KR20220079128A - Composition for preventing, treating or improving metabolic diseases containing essential oil mixture of Mentha haplocalyx, Citrus unshiu peel and Coicis Semen - Google Patents
Composition for preventing, treating or improving metabolic diseases containing essential oil mixture of Mentha haplocalyx, Citrus unshiu peel and Coicis Semen Download PDFInfo
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- KR20220079128A KR20220079128A KR1020200168587A KR20200168587A KR20220079128A KR 20220079128 A KR20220079128 A KR 20220079128A KR 1020200168587 A KR1020200168587 A KR 1020200168587A KR 20200168587 A KR20200168587 A KR 20200168587A KR 20220079128 A KR20220079128 A KR 20220079128A
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- essential oil
- derived
- dermis
- composition
- peppermint
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Abstract
본 발명은 박하, 진피 및 의이인 정유 혼합 추출물을 포함하는 대사성 질환의 예방, 치료 또는 개선용 조성물에 관한 것으로, 본 발명에 따른 박하(Mentha haplocalyx) 유래 정유, 진피(Citrus unshiu peel) 유래 정유 및 의이인(Coicis Semen) 유래 정유 혼합물은 각각의 단독 또는 2종 혼합물 대비 지방세포의 지방 축적을 억제하는 효과가 현저하고, 지방세포의 분화 및 지방의 대사 조절에 관여하는 단백질의 발현을 조절하는 효과를 나타냄으로써, 비만 등 대사성 질환의 예방, 개선 또는 치료에 유용하게 사용할 수 있으며, 셀룰라이트 감소 또는 예방을 위한 용도로도 유용하게 사용할 수 있다.The present invention relates to a composition for preventing, treating or ameliorating a metabolic disease comprising a mixed extract of mint, dermis and uiiin essential oil, and mint ( Mentha haplocalyx )-derived essential oil, dermis ( Citrus unshiu peel)-derived essential oil, and uiiin according to the present invention ( Coicis Semen )-derived essential oil mixture has a remarkable effect of inhibiting fat accumulation in adipocytes compared to either alone or a mixture of two types, and has an effect of regulating the expression of proteins involved in adipocyte differentiation and fat metabolism regulation As such, it can be usefully used for the prevention, improvement or treatment of metabolic diseases such as obesity, and can also be usefully used for cellulite reduction or prevention.
Description
본 발명은 박하, 진피 및 의이인 정유 혼합 추출물을 포함하는 대사성 질환의 예방, 치료 또는 개선용 조성물에 관한 것이다.The present invention relates to a composition for preventing, treating or ameliorating a metabolic disease, comprising a mixed extract of mint, dermis and uiyin essential oil.
비만이란 대사 장애로 인하여 지방이 체내에 과잉 축적된 상태로서, 칼로리 섭취가 성장과 신체활동에 필요한 에너지보다 초과되어 일어나는 열량 불균형 현상으로 야기되며, 대개 체내의 지방이 남자는 체중의 25%, 여자는 30% 이상인 경우를 말한다. 특히 비만은 당뇨, 고지혈증, 심혈관계 질환 및 동맥경화증 등, 각종 대사성 질환의 원인이 될 뿐만 아니라, 비만으로 인한 사망률의 증가는 대사성 질환, 담낭질환, 호르몬 감수성 암과 위장관 암 등에서 명확하게 나타나며, 비만은 또한 비치명적인 요통, 관절염, 불임 등의 신체적 질병과 정신·사회적 기능의 저하 등 정신적 질병의 위험성도 증가시킨다.Obesity is a condition in which fat accumulates excessively in the body due to metabolic disorders, and it is caused by a caloric imbalance that occurs when caloric intake exceeds energy required for growth and physical activity. is 30% or more. In particular, obesity is a cause of various metabolic diseases, such as diabetes, hyperlipidemia, cardiovascular disease and arteriosclerosis, and the increase in mortality due to obesity is clearly shown in metabolic diseases, gallbladder disease, hormone-sensitive cancer, and gastrointestinal cancer. It also increases the risk of physical diseases such as non-fatal low back pain, arthritis, and infertility, as well as mental diseases such as deterioration of mental and social functions.
AMPK(AMP-activated protein kinase)는 catalytic α subunit, regulatory β 및 γ subunit로 구성되어 있는 serine/threonine kinase이다. 에너지 균형 조절자로서 지질과 포도당을 조절하여 당뇨와 비만의 대사에 중요한 역할을 한다. AMPK는 에너지 소모로 유발된 AMP/ATP ratio의 증가에 따른 γ subunit의 AMP 결합이며, α subunit의 threonine residue Thr172 인산화와 allosteric activation 촉진에 의해 활성화가 유발된다. AMPK의 활성화는 포도당 생성 및 지방 생성을 감소시키며, ACC 및 포도당 산화의 불활성화를 통해 지방산과 같은 ATP 소비에서 활성 ATP 생산 과정으로 전환시킨다. 또한, AMPK의 활성화는 지방산 합성과 콜레스테롤 합성을 조절하는 ACC(acetyl-CoA carboxylase)를 인산화시킴으로서 ACC효소를 불활성화시킨다.AMPK (AMP-activated protein kinase) is a serine/threonine kinase composed of catalytic α subunit, regulatory β and γ subunit. As an energy balance regulator, it plays an important role in the metabolism of diabetes and obesity by regulating lipids and glucose. AMPK is an AMP binding of the γ subunit according to an increase in the AMP/ATP ratio induced by energy consumption, and its activation is induced by phosphorylation of Thr172 at the threonine residue of the α subunit and promotion of allosteric activation. Activation of AMPK reduces gluconeogenesis and adipogenesis, and through inactivation of ACC and glucose oxidation, converts ATP consumption, such as fatty acids, to active ATP production processes. In addition, activation of AMPK inactivates ACC enzyme by phosphorylating acetyl-CoA carboxylase (ACC), which regulates fatty acid synthesis and cholesterol synthesis.
전지방세포에서 지방세포로 분화가 일어나는 과정은 세포형태의 변화, 유전자 발현의 변화, 호르몬 민감성의 변화 그리고 단백질 발현의 변화가 복합적으로 작용하는 과정이다. 지방세포의 형성 과정에서 가장 중요한 조절 인자는 peroxisome proliferator-activated receptor(PPAR)s와 CAAT/enhancer binding protein(C/EBP)s로 알려져 있다. 분화가 시작되면 초기 전사 인자인 C/EBP-β, C/EBP-δ가 C/EBP-α와 PPAR-γ의 발현량을 증가시키며, 이는 최종적으로 lipoprotein lipase (LPL), leptin, adiponectin, fatty acid binding protein (FABP) 4와 같은 최종 마커들의 발현을 촉진시킨다.The process of differentiation from pre-adipocytes to adipocytes is a process in which changes in cell morphology, changes in gene expression, changes in hormone sensitivity, and changes in protein expression act in a complex way. The most important regulatory factors in the formation of adipocytes are known as peroxisome proliferator-activated receptors (PPAR) and CAAT/enhancer binding proteins (C/EBP). When differentiation begins, the early transcription factors C/EBP-β and C/EBP-δ increase the expression levels of C/EBP-α and PPAR-γ, which ultimately leads to lipoprotein lipase (LPL), leptin, adiponectin, and fatty It promotes expression of final markers such as acid binding protein (FABP) 4.
셀룰라이트(cellulite)는 과도한 지방과 노폐물의 축적에 의한 순환장애로 피부가 거친 오렌지 껍질 같이 울퉁불퉁하게 되는 현상으로, 콜라겐과 엘라스틴이 존재하는 피부의 진피층 내에서 생성되며 여성에서 특히 빈번하게 발생한다. 셀룰라이트 비만은 피하 지방조직의 이상 누적 외에도 임파체계의 이상으로 임파액이 인체 조직 간극에 정체됨으로써 그 부위에 일종의 부종현상이 나타나 마치 살이 찐 것처럼 보이는 것으로, 체형 변화의 큰 요인이 된다. 조직 간극에 임파액이 정체되면 비만증과 같은 미용상의 문제뿐만 아니라 체내에 독성물질이 누적되어 피부세포에 악영향을 끼치고 세포의 영양섭취를 위한 이온화 활동을 저해하여 피부의 노화는 물론 세포의 괴사현상까지 유발하게 된다.Cellulite is a phenomenon in which the skin becomes rough like a rough orange peel due to a circulatory disorder caused by the accumulation of excessive fat and waste products. Cellulite obesity, in addition to the abnormal accumulation of subcutaneous adipose tissue, is an abnormality in the lymphatic system, which causes lymphatic fluid to stagnate in the gaps of human tissues, resulting in a kind of edema in that area, appearing as if they are fattening, which is a major factor in body change. When lymph is stagnant in the tissue gap, not only cosmetic problems such as obesity, but also toxic substances accumulate in the body and adversely affect skin cells, inhibiting the ionization activity for nutrient intake of cells, causing skin aging as well as necrosis of cells will do
항비만 약물에 대한 연구는 전 세계적으로 다양하게 진행되고 있다. 세로토닌과 노르에피네프린의 재흡수를 억제하는 작용을 하는 sibutramine은 심혈관계 부작용으로 인하여 사용 금지되었고, 소장에서 지방분해 효소의 작용을 억제하여 지방흡수를 감소시키는 orlistat는 지방변, 복부불편감 등의 부작용이 있다. 최근에는 locaserine, bupropione-naltrexone, phentermine-topiramate가 비만치료 약물로 사용되고 있으며, 이 중 가장 빈용 처방되는 locaserine은 sibutramine과 유사한 serotonin 활성 관련 약물로서 낮은 빈도의 두통, 오심, 현훈 등의 부작용이 보고되고 있다.Research on anti-obesity drugs is being conducted in various ways around the world. Sibutramine, which inhibits the reuptake of serotonin and norepinephrine, was banned due to cardiovascular side effects, and orlistat, which inhibits the action of lipolytic enzymes in the small intestine to reduce fat absorption, has side effects such as steatorrhea and abdominal discomfort. have. Recently, locaserine, bupropione-naltrexone, and phentermine-topiramate have been used as drugs for the treatment of obesity. Among them, locaserine, which is prescribed most frequently, is a drug related to serotonin activity similar to sibutramine, and side effects such as headache, nausea, and vertigo have been reported with low frequency. .
이에, 본 발명자들은 비만과 같은 대사성 질환의 예방, 치료 또는 개선을 위한 효과적인 천연 유래 성분을 개발하기 위한 연구를 수행한 결과, 박하, 진피 및 의이인 정유 혼합 추출물이 비만과 같은 대사성 질환의 예방, 치료 또는 개선에 대한 상승 효과가 있음을 알아내고 본 발명을 완성하였다.Accordingly, the present inventors conducted a study to develop an effective natural-derived ingredient for the prevention, treatment or improvement of metabolic diseases such as obesity, and as a result, a mixed extract of mint, dermis and Uiyin essential oil prevents, treats, and treats metabolic diseases such as obesity Or, it was found that there is a synergistic effect for improvement, and the present invention was completed.
본 발명의 목적은 대사성 질환의 예방 또는 개선용 건강기능식품 조성물을 제공하는 것이다.It is an object of the present invention to provide a health functional food composition for preventing or improving metabolic diseases.
본 발명의 다른 목적은 대사성 질환의 예방 또는 개선용 건강식품 조성물을 제공하는 것이다.Another object of the present invention is to provide a health food composition for preventing or improving metabolic diseases.
본 발명의 또 다른 목적은 대사성 질환의 예방 또는 치료용 약학적 조성물을 제공하는 것이다.Another object of the present invention is to provide a pharmaceutical composition for preventing or treating metabolic diseases.
본 발명의 다른 목적은 셀룰라이트 감소 또는 예방용 화장료 조성물을 제공하는 것이다.Another object of the present invention is to provide a cosmetic composition for reducing or preventing cellulite.
본 발명의 또 다른 목적은 지방 축적 억제용 조성물을 제공하는 것이다.Another object of the present invention is to provide a composition for inhibiting fat accumulation.
본 발명의 다른 목적은 상기 지방 축적 억제용 조성물을 포함하는 다이어트 조성물을 제공하는 것이다.Another object of the present invention is to provide a diet composition comprising the composition for inhibiting fat accumulation.
상기 목적을 달성하기 위하여,In order to achieve the above object,
본 발명은 박하(Mentha haplocalyx) 유래 정유; 진피(Citrus unshiu peel) 유래 정유 및 의이인(Coicis Semen) 유래 정유를 포함하는 대사성 질환의 예방 또는 개선용 건강기능식품 조성물을 제공한다.The present invention is peppermint (Mentha haplocalyx) derived essential oil; Provided is a health functional food composition for preventing or improving metabolic diseases, including essential oil derived from Citrus unshiu peel and essential oil derived from Coicis Semen.
또한, 본 발명은 박하(Mentha haplocalyx) 유래 정유; 진피(Citrus unshiu peel) 유래 정유 및 의이인(Coicis Semen) 유래 정유를 포함하는 대사성 질환의 예방 또는 개선용 건강식품 조성물을 제공한다.In addition, the present invention is peppermint (Mentha haplocalyx) derived essential oil; Provided is a health food composition for preventing or improving metabolic diseases, including essential oil derived from Citrus unshiu peel and essential oil derived from Coicis Semen.
나아가 본 발명은 박하(Mentha haplocalyx) 유래 정유; 진피(Citrus unshiu peel) 유래 정유 및 의이인(Coicis Semen) 유래 정유를 포함하는 대사성 질환의 예방 또는 치료용 약학적 조성물을 제공한다.Furthermore, the present invention is peppermint (Mentha haplocalyx) derived essential oil; Provided is a pharmaceutical composition for preventing or treating metabolic diseases, including essential oil derived from Citrus unshiu peel and essential oil derived from Coicis Semen.
또한, 본 발명은 박하(Mentha haplocalyx) 유래 정유; 진피(Citrus unshiu peel) 유래 정유 및 의이인(Coicis Semen) 유래 정유를 포함하는 셀룰라이트 감소 또는 예방용 화장료 조성물을 제공한다.In addition, the present invention is peppermint (Mentha haplocalyx) derived essential oil; Provided is a cosmetic composition for reducing or preventing cellulite comprising essential oil derived from Citrus unshiu peel and essential oil derived from Coicis Semen.
더 나아가 본 발명은 박하(Mentha haplocalyx) 유래 정유; 진피(Citrus unshiu peel) 유래 정유 및 의이인(Coicis Semen) 유래 정유를 포함하는 지방 축적 억제용 조성물을 제공한다.Furthermore, the present invention is peppermint (Mentha haplocalyx) derived essential oil; Provided is a composition for inhibiting fat accumulation comprising an essential oil derived from Citrus unshiu peel and an essential oil derived from Coicis Semen.
또한, 본 발명은 상기 지방 축적 억제용 조성물을 포함하는 다이어트 조성물을 제공한다.In addition, the present invention provides a diet composition comprising the composition for inhibiting fat accumulation.
본 발명에 따른 박하(Mentha haplocalyx) 유래 정유, 진피(Citrus unshiu peel) 유래 정유 및 의이인(Coicis Semen) 유래 정유 혼합물은 각각의 단독 또는 2종 혼합물 대비 지방세포의 지방 축적을 억제하는 효과가 현저하고, 지방세포의 분화 및 지방의 대사 조절에 관여하는 단백질의 발현을 조절하는 효과를 나타냄으로써, 비만 등 대사성 질환의 예방, 개선 또는 치료에 유용하게 사용할 수 있으며, 셀룰라이트 감소 또는 예방을 위한 용도로도 유용하게 사용할 수 있다.The mint ( Mentha haplocalyx )-derived essential oil, Citrus unshiu peel-derived essential oil and Coicis Semen -derived essential oil mixture according to the present invention has a remarkable effect of inhibiting fat accumulation in adipocytes compared to each alone or two types of mixture. , by showing the effect of regulating the expression of proteins involved in the differentiation of adipocytes and the regulation of fat metabolism, it can be usefully used for the prevention, improvement or treatment of metabolic diseases such as obesity, and for the reduction or prevention of cellulite can also be useful.
도 1은 본 발명의 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유 혼합물의 세포독성을 측정한 결과를 나타낸 그래프이다.
도 2는 본 발명의 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유 혼합물의 3T3-L1 세포에서 지방축적정도(%)를 측정한 결과를 나타낸 그래프이다.
도 3(a)는 본 발명의 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유 혼합물의 3T3-L1 세포에서 P-AMPK 및 AMPK 단백질 발현을 western blot을 통해 분석한 결과를 나타낸 그래프이다.
도 3(b)는 본 발명의 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유 혼합물의 3T3-L1 세포에서 P-ACC 및 ACC 단백질 발현을 western blot을 통해 분석한 결과를 나타낸 그래프이다.
도 3(c)는 3T3-L1 세포에서의 CPT-1 단백질 발현을 western blot을 통해 분석한 결과를 나타낸 그래프이다.1 is a graph showing the results of measuring the cytotoxicity of the essential oil derived from peppermint, the essential oil derived from the dermis, and the essential oil derived from Uiyin of the present invention.
2 is a graph showing the results of measuring the degree of fat accumulation (%) in 3T3-L1 cells of a mixture of peppermint-derived essential oil, dermis-derived essential oil, and uiiin-derived essential oil of the present invention.
Figure 3 (a) is a graph showing the results of analyzing the expression of P-AMPK and AMPK protein in 3T3-L1 cells of a mixture of peppermint-derived essential oil, dermis-derived essential oil, and uiiin-derived essential oil of the present invention through western blot.
Figure 3 (b) is a graph showing the results of analyzing the P-ACC and ACC protein expression in 3T3-L1 cells of a mixture of peppermint-derived essential oil, dermis-derived essential oil, and uiiin-derived essential oil of the present invention through western blot.
Figure 3 (c) is a graph showing the results of analysis of CPT-1 protein expression in 3T3-L1 cells through western blot.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
대사성 질환의 예방 또는 개선용 건강기능식품 및 건강식품 조성물Health functional food and health food composition for the prevention or improvement of metabolic diseases
본 발명은 박하(Mentha haplocalyx) 유래 정유; 진피(Citrus unshiu peel) 유래 정유 및 의이인(Coicis Semen) 유래 정유를 포함하는 대사성 질환의 예방 또는 개선용 건강기능식품 또는 건강식품 조성물에 관한 것이다.The present invention is peppermint (Mentha haplocalyx) derived essential oil; It relates to a health functional food or health food composition for preventing or improving metabolic diseases, including essential oil derived from Citrus unshiu peel and essential oil derived from Coicis Semen.
본 발명에 있어서, 상기 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유는 각각 박하, 진피 및 의이인을 헥산(hexane)으로 추출하여 얻어진 것일 수 있다.In the present invention, the peppermint-derived essential oil, dermis-derived essential oil, and uiiin-derived essential oil may be obtained by extracting peppermint, dermis, and uiiin with hexane, respectively.
본 발명에 있어서, 상기 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유는 박하 유래 정유 100중량부 기준 진피 유래 정유 20 내지 250 중량부, 의이인 유래 정유 20 내지 250 중량부 포함할 수 있고, 바람직하게 박하 유래 정유 100중량부 기준 진피 유래 정유 50 내지 200 중량부, 의이인 유래 정유 50 내지 200 중량부 포함할 수 있고, 더욱 바람직하게 박하 유래 정유 100중량부 기준 진피 유래 정유 200 중량부, 의이인 유래 정유 100 중량부 포함할 수 있다.In the present invention, the peppermint-derived essential oil, dermis-derived essential oil, and uiyin-derived essential oil may include 20 to 250 parts by weight of dermis-derived essential oil and 20 to 250 parts by weight of uiyin-derived essential oil based on 100 parts by weight of mint-derived essential oil, preferably mint. Based on 100 parts by weight of the essential oil derived from the dermis, 50 to 200 parts by weight of the essential oil derived from the dermis and 50 to 200 parts by weight of the essential oil derived from Ui-in may be included, and more preferably, 200 parts by weight of the essential oil derived from the dermis, 100 parts by weight of the essential oil derived from the peppermint based on 100 parts by weight of the essential oil derived from mint may contain
본 발명에 따른 건강기능식품 또는 건강식품 조성물에 있어서, 상기 대사성 질환의 예로는 비만, 당뇨병, 고지혈증, 고콜레스테롤증, 동맥경화증 또는 지방간일 수 있고, 바람직하게는 비만, 당뇨 또는 고콜레스테롤증일 수 있으며, 더 바람직하게는 비만일 수 있다.In the health functional food or health food composition according to the present invention, examples of the metabolic disease may be obesity, diabetes, hyperlipidemia, high cholesterol, arteriosclerosis or fatty liver, and preferably obesity, diabetes or high cholesterol. , more preferably obesity.
본 발명의 조성물을 건강기능식품 조성물 또는 건강식품 조성물로 사용하는 경우, 상기 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유를 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용할 수 있고, 통상의 방법에 따라 적절하게 사용할 수 있다. 상기 조성물은 유효성분 이외에 식품학적으로 허용가능한 식품보조첨가제를 포함할 수 있으며, 유효성분의 혼합량은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다.When the composition of the present invention is used as a health functional food composition or health food composition, the peppermint-derived essential oil, dermis-derived essential oil and uiyin-derived essential oil can be added as it is or used together with other foods or food ingredients, according to a conventional method can be used appropriately. In addition to the active ingredient, the composition may include a food additive that is pharmaceutically acceptable, and the mixing amount of the active ingredient may be suitably determined according to the purpose of use (prevention, health or therapeutic treatment).
본 발명에서 사용되는 용어 "식품보조첨가제"란 식품에 보조적으로 첨가될 수 있는 구성요소를 의미하며, 각 제형의 건강기능식품 또는 건강식품을 제조하는데 첨가되는 것으로서 당업자가 적절히 선택하여 사용할 수 있다. 식품보조첨가제의 예로는 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 충진제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등이 포함되지만, 상기 예들에 의해 본 발명의 식품보조첨가제의 종류가 제한되는 것은 아니다.As used in the present invention, the term "food supplement additive" refers to a component that can be supplementally added to food, and is added to manufacture health functional food or health food of each formulation, and those skilled in the art can appropriately select and use it. Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners , pH adjuster, stabilizer, preservative, glycerin, alcohol, carbonation agent used in carbonated beverages, etc., but the above examples are not limited to the type of food supplement additive of the present invention.
또한, 본 발명의 조성물이 사용될 수 있는 건강기능식품 또는 건강식품의 종류에는 제한이 없다. 아울러 본 발명의 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유를 활성성분으로 포함하는 조성물은 당업자의 선택에 따라 건강기능식품에 함유될 수 있는 적절한 기타 보조 성분과 공지의 첨가제를 혼합하여 제조할 수 있다. 첨가할 수 있는 식품의 예로는 육류, 소세지, 빵, 쵸코렛, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 본 발명에 따른 추출물을 주성분으로 하여 제조한 즙, 차, 젤리 및 주스 등에 첨가하여 제조할 수 있다.In addition, there is no limitation on the type of health functional food or health food in which the composition of the present invention can be used. In addition, the composition comprising the essential oil derived from peppermint, essential oil derived from dermis, and essential oil derived from Uiyin of the present invention as an active ingredient can be prepared by mixing appropriate other auxiliary ingredients that may be contained in health functional foods and known additives according to the selection of those skilled in the art. have. Examples of foods that can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages and There are vitamin complexes and the like, and it can be prepared by adding the extract according to the present invention as a main component to juice, tea, jelly, juice, and the like.
대사성 질환의 예방 또는 치료용 약학적 조성물Pharmaceutical composition for the prevention or treatment of metabolic diseases
본 발명은 박하(Mentha haplocalyx) 유래 정유; 진피(Citrus unshiu peel) 유래 정유 및 의이인(Coicis Semen) 유래 정유를 포함하는 대사성 질환의 예방 또는 치료용 약학적 조성물에 관한 것이다.The present invention is peppermint (Mentha haplocalyx) derived essential oil; It relates to a pharmaceutical composition for preventing or treating metabolic diseases, including essential oil derived from Citrus unshiu peel and essential oil derived from Coicis Semen.
본 발명에 있어서, 상기 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유는 각각 박하, 진피 및 의이인을 헥산(hexane)으로 추출하여 얻어진 것일 수 있다.In the present invention, the peppermint-derived essential oil, dermis-derived essential oil, and uiiin-derived essential oil may be obtained by extracting peppermint, dermis, and uiiin with hexane, respectively.
본 발명에 있어서, 상기 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유는 박하 유래 정유 100중량부 기준 진피 유래 정유 20 내지 250 중량부, 의이인 유래 정유 20 내지 250 중량부 포함할 수 있고, 바람직하게 박하 유래 정유 100중량부 기준 진피 유래 정유 50 내지 200 중량부, 의이인 유래 정유 50 내지 200 중량부 포함할 수 있고, 더욱 바람직하게 박하 유래 정유 100중량부 기준 진피 유래 정유 200 중량부, 의이인 유래 정유 100 중량부 포함할 수 있다.In the present invention, the peppermint-derived essential oil, dermis-derived essential oil, and uiyin-derived essential oil may include 20 to 250 parts by weight of dermis-derived essential oil and 20 to 250 parts by weight of uiyin-derived essential oil based on 100 parts by weight of mint-derived essential oil, preferably mint. Based on 100 parts by weight of the essential oil derived from the dermis, 50 to 200 parts by weight of the essential oil derived from the dermis and 50 to 200 parts by weight of the essential oil derived from Ui-in may be included, and more preferably, 200 parts by weight of the essential oil derived from the dermis, 100 parts by weight of the essential oil derived from the peppermint based on 100 parts by weight of the essential oil derived from mint may contain
본 발명에 따른 약학적 조성물에 있어서, 상기 대사성 질환의 예로는 비만, 당뇨병, 고지혈증, 고콜레스테롤증, 동맥경화증 또는 지방간일 수 있고, 바람직하게는 비만, 당뇨 또는 고콜레스테롤증일 수 있으며, 더 바람직하게는 비만일 수 있다.In the pharmaceutical composition according to the present invention, examples of the metabolic disease may be obesity, diabetes, hyperlipidemia, hypercholesterolemia, arteriosclerosis or fatty liver, preferably obesity, diabetes, or high cholesterol, and more preferably may be obese.
본 발명의 일실시예에 있어서, 본 발명에 따른 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유는 지방세포의 분화 및 지방 생성을 억제하고, 지방 세포에서 AMPK, ACC 인산화를 유의적으로 증가시키고, CPT-1의 발현을 유의적으로 증가시킴으로써, 지방 축적 억제 및 지방 대사 조절 효과를 나타내므로, 비만 등 대사성 질환을 예방, 개선 또는 치료할 수 있음을 확인하였다(실험예 2 내지 3 참조).In one embodiment of the present invention, peppermint-derived essential oil, dermis-derived essential oil, and uiiin-derived essential oil according to the present invention inhibit adipocyte differentiation and adipogenesis, and significantly increase AMPK and ACC phosphorylation in adipocytes, By significantly increasing the expression of CPT-1, it has the effect of inhibiting fat accumulation and regulating fat metabolism, and thus it was confirmed that it is possible to prevent, improve or treat metabolic diseases such as obesity (see Experimental Examples 2 to 3).
본 발명의 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유는 임상 투여시에 경구 및 비경구의 여러 가지 제형으로 투여될 수 있으며, 바람직하게는 경구 투여, 경피 투여, 국소 투여, 흡입 투여, 비강내 투여, 정맥내 투여, 진피내 투여, 복강내 투여, 주사 및 피하 주사용 제형으로 이루어진 군으로부터 선택되는 어느 하나 이상의 제형으로 투여되는 것일 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 제조된다.The peppermint-derived essential oil, dermal-derived essential oil, and uiyin-derived essential oil of the present invention may be administered in various oral and parenteral formulations during clinical administration, preferably oral administration, transdermal administration, topical administration, inhalation administration, intranasal administration , intravenous administration, intradermal administration, intraperitoneal administration, may be administered in any one or more formulations selected from the group consisting of injections and subcutaneous injection formulations. In the case of formulation, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants.
경구투여를 위한 고형 제제에는 정제, 환자, 산제, 과립제, 캡슐제, 트로키제 등이 포함되며, 이러한 고형 제제는 하나 이상의 본 발명의 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유에 적어도 하나 이상의 부형제 예를 들면, 전분, 탄산칼슘, 수크로스(sucrose), 락토오스(lactose) 또는 젤라틴 등을 섞어 조제된다. 또한, 단순한 부형제 외에 마그네슘 스티레이트 탈크 같은 윤활제들도 사용된다. 경구 투여를 위한 액상 제제로는 현탁제, 내용액제, 유제 또는 시럽제 등이 해당되는데, 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다.Solid preparations for oral administration include tablets, patients, powders, granules, capsules, troches, and the like, and these solid preparations include at least one excipient in at least one essential oil derived from peppermint, dermis, and essential oil derived from Uiyin of the present invention. For example, it is prepared by mixing starch, calcium carbonate, sucrose, lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid formulations for oral administration include suspensions, solutions, emulsions, or syrups. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. can
비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁용제, 유제, 동결건조제제, 좌제, 크림, 겔, 스프레이, 점적제(drops) 등이 포함된다. 비수성용제, 현탁용제로는 프로필렌글리콜, 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세롤, 젤라틴 등이 사용될 수 있다.Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspension solutions, emulsions, freeze-dried preparations, suppositories, creams, gels, sprays, drops, and the like. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerol, gelatin, and the like can be used.
본 발명의 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유의 인체에 대한 효과적인 투여량은 환자의 나이, 몸무게, 성별, 투여형태, 건강상태 및 질환 정도에 따라 달라질 수 있으며, 일반적으로 약 0.01-1000 mg/kg/일이며, 바람직하게는 0.1-500 mg/kg/일 수 있고, 의사 또는 약사의 판단에 따라 일정시간 간격으로 1일 1회 내지 수회로 분할 투여할 수도 있다.The effective dosage of the essential oil derived from peppermint, dermis, and essential oil derived from the dermis of the present invention for the human body may vary depending on the patient's age, weight, sex, dosage form, health status and disease level, and is generally about 0.01-1000. mg/kg/day, preferably 0.1-500 mg/kg/day, may be administered in divided doses once or several times a day at regular time intervals according to the judgment of a doctor or pharmacist.
본 발명의 약학적 조성물은 유효성분으로서 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유 이외에 공지된 대사성 질환 치료제를 추가로 포함할 수 있고, 이들 질환의 치료를 위해 공지된 다른 치료와 병용될 수 있다.The pharmaceutical composition of the present invention may further include a known therapeutic agent for metabolic diseases in addition to the essential oil derived from peppermint, essential oil derived from dermis, and essential oil derived from Uiin as an active ingredient, and may be used in combination with other known treatments for the treatment of these diseases. .
셀룰라이트 감소 또는 예방용 화장료 조성물Cosmetic composition for reducing or preventing cellulite
본 발명은 박하(Mentha haplocalyx) 유래 정유; 진피(Citrus unshiu peel) 유래 정유 및 의이인(Coicis Semen) 유래 정유를 포함하는 셀룰라이트 감소 또는 예방용 화장료 조성물에 관한 것이다.The present invention is peppermint (Mentha haplocalyx) derived essential oil; It relates to a cosmetic composition for reducing or preventing cellulite comprising essential oil derived from Citrus unshiu peel and essential oil derived from Coicis Semen.
본 발명에 따른 화장료 조성물에 있어서, 상기 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유는 각각 박하, 진피 및 의이인을 헥산(hexane)으로 추출하여 얻어진 것일 수 있다.In the cosmetic composition according to the present invention, the peppermint-derived essential oil, dermis-derived essential oil, and uiiin-derived essential oil may be obtained by extracting peppermint, dermis, and uiiin with hexane, respectively.
본 발명에 있어서, 상기 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유는 박하 유래 정유 100중량부 기준 진피 유래 정유 20 내지 250 중량부, 의이인 유래 정유 20 내지 250 중량부 포함할 수 있고, 바람직하게 박하 유래 정유 100중량부 기준 진피 유래 정유 50 내지 200 중량부, 의이인 유래 정유 50 내지 200 중량부 포함할 수 있고, 더욱 바람직하게 박하 유래 정유 100중량부 기준 진피 유래 정유 200 중량부, 의이인 유래 정유 100 중량부 포함할 수 있다.In the present invention, the peppermint-derived essential oil, dermis-derived essential oil, and uiyin-derived essential oil may include 20 to 250 parts by weight of dermis-derived essential oil and 20 to 250 parts by weight of uiyin-derived essential oil based on 100 parts by weight of mint-derived essential oil, preferably mint. Based on 100 parts by weight of the essential oil derived from the dermis, 50 to 200 parts by weight of the essential oil derived from the dermis and 50 to 200 parts by weight of the essential oil derived from Ui-in may be included, and more preferably, 200 parts by weight of the essential oil derived from the dermis, 100 parts by weight of the essential oil derived from the peppermint based on 100 parts by weight of the essential oil derived from mint may contain
상기 화장료 조성물은, 예를 들면 용액, 겔, 고체 또는 반죽 무수 생성물, 수상에 유상을 분산시켜 얻은 에멀젼, 현탁액, 마이크로에멀젼, 마이크로캡슐, 미세과립구 또는 이온형(리포좀), 비이온형의 소낭 분산제의 형태, 크림, 스킨, 로션, 오일, 파우더, 연고, 스프레이 또는 콘실 스틱의 형태로 제공될 수 있다. 또한, 포말(foam)의 형태 또는 압축된 추진제를 더 함유한 에어로졸 조성물의 형태로도 제조될 수 있다.The cosmetic composition is, for example, a solution, a gel, a solid or kneaded anhydrous product, an emulsion obtained by dispersing an oil phase in an aqueous phase, a suspension, a microemulsion, a microcapsule, a microgranule or an ionic (liposome), a non-ionic vesicle dispersant It may be provided in the form of a cream, toner, lotion, oil, powder, ointment, spray or concealer stick. In addition, it may be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.
또한, 상기 화장료 조성물은 본 발명의 상기 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유에 추가로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온봉쇄제 및 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 또는 화장품에 통상적으로 사용되는 임의의 다른 성분과 같은 화장품 분야에서 통상적으로 사용되는 보조제를 함유할 수 있다.In addition, the cosmetic composition comprises a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, and a foaming agent, in addition to the essential oil derived from peppermint, the essential oil derived from the dermis, and the essential oil derived from the Uiyin of the present invention. (foaming agent), fragrance, surfactant, water, ionic or nonionic emulsifier, filler, sequestering and chelating agent, preservative, vitamin, blocking agent, wetting agent, essential oil, dye, pigment, hydrophilic or lipophilic It may contain adjuvants commonly used in the cosmetic field, such as active agents, lipid vesicles or any other ingredients commonly used in cosmetics.
본 발명의 화장료 조성물에 있어서, 통상적으로 함유되는 화장료 조성물에 본 발명의 상기 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유가 0.1 내지 50 중량%, 바람직하게는 1 내지 10 중량%의 양으로 첨가될 수 있다.In the cosmetic composition of the present invention, the peppermint-derived essential oil, dermis-derived essential oil, and uiyin-derived essential oil of the present invention are added to the commonly contained cosmetic composition in an amount of 0.1 to 50% by weight, preferably 1 to 10% by weight. can
본 발명에 따른 상기 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유를 피부 외용제로 사용하는 경우, 추가로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온봉쇄제 및 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 또는 피부용 외용제에 통상적으로 사용되는 임의의 다른 성분과 같은 피부 과학 분야에서 통상적으로 사용되는 보조제를 함유할 수 있다. 또한, 상기 성분들은 피부 과학 분야에서 일반적으로 사용되는 양으로 도입될 수 있다.When the peppermint-derived essential oil, dermis-derived essential oil, and uiyin-derived essential oil according to the present invention are used as external preparations for skin, fat substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, antioxidants, suspending agents, and stabilizing agents , foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or It may contain adjuvants commonly used in the field of dermatology, such as lipophilic active agents, lipid vesicles, or any other ingredients commonly used in external preparations for skin. In addition, the above ingredients may be introduced in an amount generally used in the field of dermatology.
본 발명에 따른 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유는 천연 식물을 원료로 하므로 약학적, 식품 또는 화장료 조성물로 사용할 경우에도 일반적인 합성 화합물에 비하여 부작용이 덜할 수 있으므로, 안전하게 약학적 조성물, 건강기능식품, 건강식품 및 화장료 조성물에 포함되어 유용하게 사용될 수 있다.Since the peppermint-derived essential oil, dermis-derived essential oil, and uiyin-derived essential oil according to the present invention are made from natural plants, even when used as a pharmaceutical, food or cosmetic composition, side effects may be less than that of general synthetic compounds. It can be usefully included in functional food, health food and cosmetic composition.
또한, 본 발명은 박하(Mentha haplocalyx) 유래 정유; 진피(Citrus unshiu peel) 유래 정유 및 의이인(Coicis Semen) 유래 정유를 포함하는 지방 축적 억제용 조성물을 제공한다.In addition, the present invention is peppermint (Mentha haplocalyx) derived essential oil; Provided is a composition for inhibiting fat accumulation comprising an essential oil derived from Citrus unshiu peel and an essential oil derived from Coicis Semen.
또한, 본 발명은 상기 지방 축적 억제용 조성물을 포함하는 다이어트 조성물을 제공한다.In addition, the present invention provides a diet composition comprising the composition for inhibiting fat accumulation.
하기의 실시예를 통하여 본 발명을 보다 상세하게 설명한다. 그러나 하기 실시예는 본 발명의 내용을 구체화하기 위한 것일 뿐 이에 의해 본 발명이 한정되는 것은 아니다.The present invention will be described in more detail through the following examples. However, the following examples are only intended to embody the contents of the present invention, and the present invention is not limited thereto.
<세포배양 및 분화유도><Cell culture and differentiation induction>
3T3-L1 세포주는 American Type Culture Collection (ATCC, CL-173TM)에서 구입하여 본 실험에 사용하였으며, 세포는 1%의 P/S가 포함된 10%의 BCS을 DMEM 배지에 37℃, 5% CO2배양기 (MCO-15AC, SANYO, Japan)에서 8일간 배양시킨 후, 배양액을 분화유도 배양액 (10% FBS, 5 ㎍/㎖ insulin, 2uM DEX, 111 ㎍/㎖ IBMX)으로 바꾸고 2일간 배양한다. 2일 후, 분화유도 배양액 (10% FBS, 5 ㎍/㎖ insulin)이 포함된 DMEM 배지로 교환해 주어 2일간 배양시킨 뒤, 마지막으로 insulin을 첨가하지 않은 10% FBS가 포함된 DMEM 배지로 배양하였다. 정유 혼합물의 각각의 농도는 (0.01, 0.02 및 0.03 ㎎/㎖) 배지 교체시기마다 함께 처리하였다.The 3T3-L1 cell line was purchased from the American Type Culture Collection (ATCC, CL-173TM) and used in this experiment. 2 After culturing for 8 days in an incubator (MCO-15AC, SANYO, Japan), the culture medium is changed to a differentiation-inducing culture medium (10% FBS, 5 μg/ml insulin, 2uM DEX, 111 μg/ml IBMX) and cultured for 2 days. After 2 days, the culture medium was exchanged with DMEM medium containing differentiation induction culture medium (10% FBS, 5 μg/ml insulin) and cultured for 2 days, and finally cultured in DMEM medium containing 10% FBS without the addition of insulin. did Each concentration of the essential oil mixture (0.01, 0.02 and 0.03 mg/ml) was treated together at each medium change interval.
<준비예 1> 박하 정유의 제조<Preparation Example 1> Preparation of essential peppermint oil
박하(Mentha haplocalyx) 건조물 500g에 추출용매로 헥산(hexane) 5L를 가하여 상온에서 24시간동안 침출시켰고, 추출액은 여과지(Advantec No.2)로 여과하여 여액을 회수한 후, 걸러진 추재에 다시 동량의 용매를 가하여 총 3회 반복 추출하였다. 상기 회수된 여액을 감압농축하여 용매를 제거하여 박하의 정유 성분이 다량 함유된 추출물(박하 유래 정유)을 수득하였다. 5L of hexane as an extraction solvent was added to 500 g of dried mint ( Mentha haplocalyx ) and leached for 24 hours at room temperature, and the extract was filtered with filter paper (Advantec No. A solvent was added and extraction was repeated three times in total. The recovered filtrate was concentrated under reduced pressure to remove the solvent to obtain an extract (mint-derived essential oil) containing a large amount of essential oil of mint.
<준비예 2> 진피 정유의 제조<Preparation Example 2> Preparation of dermis essential oil
상기 준비예 1의 방법과 동일하게, 진피(Citrus unshiu peel) 건조물 500g을 사용하여 진피의 정유 성분이 다량 함유된 추출물(진피 유래 정유)을 수득하였다. In the same manner as in Preparation Example 1, an extract (essential oil derived from dermis) containing a large amount of essential oil components of the dermis was obtained using 500 g of dried Citrus unshiu peel .
<준비예 3> 의이인 정유의 제조<Preparation Example 3> Preparation of Euiin essential oil
상기 준비예 1의 방법과 동일하게, 의이인(Coicis Semen) 건조물 500g을 사용하여 진피의 정유 성분이 다량 함유된 추출물(의이인 유래 정유)을 수득하였다. In the same manner as in Preparation Example 1, an extract (essential oil derived from Ui-in) containing a large amount of essential oil components of the dermis was obtained using 500 g of a dry product of Coicis Semen .
<실시예 1> 박하, 진피 및 의이인 정유 혼합물의 제조<Example 1> Preparation of essential oil mixture of mint, dermis and uiyiin
상기 준비예 1의 박하 유래 정유, 준비예 2의 진피 유래 정유 및 준비예 3의 의이인 유래 정유를 혼합하여, 박하, 진피 및 의이인 유래 정유의 혼합물을 제조하였다. The essential oil derived from peppermint of Preparation Example 1, the essential oil derived from dermis of Preparation Example 2, and the essential oil derived from Uiyin of Preparation Example 3 were mixed to prepare a mixture of peppermint, dermis and essential oil derived from Uiyin.
구체적으로, 하기 표 1과 같이 중량비를 달리하여, 교반하면서 혼합하여 박하, 진피 및 의이인 유래 정유의 혼합물을 수득하였다. Specifically, a mixture of essential oils derived from peppermint, dermis and uiiin was obtained by mixing with different weight ratios as shown in Table 1 below.
<실험예 1> 세포생존율 (MTT) 측정<Experimental Example 1> Cell viability (MTT) measurement
박하, 진피, 의이인의 정유가 3T3-L1 세포생존에 미치는 영향을 알아보기 위하여 96well plate에서 각 정유를 각 농도 (0.01, 0.02 및 0.03 ㎎/㎖)별로 처리하여 MTT 분석법을 이용하였다. 세포에 정유 혼합물을 처리하고 24시간 배양 후, MTT 용액 60㎕를 넣어서 암실에서 2시간 반응시킨 뒤, DMSO를 200㎕를 가하였다. 흡광도는 ELISA reader (Sunrise, TECAN, Grodig, Austria)에, 570㎚에서 흡광도를 측정하였으며, 모두 3개의 well의 평균값을 이용하여 평가하였다.In order to investigate the effect of peppermint, dermis, and Ui-in essential oils on 3T3-L1 cell survival, MTT assay was used by treating each essential oil at each concentration (0.01, 0.02, and 0.03 mg/ml) in a 96-well plate. The cells were treated with the essential oil mixture and cultured for 24 hours, then 60 μl of MTT solution was added and reacted in the dark for 2 hours, and then 200 μl of DMSO was added. Absorbance was measured in an ELISA reader (Sunrise, TECAN, Grodig, Austria) at 570 nm, and all were evaluated using the average value of three wells.
그 결과, 도 1에 나타낸 바와 같이, 0.03 ㎎/㎖을 포함한 모든 농도에서 모두 세포 생존율이 80% 이상을 나타내었다.As a result, as shown in FIG. 1 , all concentrations including 0.03 mg/ml exhibited cell viability of 80% or more.
<실험예 2> 3T3-L1 세포의 분화 및 지방축적 억제 확인<Experimental Example 2> Confirmation of 3T3-L1 cell differentiation and fat accumulation inhibition
3T3-L1 세포에서 지방 축적에 정유 혼합물이 미치는 영향을 알아보기 위하여 Oil red O 염색을 수행하였다. 세포분화 중에서 정유 혼합물을 0.03 ㎎/㎖로 이틀에 한번씩 배지를 교환하며 처리 하였다. 8일 후 세포배양액 제거 후 DPBS로 3회 세척하고 cacodylate buffer (pH 7.2)로 4℃에서 세포를 2시간 고정시킨 후, 증류수로 세척하여 지방구를 Oil red O 염색시약으로 염색하였다. 40% Isopropyl alcohol로 3회 세척을 하고 현미경(CK2, Olympus, Japan)으로 관찰하였다. 염색된 지방세포의 지방함량 측정을 위해 건조시킨 후, 100% isopropyl alcohol로 지방을 추출하여 510㎚ 파장에서 분광광도계 (Sunrise, TECAN, Grodig, Austria)로 흡광도를 측정한 다음, 처리군의 지방함량을 측정하였다.Oil red O staining was performed to investigate the effect of the essential oil mixture on fat accumulation in 3T3-L1 cells. During cell differentiation, the essential oil mixture was treated at 0.03 mg/ml, changing the medium once every two days. After 8 days, the cell culture medium was removed, washed 3 times with DPBS, fixed with cacodylate buffer (pH 7.2) at 4°C for 2 hours, washed with distilled water, and stained with Oil red O staining reagent. It was washed 3 times with 40% Isopropyl alcohol and observed under a microscope (CK2, Olympus, Japan). After drying to measure the fat content of the stained adipocytes, the fat was extracted with 100% isopropyl alcohol, and the absorbance was measured with a spectrophotometer (Sunrise, TECAN, Grodig, Austria) at a wavelength of 510 nm. Then, the fat content of the treated group was measured.
그 결과, 도 2에 나타낸 바와 같이, 비교예 1 내지 3의 박하 유래 정유, 진피 유래 정유 또는 의이인 유래 정유 단독의 경우, 각각 90%, 82% 및 87%의 지방축적억제 효과를 나타내었으며, 비교예 4의 박하 및 진피 유래 정유 혼합물의 경우, 78%의 억제 활성을 보인 반면, 실시예 1 내지 3의 따른 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유 혼합물의 경우, 각각 62%, 52% 및 60%의 억제 활성을 나타내었다. 즉, 박하 유래 정유, 진피 유래 정유 또는 의이인 유래 정유 단독 또는 2종의 혼합물 대비 본 발명의 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유 혼합물을 처리하는 경우, 현저한 지방 축적 억제 효과가 있는 것으로 나타났다.As a result, as shown in FIG. 2, in the case of the essential oil derived from peppermint, the essential oil derived from the dermis, or the essential oil derived from Uiyin of Comparative Examples 1 to 3 alone, 90%, 82%, and 87% of the fat accumulation inhibitory effect was exhibited, respectively. In the case of the mixture of peppermint and dermis-derived essential oil of Example 4, an inhibitory activity of 78% was shown, whereas in the case of the mixture of peppermint-derived essential oil, dermis-derived essential oil and Uiyin-derived essential oil according to Examples 1 to 3, 62%, 52% and It showed an inhibitory activity of 60%. That is, when the mint-derived essential oil, dermis-derived essential oil, or Ui-in-derived essential oil alone or a mixture of two types of essential oil derived from peppermint, dermis-derived essential oil, and Ui-in-derived essential oil mixture of the present invention is treated, there is a significant fat accumulation inhibitory effect.
<실험예 3> 지방세포 분화 및 지방생성 관련 단백질 발현에 미치는 영향 확인<Experimental Example 3> Confirmation of effect on adipocyte differentiation and adipogenesis-related protein expression
3T3-L1 전지방세포의 세포분화 및 성숙 지방세포의 지방생성에 관여하는 단백질 발현을 관찰하기 위하여, 대조군(CON), 양성대조군(FF) 및 비교예 1 내지 4 및 실시예 1 내지 3을 처리한 3T3-L1 세포를 대상으로 웨스턴 블랏(western blot)을 실시하였다.In order to observe the expression of proteins involved in the cell differentiation of 3T3-L1 preadipocytes and the adipogenesis of mature adipocytes, the control group (CON), the positive control group (FF) and Comparative Examples 1 to 4 and Examples 1 to 3 were treated Western blot was performed on one 3T3-L1 cell.
구체적으로, 분화유도 및 혼합물의 처치가 끝난 3T3-L1 세포에 RIPA buffer 200 ㎍/㎖을 넣어 균질화 시킨 다음 13,000 rpm, 4℃에서 20분간 원심분리하여 상층액을 분리하였다. 분리한 추출액을 Bradford법을 사용하여 단백질을 정량한 뒤 25 ㎍을 12% Sodium dodecyl sulfate (SDS, BIO-RAD, USA)-PAGE에서 전기영동을 시킨 후 전개시킨 gel을 PVDF membrane (0.45㎜, Millipore, USA)으로 transfer하였다. 단백질이 전이된 membrane을 5% skim milk 처리하여 비 특이적인 단백질에 대한 blocking을 실시한 후 p-ACC, ACC, p-AMPK, AMPK (1:1000; dilution, Cell Signaling, USA), CPT-1 (1: 1000; dilution, abcam, USA) 및 β -actin (1:3000; dilution, Santa-Cruz Biotechnology, USA) 등의 단백질 항체를 4℃에서 각각 overnight 반응 처리하여 phosphatebuffered saline (PBS)에 0.1%의 Tween 20을 함유시킨 PBS-T로 세척 후 각각의 단백질 항체에 알맞은 2차 항체로 1시간 동안 반응시켰다. 반응이 끝난 membrane은 PBS-T로 수차례 세척 후 ECL (GE Healthcare, UK)시약을 처리하여 Lugen™Sensi-Q2000 Chemidoc (Lugen Scim Inc., Korea)을 이용하여 western band를 검출하였다.Specifically, 200 μg/ml of RIPA buffer was added to 3T3-L1 cells after differentiation induction and treatment of the mixture was completed, homogenized, and then centrifuged at 13,000 rpm and 4° C. for 20 minutes to separate the supernatant. After quantifying the protein of the separated extract using the Bradford method, 25 μg was electrophoresed on 12% sodium dodecyl sulfate (SDS, BIO-RAD, USA)-PAGE. , USA). After blocking the non-specific protein by treating the protein-transferred membrane with 5% skim milk, p-ACC, ACC, p-AMPK, AMPK (1:1000; dilution, Cell Signaling, USA), CPT-1 ( 1:1000; dilution, abcam, USA) and β-actin (1:3000; dilution, Santa-Cruz Biotechnology, USA) were reacted overnight at 4°C with 0.1% of phosphate buffered saline (PBS). After washing with PBS-
에너지 대사 및 지방대사를 조절하는 pAMPK 및 AMPK의 단백 발현을 확인한 결과, 도 3(a)에 나타낸 바와 같이, 비교예 1 내지 3의 박하 유래 정유, 진피 유래 정유 또는 의이인 유래 정유 단독의 경우, 각각 0.5, 0.54 및 0.59로 pAMPK/AMPK의 발현이 유의하게 증가하였며, 비교예 4의 박하 및 진피 유래 정유 혼합물의 경우, 0.64의 단백질 발현을 증가를 보인 반면, 실시예 1 내지 3의 따른 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유 혼합물의 경우, 각각 1.05, 1.45 및 1.13으로 비교예 1 내지 3의 박하 유래 정유, 진피 유래 정유 또는 의이인 유래 정유 단독 또는 2종의 혼합물 대비 현저히 우수한 pAMPK/AMPK 발현 증가가 나타났다.As a result of confirming the protein expression of pAMPK and AMPK that regulate energy metabolism and fat metabolism, as shown in FIG. The expression of pAMPK/AMPK was significantly increased to 0.5, 0.54, and 0.59, and in the case of the mint and dermis-derived essential oil mixture of Comparative Example 4, the protein expression was increased by 0.64, whereas the mint-derived according to Examples 1 to 3 In the case of essential oil, dermis-derived essential oil, and uiiin-derived essential oil mixture, respectively, 1.05, 1.45, and 1.13, respectively, indicating significantly superior pAMPK/AMPK expression compared to peppermint-derived essential oil, dermis-derived essential oil, or uiiin-derived essential oil alone or a mixture of two of Comparative Examples 1 to 3 increase appeared.
또한, 지방산 합성과 콜레스테롤 합성을 조절하는 pACC 및 ACC의 단백 발현에 미치는 영향을 확인한 결과, 도 3(b)에 나타낸 바와 같이, 비교예 1 내지 3의 박하 유래 정유, 진피 유래 정유 또는 의이인 유래 정유 단독의 경우, 각각 0.55, 0.61 및 0.64로 pACC/ACC의 발현이 유의하게 증가하였며, 비교예 4의 박하 및 진피 유래 정유 혼합물의 경우, 0.73의 단백질 발현을 증가를 보인 반면, 실시예 1 내지 3의 따른 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유 혼합물의 경우, 각각 1.11, 1.39 및 1.27으로 비교예 1 내지 3의 박하 유래 정유, 진피 유래 정유 또는 의이인 유래 정유 단독 또는 2종의 혼합물 대비 현저히 우수한 pACC/ACC 발현 증가가 나타났다.In addition, as a result of confirming the effect on the protein expression of pACC and ACC, which regulate fatty acid synthesis and cholesterol synthesis, as shown in FIG. In the case of alone, the expression of pACC/ACC was significantly increased to 0.55, 0.61, and 0.64, respectively, and in the case of the mint and dermis-derived essential oil mixture of Comparative Example 4, the protein expression was increased by 0.73, whereas Examples 1 to In the case of the peppermint-derived essential oil, dermis-derived essential oil, and Ui-in-derived essential oil mixture according to 3, respectively, 1.11, 1.39, and 1.27, respectively, significantly compared to the peppermint-derived essential oil, dermis-derived essential oil, or Ui-iin-derived essential oil alone or a mixture of two of Comparative Examples 1 to 3 A good increase in pACC/ACC expression was observed.
또한, 지방산 산화 조절효소인 CPT(Carnitine palmitoyl transferase)-1 단백 발현에 미치는 영향을 확인한 결과, 도 3(c)에 나타낸 바와 같이, 비교예 1 내지 3의 박하 유래 정유, 진피 유래 정유 또는 의이인 유래 정유 단독의 경우, 각각 0.41, 0.49 및 0.55로 CPT-1/β-actin의 발현이 유의하게 증가하였며, 비교예 4의 박하 및 진피 유래 정유 혼합물의 경우, 0.68의 단백질 발현을 증가를 보인 반면 실시예 1 내지 3의 따른 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유 혼합물의 경우, 각각 0.84, 1.18 및 0.91로 비교예 1 내지 3의 박하 유래 정유, 진피 유래 정유 또는 의이인 유래 정유 단독 또는 2종의 혼합물 대비 현저히 우수한 CPT-1/β-actin 발현 증가가 나타났다.In addition, as a result of confirming the effect on the expression of CPT (Carnitine palmitoyl transferase)-1 protein, which is a fatty acid oxidation regulator, as shown in FIG. In the case of essential oil alone, the expression of CPT-1/β-actin was significantly increased to 0.41, 0.49, and 0.55, respectively, and the mint and dermis-derived essential oil mixture of Comparative Example 4 showed an increase in protein expression by 0.68, respectively In the case of the mixture of peppermint-derived essential oil, dermis-derived essential oil, and uiiin-derived essential oil according to Examples 1 to 3, respectively, 0.84, 1.18, and 0.91, respectively, the peppermint-derived essential oil, dermis-derived essential oil, or uiiin-derived essential oil of Comparative Examples 1 to 3 alone or two types A significantly superior increase in CPT-1/β-actin expression compared to the mixture of
즉, 박하 유래 정유, 진피 유래 정유 또는 의이인 유래 정유 단독 또는 2종의 혼합물 대비 본 발명의 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유 혼합물을 처리하는 경우, 지방세포의 분화 및 지방의 대사 조절에 관여하는 단백질의 발현을 현저하게 증가시키는 것을 확인하였다. That is, when the mint-derived essential oil, dermis-derived essential oil, or Ui-in-derived essential oil alone or a mixture of two types of peppermint-derived essential oil, dermis-derived essential oil and Ui-iin-derived essential oil mixture of the present invention is treated, adipocyte differentiation and fat metabolism regulation are improved. It was confirmed that the expression of the protein involved was significantly increased.
<통계처리><Statistics processing>
본 연구의 모든 실험 결과는 SPSS Statistics(ver. 11.0, SPSS Inc., Chicago, IL, USA) 이용하여 산출되었고, 결과는 mean±S.E. (standard error)로 표시하였다. 실험결과는 일원배치 분산분석(one-way analysis of variance, ANOVA)을 한 후, Duncan's multiple range test에 의해 p<0.05 수준에서 각 실험군의 평균치의 통계적 유의성을 검정하였다.All experimental results of this study were calculated using SPSS Statistics (ver. 11.0, SPSS Inc., Chicago, IL, USA), and the results were mean±S.E. (standard error). After one-way analysis of variance (ANOVA) was performed on the experimental results, the statistical significance of the mean value of each experimental group was tested at p<0.05 level by Duncan's multiple range test.
이제까지 본 발명에 대하여 그 바람직한 실시예들을 중심으로 살펴보았다. 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자는 본 발명이 본 발명의 본질적인 특성에서 벗어나지 않는 범위에서 변형된 형태로 구현될 수 있음을 이해할 수 있을 것이다. 그러므로 개시된 실시예들은 한정적인 관점이 아니라 설명적인 관점에서 고려되어야 한다. 본 발명의 범위는 전술한 설명이 아니라 특히 청구범위에 나타나 있으며, 그와 동등한 범위내에 있는 모든 차이점은 본 발명에 포함된 것으로 해석되어야 할 것이다.So far, with respect to the present invention, the preferred embodiments have been looked at. Those of ordinary skill in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments are to be considered in an illustrative rather than a restrictive sense. The scope of the present invention is particularly indicated in the claims rather than the foregoing description, and all differences within an equivalent scope should be construed as being included in the present invention.
Claims (17)
진피(Citrus unshiu peel) 유래 정유 및
의이인(Coicis Semen) 유래 정유를 포함하는 대사성 질환의 예방 또는 개선용 건강기능식품 조성물.essential oil from Mentha haplocalyx;
Essential oil derived from Citrus unshiu peel and
A health functional food composition for preventing or improving metabolic diseases comprising essential oil derived from Coicis Semen.
상기 대사성 질환은 비만, 당뇨병, 고지혈증, 고콜레스테롤증, 동맥경화증 및 지방간으로 이루어진 군으로부터 선택되는 어느 하나 이상인 것을 특징으로 하는 건강기능식품 조성물.The method of claim 1,
The metabolic disease is a health functional food composition, characterized in that at least one selected from the group consisting of obesity, diabetes, hyperlipidemia, hypercholesterolemia, arteriosclerosis and fatty liver.
상기 대사성 질환은 비만인 것을 특징으로 하는 건강기능식품 조성물.3. The method of claim 2,
The metabolic disease is a health functional food composition, characterized in that obesity.
상기 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유는 각각 박하, 진피 및 의이인을 헥산(hexane)으로 추출하여 얻어진 것을 특징으로 하는 건강기능식품 조성물.The method of claim 1,
The peppermint-derived essential oil, dermis-derived essential oil, and uiiin-derived essential oil is a health functional food composition, characterized in that obtained by extracting mint, dermis, and uiiin with hexane, respectively.
상기 건강기능식품은 정제, 캡슐제, 환제 또는 액제 형태의 식품인 것을 특징으로 하는 건강기능식품 조성물.The method of claim 1,
The health functional food is a health functional food composition, characterized in that the food in the form of tablets, capsules, pills or liquids.
진피(Citrus unshiu peel) 유래 정유 및
의이인(Coicis Semen) 유래 정유를 포함하는 대사성 질환의 예방 또는 개선용 건강식품 조성물.essential oil from Mentha haplocalyx;
Essential oil derived from Citrus unshiu peel and
A health food composition for preventing or improving metabolic diseases comprising essential oil derived from Coicis Semen.
상기 대사성 질환은 비만, 당뇨병, 고지혈증, 고콜레스테롤증, 동맥경화증 및 지방간으로 이루어진 군으로부터 선택되는 어느 하나 이상인 것을 특징으로 하는 건강식품 조성물.7. The method of claim 6,
The metabolic disease is a health food composition, characterized in that at least one selected from the group consisting of obesity, diabetes, hyperlipidemia, hypercholesterolemia, arteriosclerosis and fatty liver.
상기 대사성 질환은 비만인 것을 특징으로 하는 건강식품 조성물.8. The method of claim 7,
The metabolic disease is a health food composition, characterized in that obesity.
상기 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유는 각각 박하, 진피 및 의이인을 헥산(hexane)으로 추출하여 얻어진 것을 특징으로 하는 건강기능식품 조성물.7. The method of claim 6,
The peppermint-derived essential oil, dermis-derived essential oil, and uiiin-derived essential oil is a health functional food composition, characterized in that obtained by extracting mint, dermis, and uiiin with hexane, respectively.
상기 건강식품은 각종 드링크제, 육류, 소세지, 빵, 캔디류, 스넥류, 면류, 아이스크림, 유제품, 스프, 이온음료, 음료수, 알코올 음료, 껌, 차 및 비타민 복합제에서 선택되는 것을 특징으로 하는 건강식품 조성물.7. The method of claim 6,
The health food is selected from various drinks, meat, sausage, bread, candy, snacks, noodles, ice cream, dairy products, soup, ionic beverages, beverages, alcoholic beverages, gum, tea, and vitamin complexes. Health food composition, characterized in that.
진피(Citrus unshiu peel) 유래 정유 및
의이인(Coicis Semen) 유래 정유를 포함하는 대사성 질환의 예방 또는 치료용 약학적 조성물.essential oil from Mentha haplocalyx;
Essential oil derived from Citrus unshiu peel and
A pharmaceutical composition for preventing or treating metabolic diseases comprising essential oil derived from Coicis Semen.
상기 대사성 질환은 비만, 당뇨병, 고지혈증, 고콜레스테롤증, 동맥경화증 및 지방간으로 이루어진 군으로부터 선택되는 어느 하나 이상인 것을 특징으로 하는 약학적 조성물.12. The method of claim 11,
The metabolic disease is a pharmaceutical composition, characterized in that any one or more selected from the group consisting of obesity, diabetes, hyperlipidemia, hypercholesterolemia, arteriosclerosis and fatty liver.
상기 대사성 질환은 비만인 것을 특징으로 하는 약학적 조성물.13. The method of claim 12,
The metabolic disease is a pharmaceutical composition, characterized in that obesity.
상기 박하 유래 정유, 진피 유래 정유 및 의이인 유래 정유는 각각 박하, 진피 및 의이인을 헥산(hexane)으로 추출하여 얻어진 것을 특징으로 하는 약학적 조성물.12. The method of claim 11,
The peppermint-derived essential oil, dermis-derived essential oil and uiiin-derived essential oil is a pharmaceutical composition, characterized in that obtained by extracting mint, dermis, and uiiin with hexane, respectively.
진피(Citrus unshiu peel) 유래 정유 및
의이인(Coicis Semen) 유래 정유를 포함하는 셀룰라이트 감소 또는 예방용 화장료 조성물.essential oil from Mentha haplocalyx;
Essential oil derived from Citrus unshiu peel and
A cosmetic composition for reducing or preventing cellulite comprising essential oil derived from Coicis Semen.
진피(Citrus unshiu peel) 유래 정유 및
의이인(Coicis Semen) 유래 정유를 포함하는 지방 축적 억제용 조성물.essential oil from Mentha haplocalyx;
Essential oil derived from Citrus unshiu peel and
A composition for inhibiting fat accumulation comprising essential oil derived from Coicis Semen.
A diet composition comprising the composition of claim 16 .
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