활성제, 결합제, 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제 또는 보존제 등을 추가로 포함할 수 있다. The pharmaceutical composition of the present invention may further include antioxidants, buffers, bacteriostatic agents, diluents, surfactants, binders, lubricants, wetting agents, sweetening agents, flavoring agents, emulsifiers, suspending agents or preservatives in addition to the above components.
본 발명의 약학적 조성물은 경구 또는 비경구 투여할 수 있으며, 바람직하게는 경구 투여 방식으로 적용된다. 본 발명의 약학적 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다. 본 발명의 약학적 조성물의 투여량은 바람직하게는 성인 기준으로 0.001-100 ㎎/kg이다.The pharmaceutical composition of the present invention may be administered orally or parenterally, and preferably applied by oral administration. Suitable dosages of the pharmaceutical compositions of the present invention may be prescribed in various ways depending on factors such as formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be. The dosage of the pharmaceutical composition of the present invention is preferably 0.001-100 mg / kg on an adult basis.
본 발명의 약학적 조성물은 당해 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다. 이때 제형은 오일 또는 수성 매질중의 용액, 현탁액, 시럽제 또는 유화액 형태이거나 엑스제, 산제, 분말제, 과립제, 정제 또는 캅셀제 형태일 수도 있으며, 분산제 또는 안정화제를 추가적으로 포함할 수 있다.The pharmaceutical compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. The formulation may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of extracts, powders, powders, granules, tablets or capsules, and may further comprise dispersants or stabilizers.
또한, 본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 인간을 제외한 동물의 지방간 질환의 예방 또는 치료용 수의학적 조성물에 관한 것이다.The present invention also relates to a veterinary composition for preventing or treating fatty liver disease in animals other than humans, which includes rice locust extract as an active ingredient.
본 발명의 벼메뚜기 추출물을 포함하는 수의학적 조성물은 통상의 방법에 따른 적절한 부형제 및 희석제를 더 포함할 수 있다. 본 발명의 벼메뚜기 추출물을 포함하는 수의학적 조성물에 포함될 수 있는 부형제 및 희석제로는, 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트, 세탄올, 스테아릴알콜, 유동파라핀, 솔비탄모노스테아레이트, 폴리소르베이트 60, 메칠파라벤, 프로필파라벤 및 광물유를 들 수 있다. Veterinary compositions comprising the rice locust extract of the present invention may further comprise suitable excipients and diluents according to conventional methods. Excipients and diluents that may be included in the veterinary composition comprising the rice locust extract of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, Calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, cetanol, stearyl alcohol, liquid paraffin , Sorbitan monostearate,
본 발명에 따른 벼메뚜기 추출물을 포함하는 수의학적 조성물은 충진제, 항응집제, 윤활제, 습윤제, 향신료, 유화제, 방부제 등을 추가로 포함할 수 있는데, 본 발명에 의한 수의학적 조성물은 동물에 투여된 후 활성 성분의 신속, 지속 또는 지연된 방출을 제공할 수 있도록 당업계에 잘 알려진 방법을 사용하여 제형화될 수 있고, 제형은 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 용액, 시럽, 에어로졸, 연질 또는 경질 젤라틴 캅셀, 좌제, 멸균 주사용액, 멸균 외용제 등의 형태일 수 있다.The veterinary composition comprising the rice locust extract according to the present invention may further include a filler, an anticoagulant, a lubricant, a humectant, a spice, an emulsifier, an antiseptic, and the like. The veterinary composition according to the present invention is active after administration to an animal. Formulations may be formulated using methods well known in the art to provide rapid, sustained or delayed release of the components, the formulations being powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, soft Or hard gelatin capsules, suppositories, sterile injectable solutions, sterile external preparations, and the like.
본 발명에 의한 수의학적 조성물은 동물의 나이, 성별, 체중에 따라 달라질 수 있으나, 0.1~100mg/㎏의 양을 1일 1회 내지 수회 투여할 수 있고, 투여량은 투여경로, 질병의 정도, 성별, 체중, 나이 등에 따라서 증감될 수 있다. 따라서, 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다. The veterinary composition according to the present invention may vary depending on the age, sex, and weight of the animal, but the amount of 0.1-100 mg / kg may be administered once to several times a day, and the dosage may be administered by the route of administration, degree of disease, It may increase or decrease depending on gender, weight, age, and the like. Therefore, the above dosage does not limit the scope of the present invention in any aspect.
또한, 본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 인간을 제외한 동물의 지방간 질환의 예방 또는 개선용 사료 첨가제에 관한 것이다.In addition, the present invention relates to a feed additive for the prevention or improvement of fatty liver disease in animals other than humans including rice locust extract as an active ingredient.
상기 사료 첨가제는 20~90중량%의 벼메뚜기 추출물을 함유하는 고 농축액, 분말 또는 과립형태일 수 있다. The feed additive may be in the form of a high concentrate, powder or granules containing 20 to 90% by weight of rice locust extract.
본 발명의 사료 첨가제는 구연산, 후말산, 아디픽산, 젖산, 사과산 등의 유기산이나 인산나트륨, 인산칼륨, 산성피로인산염, 폴리인산염(중합인산염) 등의 인산염이나 폴리페놀, 카테킨(catechin), 알파-토코페롤, 로즈메리 추출물(rosemary extract), 비타민 C, 녹차 추출물, 감초 추출물, 키토산, 탄닌산, 피틴산 등의 천연 항산화제 중에서 선택된 하나 이상을 추가로 포함할 수 있다.The feed additive of the present invention includes organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate and polyphosphate (polyphosphate), polyphenols, catechins and alpha. It may further include one or more selected from natural antioxidants such as tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, phytic acid.
본 발명의 벼메뚜기 추출물을 함유하는 동물사료 첨가제 및 이를 포함하는 사료는 보조성분으로 아미노산, 무기염류, 비타민, 항생물질, 항균물질, 항산화, 항곰팡이 효소, 소화 및 흡수향상제, 성장촉진제, 질병예방제 등과 같은 물질과 함께 사용될 수 있다.Animal feed additives containing rice locust extract of the present invention and feed comprising the same are amino acids, inorganic salts, vitamins, antibiotics, antibacterial substances, antioxidants, antifungal enzymes, digestion and absorption enhancers, growth promoters, disease prevention agents, etc. Can be used with the same material.
상기 사료 첨가제는 동물에게 단독으로 식용 담체 중에서 다른 사료 첨가제와 조합되어 투여될 수 있다. 또한, 상기 사료첨가제는 탑 드레싱으로서 또는 이들을 동물 사료에 직접 혼합하거나 또는 사료와 별도로, 별도의 경구 제형으로, 주사 또는 경피로 또는 다른 성분과 조합하여 쉽게 투여할 수 있다. 통상적으로, 당 업계에 잘 알려진 바와 같이 단독 일일 투여량 또는 분할 일일 투여량을 사용할 수 있다. 상기 사료 첨가제를 동물 사료와 별도로 투여할 경우, 당 업계에 잘 알려진 바와 같이 추출물의 투여 형태는 이들을 비-독성 제약상 허용 가능한 식용 담체와 조합하여 즉석 방출 또는 서방성 제형으로 제조할 수 있다. 이러한 식용 담체는 고체 또는 액체, 예를 들어 옥수수 전분, 락토스, 수크로스, 콩 플레이크, 땅콩유, 올리브유, 참깨유 및 프로필렌 글리콜일 수 있다. 고체 담체가 사용될 경우, 추출물의 투여형은 정제, 캡슐제, 산제, 토로키제 또는 함당정제 또는 미분산성 형태의 탑 드레싱일 수 있다. 액체 담체가 사용될 경우, 연 젤라틴 캡슐제, 또는 시럽제 또는 액체 현탁액제, 에멀젼제 또는 용액제의 투여 형태일 수 있다. 또한, 투여 형태는 보조제, 예를 들어 보존제, 안정화제, 습윤제 또는 유화제, 용액 촉진제 등을 함유할 수 있다. 또한, 벼메뚜기 추출물이 사료 첨가제로 포함되는 동물사료는 동물의 식이 요구를 충족시키는데 통상적으로 사용되는 임의의 단백질-함유 유기 곡분일 수 있다. 이러한 단백질-함유 곡분은 통상적으로 옥수수, 콩 곡분 또는 옥수수/콩 곡분 믹스로 주로 구성되어 있다. 상기의 사료 첨가제는 침지, 분무 또는 혼합하여 상기 동물사료에 첨가하여 이용될 수 있다. 본 발명의 수의학적 조성물 또는 사료첨가제는 반려동물의 식이에 적용할 수 있다.The feed additive may be administered to the animal alone in combination with other feed additives in an edible carrier. In addition, the feed additives can be easily administered as a top dressing or directly mixed into animal feed or separately from the feed, in a separate oral formulation, by injection or transdermal or in combination with other ingredients. Typically, single or divided daily dosages can be used as is well known in the art. When the feed additives are administered separately from the animal feed, dosage forms of the extracts, as is well known in the art, can be prepared in immediate release or sustained release formulations in combination with non-toxic pharmaceutically acceptable edible carriers. Such edible carriers may be solid or liquid, for example corn starch, lactose, sucrose, soy flakes, peanut oil, olive oil, sesame oil and propylene glycol. When a solid carrier is used, the dosage form of the extract may be tablets, capsules, powders, torokies or sugar-containing tablets or top dressings in microdisperse form. If a liquid carrier is used, it may be in the form of soft gelatin capsules or syrups or liquid suspensions, emulsions or solutions. In addition, the dosage form may contain auxiliaries such as preservatives, stabilizers, wetting or emulsifying agents, solution promoters and the like. In addition, the animal feed in which the rice locust extract is included as a feed additive may be any protein-containing organic cereal meal commonly used to meet the dietary needs of the animal. Such protein-containing flours typically consist mainly of corn, soy flour, or corn / bean flour mixtures. The feed additive may be used by adding to the animal feed by dipping, spraying or mixing. The veterinary composition or feed additive of the present invention can be applied to the diet of a companion animal.
또한, 본 발명은 벼메뚜기 추출물을 인간을 제외한 개체에게 투여하는 단계를 포함하는 지방간 질환의 예방 또는 치료방법에 관한 것이다.The present invention also relates to a method for preventing or treating fatty liver disease, comprising administering rice locust extract to an individual except a human.
상기 개체는 지방간 질환을 앓는 환자를 포함하여 지방간 질환의 가능성이 있는 사람을 비롯한 동물 모두를 포함한다.
The subject includes all animals including humans with the possibility of fatty liver disease, including patients suffering from fatty liver disease.
이하, 실시예를 이용하여 본 발명을 더욱 상세하게 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로 본 발명의 범위가 이들에 의해 제한되지 않는다는 것은 당해 기술분야에서 통상의 지식을 가진 자에게 있어 자명한 것이다.
Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for explaining the present invention more specifically, it is obvious to those skilled in the art that the scope of the present invention is not limited by them.
실시예Example 1. 간암 세포주( 1. Liver cancer cell line ( HepGHepG 22 )의 배양Culture
간암 세포주(HepG2)는 ATTC(American Type Culture Collection)에서 구입하여 사용하였다. 간암 세포주(HepG2)는 10% FBS, 1% 페니실린-스트렙토마이신이 포함된 DMEM(Dulbecco's modified Eagle's medium) 배지로 배양하였다. 상기 세포는 37℃에서, 5% CO2 배양기에서 배양하였고, 배양된 세포는 24웰 플레이트에 일정한 수의 세포(3x 104 cells/ 500㎕ well)를 분주하여 실험을 하였다.Liver cancer cell line (HepG 2 ) was purchased from the American Type Culture Collection (ATTC). Liver cancer cell line (HepG 2 ) was cultured in Dulbecco's modified Eagle's medium (DMEM) medium containing 10% FBS, 1% penicillin-streptomycin. The cells were cultured in a 5% CO 2 incubator at 37 ° C., and the cultured cells were tested by dispensing a predetermined number of cells (3 × 10 4 cells / 500 μl well) in a 24-well plate.
24웰 플레이트에 세포를 분주한 후, 세포가 24웰이 완전히 붙어 세포의 모양을 갖춘 후 세포가 75%의 포화도(confluence)에 도달했을 때 사용하였다.
After dispensing cells in a 24-well plate, the cells were used when the cells reached 75% saturation (confluence) after the 24 wells were completely attached and shaped.
실시예Example 2. 유리 지방산(Free Fatty Acid; 2. Free Fatty Acid; FFAFFA ) 제조) Produce
(1) 100mM 팔미테이트(시그마 P-0500) 및 100mM 올레이트(시그마 O-75010) 저장용액의 제조(1) Preparation of 100 mM palmitate (Sigma P-0500) and 100 mM oleate (Sigma O-75010) stock solutions
70℃에서 0.1M NaOH 용액을 용매로 이용하여 제조하고, 0.2㎛의 필터를 이용하여 여과한 후, 멸균하였다.A 0.1 M NaOH solution was prepared at 70 ° C. as a solvent, filtered using a 0.2 μm filter, and then sterilized.
(2) 유리 지방산이 포함되지 않은 5%(w/v)의 BSA(시그마 A-6003) 용액의 제조(2) Preparation of 5% (w / v) BSA (Sigma A-6003) Solution Free of Free Fatty Acids
3차 증류수를 용매로 이용하여 제조한 후 멸균하고, 4℃ 냉장보관하였다.Prepared using tertiary distilled water as a solvent, sterilized, and refrigerated 4 ℃.
(3) 올레이트 및 팔미테이트의 농도가 2:1이 되도록 혼합한 5mM의 혼합지방산 저장용액 제조(3) Preparation of 5mM mixed fatty acid stock solution mixed so that the concentration of oleate and palmitate is 2: 1
60℃에서, 상기 2)에서 제조한 3차 증류수에 녹인 유리 지방산이 포함되지 않은 5%(w/v)의 BSA 용액을 용매로 이용하여 상기 1)에서 제조한 각각의 지방산의 농도가 5mM이 되도록 지방산 저장용액 10㎖을 제조하였다. 상기 1)에서 제조한 100mM 팔미테이트 및 100mM 올레이트 저장용액을 165㎕(1.65mM)의 팔미테이트(Palmitate) 및 330㎕(3.3mM)의 올레이트(oleate)를 취해 9,505㎕의 5%(w/v)의 BSA 용액에 방울방울 떨어뜨리면서 흔들며 섞어주었다(vortexing). 이후에 상온으로 식힌 다음, 0.2μm 필터로 여과하였다(상기 제조한 혼합용액은 -20℃에서 3 내지 4주 동안 보관하여도 안정하다).
At 60 ° C, the concentration of each fatty acid prepared in 1) was 5 mM using 5% (w / v) BSA solution containing no free fatty acid dissolved in tertiary distilled water prepared in 2) as a solvent. 10 ml of a fatty acid stock solution was prepared. 100 mM palmitate and 100 mM oleate stock solution prepared in 1) was taken with 165 µl (1.65 mM) of palmitate and 330 µl (3.3 mM) of oleate, and 9,505 µl of 5% (w) / v) vortexing while dropping the drops of BSA solution. Thereafter, the mixture was cooled to room temperature, and then filtered through a 0.2 μm filter (the prepared mixed solution is stable even if stored at -20 ° C. for 3 to 4 weeks).
실시예Example 3. 벼메뚜기 추출물의 제조 3. Preparation of Rice Locust Extract
벼메뚜기를 70%(v/v) 에탄올로 추출하여 에탄올 추출액을 얻고, 에탄올 추출액을 감압 증류하여 에탄올 추출물을 획득하였다.
Rice locust was extracted with 70% (v / v) ethanol to obtain an ethanol extract, and the ethanol extract was distilled under reduced pressure to obtain an ethanol extract.
실시예Example 4. 4. 나일레드Nile red (( 아디포레드Adifored ) 염색() dyeing( AdipoRedAdipoRed staining) staining) 어세이를Assay 이용한 Used 중성지방의 함량 평가Triglyceride content evaluation
간암 세포주(HepG2)에 1mM의 올레이트 및 팔미테이트 혼합(2:1) 유리 지방산을 처리하여 생성된 중성지방이 벼메뚜기 추출물에 의해 감소되는 정도를 나일레드(아디포레드) 염색 어세이로 확인하기 위하여, 간암 세포주(HepG2)를 포함하는 배지에 1mM의 올레이트 및 팔미테이트 혼합(2:1) 유리 지방산(FFA) 처리 및 유리 지방산과 본 발명의 벼메뚜기 추출물을 병용 처리하고 24시간 동안 인큐베이션 한 후, 배지를 약하게 흡입(suction)하였다. Hepatocarcinoma cell line (HepG 2 ) was treated with 1 mM oleate and palmitate mixed (2: 1) free fatty acids to determine the degree of triglyceride reduction by rice locust extract in a nired (adifored) staining assay. To this end, 1 mM oleate and palmitate mixed (2: 1) free fatty acid (FFA) treatment and a combination of free fatty acids and the rice locust extract of the present invention were treated in a medium containing a liver cancer cell line (HepG 2 ) and incubated for 24 hours. Afterwards, the medium was slightly aspirated.
이후, 37℃의 조건 하에서 아디포레드 용액(AdipoRed solution; 300㎕의 AdipoRed를 10㎖의 PBS에 녹여 제조함)을 500㎕/well씩 분주 후, 암조건 상온의 조건에서 15분 동안 약하게 shaking하여 반응하였다. 이 과정 동안 염료의 광퇴색(photobleaching)을 최소화하기 위하여 플레이트를 알루미늄 호일로 싸서 보관하였다. 반응이 종료된 후 이를 형광분도계(spectrophotometer)에 적용하여 여기파장(excitation)은 485nm, 방출파장(emission)은 572nm의 조건에서 측정하였다.Subsequently, after dispensing an adipored solution (prepared by dissolving 300 μl of AdipoRed in 10 ml of PBS) under 37 ° C., 500 μl / well was dispensed by shaking gently for 15 minutes at room temperature. Reacted. Plates were stored wrapped in aluminum foil to minimize photobleaching of the dye during this process. After the reaction was completed, it was applied to a spectrophotometer and the excitation wavelength was measured at 485 nm and emission wavelength 572 nm.
그 결과, 벼메뚜기 추출물 10, 50 및 100㎍/㎖의 농도에 대하여 중성지방이 농도 의존적으로 감소하는 효과를 확인할 수 있었다(도 1).
As a result, it was confirmed that the triglycerides concentration-dependently reduced the concentration of
실시예 5. 벼메뚜기 추출물의 처리에 따른 총 중성지방의 함량 및 지방 흡수(lipid uptake) 정도의 변화 확인 Example 5 Confirmation of Changes in Total Triglyceride Content and Lipid Uptake Levels of Rice Locust Extracts
간암 세포주(HepG2)에 1mM의 올레이트 및 팔미테이트 혼합(2:1) 유리 지방산을 처리하여 증가한 총 중성지방의 함량 및 지방 흡수(lipid uptake)가 벼메뚜기 추출물의 처리에 의해 감소 되는지를 확인하였다.Hepatocarcinoma cell line (HepG 2 ) was treated with 1 mM oleate and palmitate mixed (2: 1) free fatty acids to determine if the increased total triglyceride content and lipid uptake were reduced by treatment of rice locust extract. .
그 결과 도 2 및 도 3에 개시한 바와 같이, 벼메뚜기 추출물의 농도 의존적으로 총 중성지방의 함량 및 지방 흡수(lipid uptake)가 감소한 것을 확인하였다.
As a result, as shown in Figures 2 and 3, it was confirmed that the content of total triglycerides and lipid uptake (lipid uptake) was reduced in a concentration-dependent manner of the rice locust extract.
실시예 6. 벼메뚜기 추출물의 처리에 따른 세포 생존율 및 Caspase -3의 발현량 변화 확인 Example 6. Confirmation of Cell Survival Rate and Expression Change of Caspase- 3 According to Treatment of Rice Locust Extract
간암 세포주(HepG2)에 에탄올을 처리하여 감소한 세포 생존률 및 증진된Caspase-3 발현량이 벼메뚜기 추출물의 처리에 의해 변화가 일어나는지를 확인하였다.Hepatocarcinoma cell line (HepG 2 ) was treated with ethanol to confirm that the reduced cell viability and enhanced Caspase-3 expression were altered by treatment of rice locust extract.
그 결과 도 4 및 도 5에 개시한 바와 같이, 벼메뚜기 추출물의 농도 의존적으로 세포 생존율이 증진되었고, Caspase-3의 발현량이 현저하게 감소한 것을 확인하였다.As a result, as shown in Figs. 4 and 5, it was confirmed that the cell viability was increased depending on the concentration of the rice locust extract, and the expression level of Caspase-3 was significantly reduced.
실시예Example 7. 벼메뚜기 추출물의 투여에 따른 동물모델의 체중변화; 간, 복부피하지방, 신장 지방조직 및 소장 지방조직의 무게; 지질생화학적 지표들의 분석 7. Weight Change of Animal Model Following Administration of Rice Locust Extract; Weight of liver, abdominal subcutaneous fat, renal adipose tissue and small intestine adipose tissue; Analysis of Lipid Biochemical Indicators
(1) 실험동물(1) experimental animals
본 실시예 7에서는 10주령의 C57bl/6Jj 생쥐(마우스)를 사용하였는데, 수컷 7주령의 C57bl/6Jj 생쥐를 대한바이오링크사(Eumsung, Korea)로부터 공급받아 3주 동안 실험실 환경에 적응시킨 후, 건강상태가 양호한 마우스를 실험에 사용하였다. In Example 7, 10-week-old C57bl / 6Jj mice (mouse) were used, and male 7-week-old C57bl / 6Jj mice were supplied from Korea Biolink Co., Ltd. (Eumsung, Korea), and then adapted to a laboratory environment for 3 weeks. Mice with good health were used for the experiment.
실험동물은 실험 당일까지 고형사료(항생제 무첨가, 삼양사료 Co.)와 물을 충분히 공급하였고, 온도 23±2℃, 습도 55±15%, 12시간 명-암 주기의 환경을 유지하며, 기본사료(AIN-76A diet)와 물을 자유롭게 공급하면서 실험실 환경에 적응시켰다. 이후, 건강상태가 양호한 10주령의 마우스를 동물 사육실의 환경은 온도 23±2℃, 습도 55±15% 및 12시간 간격의 광주기(light on 07:00~19:00)로 명암으로 조절되는 SPF(Specific Pathogen Free) 환경에서 유지하면서 실험에 사용하였다. The experimental animals were supplied with solid feed (no antibiotics, Samyang Feed Co.) and water until the day of the experiment, and maintained the environment of temperature 23 ± 2 ℃, humidity 55 ± 15%, 12 hours light-dark cycle, and basic feed. (AIN-76A diet) and free water supply to adapt to the laboratory environment. Afterwards, the healthy 10-week-old mouse in the animal breeding room is controlled to light and shade with a temperature of 23 ± 2 ° C., a humidity of 55 ± 15% and a photoperiod (light on 07: 00 ~ 19: 00) at 12 hour intervals. It was used in the experiment while maintaining in SPF (Specific Pathogen Free) environment.
실험동물은 정상 식이 대조군(Nor), 고지방식이 대조군 (HFD, 60% fat), 고지방 식이를 하면서 벼메뚜기 추출물을 투여한 군(100, 200mg/kg) 및 양성대조군으로 고지방 식이를 하면서 밀크시슬 추출물(100mg/kg)을 투여한 군으로 분류하여 14주 동안 진행하였으며, 물과 각 실험군의 식이는 자유롭게 공급하였고, 벼메뚜기 추출물은 매일 14주 동안 경구투여 방식으로 공급하였다.
The experimental animals were normal diet control group (Nor), high fat diet control group (HFD, 60% fat), milk fat extract with high fat diet (100, 200mg / kg) and high control diet with rice locust extract and high fat diet. (100mg / kg) was divided into groups administered for 14 weeks, water and diet of each experimental group was freely supplied, and rice locust extract was supplied by oral administration for 14 weeks every day.
(2) 체중변화, 간 조직, 복부피하지방, 소장 및 신장의 지방조직 변화(2) changes in body weight, liver tissue, abdominal subcutaneous fat, small intestine and kidney adipose tissue
실험동물의 체중은 매주 일정한 시간에 14주 동안 측정하여 기록하였으며, 14주 후 실험동물을 간 조직 중량, 피하지방(abdominal subcutaneous fat), 소장 지방 및 신장 후 복막 지방조직(retroperitoneal adipose tissue)으로 구분하여 적출 후 지방조직의 중량을 산출하였다. The body weight of the test animals was measured and recorded for 14 weeks at a certain time every week, and after 14 weeks, the test animals were divided into liver tissue weight, subdominal subcutaneous fat, small intestine fat, and retroperitoneal adipose tissue. After extraction, the weight of the adipose tissue was calculated.
본 발명의 벼메뚜기 추출물(MTG)이 고지방식이에 의해 증가한 마우스 체중에 미치는 영향을 확인한 결과, 고지방 식이군(HFD)에 비하여 체중이 감소되는 추이를 보였다(도 6). As a result of confirming the effect of the rice locust extract (MTG) of the present invention on the mouse weight increased by the high fat diet, the weight loss was shown in comparison with the high fat diet group (HFD) (Fig. 6).
본 발명의 벼메뚜기 추출물(MTG)이 고지방 식이에 의해 증가한 간 조직 중량을 감소시킬 수 있는지 확인한 결과, 고지방 식이군(HFD)에 대비하여 고지방 식이와 벼메뚜기 추출물(MTG)을 모두 섭취한 군의 간 조직 중량이 감소하였으며, 고지방식이에 의해 마우스의 복부 피하지방, 신장 지방조직 및 소장 지방조직의 중량도 고지방 식이군(HFD)에서 증가하였으나 고지방식이와 더불어 벼메뚜기 추출물(MTG)을 섭취한 군에서는 마우스의 복부 피하지방, 신장 지방조직 및 소장 지방조직의 중량이 감소하였다(도 7).
As a result of confirming that the rice locust extract (MTG) of the present invention can reduce the liver tissue weight increased by the high fat diet, the liver tissue of the group ingesting both the high fat diet and the rice locust extract (MTG) compared to the high fat diet group (HFD) The weight was decreased and the weight of abdominal subcutaneous fat, renal adipose tissue and small intestine adipose tissue was increased in the high fat diet group (HFD), but in the group fed rice grasshopper extract (MTG) in addition to the high fat diet. The weight of abdominal subcutaneous fat, renal adipose tissue and small intestine adipose tissue of mice was reduced (FIG. 7).
(3) (3) 비알콜성Non-alcoholic 지방간( Fatty liver ( NAFLDNAFLD ) 유발 생쥐의 혈장 중 지질생화학적 지표분석Analysis of Lipid Biochemical Markers in Plasma of Induced Mice
벼메뚜기 추출물과 밀크씨슬을 14주 동안 투여 후, 각 실험동물로부터 분리한 혈장 (plasma)에서 간 기능의 지표인 ALT 및 AST를, 혈장의 지질함량의 지표인 중성지방(triglyceride), 총콜레스테롤, 저밀도 지단백, 유리지방산, 혈당의 함량을 생화학자동 분석기 (Hitachi-720, Hitachi Medical, Japan)를 이용하여 측정하였다.After 14 weeks of administration of rice locust extract and milk thistle, ALT and AST, indicators of liver function, and triglyceride, total cholesterol, and indicators of lipid content in plasma were isolated from plasma isolated from each experimental animal. Low-density lipoprotein, free fatty acid, and blood glucose content were measured using a biochemical automated analyzer (Hitachi-720, Hitachi Medical, Japan).
①① 혈중 ALT, Blood ALT, ASTAST 변화 change
벼메뚜기 추출물(MTG)이 고지방 식이에 의한 혈중 ALT와 AST 증가를 감소시킬 수 있는지를 확인한 결과, 고지방 식이군에 비하여 고지방 식이와 함께 벼메뚜기 추출물(MTG)을 섭취한 군에서의 ALT 및 AST가 현저하게 감소하였다(도 8). As a result of confirming that the rice locust extract (MTG) can reduce the blood ALT and AST increase caused by the high fat diet, the ALT and AST in the group ingesting the rice locust extract (MTG) with the high fat diet was significantly higher than the high fat diet group. Decreased (FIG. 8).
이와 같은 결과로부터 벼메뚜기 추출물(MTG)의 식이는 혈중 ALT 및 AST 효소 활성을 감소시켜, 고지방식이에 의해 유도된 비알콜성 지방간을 개선하는 것으로 나타났다.
From these results, diet of rice locust extract (MTG) was found to reduce ALT and AST enzyme activity in blood, thereby improving non-alcoholic fatty liver induced by high fat diet.
② ② C57blC57bl /6J / 6J 비알콜성Non-alcoholic 지방간 ( Fatty liver ( NAFLDNAFLD ) 유발 생쥐의 혈장 중 지질생화학적 분석Lipid Biochemical Analysis in Plasma of Induced Mice
벼메뚜기 추출물(MTG)이 고지방 식이에 의해 증가된 혈액 내 중성지방, 총 콜레스테롤, 저밀도 지단백, 유리지방산 수치를 감소시킬 수 있는지 확인한 결과, 고지방 식이(HFD) 군에서 증가된 중성지방, 총콜레스테롤, 저밀도 지단백, 유리지방산 수치가 벼메뚜기 추출물(MTG) 투여군에서 통계적으로 유의미하게 감소하였다(도 9). 따라서 벼메뚜기 추출물(MTG) 식이는 혈액 내 지질 수준을 감소시켜 고지방식이에 의해 유도된 비알콜성 지방간을 개선하는 것으로 나타났다.
Checking whether rice locust extract (MTG) can reduce the levels of triglycerides, total cholesterol, low density lipoproteins and free fatty acids in the blood, which are increased by high fat diets, and have increased triglycerides, total cholesterol, and low density in high fat diet (HFD) groups. Lipoprotein and free fatty acid levels were significantly decreased in the rice locust extract (MTG) administration group (Fig. 9). Therefore, rice locust extract (MTG) diet has been shown to reduce blood lipid levels and improve non-alcoholic fatty liver induced by high fat diet.
③ 혈청 내 혈당 농도 변화③ Change in blood glucose level in serum
고지방 식이(HFD)에 의해 혈청 내 혈당 농도는 증가하였으나, 고지방 식이와 벼메뚜기 추출물(MTG)을 모두 섭취한 군에서는 고지방식이 군에 비해 통계적으로 유의미하게 혈청 내 혈당 농도가 감소하였다(도 10).
Serum glucose level was increased by high fat diet (HFD), but serum level was significantly decreased in the high fat diet and rice locust extract (MTG) compared to the high fat diet group (Fig. 10). .
[통계 처리][Statistical processing]
상기 각 실험군의 결과는 unpaired student's T-test 통계프로그램을 사용하여 통계 처리하였으며, p<0.05 이하의 수준에서 유의성 검정을 실시하였다.The results of each experimental group were statistically processed using the unpaired student's T-test statistical program, and significance test was performed at a level of p <0.05 or less.
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| KR102270851B1 (en) | 2019-11-18 | 2021-06-28 | 세종대학교산학협력단 | Pharmaceutical composition for treating or preventing leaky gut syndrome |
| KR102561683B1 (en) * | 2021-01-29 | 2023-08-01 | 한국원자력연구원 | Composition for oral administration comprising extract of oxya chinensis sinuosa |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4753089B2 (en) | 2004-12-20 | 2011-08-17 | 株式会社岡安商店 | Composition having action of promoting degradation of body fat, and food or food compound containing the same |
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| KR20050079443A (en) * | 2004-02-05 | 2005-08-10 | 김빛나 | Process for preparing the high molecular weight antitumor fractions derived from edible grasshoppers |
| KR20160001880A (en) * | 2014-06-27 | 2016-01-07 | 대한민국(농촌진흥청장) | An Extract of Oxya chinensis sinuosa Having Anti-inflammatory and Immuno-modulating Activity and Composition Comprising the Same |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4753089B2 (en) | 2004-12-20 | 2011-08-17 | 株式会社岡安商店 | Composition having action of promoting degradation of body fat, and food or food compound containing the same |
Non-Patent Citations (1)
| Title |
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| Entomological Research. 2009, Vol.39, pp.299-303 |
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| KR20190003305A (en) | 2019-01-09 |
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