KR20190003305A - 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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- KR20190003305A KR20190003305A KR1020170181811A KR20170181811A KR20190003305A KR 20190003305 A KR20190003305 A KR 20190003305A KR 1020170181811 A KR1020170181811 A KR 1020170181811A KR 20170181811 A KR20170181811 A KR 20170181811A KR 20190003305 A KR20190003305 A KR 20190003305A
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- extract
- fatty liver
- liver disease
- rice
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—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
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- 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
-
- 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
- A23V2250/20—Natural extracts
- A23V2250/204—Animal extracts
Abstract
Description
본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 지방간 질환의 예방, 개선 또는 치료용 조성물에 관한 것이다.The present invention relates to a composition for preventing, ameliorating or treating a fatty liver disease comprising an extract of a rice plant 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 death rate of domestic liver disease is very high, 23.5 per 100,000 population. The number of deaths in the 40s is the first (41.1 / 100,000), the second in the 50s (72.4 / 100,000) (10 people / 100,000 people), including liver disease is the leading cause of death in 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 suggests that abnormal fat, which is not present in normal cells, is abnormally deposited in liver cells. In normal liver, about 5% is composed of adipose tissue, and triglyceride, fatty acid, phospholipid, cholesterol and cholesterol ester are the main components of fat. However, once fatty liver occurs, most components are replaced with triglyceride. If the liver weight is 5% or more, it is diagnosed as fatty liver. When the fatty liver becomes worse and fat mass in hepatocyte becomes larger, important components of the cells including nucleus are pushed to one side, the function of the hepatocyte is lowered, and the expanded hepatocytes due to the accumulated fat in the cells pressurize the microvascular and lymphatic vessels between the hepatocytes Resulting in obstacles to the circulation of blood and lymph in the liver. In this way, hepatocytes can not properly receive oxygen and nutrient supply, and liver function is deteriorated.
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 defined as the accumulation of fatty acids in the form of triglycerides in the parenchymal cells of the liver in excess of 5%, not by liver damage caused by alcohol. Pathologically, it is classified as simple steatosis and inflammatory fatty liver disease (steatohepatitis). It can be converted to severe liver disease such as hepatitis, liver fibrosis and cirrhosis when left for a long time. In Korea, the frequency of nonalcoholic liver disease is increasing due to changes in lifestyle.
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.Alcoholic fatty liver disease is caused by excessive alcohol consumption. Individuals have different genetic characteristics and gender. However, alcohol intake of 80g or more per day is very likely to cause liver disease such as alcoholic fatty liver disease. Women are more likely to develop alcoholic liver disease in lesser amounts. Generally, it can be calculated that one cup of shochu, one cup of beer, one cup of wine, and one cup of makkolli contain about 10 g of alcohol. The most mild form of alcoholic fatty liver disease is asymptomatic, but mild hepatic enlargement (greater than normal liver) 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, the rice-grasshopper has a body length of 30 to 38 mm, the body color is yellow-green, and the head and chest are yellowish brown. The compound eyes are egg-shaped and lustrous grayish brown. There are three thin lateral grooves on the pronotum, with brown vertical stripes on both sides. The wings are yellowish brown and longer than the end of the boat but do not fly much. Inhabits grassy areas near rice fields and cultivated areas. It occurs once a year, and it drifts with a bunch of ground in the ground. The alum is wrapped in a thin film of collagen. The full-grown nymphs have a wing crest at the fourth node and a body length of about 30 mm. The use of pesticides as a major pest of rice has decreased, but it is gradually increasing. In 1930, it originated in Hwanghae Province and bought eggs from 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. 1648812 discloses a method for producing a diet food composition containing a rice-grasshopper ingredient, Korean Patent Publication No. 2010-0002661 discloses a functional complex containing hyaluronic acid and a method for producing the same, There is no disclosure of a composition for preventing, improving or treating a fatty liver disease comprising the rice grasshopper extract of the present 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 provides a composition for preventing, ameliorating or treating a fatty liver disease comprising an extract of a rice plant as an active ingredient. The present invention provides a composition for preventing, ameliorating or treating fatty liver disease, And the weight change of the animal model according to administration of the rice grasshopper extract; Weight change of liver, subcutaneous fat, kidney fat tissue and small intestine fat tissue; Confirming that lipid biochemical indicators change significantly, thus completing the present invention.
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 health functional food composition for preventing or ameliorating fatty liver disease comprising an extract of a rice plant as an active ingredient.
또한, 본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 지방간 질환의 예방 또는 치료용 약학 조성물을 제공한다.The present invention also provides a pharmaceutical composition for preventing or treating fatty liver disease comprising an extract of a rice plant 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 the prevention or treatment of fatty liver disease in an animal other than a human, comprising a rice grasshopper 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 preventing or ameliorating fatty liver disease in an animal other than a human, comprising a rice plant 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 of preventing or treating fatty liver disease comprising administering a rice grasshopper extract to a subject other than 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 preventing, ameliorating or treating a fatty liver disease comprising a rice grasshopper extract as an active ingredient. The rice grasshopper extract as an active ingredient of the present invention is free from side effects and toxicity , Hepatocellular carcinoma cell line (HepG 2 ), and the weight change of the animal model according to the administration of the rice grasshopper extract; Weight change of liver, subcutaneous fat, kidney fat tissue and small intestine fat tissue; Since changes in lipid biochemical indicators are significant, the composition of the present invention can be useful as a health functional food, a medicine, or a feed additive for 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 a liver cancer cell line (HepG 2) on oleate and palmitate is 2: 1 was treated with 1.0mM of the free fatty acids and byeomettugi extract mixed in a concentration ratio, Nile Red, the content of triglyceride (H depot red) staining of Respectively. #### showed a statistically significant increase in nile red expression (%) in the 1.0 mM free fatty acid-treated group compared with the negative control (control) without 1.0 mM free fatty acid, and p <0.0001 , And **** indicates that nile red expression decreased statistically in the group treated with free fatty acid and rice grasshopper extract compared with free fatty acid treatment group, and p <0.0001.
FIG. 2 shows the results of measuring the content of total triglyceride after treating 1.0 mM of free fatty acid and rice grasshopper extract in which hepatocellular carcinoma cell line (HepG 2 ) was mixed at a concentration ratio of 2: 1 of oleate and palmitate. #### showed a statistically significant increase in the total triglyceride (mg / dl) in the 1.0 mM free fatty acid-treated group compared to the negative control group without the 1.0 mM free fatty acid treatment, p <0.0001, *** and **** were statistically significant reductions in total triglyceride (mg / dl) in the group treated with free fatty acid and rice husk extract compared to free fatty acid treatment group *** is p <0.001, and **** is p <0.0001.
FIG. 3 shows the result of lipid uptake after treatment of 1.0 mM free fatty acid and rice grasshopper extract in which hepatocellular carcinoma cell line (HepG 2 ) was mixed with olate and palmitate at a concentration ratio of 2: 1. #### indicates that the relative fat absorption rate was statistically significantly increased in the 1.0 mM free fatty acid-treated group compared to the negative control group without the 1.0 mM free fatty acid treatment (p <0.0001) , And **** indicates that the relative fat absorption rate was significantly decreased in the group treated with free fatty acid and rice plant extract compared to the free fatty acid treated group, and p <0.0001.
FIG. 4 shows the results of confirming the cell survival rate (%) after treatment of hepatocellular carcinoma cell line (HepG 2 ) with ethanol and rice horticultural extracts. #### showed a statistically significant decrease in cell viability (%) in the 1.0 mM free fatty acid-treated group compared to the negative control (control) without treatment with 1.0 mM of free fatty acid, and p <0.0001 ** indicates that the cell survival rate (%) was statistically significantly increased in the group treated with free fatty acid and rice plant extract compared to the free fatty acid treatment group, ** indicates that p ≪ 0.01 and **** is p < 0.0001.
FIG. 5 shows the results of confirming the expression level of Caspase-3 after treatment of hepatocellular carcinoma cell line (HepG 2 ) with ethanol and rice horticultural extracts. Compared to the negative control (control) without 1.0 mM free fatty acid, #### showed a statistically significant increase in the relative Caspase-3 expression in the 1.0 mM free fatty acid-treated group, p < 0.0001, and **** indicates that the relative expression of Caspase-3 was significantly decreased in the group treated with free fatty acid and rice plant extract compared with the group treated with free fatty acid, and p <0.0001.
FIG. 6 shows the result of confirming that the body weight increased by the high fat diet was reduced by administration of the rice plant extract.
FIG. 7 shows the results of confirming that liver, subcutaneous fat, kidney fat, and small intestine fat weight of mice increased by high fat diet decreased by the administration of the rice hatchlings extract. #### means that the weight of liver, subcutaneous fat, kidney fat tissue and small intestine fat tissue of HFD was statistically significantly increased compared to that of normal diet (p <0.0001) *, ***, and **** were statistically significantly higher than those of the high-fat diet group and the high fat diet group and the rice horticultural extract group (MTG), respectively. * Indicates p <0.05, *** indicates p <0.001, **** indicates p <0.0001, and M100 is a positive control, and is a milk seed.
FIG. 8 shows the results of confirming that ALT and AST in the blood of the mice increased by the high-fat diet were decreased by administration of the rice-grasshopper extract. ## and #### showed a statistically significant increase in the GPT (ALT) and GOT (AST) of the high-fat diet group (HFD) compared to the normal diet group (Nor) ***, ***, and **** were significantly higher than those of the high-fat diet group and the high fat diet group and the rice hunting group (MTG), respectively. (P <0.05), *** indicates p <0.001, **** indicates p <0.0001, and M100 indicates a positive control group And milk seeds.
FIG. 9 shows the results of confirming that the concentration of triglyceride, total cholesterol, low density lipoprotein, and free fatty acid in the blood of the mice increased by the high fat diet decreased by the administration of the rice hogger extract. P <0.0001, indicating that the triglyceride, total cholesterol, low density lipoprotein and free fatty acid concentrations of the high fat diet group (HFD) were statistically significantly increased compared to the normal diet (Nor) , ** and *** were statistically significantly higher than those of the high fat diet group and the high fat fat group, * Indicates p <0.05, ** indicates p <0.01, *** indicates p <0.001, and M100 is a positive control, milk seed.
FIG. 10 shows the results of confirming that the blood glucose concentration of the mice increased by the high fat diet decreased by the administration of the rice plant extract. As a result, the blood glucose level in the high-fat diet group (HFD) was statistically significantly increased compared to the norm group (Nor), indicating that p <0.0001 and * and *** were high fat diet (P <0.05), and *** was p <0.001, respectively, as compared with the high fat diet group and the high fat diet group (MTG) And M100 is a positive control, milk seeds.
본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 지방간 질환의 예방 또는 개선용 건강기능식품 조성물에 관한 것이다.The present invention relates to a health functional food composition for preventing or ameliorating fatty liver disease comprising an extract of a rice plant as an active ingredient.
상기 벼메뚜기 추출물은 하기의 단계를 포함하는 방법에 의해 제조할 수 있으나, 이에 한정하지 않는다:The rice plant extract may be produced by a method including, but not limited to, the following steps:
(1) 벼메뚜기에 추출용매를 가하여 추출하는 단계;(1) extracting the rice seedlings with an extraction solvent;
(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) The step of extracting the filtered extract of step (2) by concentration under reduced pressure and drying.
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.In the step (1), the extraction solvent is preferably selected from water, a C 1 -C 4 lower alcohol or a mixture thereof, more preferably a C 1 -C 4 lower alcohol, still more preferably 70% (v / v) ethanol, but is 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, any conventional method known in the art such as filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction may be used. The extraction solvent is preferably added by 1 to 20 times the weight of the dried rice hogger, more preferably 5 to 15 times. The extraction temperature is preferably 4 to 50 ° C, but is not limited thereto. The extraction time is preferably 0.5 to 10 hours, more preferably 0.5 to 5 hours, but not always limited thereto. In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in the step (3), but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always 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; Non-alcoholic or alcoholic fatty liver disease; And non-alcoholic or alcoholic cirrhosis, 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 plant extracts are characterized by reducing the content of triglyceride in liver tissue, reducing the content of total triglyceride in liver tissue and decreasing 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 health functional food may be prepared by adding the composition of the present invention to a food for the purpose of preventing or ameliorating fatty liver disease. The health functional food is not particularly limited, and may be a health functional food, a nutritional supplement, but it can be any type of food such as pharmafood, health food, nutraceutical, designer food, food additive, etc. Preferably, the food can be meat, sausage, bread, chocolate, candy, snack, confectionery, , Other noodles, gums, dairy products including ice cream, various soups, drinks, tea, drinks, alcoholic beverages and vitamin complexes. At this time, the method of adding is appropriately used according to a conventional method, and the amount to be added can be suitably determined according to the intended 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 component. The various components may be various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, Salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks and the like. It may also contain flesh for the production of natural fruit juices, fruit juice beverages and vegetable beverages 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 preventing or treating fatty liver disease comprising an extract of a rice plant 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 compositions of the present invention may comprise a pharmaceutically acceptable carrier. The pharmacologically acceptable carrier to be contained in the pharmaceutical composition of the present invention is one which is usually used at the time of formulation and includes saline, sterilized water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, But are not limited to, sucrose, sucrose, sucrose, sorbitol, mannitol, starch, acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methylcellulose, methylhydroxybenzoate, 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 comprise an antioxidant, a buffer, a bacteriostatic agent, a diluent, a surfactant, a binder, a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent or a preservative.
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 can be administered orally or parenterally, and is preferably administered orally. The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on such factors as formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, route of administration, excretion rate, . 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 composition of the present invention may be formulated into a unit dosage form by using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by those having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container. The formulations may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of excipients, powders, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.
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 the prevention or treatment of fatty liver disease in an animal other than a human, comprising a rice grasshopper 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. The veterinary composition comprising the rice grasshopper extract of the present invention may further comprise suitable excipients and diluents according to conventional methods. Examples of excipients and diluents that can be included in the veterinary composition containing the rice grasshopper extract of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, Calcium carbonate, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, cetanol, stearyl alcohol, , 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 grasshopper extract according to the present invention may further contain a filler, an anti-coagulant, a lubricant, a wetting agent, a spice, an emulsifier and an antiseptic agent. And may be formulated using methods well known in the art so as to provide rapid, sustained or delayed release of the composition, and the formulations may be formulated as powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, 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 body weight of the animal, but may be administered in an amount of 0.1 to 100 mg / kg once or several times a day. The dosage may vary depending on the route of administration, Sex, weight, age, and the like. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.
또한, 본 발명은 벼메뚜기 추출물을 유효성분으로 포함하는 인간을 제외한 동물의 지방간 질환의 예방 또는 개선용 사료 첨가제에 관한 것이다.The present invention also relates to a feed additive for preventing or ameliorating fatty liver disease in an animal other than a human, which comprises a rice grasshopper 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 granulate containing 20 to 90% by weight of the grasshopper 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 can be used as a feed additive containing a phosphate such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, or organic acids such as sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polymerized phosphate), polyphenol, catechin, Natural antioxidants such as tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid may be further included.
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.The animal feed additive containing the rice grasshopper extract of the present invention and the feed containing the same can be used as auxiliary ingredients such as amino acids, inorganic salts, vitamins, antibiotics, antimicrobials, antioxidants, antifungal enzymes, digestion and absorption enhancers, growth promoters, 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 alone to the animal in combination with other feed additives in the edible carrier. The feed additives can also be easily administered as top dressing or they can be mixed directly with the animal feed or separately from the feed, in separate oral formulations, by injection or transdermal, or in combination with other ingredients. Typically, a single daily dose or a divided daily dose can be used as is well known in the art. When the feed additive is administered separately from animal feed, the dosage form of the extract, as is well known in the art, may be prepared in an immediate release or sustained release formulation in combination with a non-toxic pharmaceutically acceptable food carrier. 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 a tablet, capsule, powder, troche or emulsion or top-dressing in finely divided form. When a liquid carrier is used, it may be in the form of a soft gelatin capsule, or a syrup or liquid suspension, emulsion or solution. In addition, the dosage form may contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, solution promoting agents and the like. In addition, the animal feed in which the rice grasshopper extract is included as a feed additive can be any protein-containing organic grain fraction commonly used to meet animal dietary needs. These protein-containing flours usually consist mainly of corn, soy flour or corn / soy flour mix. The feed additive may be added to the animal feed by immersion, spraying or mixing. The veterinary composition or feed additive of the present invention can be applied to the companion animal's diet.
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 the prevention or treatment of fatty liver disease comprising the step of administering a rice grasshopper extract to a subject other than a human.
상기 개체는 지방간 질환을 앓는 환자를 포함하여 지방간 질환의 가능성이 있는 사람을 비롯한 동물 모두를 포함한다.
Such an individual includes all animals, including those with a possibility of fatty liver disease, including those 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. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.
실시예Example 1. 간암 세포주( 1. Liver cancer cell line ( HepGHepG 22 )의 배양) Culture
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) were purchased from ATTC (American Type Culture Collection). Streptomycin and cultured in a DMEM (Dulbecco's modified Eagle's medium ) containing the medium-liver cancer cell line (HepG 2) is 10% FBS, 1% penicillin. The cells were cultured at 37 ° C in a 5% CO 2 incubator. The cultured cells were divided into a 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 cells were placed in a 24-well plate, the cells were used when 24 wells were completely attached to the cells and the cells reached 75% confluence.
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 Olate (Sigma O-75010) stock solution
70℃에서 0.1M NaOH 용액을 용매로 이용하여 제조하고, 0.2㎛의 필터를 이용하여 여과한 후, 멸균하였다.A 0.1 M NaOH solution was used as a solvent at 70 째 C, filtered using a 0.2 탆 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.After the third distilled water was used as a solvent, it was sterilized and stored at 4 ° C in a refrigerator.
(3) 올레이트 및 팔미테이트의 농도가 2:1이 되도록 혼합한 5mM의 혼합지방산 저장용액 제조(3) Preparation of 5 mM mixed fatty acid storage solution mixed so that the concentration of olate and palmitate was 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 5O < 0 > C, a 5% (w / v) BSA solution containing no free fatty acid dissolved in the tertiary distilled water prepared in 2) above was used as a solvent and the concentration of each fatty acid prepared in 1) 10 ml of a fatty acid storage solution was prepared. 100 μl of palmitate and 100 mM of the olate storage solution prepared in 1) above were mixed with 165 μl (1.65 mM) of palmitate and 330 μl (3.3 mM) of oleate and 9505 μl of 5% / v) in a BSA solution with vortexing. Thereafter, the mixture was cooled to room temperature, and then filtered with a 0.2 μm filter (the mixed solution was stable at -20 ° C. for 3 to 4 weeks).
실시예Example 3. 벼메뚜기 추출물의 제조 3. Manufacture of rice-grasshopper extract
벼메뚜기를 70%(v/v) 에탄올로 추출하여 에탄올 추출액을 얻고, 에탄올 추출액을 감압 증류하여 에탄올 추출물을 획득하였다.
The rice seedlings were 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 (( 아디포레드Adipo Red ) 염색() dyeing( AdipoRedAdipoRed staining) staining) 어세이를Assay 이용한 Used 중성지방의 함량 평가Evaluation of neutral fat content
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. Determine the degree of generation by the free fatty acids treated triglyceride is reduced by byeomettugi extract staining assay Nile Red (H depot red): liver cancer cell line (HepG 2) to 1mM of oleate and palmitate mixture (1: 2) to, liver cancer cell line (HepG 2) oleate of 1mM in medium containing and palmitate mixture (2: 1) combination treatment of free fatty acids (FFA) treatment and free fatty acids and byeomettugi extract of the present invention and incubated for 24 hours After that, the medium was suctioned gently.
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, 500 μl / well of AdipoRed solution (prepared by dissolving 300 μl of AdipoRed in 10 ml of PBS) was dispensed under the condition of 37 ° C., and shaken for 15 minutes under a dark condition at room temperature for 15 minutes Respectively. The plates were wrapped with aluminum foil to minimize photobleaching of the dye during this process. After the completion of the reaction, it was applied to a spectrophotometer to measure excitation at 485 nm and emission at 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 concentration-dependent reduction of triglyceride was observed at concentrations of 10, 50, and 100 μg / ml of the grasshopper extract (FIG. 1).
실시예 5. 벼메뚜기 추출물의 처리에 따른 총 중성지방의 함량 및 지방 흡수(lipid uptake) 정도의 변화 확인 Example 5: Determination of total triglyceride content and degree of lipid uptake according to the treatment of rice hull extract
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.It was confirmed that the total triglyceride content and lipid uptake decreased by the treatment of the rice grasshopper extract after treatment with hepatocellular carcinoma cell line HepG 2 (1: 2 ) with olate and palmitate mixed (2: 1) .
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 FIG. 2 and FIG. 3, it was confirmed that the total triglyceride content and lipid uptake decreased in a concentration-dependent manner of the rice plant extract.
실시예 6. 벼메뚜기 추출물의 처리에 따른 세포 생존율 및 Caspase -3의 발현량 변화 확인 Example 6 Confirmation of Cell Survival Rate and Caspase- 3 Expression by the Treatment of the Mushroom Extracts of Rice
간암 세포주(HepG2)에 에탄올을 처리하여 감소한 세포 생존률 및 증진된Caspase-3 발현량이 벼메뚜기 추출물의 처리에 의해 변화가 일어나는지를 확인하였다.Liver cancer cell line (HepG 2) the change is going on by the processes of ethanol decreased cell viability and increase the amount of Caspase-3 expression in the treatment of the extract was found to byeomettugi.
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 Fig. 4 and Fig. 5, cell survival rate was increased in a concentration-dependent manner of the rice plant extract and Caspase-3 expression was remarkably decreased.
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 by administration of rice grasshopper extract; Liver, abdominal subcutaneous fat, kidney fat tissue and small intestine fat 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. C57bl / 6Jj mice (mice) of 10 weeks of age were used in this Example 7, and male C57bl / 6Jj mice were fed from BioLink (Eumsung, Korea) for 7 weeks and adapted to the laboratory environment for 3 weeks. Mice with good health status were used in 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 fed with solid feed (antibiotics-free, Samyang Feed Co.) and water until the day of the experiment and maintained the environment of temperature 23 ± 2 ℃, humidity 55 ± 15% (AIN-76A diet) and water were freely supplied and adapted to the laboratory environment. Then, 10-week-old mice with a good health condition were kept in a dark room at 23 ± 2 ° C, humidity 55 ± 15%, and 12 hours intervals with light on (07: 00 ~ 19: 00) 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 divided into two groups: normal control group (Nor), control group (HFD, 60% fat), high fat diet group (100, 200mg / kg) (100 mg / kg). The water and the diet of each experimental group were fed freely, and the rice horticulture extract was orally administered for 14 weeks.
(2) 체중변화, 간 조직, 복부피하지방, 소장 및 신장의 지방조직 변화(2) Changes in body weight, liver tissue, abdominal subcutaneous fat, small intestine and kidney
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 weights of the experimental animals were measured and recorded for 14 weeks at a constant time every week. After 14 weeks, the experimental animals were divided into liver tissue weight, abdominal subcutaneous fat, small intestine fat and retroperitoneal adipose tissue And the weight of adipose tissue was calculated after extraction.
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 examining the effect of the rice grasshopper extract (MTG) of the present invention on the mouse body weight increased by the high fat diet method, the body weight was decreased as compared with the high fat dietary 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 whether the rice plant extract (MTG) of the present invention can reduce the liver tissue weight increased by the high fat diet, the liver tissue of the group that consumed both the high fat diet and the rice horticultural extract (MTG) The weight of the abdominal subcutaneous fat, kidney adipose tissue and small intestine adipose tissue increased in the high fat diet group (HFD) according to the high fat diet method. However, in the high fat diet group, The weight of abdominal subcutaneous fat, kidney adipose tissue and small intestine adipose tissue of mice decreased (Fig. 7).
(3) (3) 비알콜성Non-alcoholic 지방간( Fatty liver NAFLDNAFLD ) 유발 생쥐의 혈장 중 지질생화학적 지표분석) Lipid Biochemical Indicators in Plasma of Mouse
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.ALT and AST, which are indicators of liver function in plasma separated from each experimental animal after 14 weeks of rice grasshopper extract and milk seed, were measured by using triglyceride, total cholesterol, Low-density lipoprotein, free fatty acid, and blood sugar content were measured using a biochemical automatic analyzer (Hitachi-720, Hitachi Medical, Japan).
①① 혈중 ALT, Serum 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 investigating whether MTG could decrease ALT and AST in high fat diets, ALT and AST were significantly higher in high fat diet group and MTG group than high fat diet group (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, it was shown that the diet of rice hull extract (MTG) decreased the ALT and AST enzyme activities in the blood and improved the 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 분석) Lipid biochemical analysis of plasma in 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.
(MTG) decreased the triglyceride, total cholesterol, low density lipoprotein, and free fatty acid levels in the blood, which were increased by the high fat diet. As a result, the triglyceride, total cholesterol, low density Lipoprotein and free fatty acid levels were statistically significantly decreased in the rice-grasshopper extract-treated group (FIG. 9). Therefore, it has been shown that the rice plant extract (MTG) diet reduces lipid levels in the blood and improves the nonalcoholic fatty liver induced by high fat diet.
③ 혈청 내 혈당 농도 변화③ Serum glucose concentration change
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).
The blood glucose concentration in the serum was increased by the high fat diet (HFD), but the blood glucose concentration in the high fat diet and the rice horticultural extract (MTG) group was decreased statistically significantly 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 of the above experimental groups were statistically analyzed using an unpaired student's T-test statistical program, and a significance test was performed at a level of p <0.05 or less.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20210060233A (en) | 2019-11-18 | 2021-05-26 | 세종대학교산학협력단 | Pharmaceutical composition for treating or preventing leaky gut syndrome |
| KR20220109668A (en) * | 2021-01-29 | 2022-08-05 | 한국원자력연구원 | Composition for oral administration comprising extract of oxya chinensis sinuosa |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20050079443A (en) * | 2004-02-05 | 2005-08-10 | 김빛나 | Process for preparing the high molecular weight antitumor fractions derived from edible grasshoppers |
| 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 |
| 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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- 2017-12-28 KR KR1020170181811A patent/KR102041852B1/en active IP Right Grant
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20050079443A (en) * | 2004-02-05 | 2005-08-10 | 김빛나 | Process for preparing the high molecular weight antitumor fractions derived from edible grasshoppers |
| 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 |
| 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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| Entomological Research. 2009, Vol.39, pp.299-303 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20210060233A (en) | 2019-11-18 | 2021-05-26 | 세종대학교산학협력단 | Pharmaceutical composition for treating or preventing leaky gut syndrome |
| KR20220109668A (en) * | 2021-01-29 | 2022-08-05 | 한국원자력연구원 | Composition for oral administration comprising extract of oxya chinensis sinuosa |
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| Publication number | Publication date |
|---|---|
| KR102041852B1 (en) | 2019-11-07 |
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