KR20110077644A - Composition for improving obesity using wheat bran - Google Patents
Composition for improving obesity using wheat bran Download PDFInfo
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- KR20110077644A KR20110077644A KR1020090134278A KR20090134278A KR20110077644A KR 20110077644 A KR20110077644 A KR 20110077644A KR 1020090134278 A KR1020090134278 A KR 1020090134278A KR 20090134278 A KR20090134278 A KR 20090134278A KR 20110077644 A KR20110077644 A KR 20110077644A
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- composition
- extract
- wheat bran
- obesity
- water
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Abstract
Description
본 발명은 밀겨 추출물을 이용한 비만 개선제 조성물에 관한 것이다.The present invention relates to an obesity improving composition using wheat bran extract.
최근 경제 발전에 따른 생활 수준의 향상, 바쁜 생활 환경에 따른 운동 부족, 영양의 과잉 섭취 등으로 비만 인구가 급속히 늘고 있다. 우리나라의 비만 인구 비율은 성인 여성의 경우 1995년 11.7%이던 것이 2001년 29.4%, 성인 남성의 경우 1995년 18.0%이던 것이 2001년 32.6%로 빠른 증가세를 보이고 있다(보건복지부, 2007). Recently, the obese population is rapidly increasing due to the improvement of living standards according to economic development, lack of exercise due to busy living environment, and excessive intake of nutrition. The proportion of obese populations in Korea is growing rapidly from 11.7% in 1995 to 29.4% in 2001 and 18.0% in 1995 to 32.6% in 2001 (Ministry of Health and Welfare, 2007).
비만은 에너지의 섭취와 소모의 불균형으로 인하여 체내에 에너지가 과잉으로 축적되어 지방조직이 비정상적으로 증가된 상태를 말한다(Clinical Endocrinology 28:675-689, 1998; Clinical Obesity (eds. Kopelman PG, Stock MJ) 248-89 (Blackwell Science, Oxford 1998). 임상적으로는 남자는 체지방이 체중의 25%, 여자는 체중의 30% 이상일 때 비만으로 보며, 임상적으로는 BMI(Body Mass Index: 체질량지수)가 30.0 이상인 경우를 비만으로 정의한다.Obesity is an abnormal increase in fat tissue due to excessive accumulation of energy in the body due to an imbalance between energy intake and consumption (Clinical Endocrinology 28: 675-689, 1998; Clinical Obesity (eds. Kopelman PG, Stock MJ). 248-89 (Blackwell Science, Oxford 1998) Clinically, men are considered obese when their body fat is 25% or more than 30% of their body weight, and clinically they are BMI (Body Mass Index) Is more than 30.0 is defined as obesity.
비만의 원인으로는 고지방·고열량의 식생활, 바쁜 사회적 환경에 따른 운동 부족, 내분비 이상 등 환경적 요인과 유전적 요인을 들 수 있는데, 이 중 비만의 50 내지 70% 정도가 환경적 요인에 의한 것으로 알려져 있고, 나머지가 유전적 요인에 의한 것으로 알려져 있다. The causes of obesity include environmental and genetic factors such as high fat and high calorie diet, lack of exercise due to busy social environment, and endocrine disorders. Among them, about 50 to 70% of obesity is caused by environmental factors. It is known that the rest is due to genetic factors.
체내 에너지는 지방세포에 중성지방(triglyceride) 형태로 저장되었다가 체내 에너지원이 고갈되면 저장되었던 지방이 유리지방산과 글리세롤로 분해되어 에너지원으로 사용되게 되지만, 에너지의 과잉 섭취는 지방세포의 분화를 촉진하고 체내 저장 지방량을 증가시켜 비만의 직접적인 원인이 된다.Energy in the body is stored in the form of triglycerides in fat cells, and when the body's energy source is depleted, the stored fat is broken down into free fatty acids and glycerol to be used as an energy source, but excessive intake of energy leads to differentiation of fat cells. Promotes and increases the amount of fat stored in the body is a direct cause of obesity.
지방세포는 지방전구세포로부터 분화하여 만들어지는데, 이러한 지방세포의 분화에는 PPARγ(peroxisome proliferator-activated receptor γ), C/EBP family(CCAAT/enhancer binding proteins; C/EBRα, C/EBRβ 및 C/EBRδ) ADD1/SREBP1c(adipocyte determination differentiation factor 1)/(sterol regulatory element binding protein 1c)라고 불리는 전사인자들이 중추적인 역할을 담당하고 있는 것으로 알려져 있다(Genes De 2000, 14(11) 1293~1307). 이들 전사인자는 지방세포의 분화 과정 중 각기 다른 시점에서 발현이 유도되며 서로 상호작용을 통하여 여러 지방세포 특이 유전자들의 발현을 조절하고, 지방대사의 활성화와 지방세포 분화를 점진적으로 유도해 나간다(Physiol Rev 1998, 78(3):783~809; Annu Rev Biochem 2008, 77:289~312; Genes Dev 1996, 10:1096~1107).Adipocytes are produced by differentiating from adipocytes, which include peroxisome proliferator-activated receptor γ (PPARγ), C / EBP family (CCAAT / enhancer binding proteins; C / EBRα, C / EBRβ, and C / EBRδ). ) ADD1 / SREBP1c (adipocyte determination differentiation factor 1) / ( a transcription factor known as sterol regulatory element binding protein 1c) that are known to play a pivotal role (Genes De 2000, 14 (11) 1293-1307). These transcription factors induce expression at different points in the differentiation process of adipocytes, and interact with each other to regulate the expression of several adipocyte-specific genes and gradually induce activation of adipogenesis and differentiation of adipocytes ( Physiol Rev 1998, 78 (3): 783-809; Annu Rev Biochem 2008, 77: 289-312; Genes Dev 1996, 10: 1096-1107).
따라서 지방전구세포의 지방세포로의 분화 억제 및/또는 지방세포에서 지방 축적을 억제하는 활성을 갖거나, PPARγ, C/EBRα, 및/또는 ADD1/SREBP1c의 발현을 억제하는 활성을 갖는 것은 비만 개선제로 활용될 수 있다. 유효성분으로서 징코라이드 에이(ginkgolide A) 또는 빌로발라이드(bilobalide)를 개시하고 있는 대한민국 특허 제920648호, 유효성분으로서 진세노사이드(ginsenoside)-Rf 또는 화합물 K(Compound K)를 개시하고 있는 대한민국 특허 제847252호, 유효성분으로서 올티프라즈를 개시하고 있는 대한민국 특허 제576157호 등이 이러한 예에 속한다. Therefore, inhibiting the differentiation of adipocytes into adipocytes and / or inhibiting adipocyte accumulation in adipocytes, or inhibiting the expression of PPARγ, C / EBRα, and / or ADD1 / SREBP1c are antiobesity agents. Can be utilized. Republic of Korea Patent No.920648 discloses ginkgolide A or bilobalide as an active ingredient, Republic of Korea discloses ginsenoside-Rf or compound K (Compound K) as an active ingredient Patent No. 847252 and Korean Patent No. 576157, which discloses Oltipraz as an active ingredient, belong to this example.
본 발명도 밀겨 추출물이 지방세포로의 분화 억제 및 지방 축적 억제 활성을 갖고 또한 PPARγ, C/EBRα, ADD1/SREBP1c의 발현 억제 활성을 갖는다는 점에 기초하여 완성된 것이다.The present invention has also been completed based on the fact that wheat bran extract has an activity of inhibiting differentiation and adipocyte accumulation into adipocytes and also has an expression inhibitory activity of PPARγ, C / EBRα, and ADD1 / SREBP1c.
본 발명의 기술적 과제는 지방세포로의 분화 억제 및 지방 축적 억제 활성을 갖고 또한 PPARγ, C/EBRα 및 ADD1/SREBP1c의 발현을 억제하는 활성을 갖는 밀겨 추출물을 이용한 비만 개선제 조성물을 제공하는 데 있다.An object of the present invention is to provide an obesity improver composition using wheat bran extract having the activity of inhibiting the differentiation and adipogenesis of adipocytes and inhibiting the expression of PPARγ, C / EBRα and ADD1 / SREBP1c.
본 발명의 다른 목적이나 구체적인 양태는 이하에서 제시될 것이다. Other objects or specific aspects of the present invention will be presented below.
본 발명자들은 아래의 실험예에서 확인되는 바와 같이, 아래의 실시예에서 얻어진 밀겨 추출물이 인슐린에 의해 분화 촉진된 지방전구세포의 지방세포로의 분화를 억제하고 지방의 축적을 억제하며, 또한 지방세포의 분화에 중추적으로 관여하는 전사인자인 PPARγ, C/EBRα 및 ADD1/SREBP1c의 발현을 억제하는 활성을 확인할 수 있었다.The inventors of the present invention confirmed that the wheat bran extract obtained in the following examples inhibits the differentiation of adipocytes promoted by insulin into adipocytes and inhibits the accumulation of adipocytes. The activity of inhibiting the expression of PPARγ, C / EBRα and ADD1 / SREBP1c, which are centrally involved in differentiation, was identified.
본 발명은 이러한 실험 결과에 기초하여 제공되는 것으로서, 본 발명의 비만 개선제 조성물은 밀겨 추출물을 유효성분으로 포함함을 특징으로 한다.The present invention is provided based on the experimental results, the obesity improver composition of the present invention is characterized in that it comprises wheat bran extract as an active ingredient.
본 명세서에서, 상기 "밀겨"는 밀(wheat)을 도정하는 과정에서 얻어지는 부산물을 의미한다. 이러한 부산물은 통상 밀가루가 되는 배유를 제외하고 과피와 배아로 구성되게 된다.In the present specification, the "wheat bran" means a by-product obtained in the process of milling wheat. These by-products usually consist of the rind and the embryo, except for the endosperm, which becomes flour.
또한 본 명세서에서, 상기 "추출물"이란 추출 방법을 불문하고 추출 대상인 밀겨를 증류수를 포함한 물, 메탄올, 에탄올, 부탄올 등의 탄소수 1 내지 4의 저급 알콜, 또는 이들의 혼합 용매에 침지시켜 추출하여 얻어진 조추출물과 그 조추출물을 상기 열거된 용매로 분획된 추출물을 포함하는 의미로서 이해된다. 추출 방법을 불문하므로, 추출 대상을 추출 용매에 침지시키는 단계를 통하여 추출되는 한, 추출 방법은 냉침, 환류, 가온, 초음파 방사 등 임의의 방식이 모두 적용될 수 있다. 분획된 추출물의 경우 상기 조추출물을 특정 용매에 현탁시킨 후 다른 극성의 용매와 혼합·정치시켜 얻은 분획물, 상기 조추출물을 소수성 레진이 충진된 칼럼에 흡착시킨 후 상기 용매를 이동상으로 하여 얻은 분획물을 포함하는 의미이다. 사용될 수 있는 소수성 레진은 예컨대 Amberlite XAD-2™, 아래의 실시예에 사용된 Diaion HP-20™ 등 시판되는 것 중에서 적절히 선택될 수 있다. 상기 추출물의 의미에는 추출 용매가 제거된 농축된 액상의 추출물 또는 고형상의 추출물이 포함된다. In addition, in the present specification, regardless of the extraction method, the "extract" is obtained by immersing the wheat bran to be extracted by immersing it in water including distilled water, lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, butanol, or a mixed solvent thereof. Crude extracts and their crude extracts are understood as meaning including extracts fractionated with the solvents listed above. Regardless of the extraction method, as long as the extraction object is extracted through the step of immersing in the extraction solvent, the extraction method may be any method such as cooling, refluxing, heating, ultrasonic radiation, and the like. In the case of the fractionated extract, the crude extract is suspended in a specific solvent and mixed and fixed with a solvent of different polarity. The crude extract is adsorbed onto a column filled with hydrophobic resin, and the fraction obtained by using the solvent as a mobile phase is obtained. It is meant to include. Hydrophobic resins that can be used can be appropriately selected from among commercially available products such as Amberlite XAD-2 ™, Diaion HP-20 ™ used in the examples below. The meaning of the extract includes a concentrated liquid extract or solid extract from which the extraction solvent is removed.
또한 본 명세서에서, 상기 "비만"이란, 그것이 유전적 요인에 의한 비만이든 또는 환경적 요인에 의한 비만이든 지방조직이 비정상적으로 증가된 상태를 의미하며, 체질량지수의 구분에 따른 비만(BMI이 30.0 이상인 경우)과 과체중(BMI이 25~30인 경우)을 포함하는 의미이다.In addition, in the present specification, the term "obesity" refers to a state in which fat tissue is abnormally increased, whether it is caused by genetic factors or obesity due to environmental factors, and obesity according to the classification of body mass index (BMI is 30.0 Above) and overweight (if your BMI is 25-30).
또한 본 명세서에서, 상기 "유효성분"이란 단독으로 목적하는 활성을 나타내거나 또는 그 자체는 활성이 없는 담체와 함께 활성을 나타낼 수 있는 성분을 의미한다.In addition, in the present specification, the "active ingredient" alone refers to a component that can exhibit the desired activity or itself can exhibit the activity with a carrier that is not active.
또한 본 명세서에서, 상기 "비만 개선"이란 비만의 예방, 비만의 치료를 포함하여, 체지방의 감소 및/또는 체중의 감소를 포함하는 의미이다.In addition, in the present specification, "improving obesity" is meant to include the reduction of body fat and / or weight loss, including the prevention of obesity, the treatment of obesity.
본 발명의 비만 개선제 조성물은 그 유효성분인 밀겨 추출물을 비만 개선 활 성을 나타낼 수 있는 한 용도, 제형, 배합 목적 등에 따라 임의의 양(유효량)으로 포함할 수 있는데, 통상적인 유효량은 조성물 전체 중량을 기준으로 할 때 0.001 중량 % 내지 99.990 중량 % 범위 내에서 결정될 것이다. 여기서 "유효량"이란 비만 개선 효과를 유도할 수 있는 유효성분의 양을 말한다. 이러한 유효량은 당업자의 통상의 능력 범위 내에서 실험적으로 결정될 수 있다.The obesity improver composition of the present invention may include the wheat bran extract as an active ingredient in any amount (effective amount) according to the use, formulation, blending purpose, etc., as long as it can exhibit the activity to improve obesity, the typical effective amount is the total weight of the composition It will be determined in the range of 0.001% to 99.990% by weight based on. Here, the "effective amount" refers to the amount of the active ingredient that can induce the effect of improving obesity. Such effective amounts can be determined experimentally within the range of ordinary skill in the art.
본 발명의 조성물이 적용(처방)될 수 있는 대상은 포유동물 및 사람이며, 특히 사람인 경우가 바람직하다.Subjects to which the compositions of the invention can be applied (prescribed) are mammals and humans, in particular humans.
다른 측면에 있어서, 본 발명은 밀겨 추출물을 유효성분으로 포함하는 다이어트용 조성물에 관한 것이다.In another aspect, the present invention relates to a diet composition comprising wheat bran extract as an active ingredient.
본 명세서에서 "다이어트"란 체중 상태가 비만이나 과체중은 아니지만 미용이나 건강 목적으로 체중/체지방의 감소가 바람직하거나 필요한 상태로서 정의된다. 통상 본 발명의 다이어트용 조성물은 정상인의 미용이나 건강 목적으로 사용하기 위하여 제조·사용될 것이다. As used herein, "diet" is defined as a condition in which a weight condition is not obese or overweight, but for which a reduction in weight / body fat is desirable or necessary for cosmetic or health purposes. Usually, the composition for a diet of the present invention will be manufactured and used for use for normal beauty or health purposes.
본 발명의 다이어트용 조성물에 있어서, 그 유효성분인 밀겨 추출물의 의미, 그것의 유효량 등과 관련하여서는 상기 본 발명의 비만 개선제 조성물과 관련하여 전술한 바가 그대로 유효하다.In the composition for a diet of the present invention, with respect to the meaning of the wheat bran extract, the effective amount thereof, and the like as its active ingredient, the above-mentioned bar is effective as it relates to the obesity improving composition of the present invention.
또 다른 측면에 있어서, 본 발명은 밀겨 추출물을 유효성분으로 포함하는 인슐린 저항성 개선제 조성물에 관한 것이다.In another aspect, the present invention relates to an insulin resistance improving composition comprising wheat bran extract as an active ingredient.
비만은 인슐린 저항성의 여러 원인 중 하나에 해당한다. 고도비만에서 인슐린 저항성이 발병하지 않는 경우도 있지만, 인슐린 저항성과 비만은 밀접한 상관관 계가 있어, 일반적으로 비만이 심할수록 특히 내장 비만(visceral obesity)이 심할수록 인슐린 저항성도 심해진다고 알려져 있다.Obesity is one of several causes of insulin resistance. Although insulin resistance does not develop at high obesity, insulin resistance and obesity have a close correlation.
지방세포는 생리활성물질인 아디포사이토카인을 분비하는데, 이 아디포사이토카인은 대사의 항상성을 유지하는데 중요한 역할을 하지만, 비만 즉 지방의 과도한 축적 상태가 되면 이 아디포사이토카인의 생성·분비가 과잉 또는 과소가 되어 균형이 파괴됨으로써 인슐린 저항성이 야기되는 것으로 이해되고 있다.Adipocytes secrete the bioactive adipocytokine, which plays an important role in maintaining metabolic homeostasis. However, when adipocytokine is produced, the adipocytokine is produced and secreted. It is understood that insulin resistance is caused by the excess or underestimation resulting in a breakdown of the balance.
아디포사이토카인 중에는 인슐린 감수성을 항진시키는 것과 인슐린 저항성을 야기시키는 것이 있으며, 전자의 대표적인 경우가 아디포넥틴(adiponectin)과 렙틴(leptin), AMPK(AMP-의존성 단백질 키나아제)이며, 후자의 대표적인 경우가 TNF-α, lL-6, 레지스틴(resistin) 등을 들 수 있다.Among the adipocytokines, there are those that promote insulin sensitivity and cause insulin resistance. The former are adiponectin, leptin, and AMPK (AMP-dependent protein kinase), and the latter is TNF. -α, lL-6, a resistin, etc. are mentioned.
또 지방세포에서 발현되는 Fas(fatty acid synthase)나 aP2(adipocyte fatty-acid-binding protein 2)도 인슐린 저항성을 야기시키는 것으로 알려져 있다. Fatty acid synthase (Fas) and adiocytes (adipocyte fatty acid-binding protein 2), which are expressed in adipocytes, are also known to cause insulin resistance.
Fas는 지방세포의 분화에 있어 초기에 발현되며(J. Biol. Chem., 255, 4745 - 4750 (1980)) 아세틸 코에이 (acetyl-CoA)와 말로닐 코에이 (malonyl-CoA)로부터 팔미테이트 (palmitate)를 생산하는 효소인데, Fas는 과도한 에너지를 중성지방의 형태로 저장하여 비만을 일으키는 한편, 그것에 의해 생산된 중성지방인 팔미테이트는 췌장의 β세포를 파괴하여 인슐린 저항성을 야기시킨다고 보고되어 있으며(Proc. Natl. Acad. Sci. 95, 2498 ∼ 2502(1998)), aP2는 그 유전자의 결손 쥐에서 인슐린 저항성 발병률이 낮고 aP2 저해제가 인슐린 저항성을 감소시킨다는 결 과로부터 인슐린 저항성 치료제 개발에 강력한 표적 단백질로 제안되어 있다(Nature 447, 959-965 (21 July 2007)).Fas is initially expressed in the differentiation of adipocytes (J. Biol. Chem., 255, 4745-4750 (1980)) and palmitate from acetyl-CoA and malonyl-CoA It is an enzyme that produces (palmitate), and Fas stores excess energy in the form of triglycerides to cause obesity, while the triglycerides produced by it, palmitate, are reported to destroy the pancreatic β cells causing insulin resistance. (Proc. Natl. Acad. Sci. 95, 2498-2502 (1998)), aP2 is a potent in the development of insulin-resistant therapies due to the low incidence of insulin resistance in mice lacking the gene and the aP2 inhibitors that decrease insulin resistance. It is proposed as a target protein (Nature 447, 959-965 (21 July 2007)).
본 발명자들은 본 발명의 비만 개선제 조성물의 유효성분인 밀겨 추출물이 인슐린 저항성 개선제로서의 활용 가능성을 고려하여, 밀겨 추출물이 처리된 지방세포에서 인슐린 저항성을 야기하는 아디포사이토카인인 레지스틴, 그리고 Fas와 aP2를 정량하였는데, 이들의 발현이 현저히 억제됨을 확인할 수 있었다. 이러한 실험 결과는 밀겨 추출물이 비만 개선제로서 뿐만 아니라 인슐린 저항성 개선제로서도 유용하게 활용될 수 있음을 보여주는 것이라 할 수 있다.The present inventors consider that the bran extract, an active ingredient of the obesity improver composition of the present invention, can be utilized as an insulin resistance improving agent, resulting in insulin resistance in adipocytes treated with wheat bran extract, Resistine, and Fas and aP2. It was confirmed that their expression is significantly suppressed. These experimental results show that wheat bran extract can be useful as an anti-obesity enhancer as well as an obesity improver.
본 명세서에서, 상기 "인슐린 저항성"이란 세포, 장기, 인체 등 개체 등이 정상적인 대사 활동을 위하여 통상적인 양 이상의 인슐린을 필요로 하는 상태, 즉 인슐린의 작용력이 저하된 인슐린의 작용 부전 상태를 의미하며, 인슐린 비의존형 당뇨병 또는 제2형 당뇨병과 동일한 의미로 받아들여진다. 당뇨병은 췌장의 β-세포가 파괴된 결과 그 치료를 위하여 인슐린이 필수적으로 요구되는 인슐린 의존형 당뇨병(제1형 당뇨병)과 인슐린의 작용력이 저하되어 그 치료에 인슐린이 요구되지 않은 인슐린 비의존형 당뇨병(제2형 당뇨병)으로 구분되기 때문에, 당업계에서 인슐린 저항성, 인슐린 비의존형 당뇨병 그리고 제2형 당뇨병은 동일한 의미로 받아들여지고 있다. In the present specification, the term "insulin resistance" refers to a state in which an individual such as a cell, an organ, or a human body needs more than a usual amount of insulin for normal metabolic activity, that is, an insufficiency state of insulin in which insulin's action force is reduced. , Insulin-independent diabetes or
본 발명의 인슐린 저항성 개선제 조성물에 있어서, 그 유효성분인 밀겨 추출물의 의미, 그것의 유효량 등과 관련하여서는 상기 본 발명의 비만 개선제 조성물과 관련하여 전술한 바가 그대로 유효하다.In the insulin resistance improver composition of the present invention, the meaning of wheat bran extract as an effective ingredient, its effective amount, and the like, as described above with respect to the obesity improver composition of the present invention, is effective as it is.
본 발명의 조성물은 다른 구체적인 양태에 있어서, 식품 조성물로서 파악할 수 있다.The composition of this invention can be grasped | ascertained as a food composition in another specific aspect.
본 발명의 식품 조성물은 건강 보조식품, 특수 영양 보충용 식품, 기능성 음료 등으로 제조될 수 있다.The food composition of the present invention may be prepared as a dietary supplement, a special nutritional supplement, a functional beverage, and the like.
본 발명의 식품 조성물에는 그 유효성분 이외에 감미제, 풍미제, 생리활성 성분, 미네랄 등이 포함될 수 있다.The food composition of the present invention may include sweeteners, flavoring agents, bioactive ingredients, minerals, etc. in addition to the active ingredients.
감미제는 식품이 적당한 단맛을 나게 하는 양으로 사용될 수 있으며, 천연의 것이거나 합성된 것일 수 있다. 바람직하게는 천연 감미제를 사용하는 경우인데, 천연 감미제로서는 옥수수 시럽 고형물, 꿀, 수크로오스, 프룩토오스, 락토오스, 말토오스 등의 당 감미제를 들 수 있다. Sweeteners may be used in amounts that give the food a suitable sweet taste, and may be natural or synthetic. Preferably, natural sweeteners are used. Examples of natural sweeteners include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose and maltose.
풍미제는 맛이나 향을 좋게 하기 위하여 사용될 수 있는데, 천연의 것과 합성된 것 모두 사용될 수 있다. 바람직하게는 천연의 것을 사용하는 경우이다. 천연의 것을 사용할 경우에 풍미 이외에 영양 강화의 목적도 병행할 수 있다. 천연 풍미제로서는 사과, 레몬, 감귤, 포도, 딸기, 복숭아 등에서 얻어진 것이거나 녹차잎, 둥굴레, 대잎, 계피, 국화 잎, 자스민 등에서 얻어진 것일 수 있다. 또 인삼(홍삼), 죽순, 알로에 베라, 은행 등에서 얻어진 것을 사용할 수 있다. 천연 풍미제는 액상의 농축액이나 고형상의 추출물일 수 있다. 경우에 따라서 합성 풍미제가 사용될 수 있는데, 합성 풍미제는 에스테르, 알콜, 알데하이드, 테르펜 등이 이용될 수 있다. Flavoring agents can be used to enhance the taste or aroma, both natural and synthetic. Preferably, a natural one is used. When using natural ones, the purpose of nutritional fortification can be performed in addition to the flavor. The natural flavor may be obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, and the like, or may be obtained from green tea leaves, round leaves, jujube leaves, cinnamon, chrysanthemum leaves, jasmine and the like. Also, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, banks and the like can be used. The natural flavoring agent may be a liquid concentrate or a solid form of extract. In some cases, synthetic flavoring agents may be used, and synthetic flavoring agents may include esters, alcohols, aldehydes, terpenes, and the like.
생리 활성 물질로서는 카테킨, 에피카테킨, 갈로가테킨, 에피갈로카테킨 등 의 카테킨류나, 레티놀, 아스코르브산, 토코페롤, 칼시페롤, 티아민, 리보플라빈 등의 비타민류 등이 사용될 수 있다.As the physiologically active substance, catechins such as catechin, epicatechin, gallocatechin, epigallocatechin, vitamins such as retinol, ascorbic acid, tocopherol, calciferol, thiamine, riboflavin, and the like can be used.
미네랄로서는 칼슘, 마그네슘, 크롬, 코발트, 구리, 불소화물, 게르마늄, 요오드, 철, 리튬, 마그네슘, 망간, 몰리브덴, 인, 칼륨, 셀레늄, 규소, 나트륨, 황, 바나듐, 아연 등이 사용될 수 있다.As minerals, calcium, magnesium, chromium, cobalt, copper, fluoride, germanium, iodine, iron, lithium, magnesium, manganese, molybdenum, phosphorus, potassium, selenium, silicon, sodium, sulfur, vanadium, zinc and the like can be used.
또한 본 발명의 식품 조성물은 상기 감미제 등 이외에도 필요에 따라 보존제, 유화제, 산미료, 점증제 등을 포함할 수 있다. In addition, the food composition of the present invention may contain a preservative, an emulsifier, an acidulant, a thickener, and the like, in addition to the sweetener.
이러한 보존제, 유화제 등은 그것이 첨가되는 용도를 달성할 수 있는 한 극미량으로 첨가되어 사용되는 것이 바람직하다. 극미량이란 수치적으로 표현할 때 식품 조성물 전체 중량을 기준으로 할 때 0.0005중량% 내지 약 0.5중량% 범위를 의미한다.Such preservatives, emulsifiers and the like are preferably added and used in very small amounts as long as the use to which they are added can be achieved. By trace amount is meant numerically expressed in the range of 0.0005% to about 0.5% by weight based on the total weight of the food composition.
사용될 수 있는 보존제로서는 소듐 소르브산칼슘, 소르브산나트륨, 소르브산칼륨, 벤조산칼슘, 벤조산나트륨, 벤조산칼륨, EDTA(에틸렌디아민테트라아세트산) 등을 들 수 있다. Examples of preservatives that can be used include sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), and the like.
사용될 수 있는 유화제로서는 아카시아검, 카르복시메틸셀룰로스, 잔탄검, 펙틴 등을 들 수 있다.Emulsifiers that can be used include acacia gum, carboxymethylcellulose, xanthan gum, pectin and the like.
사용될 수 있는 산미료로서는 연산, 말산, 푸마르산, 아디프산, 인산, 글루콘산, 타르타르산, 아스코르브산, 아세트산, 인산 등을 들 수 있다. 이러한 산미료는 맛을 증진시키는 목적 이외에 미생물의 증식을 억제할 목적으로 식품 조성물이 적정 산도로 되도록 첨가될 수 있다.Examples of acidulants that may be used include lead acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, and the like. Such acidulant may be added so that the food composition is at an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to the purpose of enhancing taste.
사용될 수 있는 점증제로서는 현탁화 구현제, 침강제, 겔형성제, 팽화제 등을 들 수 있다. Thickeners that can be used include suspending implements, sedimenters, gel formers, swelling agents and the like.
또한 본 발명의 식품 조성물은 향미나 기호성을 향상시키고 그 기능성을 상승 보강하거나 다른 기능성(예컨대 지방간 개선 활성, 숙취 해소 활성 등)을 부가하기 위하여 이미 비만 개선 활성이나 간 기능 개선 활성 등이 있다고 알려진 임의의 천연물 유래의 분말 또는 추출물을 포함할 수 있는데, 선지 분말 또는 추출물, 콩나물 분말 또는 추출물, 조개 분말 또는 추출물, 굴 분말 또는 추출물, 산미나리 분말 또는 추출물, 무 즙 또는 추출물, 오이 즙 또는 추출물, 부추 즙 또는 추출물, 시금치 즙 또는 추출물, 연근 즙 또는 추출물, 칡 즙 또는 추출물, 솔잎 즙 또는 추출물, 인삼 즙 또는 추출물, 백화사설초 분말 또는 추출물, 감초 분말 또는 추출물, 갈화 분말 또는 추출물, 갈근 분말 또는 추출물, 사인 분말 또는 추출물, 박 분말 또는 추출물, 생강 분말 또는 추출물, 대추 분말 또는 추출물, 인진 분말 또는 추출물, 지구자 분말 또는 추출물, 수비계 분말 또는 추출물, 백출 분말 또는 추출물, 저령 분말 또는 추출물, 진피(진귤의 껍질) 분말 또는 추출물, 구기자 분말 또는 추출물, 녹차 분말 또는 추출물, 오미자 분말 또는 추출물, 헛개나무 추출물, 지치 추출물, 노근 추출물, 계피 추출물, 데커시놀 등이 예시될 수 있다. 상기에서 추출물의 의미는 밀겨 추출물과 관련하여 앞서 설명한 바와 같다. 이러한 추출물은 추출 대상을 혼합하여 얻어질 수도 있다.In addition, the food composition of the present invention is already known to have an activity for improving obesity or improving liver function in order to improve flavor and palatability, to enhance or enhance its functionality, or to add other functionalities (such as fatty liver improving activity and hangover relieving activity). It may include powders or extracts derived from natural products of, ointment powder or extracts, bean sprouts powder or extracts, shellfish powder or extracts, oyster powder or extracts, parsley powder or extracts, radish juice or extracts, cucumber juice or extracts, leek Juice or extract, spinach juice or extract, lotus root juice or extract, sesame juice or extract, pine needle juice or extract, ginseng juice or extract, birch powder or extract, licorice powder or extract, brownish powder or extract, brown root powder or extract, Sine powder or extract, gourd powder or extract, ginger powder Silver extract, jujube powder or extract, phosphorus powder or extract, spermatozoon powder or extract, hydrophobic powder or extract, white extract powder or extract, spirit powder or extract, dermis (peel of tangerine) powder or extract, wolfberry powder or extract, Green tea powder or extract, Schisandra chinensis powder or extract, Hawthorn extract, Chichi extract, Root extract, Cinnamon extract, Decusinol and the like can be exemplified. The meaning of the extract in the above is as described above with respect to wheat bran extract. Such extracts may be obtained by mixing the extraction objects.
본 발명의 조성물은 구체적인 양태에 있어서는 약제학적 조성물로 이용될 수 있다.The composition of the present invention can be used as a pharmaceutical composition in a specific embodiment.
본 발명의 약제학적 조성물은 유효물질인 밀겨 추출물을 이외에 약제학적으로 허용되는 담체, 부형제 등을 포함하여, 경구용 제형(정제, 현탁액, 과립, 에멀젼, 캡슐, 시럽 등), 비경구형 제형(멸균 주사용 수성 또는 유성 현탁액), 국소형 제형(용액, 크림, 연고, 겔, 로션, 패치) 등으로 제조될 수 있다.The pharmaceutical composition of the present invention includes oral formulations (tablets, suspensions, granules, emulsions, capsules, syrups, etc.), parenteral formulations (sterile), in addition to bran extract, which is an active substance, in addition to pharmaceutically acceptable carriers, excipients, and the like. Aqueous or oily suspensions for injection), topical formulations (solutions, creams, ointments, gels, lotions, patches) and the like.
상기에서 "약제학적으로 허용되는" 의미는 유효성분의 활성을 억제하지 않으면서 적용(처방) 대상이 적응가능한 이상의 독성(충분히 낮은 독성)을 지니지 않는다 의미이다.As used herein, "pharmaceutically acceptable" means that the subject of application (prescription) does not have more toxicity (adequately low toxicity) to which the subject of application (prescription) is adaptable without inhibiting the activity of the active ingredient.
약제학적으로 허용되는 담체의 예로서는 락토스, 글루코스, 슈크로스, 전분(예컨대 옥수수 전분, 감자 전분 등), 셀룰로오스, 그것의 유도체(예컨대 나트륨 카르복시메틸 셀룰로오스, 에틸셀룰로오스, 등) 맥아, 젤라틴, 탈크, 고체 윤활제(예컨대 스테아르산, 스테아르산 마그네슘 등), 황산 칼슘, 식물성 기름(예컨대 땅콩 기름, 면실유, 참기름, 올리브유 등), 폴리올(예컨대 프로필렌 글리콜, 글리세린 등), 알긴산, 유화제(예컨대 TWEENS), 습윤제(예컨대 라우릴 황산 나트륨), 착색제, 풍미제, 정제화제, 안정화제, 항산화제, 보존제, 물, 식염수, 인산염 완충 용액 등을 들 수 있다. 이러한 담체는 본 발명의 약제학적 조성물의 제형에 따라 적당한 것을 하나 이상 선택하여 사용할 수 있다.Examples of pharmaceutically acceptable carriers include lactose, glucose, sucrose, starch (such as corn starch, potato starch, etc.), cellulose, derivatives thereof (such as sodium carboxymethyl cellulose, ethylcellulose, etc.) malt, gelatin, talc, solids Lubricants (e.g. stearic acid, magnesium stearate, etc.), calcium sulfate, vegetable oils (e.g. peanut oil, cottonseed oil, sesame oil, olive oil, etc.), polyols (e.g. propylene glycol, glycerin, etc.), alginic acid, emulsifiers (e.g. TWEENS), wetting agents (e.g. Sodium lauryl sulfate), colorants, flavoring agents, tableting agents, stabilizers, antioxidants, preservatives, water, saline, phosphate buffer solutions and the like. The carrier may be selected from one or more of suitable pharmaceutical formulations according to the formulation of the pharmaceutical composition of the present invention.
부형제도 본 발명의 약제학적 조성물의 제형에 따라 적합한 것을 선택하여 사용할 수 있는데, 예컨대 본 발명의 약제학적 조성물이 수성 현탁제로 제조될 경우에 적합한 부형제로서는 나트륨 카르복시메틸 셀룰로오스, 메틸 셀룰로오스, 히드로프로필메틸셀룰로오스, 알긴산 나트륨, 폴리비닐피롤리돈 등의 현탁제나 분산 제 등을 들 수 있다. 주사액으로 제조되는 경우 적합한 부형제로서는 링거액, 등장 염화나트륨 등을 들 수 있다.Excipients may be selected and used according to the formulation of the pharmaceutical composition of the present invention, for example, when the pharmaceutical composition of the present invention is prepared with an aqueous suspending agent, suitable excipients are sodium carboxymethyl cellulose, methyl cellulose, hydropropylmethylcellulose And suspending agents and dispersing agents such as sodium alginate and polyvinylpyrrolidone. Suitable excipients when prepared from injection solutions include Ringer's solution, isotonic sodium chloride, and the like.
본 발명의 약제학적 조성물은 경구 또는 비경구로 투여될 수 있고, 경우에 따라서는 국소적으로 투여될 수 있다.The pharmaceutical composition of the present invention can be administered orally or parenterally, and in some cases, can be administered topically.
본 발명의 약제학적 조성물은 그 1일 투여량이 통상 0.001 ~ 150 mg/kg 체중 범위이고, 1회 또는 수회로 나누어 투여할 수 있다. 그러나, 본 발명의 약제학적 조성물의 투여량은 투여 경로, 환자의 연령, 성별, 체중, 환자의 중증도 등의 여러 관련 인자에 비추어 결정되는 것이므로 상기 투여량은 어떠한 측면으로든 본 발명의 범위를 제한하는 것으로 이해되어서는 아니 된다. The daily dose of the pharmaceutical composition of the present invention is usually 0.001 to 150 mg / kg body weight, and may be administered once or several times. However, since the dosage of the pharmaceutical composition of the present invention is determined in view of various related factors such as the route of administration, the age, sex, weight of the patient, the severity of the patient and the like, the dosage may limit the scope of the present invention in any aspect. It should not be understood as.
전술한 바와 같이, 본 발명에 따르면 비만 개선제 조성물을 제공할 수 있다. 또한 본 발명에 따르면 인슐린 저항성 개선제 조성물을 제공할 수 있다. 본 발명의 비만 개선제 조성물과 인슐린 개선제 조성물 등은 기능성 식품, 약품 등으로 제품화될 수 있다.As described above, the present invention can provide an obesity improving composition. In addition, the present invention can provide an insulin resistance improving agent composition. Obesity improver composition and insulin improver composition of the present invention can be commercialized as a functional food, drugs and the like.
이하 본 발명을 실시예 및 실험예를 참조하여 설명한다. 그러나 본 발명의 범위가 이러한 실시예 및 실험예에 한정되는 것은 아니다.Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these examples and experimental examples.
<< 실시예Example > > 밀겨Push 추출물의 제조 Preparation of Extract
밀겨(Red-dog) 200g을 2ℓ물에 넣고 6시간 열수 추출하였다. 열수 추출물을 여과시키고 그 여과액을 Diaion HP-20 레진이 충진된 칼럼에 넣고 100% 에탄올과 물을 이동상으로 하여 100% 에탄올을 통과한 샘플을 Red-dog A라 하였고, 물을 통과한 샘플을 Red-dog B라 명명하여 실험에 사용하였다.200 g of Red-dog was placed in 2 L of water and extracted with hot water for 6 hours. The hot water extract was filtered and the filtrate was placed in a column filled with Diaion HP-20 resin, and 100% ethanol and water were used as the mobile phase, and the sample passed through 100% ethanol was called Red-dog A. Red-dog B was used for the experiment.
<< 실험예Experimental Example > > 비만 개선 활성 실험 및 인슐린 저항성 개선 활성 실험Obesity improvement activity experiment and insulin resistance improvement activity experiment
<< 실험예Experimental Example 1> 1> 비만 개선 활성 실험Obesity Improvement Active Experiment
<실험예 1-1> 전구지방세포의 지방세포로의 분화 억제 활성 평가 Experimental Example 1-1 Evaluation of Differentiation Inhibitory Activity of Preadipocytes into Adipocytes
마우스 전구지방세포인 3T3-L1은 10 % BCS DMEM 배지를 넣고 37 ℃, 5 % CO2 의 조건에서 배양하였다. 3T3-L1, a mouse precursor cell, was added with 10% BCS DMEM medium and cultured at 37 ° C. and 5% CO 2 .
3T3-L1 전구지방세포를 24 well plate에 5X104/well의 세포수로 분주한 후 100% confluency 시점이 되면 2일 동안 더 유지시켰다. 전구지방세포는 MDI (0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 1 uM dexamethasone, 1 ug/ml insulin) 를 포함하는 10% FBS DMEM 배지로 지방세포 분화를 2일 동안 유도하였고, 배양 48 시간 후, 1 ug/ml insulin이 함유된 10% FBS DMEM으로 이틀 동안 배양하였다. 그 후 2일 마다 4일 동안 10% FBS DMEM 배양액으로 교체하였다. 지방세포 분화 유도 동안 Red-dog A 및 B는 25, 50, 100 ug/ml의 농도로 각 배양액에 처리하였고, 분화가 완성되는 시점인 8일째에 지방세포 분화 정도를 관찰하였다. 지방세포 분화 정 도는 Oil Red O 염색을 통해 1차적으로 현미경을 통해 확인하였고, 지방세포 염색 정도는 510 nm 흡광도에서 세포의 분화된 지방량을 측정하였다.After dispensing 3T3-L1 progenitor cells with a cell number of 5 × 10 4 / well in a 24 well plate, 100% confluency was maintained for 2 more days. Preadipocytes induced adipocyte differentiation for 2 days with 10% FBS DMEM medium containing MDI (0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 1 uM dexamethasone, 1 ug / ml insulin). After hours, the cells were incubated for 2 days with 10% FBS DMEM containing 1 ug / ml insulin. It was then replaced with 10% FBS DMEM culture for 4 days every 2 days. During induction of adipocyte differentiation, Red-dog A and B were treated in each culture at concentrations of 25, 50, and 100 ug / ml, and the degree of adipocyte differentiation was observed on
실시예 각 시료의 Oil Red O 염색 결과 및 지방량의 정량 분석 결과를 <도 1>에 나타내었는데, <도 1>를 참조하여 보면 실시예의 시료 모두 농도 의존적으로 전구지방세포가 지방세포로의 분화 억제 활성을 가짐을 알 수 있다. Example The results of quantitative analysis of Oil Red O staining and fat amount of each sample are shown in <Figure 1>. Referring to <Figure 1>, all of the samples of the Examples showed concentration-dependent proliferation inhibitory activity of adipocytes into adipocytes. It can be seen that it has.
<실험예 1-2> PPAR γ의 전사 촉진 억제 활성 평가 <Experimental Example 1-2> inhibition promotes transcription activity of PPAR γ evaluation
pGL3-basic luciferase expression 벡터(Promega)에 PPRE(PPARγ response element) 유전자와 PPARγ 유전자를 삽입하여 얻은 재조합 벡터와 control reporter로서 Renilla luciferase의 cDNA를 가지는 pRL-SV-40 프라스미드(Promega)를 HEK 293T cell에 co-transfection 하였다. 유전자들은 하루 동안 발현시킨 후 세포에 50, 100 ug/ml 실시예의 시료를 단독으로 또 PPARγ 유도물질인 4ug/ml troglitazone와 함께 같이 24 시간 동안 처리한 후 시료에 의한 PPARγ의 전사 활성 정도를 luciferase 발현 강도를 통해 관찰하였다. Renilla is a recombinant vector and control reporter obtained by inserting the PPARγ response element (PPRE) gene and the PPARγ gene into the pGL3-basic luciferase expression vector (Promega). pRL-SV-40 Pramide (Promega) containing luciferase cDNA was co-transfected into HEK 293T cells. After expressing the genes for one day, the cells were treated with 50, 100 ug / ml of the sample alone or with Pugγ-inducing substance 4ug / ml troglitazone for 24 hours, and then luciferase expression of the PPARγ transcriptional activity by the sample. Observed through the intensity.
실시예 각 시료의 PPARγ 전사 활성에 미치는 영향을 <도 2>에 정량적으로 나타내었는데, 실시예의 시료 모두 농도 의존적으로 PPARγ 전사 촉진 활성을 억제함을 알 수 있다. EXAMPLES The effect on the PPARγ transcriptional activity of each sample is shown quantitatively in FIG. 2, but it can be seen that all the samples of the Example inhibit PPARγ transcriptional promoting activity in a concentration-dependent manner.
<실험예 1-3> 지방세포 분화에 관여하는 전사인자의 발현 억제 활성 평가 Experimental Example 1-3 Evaluation of Expression Inhibitory Activity of Transcription Factors Involved in Adipocyte Differentiation
<실험예 1-3-1> 유전자 수준에서의 발현 억제 활성 평가 Experimental Example 1-3-1 Evaluation of Expression Inhibitory Activity at the Gene Level
지방세포로 분화하는 단계에서는 PPARγ, C/EBRα 및 ADD1/SREBP1c의 발현의 증가한다. In the stage of differentiation into adipocytes, expression of PPARγ, C / EBRα and ADD1 / SREBP1c is increased.
상기 <실험예 1-1>과 동일하게 Red-dog A 및 B를 50, 100 ug/ml 농도로 처리 한 후 8일 동안 지방세포를 분화를 시켜 PPARγ, C/EBRα 및 ADD1/SREBP1c의 발현 정도를 Real-time PCR로 확인하였다. 8일 동안 분화된 3T3-L1 cell을 PBS로 두 번 wash 한 후 cell pellet을 모아 RNA prep kit(Qiagen)로 RNA를 분리했다. 추출 된 RNA를 1 ug 사용하여 cDNA를 합성한 후 SYBR green(Takara)과 아래 [표 1]의 primer를 이용하여 Real-time PCR을 했고 대조군 유전자는 GAPDH를 사용하였다. PCR 조성은 RNA 1ug으로 합성된 cDNA를 증류수로 1/50로 희석하여 5ul를 사용하였고, 10 pmole primer를 각각 0.5ul, 2X SYBR green 10ul, 4ul는 증류수를 사용하여 total volum은 20ul 맞추었다. Real-time PCR cycle은 초기변성은 95℃ 30초, 변성은 95℃ 5초, annealing은 60℃ 15초, 신장반응은 72℃ 10초로 하여 40 cycle을 진행했고 융해곡선은 55℃ 시작해서 95℃를 끝으로 하여 0.5℃씩 상승 시키며 80번을 반응하여 원하는 형광값을 검출했다(형광신호 정량은 Bio-Rad MyiQ program을 사용하였다). In the same manner as in <Experimental Example 1-1>, after treating Red-dog A and B at a concentration of 50 and 100 ug / ml, the expression of PPARγ, C / EBRα and ADD1 / SREBP1c by differentiating adipocytes for 8 days. Was confirmed by Real-time PCR. After washing the differentiated 3T3-L1 cells with PBS twice for 8 days, cell pellets were collected and RNA was isolated by RNA prep kit (Qiagen). After synthesis of cDNA using 1 ug of the extracted RNA, real-time PCR was performed using SYBR green (Takara) and the primers shown in [Table 1], and the control gene was GAPDH. PCR composition was 5ul by diluting cDNA synthesized with 1ug of RNA to 1/50 with distilled water, and 0.5ul for 10 pmole primer, 10ul for 2X SYBR green, and 4ul for distilled water to adjust the total volum to 20ul. In the real-time PCR cycle, the initial degeneration was 95 ℃ 30 seconds, the degeneration was 95
[표 1]TABLE 1
프라이머primer
결과를 <도 3a 내지 3c>에 나타내었다. <도 3a 내지 3c>를 참조하여 보면 실시예의 각 시료 모두 농도 의존적으로 PPARγ, C/EBRα 및 ADD1/SREBP1c의 발현을 억제함을 보여준다.The results are shown in FIGS. 3A to 3C. 3A to 3C show that each sample of the example inhibits the expression of PPARγ, C / EBRα and ADD1 / SREBP1c in a concentration-dependent manner.
<실험예 1-3-2> 단백질 수준에서의 발현 억제 활성 평가 Experimental Example 1-3-2 Evaluation of Expression Inhibitory Activity at the Protein Level
지방세포 분화 유도 방법과 같이 3T3-L1 세포에 시료를 이틀 간격으로 처리한 후 세포를 PBS로 두 번 wash하고 RIPA buffer (50 mM Tris-HCl, pH 8.0, 150mM sodium chloride, 1 % NP-40, 0.5 % sodium dexycholate, 0.1 % sodium dodecyl sulfate, protease inhibitor)를 이용하여 세포를 용해시켰다. 13000 rpm에서 30분간 원심 분리하여 단백질을 추출 정량하여 8 % SDS-PAGE gel을 이용하여 전기 영동했다. 전기 영동된 protein 들을 membrane에 이동시킨 후 5% skim milk를 포함하는 TBST로 blocking 했다. 1차 항체(PPARγ, C/EBPα)(SantaCruz)와 2차 항체(SantaCruz)로 반응시킨 후 ECL(Thermo scientific)을 통해 PPARγ 및 C/EBPα 단백질의 발현 정도를 β-actin의 발현 정도와 비교하여 평가하였다.After treatment with 3T3-L1 cells at two-day intervals as in the adipocyte differentiation method, cells were washed twice with PBS and RIPA buffer (50 mM Tris-HCl, pH 8.0, 150 mM sodium chloride, 1% NP-40, Cells were lysed using 0.5% sodium dexycholate, 0.1% sodium dodecyl sulfate, protease inhibitor). Protein was extracted and quantified by centrifugation at 13000 rpm for 30 minutes and electrophoresed using 8% SDS-PAGE gel. Electrophoretic proteins were transferred to the membrane and blocked with TBST containing 5% skim milk. After reacting with primary antibodies (PPARγ, C / EBPα) (SantaCruz) and secondary antibodies (SantaCruz), the expression levels of PPARγ and C / EBPα proteins were compared with the expression levels of β-actin through ECL (Thermo scientific). Evaluated.
결과를 <도 4>에 나타내었다. <도 3>의 결과와 유사하게 실시예의 각 시료는 모두 농도 의존적으로 PPARγ 및 C/EBRα의 발현을 억제함을 알 수 있다.The results are shown in FIG. 4. Similar to the results of FIG. 3, it can be seen that each sample of the example inhibits the expression of PPARγ and C / EBRα in a concentration-dependent manner.
<< 실험예Experimental Example 2> 2> 인슐린 저항성 개선 활성 실험Insulin Resistance Improvement Activity Experiment
실시예의 추출물의 인슐린 저항성 개선제로의 가능성을 평가하기 위하여 <실험예 1>에서 동일한 방법으로 아래 [표 2]의 프라이머를 이용하여 8일 동안 분화된 지방세포에서 레지스틴, aP2 및 Fas의 발현 정도를 정량 평가하였다.In order to evaluate the potential of the extract of the Example as an insulin resistance improving agent in <Experimental Example 1> using the primers of the following [Table 2] by the same method in the expression level of the resistin, aP2 and Fas in the differentiated fat cells for 8 days Quantitative evaluation.
[표 2]TABLE 2
프라이머primer
결과를 <도 5a 내지 도 5c>에 나타내었는데, 실시예의 시료 모두 농도 의존적으로 레지스틴, aP2 및 Fas의 발현을 억제함을 알 수 있다.The results are shown in FIGS. 5A to 5C, but it can be seen that the samples of the examples inhibit the expression of resistin, aP2 and Fas in a concentration-dependent manner.
도 1은 밀겨 추출물이 농도 의존적으로 지방세포의 지방 축적 억제 활성을 가짐을 보여주는 결과이다.1 is a result showing that wheat bran extract has a fat accumulation inhibitory activity of fat cells in a concentration-dependent manner.
도 2는 밀겨 추출물이 농도 의존적으로 PPARγ 전사 활성을 억제함을 보여주는 결과이다.Figure 2 shows that wheat bran extract inhibits PPARγ transcriptional activity in a concentration-dependent manner.
도 3은 밀겨 추출물이 농도 의존적으로 유전자 수준에서 PPARγ, C/EBRα 및 ADD1/SREBP1c의 발현을 억제함으로 보여주는 결과이다.Figure 3 shows that wheat bran extract inhibits the expression of PPARγ, C / EBRα and ADD1 / SREBP1c at the gene level in a concentration-dependent manner.
도 4는 밀겨 추출물이 농도 의존적으로 단백질 수준에서 PPARγ 및 C/EBRα의 발현을 억제함을 보여주는 결과이다.4 shows that wheat bran extract inhibits the expression of PPARγ and C / EBRα at a protein level in a concentration-dependent manner.
도 5는 밀겨 추출물이 농도 의존적으로 레지스틴, aP2 및 Fas의 발현을 억제함을 보여주는 결과이다.5 is a result showing that wheat bran extract inhibits the expression of resistin, aP2 and Fas in a concentration-dependent manner.
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