KR20110053141A - Composition for anti-diabetes comprising germinated brown rice extracts - Google Patents
Composition for anti-diabetes comprising germinated brown rice extracts Download PDFInfo
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- KR20110053141A KR20110053141A KR1020090109981A KR20090109981A KR20110053141A KR 20110053141 A KR20110053141 A KR 20110053141A KR 1020090109981 A KR1020090109981 A KR 1020090109981A KR 20090109981 A KR20090109981 A KR 20090109981A KR 20110053141 A KR20110053141 A KR 20110053141A
- Authority
- KR
- South Korea
- Prior art keywords
- brown rice
- germinated brown
- extract
- organic solvent
- diabetes
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Abstract
Description
본 발명은 발아현미 추출물을 유효성분으로 포함하는 혈청 글루코즈 수준을 조절하기 위한 조성물에 관한 것으로 보다 구체적으로는 당뇨, 특히 인슐린 비의존성 당뇨의 예방 또는 치료용 조성물에 관한 것이다. The present invention relates to a composition for controlling serum glucose level comprising germinated brown rice extract as an active ingredient, and more particularly to a composition for preventing or treating diabetes, in particular insulin-independent diabetes.
당뇨는 혈액 내에 존재하는 포도당이 소변을 통해 배출되는 질환으로서, 근본적인 치유가 되지 않는 만성퇴행성질환 중의 하나이다. 급속한 경제발전에 따른 식생활의 변화로 평균수명은 연장되는 현상에 반하여 당뇨병 등의 만성퇴행성질환은 증가하는 현상을 보이고 있다. 국내에서 당뇨의 유병율은 약 5~10%에 달하는 것으로 조사되어 있으며, 2007년 당뇨 환자는 400만명을 넘어서 인구 100명 중 8.3명이 당뇨 질환자이지만 2025년에는 당뇨 환자가 약 680만명으로 늘어날 것으로 예측되고 있다. 당뇨는 전 세계적으로 3번째로 심각한 질병으로 다양한 합병증으로 인해 환자의 수명이 5~10년 정도 단축되며, 합병증을 포함하여 현재 우리나라의 경우 사망원인 중 4순위에 있다.Diabetes is a disease in which glucose in the blood is released through urine and is one of the chronic degenerative diseases that does not cure fundamentally. As life expectancy changes due to rapid economic development, life expectancy is increasing, whereas chronic degenerative diseases such as diabetes are increasing. The prevalence of diabetes in Korea is estimated to be about 5 to 10%. In 2007, the number of diabetics exceeded 4 million, 8.3 out of every 100 people with diabetes, but the number of diabetics is expected to increase to about 6.8 million by 2025. have. Diabetes is the third most serious disease in the world and the life span of patients is shortened by 5 to 10 years due to various complications, including complications.
당뇨 치료는 식이요법, 운동요법 및 약물요법으로 대별되며 환자의 병태에 따라 결정되고 있다. 당뇨는 질환 그 자체보다 합병증의 예방 및 치료가 매우 중요하므로 관리의 목적은 정상 혈당유지와 합병증을 예방하는 데 있다고 할 수 있다. 한편 당뇨 치료제는 크게 3 그룹으로 구분되며, ① 당이 소장에 흡수되지 못하도록 저해하는 당질분해 저해제, ② 인슐린의 작용을 도와주는 인슐린 저항 개선제, ③ 인슐린 분비 촉진제 등으로 나눌 수 있다. 이들 중 당질분해 저해성 경구 혈당강하제는 식이요법과 운동요법으로 혈당조절이 이루어지지 않는 제2형 당뇨환자들에게 주로 투약되고 있다. 그동안 수많은 약제들이 개발되어 왔으나, 대부분은 부작용이나, 유효성 혹은 안전성이 문제되어 현재 임상에 사용되고 있는 약제는 소수에 불과하다. Diabetes treatment is roughly divided into diet, exercise therapy and drug therapy and is determined by the patient's condition. Diabetes is more important to prevent and treat complications than the disease itself, so management can be said to maintain normal blood sugar and prevent complications. Meanwhile, diabetic drugs can be divided into three groups. ① Glycolytic inhibitors that inhibit sugar from being absorbed into the small intestine, ② Insulin resistance improving agent to help the action of insulin, ③ insulin secretagogues. Among these, glycolytic inhibitor oral hypoglycemic agents are mainly administered to patients with type 2 diabetes who do not have glycemic control by diet and exercise therapy. Many drugs have been developed in the meantime, but most of them have side effects, but only a few drugs are currently being used in the clinic because of their effectiveness or safety.
현재 혈당강하제는 설폰요소제인 글리메피라이드(glimepiride)가 주로 사용되고 있으나, 저혈당증 유발 인슐린 분비기능 상실이 부작용으로 알려져 있다. 비구아니드계 약물인 글루코파지(glucophage)는 국내에서 많이 사용되는 약제로 주로 간에서 인슐린 작용을 도와 혈당을 낮추지만 부작용으로 위장장애, 설사, 젖산증 등을 나타낸다. Currently, glycemic hypoglycemic agents, glymepiride, a sulfone urea agent, are mainly used, but hypoglycemia-induced loss of insulin secretion function is known as a side effect. Glucophage (glucophage), a biguanide-based drug, is a drug that is widely used in Korea, mainly to help insulin function in the liver to lower blood sugar, but side effects include gastrointestinal disorders, diarrhea, and lactic acidosis.
알파-글루코시데이즈 저해제는 소장에서 탄수화물의 분해를 저해함으로써 포도당의 흡수를 감소시켜며, 해당 약제로는 아카보즈(acarbose), 보글리보즈(voglibose), 미글리톨(miglitol) 등이 대표적이며 아카보즈(acarbose)는 현재 국내에서도 사용되고 있고, 보글리보즈(voglibose), 미글리톨(miglitol) 등도 최근 에 국내에 소개되기 시작하였으나 부작용으로는 설사와 배탈 등을 나타낸다. Alpha-glucosidase inhibitors reduce glucose uptake by inhibiting the breakdown of carbohydrates in the small intestine, and the drugs include acarbose, avoglibose, and miglitol. Boaz (acarbose) is currently used in Korea, boglibose (voglibose), miglitol (miglitol), etc. have recently been introduced in Korea, but side effects include diarrhea and upset stomach.
이와 같이 약물요법에 따른 부작용으로 인하여 식품소재를 이용한 항당뇨 천연물질에 대한 관심이 증가되고 있는 추세이다. 약초식물인 뽕잎, 달개비풀, 인삼, 연과육, 달맞이꽃, 결명자, 구기자 등이, 대두, 해조류, 우엉, 메밀 등의 식품들이 당뇨병에 효능을 갖는 것으로 보고, 또는 알려짐으로써 민간요법으로 이용되고 있다. As a result of the side effects of the drug therapy, there is a growing trend to increase the anti-diabetic natural substances using food materials. Herb plants, such as mulberry leaves, coriander, ginseng, lotus flesh, evening primrose, gwanjaja, gojija, etc., soybeans, seaweeds, burdock, buckwheat and other foods are reported to have an effect on diabetes, or is being used as a folk remedy.
현미(Oryza sativa)는 도정에 의한 영양분의 손실이 거의 없어 백미에 비해 비타민, 무기질 및 식이섬유가 풍부함에도 불구하고 소화성이 낮고 식감이 좋지 않아 선호도가 낮다. 최근에는 이를 개선하기 위한 발아현미 제품이 출시되고 있다. 발아현미란 적당한 온도와 수분, 산소 등을 공급해 현미의 싹을 1~5 mm까지 발아시킨 것으로 현미의 영양과 기능을 극대화시키면서 식미는 현미보다 훨씬 부드러운 쌀이다. 발아현미는 현미의 발아 과정에서 세포벽 분해효소의 작용에 의해 현미 외피가 분해되어 수분흡수가 용이하게 됨에 따라 취반성이 개선되는 것으로 알려져 있다. 발아현미에는 GABA, 비타민 E, 스쿠알렌(squalene), 시토스테롤(sitosterol), 캄페스테롤(campesterol) 및 스티그마스테롤(stigmasterol) 등이 주요한 구성 성분으로 밝혀져 있고, 탄수화물의 인체 내 소화율 증가, 반영양적 요소의 감소나 제거 등 다양한 기능성 식품 소재로의 활용 연구가 활발히 이루어지고 있다. 한편 생리·약리활성으로는 고혈압 예방, 신경안정 등이 보고되고 있다. 그 이외 발아 중 저장 단백질의 질적 변화, 미량 영양소의 증가, 구성 탄수화물의 인체 내 소화율 증가 및 반영양적 요소의 감소나 제거 등 다양한 기능성 식품소재로 의 활용 연구가 활발히 이루어지고 있으나 발아 현미 추출물 자체가 소장에서 포도당의 흡수를 저해하는 탁월한 효과가 있음이 보고된 바는 없다. Brown rice ( Oryza sativa ) has less nutrients due to milling and is less preferred because of its low digestibility and poor texture, despite its richer vitamins, minerals and fiber than white rice. Recently, germinated brown rice products have been released to improve this. Germinated brown rice is a germination of brown rice to 1 ~ 5 mm by supplying proper temperature, moisture and oxygen, while maximizing the nutrition and function of brown rice, the taste is much softer than brown rice. Germinated brown rice is known to improve the cohesiveness as the brown rice shell is decomposed by the action of cell wall degrading enzymes during the germination process of brown rice to facilitate the absorption of moisture. Germinated brown rice contains GABA, vitamin E, squalene, sitosterol, campesterol, and stigmasterol as major constituents. Research on the utilization of various functional foods such as removing or removing them is being actively conducted. On the other hand, physiological and pharmacological activities have been reported to prevent hypertension and neurostable. In addition, research on the use of various functional food materials such as qualitative changes of stored protein during the germination, an increase in micronutrients, an increase in digestibility of constituent carbohydrates, and a reduction or elimination of semi-quantitative factors have been actively conducted. It has not been reported that there is an excellent effect of inhibiting the absorption of glucose in.
이러한 배경 하에 본 발명자들은 발아현미로부터 다양한 생리·약리활성을 갖는 미량소재에 대해 연구하던 중 발아현미의 수, 추출물, 또는 유기용매 추출물이 소장 내에서 포도당 흡수를 저해하여 혈당강하작용을 나타낸다는 점을 확인함으로써 본 발명을 완성하게 되었다.Under these backgrounds, the present inventors studied micromaterials having various physiological and pharmacological activities from germinated brown rice, and the number, extract, or organic solvent extract of germinated brown rice showed hypoglycemic action by inhibiting glucose absorption in the small intestine. By confirming that the present invention was completed.
본 발명의 목적은 발아현미 수, 유기용매 또는 이들의 혼합용매 추출물을 유효성분으로 포함하는 혈청 글루코즈 수준을 조절하기 위한 조성물을 제공하는 것이다. 여기에서 상기 추출에 사용되는 물은 냉수 또는 열수일 수 있다.It is an object of the present invention to provide a composition for regulating serum glucose levels comprising germinated brown rice water, an organic solvent or a mixed solvent extract thereof as an active ingredient. Here, the water used for the extraction may be cold or hot water.
본 발명의 다른 목적은 발아현미 추출물을 유효성분으로 하는 당뇨병, 특히 인슐린 비의존성 당뇨병의 예방 또는 치료용 조성물을 제공하는 것이다.Another object of the present invention is to provide a composition for the prevention or treatment of diabetes, in particular, insulin-independent diabetes, which has germinated brown rice extract as an active ingredient.
본 발명의 다른 목적은 발아현미 추출물을 유효성분으로 하는 소장에서의 포도당 흡수 저해용 조성물을 제공하는 것이다. It is another object of the present invention to provide a composition for inhibiting glucose absorption in the small intestine having a germinated brown rice extract as an active ingredient.
본 발명의 다른 목적은 발아현미 추출물을 유효성분으로 포함하는 당뇨 질환 개선을 위한 건강기능식품을 제공하는 것이다. Another object of the present invention is to provide a health functional food for diabetic disease improvement comprising the germinated brown rice extract as an active ingredient.
본 발명은 발아현미 추출물을 유효성분으로 포함하는 혈청 글루코즈 수준을 조절하기 위한 조성물에 관한 것으로서, 구체적으로 발아현미 수, 유기용매 또는 이들의 혼합용매 추출물을 유효성분으로 포함하는 혈당강하제용 당뇨병 예방 또는 치료용 조성물에 관한 것이다. The present invention relates to a composition for controlling serum glucose level comprising the germinated brown rice extract as an active ingredient, specifically preventing the hypoglycemic agent diabetic comprising a germinated brown rice water, an organic solvent or a mixed solvent extract thereof as an active ingredient A therapeutic composition.
본 발명의 발아현미 추출물은 독성, 부작용 등의 우려가 없는 식품소재로, 특히 소장에 포도당의 흡수를 저해함으로써, 당뇨를 효과적으로 예방 또는 치료할 수 있을 뿐만 아니라 당뇨 질환으로 인한 각종 증상의 개선을 위한 건강기능식품용으로도 유용하게 사용될 수 있다. Germinated brown rice extract of the present invention is a food material that does not have toxicity, side effects, and the like, in particular, by inhibiting the absorption of glucose in the small intestine, not only can effectively prevent or treat diabetes, but also improve the health of various symptoms due to diabetes diseases. It can also be usefully used for nutraceuticals.
본 발명은 발아현미 수, 유기용매 또는 이들의 혼합용매 추출물을 유효성분으로 포함하는 혈청 글루코즈 수준을 조절하기 위한 조성물에 관한 것이다. 상기 추출에 사용되는 물은 냉수 또는 열수일 수 있다.The present invention relates to a composition for regulating serum glucose level comprising germinated brown rice water, organic solvent or mixed solvent extract thereof as an active ingredient. The water used for the extraction may be cold or hot water.
본 발명의 발아현미 추출물은 구체적으로 유기용매, 보다 구체적으로 C1 -10 유기용매로 추출할 수 있으며, 메탄올, 에탄올, 이소프로필 알코올, 부탄올, 펜탄올, 헥산올, 시클로헥산올, 디에틸에테르, 테트라하이드로퓨란, 에틸아세테이트, 에틸 아세토아세테이트, 에틸 벤조에이트, 디클로로메탄, 헥산, 클로로포름 아세톤 또는 이들의 혼합용매가 사용될 수 있다. Germinated brown rice extract is specifically an organic solvent of the present invention, more specifically C 1 -10 can be extracted with an organic solvent, methanol, ethanol, isopropyl alcohol, butanol, pentanol, hexanol, cyclohexanol, diethyl ether , Tetrahydrofuran, ethyl acetate, ethyl acetoacetate, ethyl benzoate, dichloromethane, hexane, chloroform acetone or a mixed solvent thereof may be used.
본 발명의 발아현미 추출물은 당뇨병으로 인해 상승된 혈청 글루코즈 수준을 조절할 수 있으며, 구체적으로 상기 당뇨병은 인슐린 비의존성 당뇨병이다. Germinated brown rice extract of the present invention can regulate elevated serum glucose levels due to diabetes, specifically, the diabetes is insulin-independent diabetes.
본 발명의 발아현미 추출물은 알파-글루코시다아제 저해 활성 또는 소장에서의 포도당 흡수 저해 활성을 통해 혈청 글루코즈 수준을 조절할 수 있다. Germinated brown rice extract of the present invention can regulate serum glucose levels through alpha-glucosidase inhibitory activity or glucose uptake inhibitory activity in the small intestine.
본 발명은 또한 발아현미 수, 유기용매 또는 이들의 혼합용매 추출물을 유효성분으로 포함하는 혈청 글루코즈 수준 개선용 분말, 식품, 건강기능식품 또는 음 료에 관한 것이다. 여기에서 상기 추출에 사용되는 물은 냉수 또는 열수일 수 있다.The present invention also relates to a powder for improving serum glucose level, food, health functional food or beverage comprising germinated brown rice water, organic solvent or mixed solvent extract thereof as an active ingredient. Here, the water used for the extraction may be cold or hot water.
본 발명은 또한 발아현미 수, 유기용매 또는 이들의 혼합용매 추출물을 유효성분으로 포함하는 조성물을 섭취함으로써, 인간을 제외한 포유동물에서 혈청 글루코즈 수준을 감소시키기 위한 방법에 관한 것이다. 여기에서 상기 추출에 사용되는 물은 냉수 또는 열수일 수 있다.The present invention also relates to a method for reducing serum glucose levels in mammals other than humans by ingesting a composition comprising germinated brown rice water, an organic solvent or a mixed solvent extract thereof as an active ingredient. Here, the water used for the extraction may be cold or hot water.
본 발명은, The present invention,
(a) 발아현미를 건조시키는 단계;(a) drying the germinated brown rice;
(b) 건조된 발아현미를 수, 유기용매 또는 이들의 혼합용매로 추출하는 단계;(b) extracting the dried germinated brown rice with water, an organic solvent or a mixed solvent thereof;
(c) 비극성 용매로부터 극성 용매 순으로 분획하는 단계; 및(c) fractionating from nonpolar solvents to polar solvents in order; And
(d) 분획 후 남은 분획을 건조시키는 단계를 포함하는(d) drying the remaining fraction after fractionation.
발아현미 추출물을 제조하는 방법에 관한 것이다. 여기에서 상기 추출에 사용되는 물은 냉수 또는 열수일 수 있다.A method for preparing germinated brown rice extracts. Here, the water used for the extraction may be cold or hot water.
본 발명의 발아현미 추출물은 통상의 추출방법이 제한 없이 사용될 수 있으나, 바람직하게 본 발명은 건조된 상태의 발아현미를 분쇄한 후 실온에서 추출하는 한편, 상기 추출물은 물 또는 유기용매, 유기용매의 경우 특히 C1 -10 유기용매, 구체적으로 메탄올, 에탄올, 이소프로필 알코올, 부탄올, 헥산, 클로로포름, 에틸아세테이트, 아세톤 등의 유기용매로 분획할 수 있으며, 비극성 용매로부터 극성 용매 순으로 분획할 수 있다. 보다 구체적으로 본 발명의 추출물은 메탄올 추출 후, 그 잔사를 헥산, 클로로포름, 메틸아세테이트 순으로 분획하고. 남은 것을 메탄올 분획으로 하여 여과함으로써 수득할 수 있다 (실시예 1 및 도 1 참조).Germinated brown rice extract of the present invention can be used without any conventional extraction method, but preferably the present invention is extracted at room temperature after grinding the germinated brown rice in a dry state, the extract is water or organic solvent, organic solvent when a particular fraction may be C 1 -10 organic solvent, specifically, methanol, ethanol, isopropyl alcohol, butanol, hexane, chloroform, ethyl acetate, can be fractionated in an organic solvent such as acetone, and polar solvent from the non-polar solvent in order . More specifically, the extract of the present invention, after methanol extraction, the residue is fractionated in the order of hexane, chloroform, methyl acetate. The remaining can be obtained by filtration with methanol fraction (see Example 1 and FIG. 1).
당뇨는 인슐린 의존성 당뇨(제1형 당뇨)와 인슐린 비의존성 당뇨(제2형 당뇨)로 구분되며 선진국에서는 제2형 당뇨가 크게 증가하고 있다. 제2형 당뇨는 에너지의 과잉섭취와 운동부족 등의 외부 환경인자에 영향을 받고 있으며 인슐린저항성의 증가와 베타세포 기능의 감소로 인하여 혈액에서 고혈당 증상을 나타내면서, 여러 합병증을 동반하게 된다. Diabetes is divided into insulin-dependent diabetes (
본 발명에서의 혈당강하작용이란 당뇨의 치료방법 중 소장에서 포도당의 흡수에 대하여 저해능을 가짐으로써, 혈액 중에 혈당수치를 낮추어 주면서 인슐린 비의존성 당뇨에 크게 효과적임을 밝혀냈다.In the present invention, the hypoglycemic effect has been found to be highly effective for insulin-independent diabetes while lowering the blood glucose level in blood by inhibiting glucose absorption in the small intestine.
혈당강하 작용의 하나로 위장관내 알파-글루코시디아제의 저해가 있다. 알파-글루코시다아제의 혈당강하 기전은 소장 내에서 탄수화물의 분해를 저해하여 포도당의 흡수를 지연시킴으로써 소장에서 혈액으로 포도당 흡수를 저해하는 것이다. 본 발명의 핵심인 소장에서 포도당의 흡수저해에 앞서 발아현미 추출물은 알파-글루코시다아제를 저해함으로써 혈당강하에 효과가 있음을 1차적으로 확인하였다. (실험예 1 및 도 4) One hypoglycemic action is the inhibition of alpha-glucosidase in the gastrointestinal tract. The hypoglycemic mechanism of alpha-glucosidase inhibits glucose uptake into the blood from the small intestine by inhibiting the breakdown of carbohydrates in the small intestine and delaying the absorption of glucose. Prior to the inhibition of glucose absorption in the small intestine, which is the core of the present invention, the germinated brown rice extract was primarily confirmed to have an effect on hypoglycemia by inhibiting alpha-glucosidase. Experimental Example 1 and FIG. 4
섭취된 식이 탄수화물은 아밀라아제(amylase)와 글루코시다아제 (glucosidase)에 의해 소화되어 포도당으로 분해된 후 소장 상피세포에서 흡수, 수 송된다. 이 수송에 관여하는 운반체는 GLUT(sodium-independent glucose transporters) 계열과 SGLT(sodium-dependent glucose transporters) 계열로 대별되며, 포도당은 SGLT1의 작용에 의해 소장 내에서 흡수된다. SGLT1은 소장 내 가장 함량이 높은 포도당 운반체로서 상피세포의 BBMV(brush border membrane vesicle)에 존재하기 때문에 포도당 과다 흡수에 의한 당뇨의 예방용 소재개발에 있어 주목을 받아온 운반체 중의 하나이다. Ingested dietary carbohydrates are digested by amylase and glucosidase, broken down into glucose, and then absorbed and transported by small intestinal epithelial cells. Carriers involved in this transport are classified into the GLUT (sodium-independent glucose transporters) family and the SGLT (sodium-dependent glucose transporters) family, and glucose is absorbed in the small intestine by the action of SGLT1. SGLT1 is one of the carriers with the highest content of glucose in the small intestine and has been attracting attention in developing a material for preventing diabetes due to excessive absorption of glucose because it is present in BBMV (brush border membrane vesicle) of epithelial cells.
따라서 SGLT1의 생합성 억제 또는 활성저해에 의한 당뇨의 예방 및 치료제 개발 전략은 기존의 전분가수분해효소 저해제에 의한 방법에 비해 음식물의 섭취량과 종류에 관계없이 포도당의 흡수를 직접적으로 제어할 수 있다는 관점에서 유효성이 높다. 이들 중 GLUT은 글루코즈(glucose)의 흡수 연구와 인슐린 액션(insulin action)에 연관되어 있고 특히 GLUT1과 3은 베이절(basal)에 존재하여 소장에서 혈액으로 포도당 흡수에 관계한다. 이들 트랜스포터(transporter)를 저해하는 물질에 대한 연구가 플라보노이드(flavonoid)계 물질을 중심으로 진행되고 있으나 저해효과만을 확인하였을 뿐 저해 기작은 아직 밝혀지지 않은 상태이다. 최근 플라보노이드류 쿼세틴(quercetin)의 소장에서 혈액으로 포도당 흡수의 저해효과가 밝혀지면서 식용·약용 생물자원에도 저해활성 성분이 함유되어 있을 것으로 예상되며, 이 성분들을 소재화하여 장에서의 포도당 흡수를 낮출 수 있다면 당뇨를 예방할 수 있는 최선의 방안이라 하겠다. Therefore, the strategy of preventing and treating diabetes by inhibiting or inhibiting biosynthesis of SGLT1 can be directly controlled in terms of glucose uptake regardless of food intake and type, compared to conventional starch hydrolase inhibitors. Highly effective. Of these, GLUT is involved in glucose uptake studies and insulin action, and GLUT1 and 3, in particular, are present in the basal and are involved in glucose uptake into the blood from the small intestine. Studies on the substances that inhibit these transporters (transporters) are being conducted mainly on flavonoid-based substances, but only the inhibitory effect has been confirmed, but the mechanism of inhibition is still unknown. Recently, as the inhibitory effect of glucose absorption into the blood from the small intestine of the flavonoids quercetin has been revealed, it is expected that edible and medicinal biological resources will contain inhibitory active ingredients. If you can, it is the best way to prevent diabetes.
본 발명에서는, 발아현미 추출물에서 소장에서 혈액으로 포도당의 흡수를 저해효과를 확인하기 위하여 Caco-2 세포를 배양하여, 본 발명의 추출물을 농도별로 처리한 결과, 발아현미 추출물이 대조군으로 사용된 현미에 비하여 높은 혈당강하 효과를 가짐을 확인하였다 (실험예 2). In the present invention, in order to confirm the inhibitory effect of glucose uptake into the blood from the small intestine extract from the germinated brown rice, Caco-2 cells were cultured, and the extracts of the present invention were treated according to the concentration. It was confirmed that it has a high blood sugar lowering effect compared to (Experimental Example 2).
또 다른 하나의 양태로서, 본 발명은 발아현미 추출물을 유효성분으로 하는 SGLT1 저해용 조성물에 관한 것이다. As another aspect, the present invention relates to a composition for inhibiting SGLT1 using the germinated brown rice extract as an active ingredient.
본 발명자들은 소장에서 혈액으로 포도당의 흡수 주요 인자인 SGLT1(sodium-dependent glucose transporters1), HNF-1α(hepatocyte nuclear factor-1α), HNF-1β(hepatocyte nuclear factor-1β)의 발현이 감소됨을 측정함으로써 본 발명의 발아현미 추출물, 구체적으로는 발아현미 메탄올 추출물이 소장에서 혈액으로 포도당의 흡수 저해 활성을 가짐을 확인하였다 (실험예 3 및 도 6 내지 8 결과 참조). By measuring the expression of sodium-dependent glucose transporters1 (SGLT1), hepatocyte nuclear factor-1α (HNF-1α), and hepatocyte nuclear factor-1β (HNF-1β), which are major factors for glucose uptake into the blood in the small intestine, Germinated brown rice extract of the present invention, specifically, germinated brown rice methanol extract was confirmed to have the activity of inhibiting the absorption of glucose into the blood in the small intestine (see Experimental Example 3 and Figures 6 to 8 results).
본 발명에서의 용어 “HNF-1α(hepatocyte nuclear factor-1α)”란 TCF1(transcription facter1)로 불리며, 이것은 호메오도메인-함유 트랜스크립션 팩터(homeodomain-contatining transcription facter)로서 전사인자에 중요한 물질로 간, 신장, 췌장 그리고 소장에서 대사작용에 관여한다. 이것은 호메오박스 프로틴 2(homeobox protein 2, CDX2) 와 GATA-바인딩 단백질 그룹과 상호작용을 하여 소장에서의 SGLT1 발현을 조절하는 것으로 보고되어 있다. 또한 본 발명에서의 용어, “HNF-1β(hepatocyte nuclear factor-1β)”란, TCF2(transcription facter2)로 불리며 HNF-1α의 호메오도메인-함유(homeodomain-contatining)의 상위 역할을 한다. 또한, 간, 신장, 췌장 그리고 소장에서 대사 작용을 돕는 역할을 하고, 이것은 HNF-1α와 상호작용을 하여 소장에서 SGLT1 발현을 조절하는 유전자로 보고되어 있다. The term “hepatocyte nuclear factor-1α” in the present invention is called a transcription facter1 (TCF1), which is a homeodomain-contatining transcription facter and is an important substance for transcription factors. It is involved in metabolism in the liver, kidneys, pancreas and small intestine. It has been reported to interact with homeobox protein 2 (CDX2) and GATA-binding protein groups to regulate SGLT1 expression in the small intestine. In addition, the term "hepatocyte nuclear factor-1β" in the present invention is referred to as transcription facter2 (TCF2) and plays an upper role of homeodomain-contatining of HNF-1α. It also plays a role in helping metabolism in the liver, kidney, pancreas and small intestine, which has been reported to interact with HNF-1α to regulate SGLT1 expression in the small intestine.
이와 같이 상기 전사인자들은 소장에서 포도당 흡수의 수송단백질인 SGLT1 발현을 조절하여 소장에서 포도당의 흡수에 관여하는 일련의 유전자 발현을 조절하는 역할을 하는데, 특히, HNF-1α와 HNF-1β의 발현에 의해서 SGLT1 발현을 조절하는 중요한 전사인자로, 소장에 포도당 흡수를 총괄적으로 조절하는 역할을 한다. As such, the transcription factors regulate SGLT1 expression, a transport protein for glucose uptake in the small intestine, thereby regulating a series of gene expressions involved in glucose uptake in the small intestine, particularly in the expression of HNF-1α and HNF-1β. It is an important transcription factor that regulates SGLT1 expression, and it is responsible for regulating glucose uptake in the small intestine.
본 발명의 발아현미 추출물이 소장에서 포도당의 흡수저해를 하는지 여부를 확인하기 위하여, SGLT1 및 SGLT1 발현에 관련되는 전사인자인 HNF-1α(hepatocyte nuclear factor-1α), HNF-1β(hepatocyte nuclear factor-1β)의 발현을 확인한 결과, 상기 유전자 모두 발현이 감소됨을 확인하였으며, 이로부터 본 발명의 추출물이 소장에서 포도당의 흡수저해 활성을 가짐을 확인하였다 (실험예 3 및 도 6 내지 8 결과 참조).In order to determine whether the germinated brown rice extract of the present invention inhibits glucose absorption in the small intestine, hepatocyte nuclear factor-1 (HNF-1α) and HNF-1β (hepatocyte nuclear factor-), which are transcription factors related to SGLT1 and SGLT1 expression As a result of confirming the expression of 1β), it was confirmed that the expression of all of the genes was reduced, and from this it was confirmed that the extract of the present invention has the inhibitory activity of glucose absorption in the small intestine (see Experimental Example 3 and Figures 6 to 8 results).
이하, 본 발명에 따르는 실시예 및 본 발명에 따르지 않는 비교 예를 통하여 본 발명을 보다 상세히 설명하나, 본 발명의 범위가 하기 제시된 실시예에 의해 제한되는 것은 아니다.Hereinafter, the present invention will be described in more detail through examples according to the present invention and comparative examples not according to the present invention, but the scope of the present invention is not limited to the examples given below.
실시예Example 1 : 발아현미로부터 검색 시료의 조제 1: Preparation of Search Samples from Germinated Brown Rice
여러 가지 곡류, 양념채소류, 향신료, 식용해조류자원에서 추출한 추출물을 대상으로 항당뇨 활성을 검색하던 중 발아현미 추출물이 높은 포도당 흡수 저해능을 나타냄을 확인하였다. 이에 발아현미에 포함된 포도당 흡수 저해활성물질을 분리하고자 건조된 상태의 발아현미 40 kg를 분쇄한 후, 실온에서 100% 메탄올 40 L로 추출하고, 그 추출물을 농축, 건조한 44.5 g을 헥산, 클로로포름, 에틸아세테이트 순으로 분획하고 남은 것을 메탄올 분획으로 하여 각각을 여과포로 여과하고 여과물을 감압 농축한 후 건조하여 시료를 조제하였다 (도 1). 물로 추출한 경우에는 상기의 100% 메탄올을 대신에 열수 또는 냉수로 추출한 후 추출물을 상기의 방법과 같이 동일하게 분획하여 시료를 조제하였다.Germinated brown rice extract showed high glucose uptake inhibitory activity during the anti-diabetic activity of extracts from various grains, spices, spices and edible algae. Thus, 40 kg of dried germinated brown rice was pulverized to separate the glucose absorption inhibiting active material contained in the germinated brown rice, extracted with 40 L of 100% methanol at room temperature, and the extract was concentrated and dried 44.5 g of hexane and chloroform. After fractionation in the order of ethyl acetate, the remainder was methanol fraction, and each was filtered through a filter cloth, and the filtrate was concentrated under reduced pressure and dried to prepare a sample (FIG. 1). In the case of extraction with water, 100% methanol was extracted with hot or cold water instead, and the extract was fractionated in the same manner as in the above method to prepare a sample.
<< 항당뇨Antidiabetic 활성의 측정> Measurement of Activity
실험예Experimental Example 1 : α- 1: α- 글루코시다아제Glucosidase 저해활성 실험방법 Inhibitory Activity Test Method
α-글루코시다아제 저해활성 측정용 실험방법은 기질인 400 mM PNP-글루코사이드(glycoside)와 1 유니트 α-글루코시다아제(α-glucosidase, from Sacchromyces cerevisiae)를 이용하여 수행하였다. 각 분획물에 대한 실험을 위해 증류수 750 ul, 1 M KPB (pH 7.5) 50 ul, α-글루코시다아제(from sacchromyces cerevisiae) 50 ul, 시료 (1 mg/ml) 100 ul로 구성된 반응액을 실온에서 1분간 반응시켰다. 이 반응액에 PNP- 글루코사이드 50 ul를 첨가하여 4분 후의 흡광도를 분광광도계(spectrophotometer, 405nm)로 측정하였다. The experimental method for measuring α-glucosidase inhibitory activity was carried out using a substrate 400 mM PNP-glucoside (glycoside) and 1 unit α-glucosidase (α-glucosidase, from Sacchromyces cerevisiae ). For experiments on each fraction, the reaction solution consisting of 750 ul of distilled water, 50 ul of 1 M KPB (pH 7.5), 50 ul of α-glucosidase (from sacchromyces cerevisiae ) and 100 ul of sample (1 mg / ml) at room temperature The reaction was carried out for 1 minute. 50 ul of PNP-glucoside was added to the reaction solution, and the absorbance after 4 minutes was measured with a spectrophotometer (405 nm).
<수학식 2><Equation 2>
저해능 (Inhibitory effect, %) = [(Cc - Cs) / Cc] × 100Inhibitory effect (%) = [(Cc-Cs) / Cc] × 100
Cs : 시료를 처리한 실험구의 흡광도Cs: absorbance of the experimental zone treated with the sample
Cc : 시료를 처리하지 않은 실험구의 흡광도Cc: absorbance of the test zone without sample
시료를 처리하지 않은 대조군을 기준으로 비교하였을 때, 농도별로 처리한 발아현미 메탄올 추출물에서 소장 내에 존재하는 α-글루코시다아제를 현저하게 저해함을 확인하였다. (표 1 및 도 4)Compared to the control group without the sample, it was confirmed that the germinated brown rice methanol extract treated by concentration significantly inhibited α-glucosidase present in the small intestine. Table 1 and FIG. 4
<표 1> α-글루코시다아제 저해능 측정 결과Table 1 Results of α-glucosidase inhibitory activity
실험예Experimental Example 2 : 소장에서 혈액으로 포도당 흡수 실험방법 2: Experimental Method of Glucose Absorption into Blood from Small Intestine
소장에서 혈액으로 포도당 흡수의 저해활성 측정용 실험방법은 Caco-2 세포와 2-NBDG(2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino-2-deoxygluocose)를 이용하여 측정하였다. 즉, 각 분획물에 대한 포도당 흡수저해 실험을 위해 Caco-2(human colonic cancer cell)을 6×104 cells/well 농도로 24 웰-플레이트, 37℃, CO2 인큐베이터에서 13~14일 동안 배양한 후 PBS로 2회 세척한 다음 PBS 400 ㎕, 100 μM 2-NBDG 500 ㎕, 샘플 (1 mg/ml) 100 ㎕를 첨가하고 37℃에서 2시간 반응시켰다. 이 반응액을 제거하고 세포를 아이스-콜드 PBS로 3회 세척한 후 2-NBDG의 형 광세기를 형광분광광도계(spectrophotofluorometer, 여기 : 485nm, 발광 : 535nm)로 측정하였다. 글루코즈 업테이크(Glucose uptake) 양 측정에 미치는 세포농도의 영향을 보정해 주기 위해 MTT 어세이를 다음과 같이 실시하였다. 형광세기 측정이 끝난 웰 플레이트에 MTT 용액 (5 mg/ml) 50 ㎕를 가하여 37℃에서 3시간 반응시킨 후 570 nm에서 흡광도를 측정하였다. 시료에 의한 포도당 흡수 저해활성은 아래 식에 따라 환산하였다. Experimental methods for measuring the inhibitory activity of glucose uptake into the blood in the small intestine were measured using Caco-2 cells and 2-NBDG (2- (N- (7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino-2 -deoxygluocose), ie, human colonic cancer cells (CaCO-2) at a concentration of 6 × 10 4 cells / well, 24 well-plates, 37 ° C., CO 2 for glucose uptake experiments for each fraction. After incubation for 13-14 days in the incubator and washed twice with PBS, 400 μl of PBS, 500 μl of 100 μM 2-NBDG, 100 μl of sample (1 mg / ml) were added and reacted at 37 ° C. for 2 hours. After removing the reaction solution and washing the cells three times with ice-cold PBS, the fluorescence intensity of 2-NBDG was measured with a spectrophotofluorometer (excitation: 485 nm, emission: 535 nm). In order to correct the effect of cell concentration on both measurements, the MTT assay was performed as follows: MTT on the well plate after fluorescence intensity measurement 50 μl of solution (5 mg / ml) was added and reacted for 3 hours at 37 ° C. and then absorbance was measured at 570 nm The glucose absorption inhibitory activity by the sample was calculated according to the following equation.
<수학식 1>&Quot; (1) "
저해능(Inhibitory effect, %) = [(Fc - Fs) / Fc] × 100 Inhibitory effect (%) = [(Fc-Fs) / Fc] × 100
Fs : 시료를 처리한 실험구의 형광세기Fs: Fluorescence intensity of the experimental group treated with the sample
Fc : 시료를 처리하지 않은 실험구의 형광세기 Fc: Fluorescence intensity of the experimental group without sample treatment
발아현미 메탄올 추출물을 농도별로 처리한 결과, 시료를 처리하지 않은 대조군 100%를 기준으로 비교하였을 때, 높은 포도당 흡수 저해효과가 나타났음을 확인하였다. (표2 및 도 5)As a result of treatment of the germinated brown rice methanol extract by concentration, it was confirmed that the high glucose absorption inhibitory effect appeared when compared with the
<표 2> 포도당 흡수 저해능 측정 결과Table 2 Measurement Results of Glucose Absorption Inhibition
실험예Experimental Example 3 : 발아현미의 3: germinated brown rice SGLT1SGLT1 , , HNFHNF α-1, α-1, HNFHNF β-1 발현 억제Inhibition of β-1 Expression
본 발명 메탄올 추출물을 Caco-2 세포주의 배양 6일째 배지에 첨가, 처리하고 14일째 배양을 종료한 후 RNA을 추출하여 소장흡수에 관여하는 핵심전사인자인 SGLT1와 HNFα-1, HNFβ-1 프라이머 (primer)를 이용하여 RT-PCR(reverse transcraptase-PCR)을 실시한 다음 아가로즈(agarose)에서 전기영동을 실시하여 그 발현의 정도를 비교하였다. 그 결과, 시료를 처리하지 않은 대조군을 100%로 기준하여 비교하였을 때, SGLT1, HNFα-1, HNFβ-1 유전자 발현이 현저하게 억제되었음을 확인할 수 있었다. (도 6 내지 8)The methanol extract of the present invention was added to the culture medium of the Caco-2 cell line on day 6, and after completion of the culture on day 14, RNA was extracted and key transcription factors SGLT1, HNFα-1 and HNFβ-1 primers involved in intestinal absorption. RT-PCR (reverse transcraptase-PCR) was performed using primers, followed by electrophoresis on agarose to compare the expression levels. As a result, it was confirmed that SGLT1, HNFα-1, HNFβ-1 gene expression was significantly suppressed when compared to the control group was not treated as a
도 1은 발아현미로부터 시료를 수득하는 과정을 도시한 것이다. Figure 1 shows the process of obtaining a sample from germinated brown rice.
도 2는 발아현미 추출 방법에 따라 α-글루코시다아제 활성(Glucosidase activity) 억제활성을 보인 도이다.2 is a diagram showing α-glucosidase activity inhibitory activity according to the germination brown rice extraction method.
도 3은 발아현미 추출 방법에 따라 글루코즈 업테이크 활성(Glucose uptake activity) 억제활성을 보인 도이다. Figure 3 is a diagram showing the glucose uptake activity (Glucose uptake activity) inhibitory activity according to the germination brown rice extraction method.
도 4는 발아현미 메탄올 추출물 농도에 의해 α- 글루코시다아제 활성 억제활성을 보인 도이다.Figure 4 shows the inhibitory activity of α-glucosidase activity by the concentration of germinated brown rice methanol extract.
도 5는 발아현미 메탄올 추출물 농도에 의해 글루코즈 업테이크 활성 억제활성을 보인 도이다.5 is a diagram showing the glucose uptake inhibitory activity by the germinated brown rice methanol extract concentration.
도 6은 발아현미 메탄올 추출물의 SGLT1 mRNA 발현 억제 활성을 보인 도이다.6 is a diagram showing the SGLT1 mRNA expression inhibitory activity of the germinated brown rice methanol extract.
도 7은 발아현미 메탄올 추출물의 HNFα-1 mRNA 발현 억제 활성을 보인 도이다.Figure 7 shows the inhibitory activity of HNFα-1 mRNA expression of germinated brown rice methanol extract.
도 8은 메탄올 추출물의 HNFβ-1 mRNA 발현 억제 활성을 보인 도이다.8 is a diagram showing the inhibitory activity of HNFβ-1 mRNA expression of methanol extract.
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KR101485575B1 (en) * | 2013-02-13 | 2015-01-22 | 서선유 | An method of Extracts for A GABA ingredient from germinated brown rice and that of using health function composition for a diabetic |
KR20240092261A (en) | 2022-12-14 | 2024-06-24 | 박동기 | Method of manufacturing anti-diabetes sprouted grains using herb medicine and additives and method of manufacturing artificial food using the same |
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KR20240092261A (en) | 2022-12-14 | 2024-06-24 | 박동기 | Method of manufacturing anti-diabetes sprouted grains using herb medicine and additives and method of manufacturing artificial food using the same |
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