KR100787175B1 - A herbal mixture extract comprising Atractylodis Rhizoma Alba, Gastrodia elata, Aurantii nobilis pericarpium, Poria cocos, Crataegur, Siegesbeckia glabrescens Makino, and food supplement comprising the same for prevention and treatment of Arteriosclerosis - Google Patents
A herbal mixture extract comprising Atractylodis Rhizoma Alba, Gastrodia elata, Aurantii nobilis pericarpium, Poria cocos, Crataegur, Siegesbeckia glabrescens Makino, and food supplement comprising the same for prevention and treatment of Arteriosclerosis Download PDFInfo
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- KR100787175B1 KR100787175B1 KR1020060050066A KR20060050066A KR100787175B1 KR 100787175 B1 KR100787175 B1 KR 100787175B1 KR 1020060050066 A KR1020060050066 A KR 1020060050066A KR 20060050066 A KR20060050066 A KR 20060050066A KR 100787175 B1 KR100787175 B1 KR 100787175B1
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- extract
- smooth muscle
- vascular smooth
- prevention
- treatment
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Abstract
본 발명은 백출, 천마, 진피, 복령, 산사 및 희렴을 포함하는 한약제제 혼합물의 동맥경화 및 관련 질환의 예방 및 치료용 추출물과, 이를 유효 성분으로 포함하는 동맥경화 및 관련 질환의 예방 및 치료용 건강식품 조성물에 관한 것으로, 상기 추출물의 제조시 용매로서는 물 또는 에탄올을 이용하여 추출하는 것이 바람직하다. 또한, 본 발명은 상기 한약제제 혼합물에 반하, 황련으로 이루어진 군중에서 1이상을 선택하여 추가한 한약제제 혼합물의 동맥경화 및 관련 질환의 예방 및 치료용 추출물과, 이를 유효 성분으로 포함하는 동맥경화 및 관련 질환의 예방 및 치료용 건강식품 조성물을 제공한다.The present invention is an extract for the prevention and treatment of atherosclerosis and related diseases of herbal mixtures, including baekchul, cheonma, dermis, bokyeong, hawthorn and haejugo, and for the prevention and treatment of atherosclerosis and related diseases comprising the same as an active ingredient. It relates to a health food composition, it is preferable to extract using water or ethanol as a solvent in the preparation of the extract. In addition, the present invention is an extract for preventing and treating atherosclerosis and related diseases of the Chinese herbal medicine mixture, which is selected by adding one or more selected from the group consisting of rhubarb, and the arteriosclerosis comprising the same as an active ingredient It provides a health food composition for the prevention and treatment of related diseases.
본 발명의 추출물에 대해 실험한 결과, 상기 추출물은 혈소판유래성장인자(Platelet derived growth factor; PDGF)에 의해 증가된 혈관 평활근 세포의 증식을 양성 대조군인 PDGF 단독 투여군에 비해 약 2배 정도 억제하였으며, 또한 상기 추출물을 1 ㎎/㎖ 농도로 24시간동안 혈관 평활근 세포에 처치하였을 때 양성 대조군에 비해 약 50% 정도의 현저한 세포 이주 억제 효과를 나타내었다. 따라서, 본 발명에 따른 추출물은 동맥경화 및 관련 질환의 예방 및 치료용 제재로 유용하게 사용될 수 있다. As a result of experiments on the extract of the present invention, the extract inhibited about 2 times the proliferation of vascular smooth muscle cells increased by platelet derived growth factor (PDGF) compared to the positive control PDGF alone group, In addition, when the extract was treated with vascular smooth muscle cells at a concentration of 1 mg / ml for 24 hours, it showed a significant cell migration inhibition effect of about 50% compared to the positive control. Therefore, the extract according to the present invention can be usefully used as an agent for preventing and treating atherosclerosis and related diseases.
백출, 천마, 혈관 평활근 세포(SMC), 혈소판유래성장인자(Platelet derived growth factor; PDGF), 증식(proliferation), 이주(migration), 동맥경화 Eruption, shunt, vascular smooth muscle cell (SMC), platelet derived growth factor (PDGF), proliferation, migration, arteriosclerosis
Description
도 1은 본 발명에 따른 추출물을 혈관 평활근 세포에 농도별로 투여하고 세포생존율에 미치는 효과를 측정하여 나타낸 그래프.1 is a graph showing the effect of the extract on the vascular smooth muscle cells according to the concentration of the extract according to the present invention and measuring the effect on cell viability.
도 2는 혈소판유래성장인자에 의해 증가된 혈관 평활근 세포 증식을 본 발명에 따른 추출물이 억제하는 효과를 측정하여 나타낸 그래프.Figure 2 is a graph showing the effect of inhibiting the extract according to the present invention vascular smooth muscle cell proliferation increased by platelet derived growth factor.
도 3는 본 발명에 따른 추출물이 혈관 평활근의 이주에 미치는 효과를 트랜스웰 이주능 분석법으로 측정하여 나타낸 그래프. Figure 3 is a graph showing the effect of the extract according to the invention on the migration of vascular smooth muscle measured by Transwell migration ability analysis method.
도 4는 혈소판유래성장인자에 의해 증가된 혈관 평활근 세포 이주를 본 발명에 따른 추출물이 억제하는 효과를 나타낸 사진.Figure 4 is a photograph showing the effect of the extract according to the present invention to inhibit vascular smooth muscle cell migration increased by platelet-derived growth factor.
본 발명은 백출, 천마, 진피, 복령, 산사 및 희렴을 포함하는 한약제제 혼합물의 동맥경화 및 관련 질환의 예방 및 치료용 추출물, 및 이를 유효 성분으로 포함하는 동맥경화 및 관련 질환의 예방 및 치료용 건강식품 조성물에 관한 것이다.The present invention is an extract for the prevention and treatment of atherosclerosis and related diseases of the herbal medicine mixture including baekchul, cheonma, dermis, bokyeong, hawthorn and haejugo, and for the prevention and treatment of atherosclerosis and related diseases comprising the same as an active ingredient It relates to a health food composition.
동맥경화증은 동맥벽에 비후(肥厚)나 조직의 변성(變性)이 일어나서 경화(硬化)하는 질환으로, 동맥의 내막이 두터워져 내경이 좁아지면서 산소와 각종 영양분을 공급하는 관동맥과 뇌 및 하지로 가는 동맥에 혈류장애가 나타나는데, 노화와 더불어 발생하는 주요 질환 중의 하나이다. 동맥의 내막에 과도한 세포증식에 의해 생겨난 죽종 주위에 섬유화가 일어나 혈관이 점차 단단해지는 현상을 죽상동맥경화라 하며, 죽상이란 용어를 생략하여 동맥경화라 하기도 한다. Atherosclerosis is a disease in which thickening or tissue degeneration occurs in the artery wall, causing it to stiffen. The inner lining of the arteries becomes thick, narrowing the inner diameter, and supplying oxygen and various nutrients to the coronary arteries, brain, and lower limbs. Blood flow disorders appear in the arteries, one of the major diseases that occur with aging. Atherosclerosis is a process in which fibrosis occurs around an atherosclerosis caused by excessive cell proliferation in the artery's lining, thereby stiffening blood vessels. Atherosclerosis is sometimes referred to as atherosclerosis.
동맥경화증은 10세 전후에서 시작되며 40세 후반에 혈류장애가 나타나는데, 때에 따라 섬유막이 파열되어 혈관 안에 혈전이 생길 수 있고, 죽종 안에 출혈이 일어나 혈관내경이 급격하게 좁아져 각종 질병을 유발한다. 동맥경화는 뇌동맥 또는 관상동맥에서 일어나기 쉬운데, 뇌동맥경화증의 경우에는 두통, 현기증, 정신장애를 나타내고 뇌연화증의 원인이 되며, 관상동맥경화증의 경우에는 심장부에 동통과 부정맥을 일으켜 협심증, 심근경색 등의 원인이 되는 것으로 알려져 있다. 또한 이로 인해 심장병, 뇌일혈 등이 유발되어, 동맥경화증으로 인한 질병이 현대 사회에 있어, 특히 50∼60대의 남성들에게 가장 큰 사망요인으로 부각되고 있다. 서구에서는 예전부터 주된 사망원인이 되는 흔한 질병이며, 국내 통계청발표에 따르면 1995년 20%에서 2000년에는 73%로 증가하여 국내에서도 동맥경화와 관련된 사망률 이 약 50%정도 증가한 것으로 나타났다. 또한, 동맥경화는 중심성 비만을 포함하여 비만(과체중), 혈전, 과응고 및 혈전유발 상태(동맥과 정맥) 등의 대사 증후군이나 지질이상혈증 등과 같은 질환의 발생 및 진행과 깊이 관련되고, 당뇨병성 만성합병증을 유발하거나 악화시킨다.Atherosclerosis begins around the age of 10 years and blood flow disorders appear in late 40 years. Sometimes, the fibrous membrane ruptures to cause blood clots in the blood vessels, and bleeding occurs within the atherosclerosis, which causes the disease to narrow rapidly. Atherosclerosis is more likely to occur in the cerebral artery or coronary arteries. In the case of cerebral arteriosclerosis, headache, dizziness, and mental disorders are indicated and cause encephalopathy, and in the case of coronary arteriosclerosis, pain and arrhythmia in the heart cause angina and myocardial infarction. It is known to become. In addition, this causes heart disease, cerebral hemorrhage, and diseases caused by arteriosclerosis are emerging as the leading cause of death in modern society, especially among men in their 50s and 60s. In the West, it is a common cause of mortality, and according to the National Statistical Office's announcement, it increased from 20% in 1995 to 73% in 2000, and the atherosclerosis-related mortality rate increased about 50% in Korea. In addition, atherosclerosis is deeply related to the development and progression of diseases such as metabolic syndrome such as obesity (overweight), blood clots, hypercoagulation and thrombolytic conditions (arteries and veins), including hypertrophy, and dyslipidemia. Causes or worsens chronic complications.
동맥은 대동맥, 중동맥, 소동맥으로 구분되며, 내막, 중막, 및 외막의 세 층으로 구성되며 특히, 죽상동맥경화증은 주로 탄력성 및 근육성 동맥에서 잘 발생한다. 혈관 수축과 확장기능을 하는 혈관 평활근 세포는 자극을 받으면 내막으로 이주하여 증식할 수 있으며, LDL에 대한 수용체를 가지고 있다. 정상적인 상태에서는 매우 엄격한 기작에 의하여 증식이 조절되지만, 비정상적인 경우 과다 증식되어 혈관계 질환을 야기 시킬 수 있다. 또한, 혈관 평활근 세포는 협심증과 심근 경색증으로 대표되는 허혈성 심질환 치료시 혈관 재협착을 유발하는데, 이를 방지하기 위해 평활근 세포 증식을 억제하는 물질을 사용하고 있으며, 항산화제, 항혈소판제, 혈관확장제, 항응고제 등의 투여가 실시되고 있지만, 임상적으로 충분한 효과를 나타내고 있지 않다.Arteries are divided into aorta, middle and small arteries, and are composed of three layers of inner, middle, and outer membranes. In particular, atherosclerosis occurs mainly in elastic and muscular arteries. Vascular smooth muscle cells that function as vasoconstrictors and dilates can migrate and proliferate into the lining when stimulated and have receptors for LDL. In normal conditions, the proliferation is controlled by a very strict mechanism, but in abnormal cases it can overproliferate and cause vascular disease. In addition, vascular smooth muscle cells cause vascular restenosis in the treatment of ischemic heart disease such as angina pectoris and myocardial infarction. To prevent this, vascular smooth muscle cells are used to inhibit smooth muscle cell proliferation.Antioxidants, antiplatelets, vasodilators, anticoagulants Although administration of the back is performed, it does not show clinically sufficient effect.
반하, 백출, 천마는 반하백출천마탕의 주요 구성성분이며, 족태음의 담궐두통을 주치로 한 방제로 비위내상으로 인한 두통여렬, 신중여산, 오심번민, 사지궐냉, 구토, 현훈 증상에 처방하며, 그 제습화담의 효능으로 최근 담습의 조체로 인한 심혈관계질환의 예방과 치료에 응용되고 있다. 병리적으로 비위의 허약은 위내정수와 수독에 의한 담음을 발생시키며 또한, 담음은 혈류의 장애를 초래하여 심혈관계질환의 병태와 관련되어 있으므로 반하백출천마탕은 담음이 관여하는 혈류장애 나 심혈관계 질환에 대한 개선과 혈압강하의 효과가 기대된다. Banha, Baekchul, Cheonma are the main components of Banha Baekchulcheonmatang, and they are prescribed for the symptom of headaches caused by nasal injuries, prudent acid, nausea, cold limbs, vomiting, and vertigo. In recent years, it has been applied to the prevention and treatment of cardiovascular diseases due to the dehumidification. Pathologically, weakness of the stomach is caused by intragastric infiltration and venom. Also, because of the impairment of blood flow, it is associated with the condition of cardiovascular disease. It is expected to improve the disease and lower blood pressure.
이에 본 발명자는 백출, 천마, 진피, 복령, 산사 및 희렴을 포함하는 한약제제 혼합물을 추출하여 동맥경화 치료 효능을 연구한 결과, 본 발명에 따른 추출물이 혈소판유래성장인자(Platelet derived growth factor; PDGF)에 의해 증가된 혈관 평활근 세포의 증식 및 유주 활성을 억제하는 효과가 있음을 발견하고 본 발명을 완성하였다.Therefore, the present inventors have studied the efficacy of treating arteriosclerosis by extracting a mixture of herbal preparations including baekchul, cheonma, dermis, bokyeong, hawthorn and haejugo, the extract according to the present invention is a platelet derived growth factor (PDGF) The present invention was found to have an effect of inhibiting the growth and vascular activity of vascular smooth muscle cells increased by).
따라서, 본 발명의 목적은 백출, 천마, 진피, 복령, 산사 및 희렴을 포함하는 한약제제 혼합물의 추출물에 대해 혈관 평활근 세포의 증식 및 유주 활성을 억제하는 객관적인 효능을 밝혀 동맥경화 및 관련 질환의 예방 및 치료용 추출물을 제공하는 것이다.Accordingly, it is an object of the present invention to uncover the objective efficacy of inhibiting the proliferation and lipogenic activity of vascular smooth muscle cells against extracts of herbal mixtures including baekchul, cheonma, dermis, bokyeong, hawthorn and hunger, preventing the prevention of atherosclerosis and related diseases. And to provide a therapeutic extract.
본 발명의 다른 목적은 백출, 천마, 진피, 복령, 산사 및 희렴을 포함하는 한약제제 혼합물의 추출물을 포함하는 동맥경화 및 관련 질환의 예방 및 치료용 건강식품 조성물을 제공하는 것이다.Another object of the present invention is to provide a health food composition for the prevention and treatment of atherosclerosis and related diseases, including extracts of herbal mixtures including baekchul, cheonma, dermis, bokyeong, hawthorn and hunger.
본 발명의 또 다른 목적은 상기 한약제제 혼합물에 반하, 황련으로 이루어진 군중에서 1이상을 선택하여 추가로 혼합한 한약제제 혼합물의 동맥경화 및 관련 질환의 예방 및 치료용 추출물과, 이를 유효성분으로 포함하는 동맥경화 및 관련 질환의 예방 및 치료용 건강식품 조성물을 제공하는 것이다.Another object of the present invention is an extract for the prevention and treatment of atherosclerosis and related diseases of the Chinese herbal medicine mixture, in addition to the herbal medicine mixture, by selecting one or more from the group consisting of rhubarb, and further comprising the same as an active ingredient To provide a health food composition for the prevention and treatment of atherosclerosis and related diseases.
상기 목적을 달성하기 위하여, 본 발명은 백출, 천마, 진피, 복령, 산사 및 희렴을 포함하는 한약제제 혼합물의 동맥경화 및 관련 질환의 예방 및 치료용 추출물을 제공한다. 또한, 본 발명은 상기 추출물을 유효성분으로 포함하는 것을 특징으로 하는 동맥경화 및 관련 질환의 예방 및 치료용 건강식품 조성물을 제공한다.In order to achieve the above object, the present invention provides an extract for the prevention and treatment of atherosclerosis and related diseases of the herbal medicine mixture, including baekchul, cheonma, dermis, bokyeong, hawthorn and hunger. The present invention also provides a health food composition for the prevention and treatment of atherosclerosis and related diseases comprising the extract as an active ingredient.
또한, 본 발명은 상기 한약제제 혼합물에 반하, 황련으로 이루어진 군중에서 1이상을 선택하여 추가로 혼합한 한약제제 혼합물의 동맥경화 및 관련 질환의 예방 및 치료용 추출물과, 이를 유효성분으로 포함하는 동맥경화 및 관련 질환의 예방 및 치료용 건강식품 조성물을 제공한다.In addition, the present invention is an extract for preventing and treating atherosclerosis and related diseases of the Chinese herbal medicine mixture, which is further selected by selecting one or more from the crowd consisting of rhubarb, and an artery comprising the same as an active ingredient, in contrast to the herbal medicine mixture. It provides a health food composition for the prevention and treatment of sclerosis and related diseases.
상기 추출물들의 제조시 용매로서 물 또는 에탄올을 이용하여 추출하는 것이 바람직하다. It is preferable to extract using water or ethanol as a solvent in the preparation of the extracts.
이하, 본 발명의 구성을 구체적으로 설명한다. EMBODIMENT OF THE INVENTION Hereinafter, the structure of this invention is demonstrated concretely.
본 발명은 백출, 천마, 진피, 복령, 산사 및 희렴을 포함하는 한약제제 혼합물의 동맥경화 및 관련 질환의 예방 및 치료용 추출물을 제공한다.The present invention provides extracts for the prevention and treatment of atherosclerosis and related diseases of herbal mixtures, including baekchul, cheonma, dermis, Bokryeong, hawthorn and hunger.
본 발명의 조성물의 각 성분 약재별 약리학적 특성은 다음과 같다. The pharmacological properties of each ingredient medicine of the composition of the present invention are as follows.
본 발명에 사용되는 백출(Atractylodis Rhizoma Alba)은 국화과에 속하는 다년생 식물 삽주의 근경을 일으키며, 방향성건위제로 한방에서는 비장을 보강하는 건비, 보비 처방에 빈용되며 진정, 이뇨, 지한, 자양, 안테효과, 진통작용, 항염증작용, 혈당치저해효능, 혈압강하, 이뇨작용등의 효과가 있다.Atractylodis Rhizoma Alba used in the present invention causes rooting of perennial plant inserts belonging to the Asteraceae family, and is a fragrant conditional agent used in oriental medicine to supplement the spleen, supplemented with sputum and bobby prescription, soothing, diuretic, cold, nourishing, ante effect, Analgesic, anti-inflammatory, hypoglycemic effect, lowering blood pressure, diuretic effect is effective.
천마(Gastrodia elata)는 난초과의 여러해살이풀로 한방에서 뇌신경쇠약증, 진통, 두통, 중풍, 고혈압 등에 효과가 있다고 알려져 있으며 또한 콩팥염 및 당뇨병 등에도 사용된다. Gastrodia elata is a perennial herb of Orchidaceae. It is known for its effects on neurasthenia, pain, headache, stroke and hypertension. It is also used for kidney disease and diabetes.
진피(Aurantii nobilis pericarpium)는 귤껍질로서, 산초과 또는 운향과(Rutaceae)에 속하는 광귤나무(Citrus Auranticum L.) 또는 제주도산 귤나무(C. Junos), 귤나무(C. nobilis) 등의 익은 열매의 껍질을 벗겨 햇볕에 말린것인데, 동의치료에서는 오래 묵인것일수록 좋다고 알려져 있으며, 성분으로는 정유와 플라보노이드인 헤스페리딘(hesperidin)이 함유되어 있어 방향성 건위약 또는 진해, 거담, 복만, 복통 등에 널리 사용되고 있는 안정성이 확보된 생약의 일종이다.The dermis (Aurantii nobilis pericarpium) is a tangerine peel, ripened such as Citrus Auranticum L. or C. Junos and C. nobilis belonging to the Sanaceae or Rutaceae family. It is peeled and dried in the sun, and it is known that the longer it stays in motion therapy, the ingredients contain essential oils and flavonoids, hesperidin, which are widely used in fragrant dry stomach, Jinhae, sputum, stomachache, abdominal pain, etc. It is a kind of herbal medicine with stability.
복령(Poria cocos)은 나무뿌리에 기생하며, 균핵(菌核) 크기는 10∼30cm이며 백색인 것을 백복령(白茯笭), 적색인 것을 적복령(赤茯笭)이라 한다. 한약재로 강장 ·이뇨 ·진정 등에 효능이 있어 신장병 ·방광염 ·요도염에 이용한다. Poor cocos (Poria cocos) is parasitic in the roots of the tree, the size of the fungal nucleus (菌核) is 10 ~ 30cm and white is called the white baekbokyeong (白 茯 笭), red is called the red bokbokyeong (赤 茯 笭). It is an herbal medicine and is effective in tonic, diuretic and soothing. It is used for kidney disease, cystitis and urethritis.
산사(Crataegur)는 장미과에 속하는 낙엽성 소교목으로 그 성분으로는 히페로사이드(Hyperoside), 케르세틴(quercetin), 올레아놀(oleanol)이 있으며 과실은 이과로서 구형 또는 도란형이며 산사자라 한다. 약효는 건위, 소화, 강장, 고혈압, 월경통 등에 사용한다. Crataegur is a deciduous arborescent belonging to the family Rosaceae, and its components include hyperside, quercetin, and oleanol. Fruits are spherical or obovate, and hawthorn. The drug is used in stomach, digestion, tonic, high blood pressure, dysmenorrhea, etc.
희렴(Siegesbeckia glabrescens Makino)은 국화과의 한해살이 초본으로서 성분으로는 디테르펜다투로시드, 다투린, 알칼로이드, 사포닌, 정유 등이 있으며 혈압내림, 악창, 풍습, 중풍에 효과가 있다.Siegesbeckia glabrescens Makino is an annual herb of Asteraceae, and its components include diterpentauroside, daturin, alkaloids, saponins, and essential oils, and are effective in lowering blood pressure, swelling, customs, and stroke.
본 발명에 따른 한약제제 혼합 추출물의 혈관 평활근 세포의 이주 활성 억제 효과를 규명하기 위해 세포 실험을 하였다. 표 1은 본 발명의 추출물이 혈관 평활근 세포의 이주능을 억제하는 효능에 대해 관찰한 결과이다. 그 결과, PDGF에 의해 95%까지 혈관 평활근 세포가 이주하였으나, 본 발명의 추출물은 1㎎/㎖의 농도에서 42%의 혈관 평활근 세포의 이주를 나타내어 PDGF에 의한 세포이주능을 억제하는 효과를 나타냈다.Cell experiments were carried out to investigate the inhibitory effect of vascular smooth muscle cells of the medicinal herb extract according to the present invention. Table 1 shows the results of the extract of the present invention observed the efficacy of inhibiting the migration ability of vascular smooth muscle cells. As a result, vascular smooth muscle cells migrated up to 95% by PDGF, but the extract of the present invention exhibited 42% vascular smooth muscle cell migration at a concentration of 1 mg / ml, indicating an effect of inhibiting cell migration by PDGF.
본 발명은 상기 한약제제 혼합물에 반하, 황련으로 이루어진 군중에서 1이상을 선택하여 추가로 혼합한 한약제제 혼합물의 동맥경화 및 관련 질환의 예방 및 치료용 추출물을 제공한다.The present invention provides an extract for the prevention and treatment of atherosclerosis and related diseases of the Chinese herbal medicine mixture in which at least one selected from the group consisting of rhubarb is further mixed with the herbal medicine mixture.
반하(Pinellia termate)는 청남성과의 다년초로 끼무릇의 코르크층을 제거하고 덩이줄기를 사용하는 생약으로 진토, 이뇨, 안압강하, 진해거담, 분비억제, 항부정맥, 항암작용 등의 효과가 알려져 있다. Vanellia (Pinellia termate) is a perennial herb that removes all cork layers and uses tubers. It is known to have the effects of clay, diuresis, intraocular pressure drop, antitussive expectoration, anti-secretion, antiarrhythmic and anticancer activity.
황련 (Coptis Rhizoma)은 미나리아제비과(Ranunculaceae)에 속하는 다년생 초본으로, 주성분으로는 베르베린(berberine), 콥티신(Coptisine), 보레닌(worenine), 팔마틴(palmatine) 및 야테오리진(Jateorrhizine) 등을 함유하며, 고미건위정장약등으로 쓰이고, 주성분인 베르베린은 장내세균의 발육을 억제하는 항균작용이 있으며, 한방에서는 위장병, 출혈, 동맥경화, 염증, 정신불안 등에 사용되고 있는 생약이다.Coptis Rhizoma is a perennial herb belonging to the Ranunculaceae, and its main ingredients include berberine, copticine, borenine, palmin, and yaterorhizine. It is used as a high-density gastrointestinal pill, and the main ingredient is berberine, which has an antibacterial effect of inhibiting the growth of intestinal bacteria, and is used in herbal medicine for gastrointestinal diseases, bleeding, arteriosclerosis, inflammation, and mental anxiety.
반하 및 황련을 모두 포함하는 본 발명의 한약제제 혼합 추출물의 혈관 평활근 세포의 증식 및 이주 활성 억제 효과를 규명하기 위해 세포 실험을 하였다. 먼저, 혈관 평활근 세포 증식에 관한 실험에서는, 음성 대조군에서 48시간에 증식된 세포를 100%라 하면 PDGF 단독 투여시 세포 증식 정도는 160%로 60% 증가하였다. 반면에, PDGF와 반하 및 황련을 모두 포함하는 본 발명의 추출물을 병용 투여하였을 경우는 약 90-100% 정도로 음성 대조군과 비슷한 혈관 평활근 세포의 증식을 나타내므로서, 양성 대조군인 PDGF 단독 투여군에 비해 혈관 평활근 세포 증식을 현저하게 억제하였다(도2).Cell experiments were carried out to investigate the effect of inhibiting the proliferation and migration activity of vascular smooth muscle cells of the medicinal herb extract of the present invention containing both halves and barberry. First, in the experiment on vascular smooth muscle cell proliferation, the percentage of cell proliferation increased by 60% to 160% when PDGF alone was administered when 100% of cells proliferated at 48 hours in the negative control group. On the other hand, when the co-administration of the extract of the present invention including both PDGF and halves and rhubarb shows about 90-100% proliferation of vascular smooth muscle cells similar to the negative control, compared with the positive control PDGF alone group. Vascular smooth muscle cell proliferation was significantly suppressed (FIG. 2).
그리고, 트랜스웰 이주능 분석법을 이용하여 반하 및 황련을 모두 포함하는 본 발명의 추출물이 혈관 평활근 세포의 이주에 미치는 효능을 측정한 결과, PDGF를 단독 투여하여 48시간 경과 후 평활근 세포의 유주활성을 100%로 나타내었을 때, PDGF와 반하 및 황련을 모두 포함하는 본 발명의 추출물을 농도별로 병용투여한 군은 80%-40%로 나타났다. 따라서, 반하 및 황련을 모두 포함하는 추출물은 농도 의존적으로 PDGF에 의한 혈관 평활근세포의 이주를 억제하는 것으로 나타났다(도3).In addition, as a result of measuring the efficacy of the extract of the present invention containing both halves and rhubarb on the migration of vascular smooth muscle cells by using the transwell migration assay, PDGF alone was administered 48 hours after the smooth muscle cell migration activity. When expressed as 100%, the group administered with the concentration of the extract of the present invention containing both PDGF and half and rhubarb appeared in 80% -40%. Therefore, the extract containing both halves and rhubarb was shown to inhibit the migration of vascular smooth muscle cells by PDGF in a concentration-dependent manner (Fig. 3).
또한, 반하 및 황련을 모두 포함하는 본 발명의 한약제제 추출물에 대해 혈관 평활근 세포의 이주 활성 억제 효과를 상처치유 (Wound healing) 분석법으로 측정한 결과, PDGF를 단독 투여하여 24시간 경과후 혈관 평활근 세포의 유주활성을 100%로 나타내었을 때, PDGF와 반하 및 황련을 모두 포함하는 본 발명의 추출물을 병용 투여한 군은 약 20-50% 정도의 유주활성을 나타내어 PDGF에 의한 혈관 평활근 세포의 이주를 억제하는 것으로 나타났다(표2, 도4). 표 2는 반하 및 황련을 모두 포함하는 본 발명의 추출물을 투여하고 혈소판유래성장인자에 의해 증가된 혈관 평활근 세포 이주 억제에 미치는 효과를 3-4회 반복실험한 후 통계처리한 결과를 정량화하여 나타낸 표이다.In addition, the effect of inhibiting the migration activity of vascular smooth muscle cells in the herbal extracts of the present invention including both halves and rhubarb was measured by wound healing assay, and vascular
따라서, 상기 본 발명에 따른 추출물들은 혈관 평활근 세포의 이주 및 증식을 효과적으로 억제하므로, 혈관 평활근 세포의 이주 및 증식에 관련된 동맥경화 및, 이와 관련된 질환으로서 관상동맥 심장병, 심근경색증, 협심증, 고지혈증, 뇌연화증, 뇌일혈, 뇌졸중, 뇌경색, 뇌출혈, 혈관 재협착 및 폐색 뿐만 아니라 당뇨병과 그 합병증, 비만, 지질이상혈증 등과 같은 잠재성 대사질환의 예방 및 치료에 사용될 수 있다.Therefore, the extracts according to the present invention effectively inhibit the migration and proliferation of vascular smooth muscle cells, and thus, arteriosclerosis and the related diseases as coronary heart disease, myocardial infarction, angina pectoris, hyperlipidemia, encephalopathy It can be used for the prevention and treatment of latent metabolic diseases such as cerebral hemorrhage, stroke, cerebral infarction, cerebral hemorrhage, vascular restenosis and obstruction as well as diabetes and its complications, obesity, and dyslipidemia.
본 발명은 상기 한약제제 혼합물들을 물, 유기용매로 이루어진 군중에서 1이상의 화합물을 선택하여 추출한 동맥경화 예방 및 치료용 추출물을 제공한다.The present invention provides an extract for preventing and treating atherosclerosis, wherein the herbal mixture is selected by extracting at least one compound from a crowd consisting of water and an organic solvent.
본 발명에 따른 한약제제 혼합 추출물들은 상기 한약제제 혼합물들을 물 또는 유기용매, 또는 물과 유기용매의 혼합용매로 추출하여 얻을 수 있는데, 유기용매로는 저급 알콜, 아세톤, 클로로포름, 메틸렌클로라이드, 에테르, 에틸아세테이트, 헥산 등을 예시할 수 있다. 상기 저급 알콜로는 메탄올, 에탄올, 프로판올 및 부탄올을 예시할 수 있으며, 에탄올이 가장 바람직하다. Herbal medicine mixture extracts according to the present invention can be obtained by extracting the herbal medicine mixture with water or an organic solvent, or a mixed solvent of water and an organic solvent, the organic solvent is lower alcohol, acetone, chloroform, methylene chloride, ether, Ethyl acetate, hexane, etc. can be illustrated. The lower alcohol may be exemplified by methanol, ethanol, propanol and butanol, with ethanol being most preferred.
물을 용매로 하여 상기 한약제제 혼합물들을 추출하는 경우에는, 상기 한약제제 혼합물 총 무게의 1 내지 2 배, 바람직하게는 1.5 배의 물을 첨가하고 80 내지 100℃, 바람직하게는 90 내지 100℃의 온도에서 1 내지 24시간, 바람직하게는 3 내지 10시간동안 추출한 후 여과하여 본 발명에 따른 한약제제 혼합물들의 열수 추출물을 제조할 수 있다. 유기용매를 이용하여 상기 한약제제 혼합물들을 추출하는 경우는 상기 한약제제 혼합물 총 무게의 1 내지 10배, 바람직하게는 5배의 유기용매를 첨가하고 실온에서 추출한 후, 여과하여 얻어진 여액을 감압 농축하여 제조할 수 있다. 상기 추출방법들에서 추출공정은 필요에 따라 2회 이상 반복하여 실시할 수 있다. 또한, 본 발명의 한약제제 혼합물들의 추출물은 여액을 농축시킨 엑기스 상태 또는 상기 여과된 여액을 농축시키고 건조하여 분말 상태로 제조하는 것을 포함한다. 또한, 상기 건조는 통상적인 모든 방법이 사용될 수 있으나 바람직하게는 동결건조 방법으로 수행된다.When extracting the herbal mixtures using water as a solvent, 1 to 2 times, preferably 1.5 times, water of the total weight of the herbal mixture is added and 80 to 100 ° C, preferably 90 to 100 ° C. Extraction for 1 to 24 hours, preferably 3 to 10 hours at a temperature and then filtered to prepare a hydrothermal extract of the herbal mixtures according to the present invention. When extracting the herbal mixtures using an organic solvent, an organic solvent of 1 to 10 times, preferably 5 times the total weight of the herbal mixture is added, extracted at room temperature, and the filtrate obtained by filtration is concentrated under reduced pressure. It can manufacture. In the extraction methods, the extraction process may be repeated two or more times as necessary. In addition, the extracts of the herbal mixtures of the present invention include extracting the concentrated filtrate or preparing the powdered filtrate by concentrating and drying the filtrate. In addition, the drying may be any conventional method may be used but is preferably carried out by a lyophilization method.
상기 본 발명에 사용되는 한약제제들은 오랫동안 생약으로 사용되어 오던 약제로서 독성 및 부작용 등의 문제가 없다. 또한, 본 발명은 상기 혼합 한약제제 추출물들을 유효성분으로 포함하는 것을 특징으로 하는 동맥경화 및 관련 질환의 예방 및 치료용 건강식품 조성물들을 제공한다. 본 발명에 따른 한약제제 혼합 추출물들은 흰쥐에 투여해도 부작용이나 독성 등의 문제가 없어 동맥경화 및 관련 질환의 예방 및 치료용 건강식품 조성물의 유효성분으로 이용될 수 있다. 본 발명에 따른 추출물들을 식품 또는 음료 첨가물로 사용할 경우, 상기 추출물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용되고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 일반적으로, 식품 또는 음료의 제조시에는 본 발명의 한약제제 혼합 추출물들이 총 원료에 대하여 0.0001 내지 50 중량%로, 바람직하게는 0.001 내지 20 중량%의 양으로 첨가된다. 그러나, 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다. 또한, 유효 성분은 안전성 면에서 아무런 문제가 없기 때문에 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간 복용이 가능하다. Chinese herbal medicines used in the present invention have been used as a herbal medicine for a long time there is no problem such as toxicity and side effects. The present invention also provides health food compositions for the prevention and treatment of atherosclerosis and related diseases, characterized in that the mixed herbal extracts as an active ingredient. Herbal medicine mixture extract according to the present invention can be used as an active ingredient of the health food composition for the prevention and treatment of atherosclerosis and related diseases do not have problems such as side effects or toxicity even when administered to rats. When the extracts according to the present invention are used as food or beverage additives, the extracts may be added as is or used together with other food or food ingredients, and may be appropriately used according to conventional methods. Generally, in the preparation of food or beverages, the herbal extracts of the present invention are added in an amount of 0.0001 to 50% by weight, preferably 0.001 to 20% by weight, based on the total raw material. However, in the case of long-term intake for health and hygiene or health control, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety. . In addition, since the active ingredient has no problem in terms of safety, it can be taken for a long time for health and hygiene purposes or for health control purposes.
상기 식품의 종류에는 특별한 제한은 없다. 본 발명에 따른 바람직한 건강식품 조성물은 통상의 방법에 따라 제형화 하여 투여될 있는데, 열수 추출물을 그대로 탕제로, 또는 농축하여 엑기스로 하거나 건조하여 분말화 할 수 있고, 열수 추출물에 담체를 넣어 건조한 후 캡슐화 하거나 정제, 과립, 음료, 분말 등의 형태로 제형화 할 수 있다. 상기 물질을 첨가할 수 있는 식품의 예로는 쥬스, 즙, 젤리, 육류, 소세지, 빵, 쵸코렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 파우치, 앰플, 차, 드링크제 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다.There is no particular limitation on the kind of food. The preferred health food composition according to the present invention can be formulated and administered according to a conventional method, and the hot water extract can be powdered by extracting it with water, or concentrated to extract or dried, and then dried by putting a carrier in the hot water extract. It may be encapsulated or formulated in the form of tablets, granules, beverages, powders and the like. Examples of the food to which the substance may be added include juice, juice, jelly, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream including ice cream, various soups, Drinks, pouches, ampoules, teas, drinks and vitamin complexes, and the like, and includes all of the health food in the usual sense.
본 발명의 추출물의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 그러나, 바람직한 효과를 위해서, 본 발명의 추출물은 1일 0.001 내지 300mg/kg으로, 바람직하게는 0.001 내지 100mg/kg으로 투여하는 것이 좋다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다.Preferred dosages of the extracts of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.001 to 300 mg / kg, preferably 0.001 to 100 mg / kg per day. Administration may be administered once a day or may be divided several times.
이하, 본 발명을 하기 실시예에 의거하여 좀 더 구체적으로 설명하고자 한다. 단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 범위가 이들만으로 제한되는 것은 아니다. Hereinafter, the present invention will be described in more detail based on the following examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.
[실시예 1] 본 발명에 따른 동맥경화 및 관련 질환의 예방 및 치료용 추출물의 제조Example 1 Preparation of Extracts for Preventing and Treating Arteriosclerosis and Related Diseases According to the Present Invention
백출 12 g, 천마 6 g, 진피 6 g, 복령 9 g, 산사 9 g, 희렴 9 g을 잘 씻은 후 추출용기에 넣고 물 500 ㎖을 가하여 100℃에서 3시간 열수 추출하였다. 얻어진 용액을 실온에서 식힌 후 거름종이로 여과하였다. 상기 추출액은 진공회전증발기를 이용하여 40℃ 이하에서 감압농축하고 동결건조하여 분말상태의 본 발명에 따른 혼합 한약제제 추출물을 얻었다(수율 : 15 w/v%). 상기 한약제제 혼합 추출물을 이하의 실험예 2 및 제제예에 사용하였다.12 g of Baekchul, 6 g of Cheonma, 6 g of Dermis, 9 g of Bokryeong, 9 g of Sansa, 9 g of Happiness were washed well, and then put into an extraction container and 500 ml of water was added to extract hot water at 100 ° C. for 3 hours. The resulting solution was cooled at room temperature and then filtered through a filter paper. The extract was concentrated under reduced pressure at 40 ° C. or lower using a vacuum rotary evaporator and lyophilized to obtain a mixed herbal extract according to the present invention in powder form (yield: 15 w / v%). The herbal mixture extract was used in the following Experimental Example 2 and Formulation Example.
[실시예 2] 반하 및 황련을 모두 포함하는 한약제제 혼합물의 동맥경화 및 관련 질환의 예방 및 치료용 추출물의 제조Example 2 Preparation of Extracts for Preventing and Treating Arteriosclerosis and Related Diseases
반하 9 g, 백출 12 g, 천마 6 g, 진피 6 g, 복령 9 g, 산사 9 g, 희렴 9 g, 황련 9 g 을 잘 씻은 후 추출용기에 넣고 물 500 ㎖을 가하여 100℃에서 3시간 열수 추출하였다. 얻어진 용액을 실온에서 식힌 후 거름종이로 여과하였다. 상기 추출액은 진공회전증발기를 이용하여 40℃ 이하에서 감압농축하고 동결건조하여 분말상태의 본 발명에 따른 혼합 한약제제 추출물을 얻었다(수율 : 15 w/v%). 상기 한약제제 혼합 추출물을 이하의 실험예에 사용하였다.After washing 9g, Baekchul 12g, Cheonma 6g, Dermis 6g, Bokryeong 9g, Sansa 9g, Happiness 9g, 9g of rhubarb, wash well and put into an extraction container and add 500ml of water and heat it at 100 ℃ for 3 hours. Extracted. The resulting solution was cooled at room temperature and then filtered through a filter paper. The extract was concentrated under reduced pressure at 40 ° C. or lower using a vacuum rotary evaporator and lyophilized to obtain a mixed herbal extract according to the present invention in powder form (yield: 15 w / v%). The herbal mixture extract was used in the following experimental example.
[실험예 1] 실시예2에서 얻은 추출물의 농도에 따른 혈관 평활근 세포의 독성 실험 Experimental Example 1 Toxicity Test of Vascular Smooth Muscle Cells According to the Concentration of the Extract Obtained in Example 2
실시예2에서 얻은 추출물의 혈관 평활근세포에 대한 독성 실험은 MTT법을 통해 관찰하였다. 혈관 평활근 세포를 96웰-플레이트에 90-100% 자랄 때까지 배양한 후 혈청이 없는 배지로 기존 배지를 갈아 24시간 동안 배양하여 혈청을 고갈시켰다. 그 다음, 실시예2에서 얻은 추출물을 혈청이 없는 배지에 녹여 96웰-플레이트의 세포에 농도별(㎎/㎖)로 처치하였다. 상기 추출물을 처치하여 24시간 경과 후, 2㎎/㎖ 농도로 MTT 용액을 첨가하였다. 약 4시간 후 플레이트에 형성된 포르마겐(formagen)을 DMSO 200㎕에 녹여 ELISA 리더로 540nm에서 흡광도를 측정하였다. 시료에 의한 세포의 생존률은 하기식을 이용하여 계산하였고, 각각 농도별 실험군에서 대조군에 대하여 70 % 이하의 생존률을 나타낸 시료를 세포독성을 나타내는 시료로 판단하였다.Toxicity experiments on vascular smooth muscle cells of the extract obtained in Example 2 were observed through the MTT method. Vascular smooth muscle cells were incubated in 96 well-plates until they grew 90-100%, and then serum was depleted by grinding the existing medium with serum-free medium for 24 hours. Then, the extract obtained in Example 2 was dissolved in medium without serum and treated to cells of 96 well-plates by concentration (mg / ml). After 24 hours of treatment with the extract, MTT solution was added at a concentration of 2 mg / ml. After about 4 hours, the formalin formed on the plate was dissolved in 200 µl of DMSO, and the absorbance was measured at 540 nm with an ELISA reader. The survival rate of the cells by the sample was calculated using the following equation, and the samples showing survival rates of 70% or less with respect to the control group in each experimental group for each concentration were determined as samples showing cytotoxicity.
세포의 생존률(%) = (실험군의 흡광도 / 대조군의 흡광도) ×100% Survival rate of cells = (absorbance of experimental group / absorbance of control group) × 100
상기 실험의 결과를 하기 도1에 나타내었다. 도1에 도시한 바와 같이 혈관 평활근 세포는 실시예2에서 얻은 추출물 1㎎/㎖에서 세포 생존율이 약 20% 증가하는 것으로 나타나 독성이 없는 것으로 관찰되었으나, 실시예2에서 얻은 추출물 5㎎/㎖의 농도에서 혈관 평활근 세포의 생존율이 약 50% 정도 감소하는 것으로 관찰되었다. 따라서, 실시예2에서 얻은 추출물 1㎎/㎖ 이상의 농도에서는 혈관 평활근 세포에 대한 독성이 관찰되었다.The results of the experiment are shown in FIG. 1. As shown in FIG. 1, the vascular smooth muscle cells were found to be non-toxic because the cell viability was increased by about 20% in 1 mg / ml of the extract obtained in Example 2, but 5 mg / ml of the extract obtained in Example 2 was observed. It was observed that the survival rate of vascular smooth muscle cells decreased by about 50% at the concentration. Therefore, toxicity against vascular smooth muscle cells was observed at the concentration of 1 mg / ml or more of the extract obtained in Example 2.
[실험예 2] 실시예1에서 얻은 추출물의 농도에 따른 혈관 평활근 세포의 이주 억제 효과Experimental Example 2 Effect of Inhibiting Migration of Vascular Smooth Muscle Cells According to the Concentration of the Extract Obtained in Example 1
혈관 평활근 세포의 이주 억제 효과를 상처 치유 분석(wound healing assay)으로도 관찰하였다. 혈관 평활근 세포를 24웰-플레이트에서 약 100% 자랄 때까지 배양하고, 혈관 평활근 세포의 증식을 정지시키기 위해 증식 억제 물질인 마이토마이신(Mitomycin)을 25㎍/㎖ 농도로 투여하였다. 1시간 후 200㎕용 피펫팁을 이용하여 웰-플레이트 바닥에 일정한 굵기의 선을 긋고 배지를 제거하여, PBS로 세척하였다. 그리고 나서, 1% FBS가 첨가된 배지에 실시예1에서 얻은 추출물을 투여하지 않거나 1 mg/ml의 농도로 투여하고 30분 후 5ng/㎖ PDGF를 첨가하고, 12시간 경과후 혈관 평활근 세포의 유주 거리를 측정하였다.The inhibitory effect of vascular smooth muscle cell migration was also observed by wound healing assay. Vascular smooth muscle cells were incubated until growth was about 100% in 24 well-plates, and a growth inhibitory substance, mitomycin, was administered at a concentration of 25 µg / ml to stop the proliferation of vascular smooth muscle cells. After 1 hour, using a 200 μl pipette tip, a line of constant thickness was drawn at the bottom of the well-plate, and the medium was removed and washed with PBS. Then, in the medium to which 1% FBS was added, the extract obtained in Example 1 was not administered or administered at a concentration of 1 mg / ml, and 5 ng / ml PDGF was added 30 minutes later. The distance was measured.
상기 실험 결과를 하기 표3에 나타내었다.The experimental results are shown in Table 3 below.
상기 표3은 이주한 거리를 백분율로 나타낸 표이다. 표1에서 나타낸 바와 같이 PDGF 단독 투여한 후 12시간째 측정한 결과 95% 정도의 혈관 평활근 세포의 이주가 관찰되었으나, 실시예1에서 얻은 추출물을 전투여 하였을 경우에는 42% 정도의 세포 이주를 나타내어 PDGF에 의한 세포 이주를 억제함을 관찰할 수 있었다.Table 3 is a table showing the migrated distance in percentage. As shown in Table 1, as measured by 12 hours after PDGF alone administration, about 95% of vascular smooth muscle cell migration was observed.However, when the extract obtained in Example 1 was subjected to combat, the cell migration was about 42%. It was observed that cell migration by PDGF was inhibited.
[실험예 3] 실시예2에서 얻은 추출물의 혈관 평활근 세포의 증식 억제 효과Experimental Example 3 Inhibition of Proliferation of Vascular Smooth Muscle Cells of the Extract Obtained in Example 2
혈관 평활근 세포를 96웰-플레이트에서 약 50% 자랄 때까지 배양하고, 혈청이 없는 배지로 기존 배지를 갈아 24시간 동안 배양하여 혈청을 고갈시켰다. 그 다음, 음성 대조군에는 배지만 100㎕, 실험군에는 실시예2에서 얻은 추출물을 1㎎/㎖의 농도로 배지에 녹여 100㎕씩 전투여하고. 1시간 후 PDGF를 양성 대조군과 실험군에 5ng/㎖ 농도로 투여하였다. 각 대조군과 실험군을 12시간에서 72시간까지 배양하고, 각 배양 시간이 경과하면 2 ㎎/㎖의 MTT 용액 50㎕를 첨가하고 각 플레이트에 형성된 포르마겐(formagen)을 DMSO 200㎕에 녹여 ELISA 리더로 540nm에서 흡광도를 측정하였다.Vascular smooth muscle cells were cultured in 96 well-plates until about 50% growth, and the serum was depleted by grinding the existing medium with serum-free medium for 24 hours. Next, 100 μl of the medium in the negative control group, and 100 μl of the extract obtained in Example 2 in the medium at a concentration of 1 mg / ml in the experimental group. After 1 hour, PDGF was administered to the positive control group and the experimental group at a concentration of 5 ng / ml. Each control group and the experimental group were incubated for 12 hours to 72 hours, and after each incubation time, 50 µl of 2 mg / ml MTT solution was added, and formagen formed in each plate was dissolved in 200 µl of DMSO, followed by ELISA reader. Absorbance was measured at 540 nm.
상기 실험 결과를 하기 도2에 나타내었다. 도2에 도시한 바와 같이 혈관 평활근 세포는 PDGF 단독 투여하였을 경우에는 음성 대조군에 비해 2배 정도까지 세포 생존율이 관찰되었으나, 실시예2에서 얻은 추출물과 PDGF 병용 투여 군에서는 음성 대조군과 비슷한 생존율을 나타내었다.The experimental results are shown in FIG. 2. As shown in FIG. 2, when PDGF alone was administered, vascular smooth muscle cells showed a cell survival rate of about 2 times compared to that of the negative control group, but the extract obtained in Example 2 and the PDGF combination group showed similar survival rates as the negative control group. It was.
[실험예 4] 실시예2에서 얻은 추출물이 혈관 평활근의 이주에 미치는 효과를 트랜스웰 이주능 분석법으로 나타낸 결과 Experimental Example 4 The effect of the extract obtained in Example 2 on the migration of vascular smooth muscle
혈관 평활근 세포의 이주 억제 효과를 타입 I 콜라젠 (type I collagen) 으로 코팅된 트랜스웰을 이용하여 관찰하였다. 혈관 평활근 세포를 3개의 플레이트에서 약 100% 자랄 때까지 배양하고, 혈청이 없는 배지로 기존 배지를 갈아 24시간동안 배양하여 혈청을 고갈시켰다. 업퍼 챔버 (Upper chamber)에 혈관 평활근 세포 100㎕(1×106) 와 본 발명에 따른 추출물을 0, 0.1, 1㎎/㎖로 각각 투여한 후, 로어 챔버 (lower chamber)에 600㎕의 혈청이 없는 배지를 첨가하였다. 1시간 후 50 ng/㎖의 PDGF를 로어 챔버 (lower chamber)에 첨가하고 37℃에서 배양하였다. 48시간 경과 후, 업퍼 서피시스 (upper surfaces)를 세척하고 메탄올로 30분간 고정하여 김사액 (Giemsa solution)으로 1시간 동안 염색한 후, 이주된 혈관 평활근 세포수를 측정하였다.The migration inhibitory effect of vascular smooth muscle cells was observed using a transwell coated with type I collagen. Vascular smooth muscle cells were incubated until the growth of about 100% in three plates, the existing medium was changed to a medium without serum and cultured for 24 hours to deplete the serum. 100 μl (1 × 10 6 ) of vascular smooth muscle cells and the extract according to the present invention were administered to the upper chamber at 0, 0.1 and 1 mg / ml, respectively, and then 600 μl of serum was added to the lower chamber. No medium was added. After 1
상기 실험 결과를 하기 도3에 나타내었다.The experimental results are shown in FIG. 3.
도3에 도시된 바와 같이 PDGF를 단독 투여하여 48시간 경과 후 혈관 평활근 세포의 이주를 100%로 나타내었을 때, 실시예2에서 얻은 추출물을 0.1㎎/㎖과 1㎎/㎖의 농도로 병용 투여하였을 경우 각각 약 80%와 40%로 나타나 혈관 평활근 세포의 이주 억제 효능을 관찰할 수 있었다.As shown in FIG. 3, when the PDGF alone was administered for 48 hours and the migration of vascular smooth muscle cells was represented as 100%, the extract obtained in Example 2 was administered in combination of 0.1 mg / ml and 1 mg / ml. In this case, about 80% and 40%, respectively, were observed to inhibit the migration of vascular smooth muscle cells.
[실험예 5] 실시예2에서 얻은 추출물의 농도에 따른 혈관 평활근 세포의 이주 억제 효과Experimental Example 5 Inhibition of Migration of Vascular Smooth Muscle Cells According to the Concentration of the Extract Obtained in Example 2
혈관 평활근 세포의 이주 억제 효과를 상처 치유 분석(wound healing assay)으로도 관찰하였다. 혈관 평활근 세포를 24웰-플레이트에서 약 100% 자랄 때까지 배양하고, 혈관 평활근 세포의 증식을 정지시키기 위해 증식 억제 물질인 마이토마이신(Mitomycin)을 25㎍/㎖ 농도로 투여하였다. 1시간 후 200㎕용 피펫팁을 이용하여 웰-플레이트 바닥에 일정한 굵기의 선을 긋고 배지를 제거하여, PBS로 세척하였다. 그리고 나서, 1% FBS가 첨가된 배지에 실시예2에서 얻은 추출물을 농도별로 투여하고 30분 후 5ng/㎖ PDGF를 첨가하여 12시간 간격으로 혈관 평활근 세포의 유주 거리를 측정하였다.The inhibitory effect of vascular smooth muscle cell migration was also observed by wound healing assay. Vascular smooth muscle cells were incubated until growth was about 100% in 24 well-plates, and a growth inhibitory substance, mitomycin, was administered at a concentration of 25 µg / ml to stop the proliferation of vascular smooth muscle cells. After 1 hour, using a 200 μl pipette tip, a line of constant thickness was drawn at the bottom of the well-plate, and the medium was removed and washed with PBS. Then, the concentration of vascular smooth muscle cells was measured at 12 hour intervals by administering the extract obtained in Example 2 to the medium to which 1% FBS was added at different concentrations, and adding 5 ng / ml PDGF after 30 minutes.
상기 실험 결과를 하기 표4와 도4에 나타내었다.The experimental results are shown in Table 4 and FIG. 4.
도4는 혈관 평활근 세포의 유주 사진이며 상기 표4는 이주한 거리를 백분율로 나타낸 표이다. 도4에 도시된 바와 같이 PDGF 투여하여 세포의 이주를 유도한 결과 24시간째 혈관 평활근 세포의 이주가 100% 진행된 것을 관찰하였고, 실시예2에서 얻은 추출물과 PDGF를 병용투여 하였을 경우 실시예2에서 얻은 추출물에 의해 혈관 평활근 세포의 이주가 억제된 것을 관찰할 수 있었다. 또한, 표4에 나타낸 바와 같이 PDGF 단독 투여한 후 12시간째 측정한 결과 95% 정도의 혈관 평활근 세포의 이주가 관찰되었으나, 실시예2에서 얻은 추출물을 전투여하였을 경우에는 59.3% 와 35.5%로 약 40-60% 정도의 세포 이주를 나타내어 PDGF에 의한 세포 이주를 억제함을 관찰할 수 있었다. 또, 24시간째에는 PDGF 투여군에서 100% 이주가 진행된 반면, 실시예2에서 얻은 추출물을 전투여 하였을 경우에는 약 50% 정도 혈관 평활근 세포의 이주가 관찰되어 PDGF에 의한 세포 이주를 상기 추출물이 억제함을 관찰할 수 있었다.4 is a photograph of vascular smooth muscle cells, and Table 4 is a table showing the migrated distance as a percentage. As shown in FIG. 4, as a result of inducing the migration of cells by PDGF administration, the migration of vascular smooth muscle cells was observed 100% at 24 hours, and when the extract obtained in Example 2 and PDGF were co-administered in Example 2 It was observed that the obtained extract suppressed the migration of vascular smooth muscle cells. In addition, as shown in Table 4, as measured by 12 hours after PDGF alone administration, about 95% of vascular smooth muscle cell migration was observed.However, when the extract obtained in Example 2 was subjected to combat, it was 59.3% and 35.5%. About 40-60% of the cell migration was observed to inhibit cell migration by PDGF. In addition, at 24 hours, 100% migration progressed in the PDGF-administered group, whereas about 50% migration of vascular smooth muscle cells was observed when the extract obtained in Example 2 was battled, and the extract inhibited cell migration by PDGF. Could be observed.
제제예Formulation example 1. One. 츄잉껌의Chewing gum 제조 Produce
실시예1에서 얻은 추출물 0.24 ∼ 0.64%0.24 to 0.64% of the extract obtained in Example 1
껌베이스 20%
설탕 76.36 ∼ 76.76%Sugar 76.36-76.76%
후르츠향 1%1% of fruit flavor
물 2%Water 2%
상기의 조성 및 함량으로 통상적인 방법을 사용하여 츄잉껌을 제조하였다.Chewing gum was prepared using a conventional method with the above composition and content.
제제예Formulation example 2. 음료의 제조 2. Manufacture of beverage
실시예1에서 얻은 추출물 0.5g ∼ 1.3g0.5 g to 1.3 g extract obtained in Example 1
꿀 522㎎522mg of honey
치옥토산아미드 5㎎Chioctosanamide 5mg
니코틴산아미드 10㎎Nicotinamide 10mg
염산리보플라빈나트륨 3㎎Riboflavin Sodium Hydrochloride 3mg
염산피리독신 2㎎Pyridoxine hydrochloride 2mg
이노시톨 30㎎Inositol 30mg
오르트산 50㎎Ortic Acid 50mg
물 300㎖300 ml of water
상기의 조성 및 함량으로 하여 통상적인 방법을 사용하여 음료를 제조하였다.With the above composition and content, beverages were prepared using conventional methods.
제제예Formulation example 3. 사탕의 제조 3. Manufacture of Candy
사탕의 제조시 사용되는 재료는 다음과 같다.The materials used in the manufacture of the candy are as follows.
실시예1에서 얻은 추출물 Extract obtained in Example 1
결정질 락티톨(Danisco Sweeteners)Crystalline Lactitol (Danisco Sweeteners)
물water
아세설팜 K(Hoechst)Acesulfame K (Hoechst)
블루베리 향(DI 27328)Blueberry Scent (DI 27328)
시트르산(Bahrat Starch Industries, Ltd)Citric Acid (Bahrat Starch Industries, Ltd)
20%의 실시예1에서 얻은 추출물 및 80%의 결정질 락티톨(Danisco Sweeteners)의 혼합물을 물로 희석하고 소스 팬에 두었다. 모든 물질이 가용화될 때까지 배치를 먼저 고온 플레이트상에서 가열하였다. 이어서 배치를 진공 조리기로 옮기고 추가로 가열하였다. 형성된 블렌드는 심지어 비교적 낮은 온도에서 점성의 매쓰를 형성하였다. 이어서 시럽을 조리기에서 후판으로 옮기고 낙하 롤링을 위해 적합한 조직이 성취될 때까지 경화시켰다. 경화된 매쓰를 낙하 롤러를 통해 공급하였다. 일단 각인된 사탕은 허용되는 품질을 가졌다.A mixture of 20% of the extract obtained in Example 1 and 80% of crystalline lactitol (Danisco Sweeteners) was diluted with water and placed in a sauce pan. The batch was first heated on a hot plate until all material was solubilized. The batch was then transferred to a vacuum cooker and further heated. The blend formed formed a viscous mass even at relatively low temperatures. The syrup was then transferred from the cooker to the thick plate and cured until a suitable tissue for drop rolling was achieved. The cured mass was fed through a drop roller. Once imprinted, the candy had an acceptable quality.
제제예Formulation example 4. 술(주정)의 제조 4. Production of alcohol
백출 240 g, 천마 120 g, 진피 120 g, 복령 180 g, 산사 180 g, 희렴 180 g 을 잘 씻은 후 추출용기에 넣고 물 10,000 ㎖을 가하여 100℃에서 약3시간동안 달여 농축시켜 추출액을 얻는다. 그와 별도로 쌀 4 kg로 만든 증미, 전통누룩 1 kg 및 주모 0.05 kg을 균일하게 혼합한 후 상기 추출액을 붓고 10∼25℃에서 약 3일간 발효시켰다. 발효액을 여과하여 액상부분과 고형부분을 분리하고 55∼60℃로 가열한 후 여액에 정제수를 가하여 10,000 ㎖로 조절하여 주제로 제조하였다.Wash out 240 g, Cheonma 120 g, dermis 120 g, Bokryeong 180 g, Sansa 180 g, Happiness 180 g, put it in an extraction container, add 10,000 ml of water, and concentrate it for about 3 hours at 100 ° C to obtain an extract. Separately, the mixture was gradually mixed with steamed rice made from 4 kg of rice, 1 kg of traditional yeast, and 0.05 kg of jujube. The extract was poured and fermented at 10-25 ° C. for about 3 days. The fermentation broth was filtered to separate the liquid and solid portions, heated to 55-60 ° C., and purified water was added to the filtrate to adjust 10,000 ml to prepare a subject.
이상과 같이, 본 발명에 따른 추출물은 1㎎/㎖의 농도에서 혈소판유래성장인자(Platelet derived growth factor; PDGF)에 의해 증가된 혈관 평활근 세포의 증식을 크게 억제시켰고, 유주 활성을 억제시켜 동맥경화 및 관련 질환의 예방 및 치료에 탁월한 효과를 나타내었다.As described above, the extract according to the present invention significantly inhibited the proliferation of vascular smooth muscle cells increased by platelet derived growth factor (PDGF) at a concentration of 1 mg / ml, and inhibited lactose activity to atherosclerosis. And excellent effects in the prevention and treatment of related diseases.
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| KR101471048B1 (en) * | 2013-03-05 | 2014-12-08 | 동의대학교 산학협력단 | A composition comprising Siegesbeckia pubescens extracts having anti-obesity activity |
| KR20200025254A (en) * | 2018-08-29 | 2020-03-10 | 윤화순 | Preparation for improving blood circulation comprising mixed oriental medicinal extracts |
| KR20200047010A (en) | 2018-10-26 | 2020-05-07 | 대전대학교 산학협력단 | Compositions comprising extract of Crataegi fructus and Lycopus lucidus for prevention or treatment of lipid-related cardiovascular diseases or obesity |
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| KR101415697B1 (en) * | 2012-08-28 | 2014-07-04 | (주)뉴메드 | A pharmaceutical composition comprising the combined extract of Crataegi Fructus and Citri Pericarpium for treating or preventing obesity or lipid-related metabolic disorder |
| KR101894540B1 (en) * | 2016-11-10 | 2018-09-05 | 한약진흥재단 | Composition for improving coenzyme Q10 with extract of Pinellia ternata, Atractylodes macrocephala, Gastrodia elata, Citrus unshiu peel, Poria cocos, Crataegus pinnatifida, Siegesbeckia pubescens Makino and Coptis japonica Makino |
| KR102117896B1 (en) * | 2018-02-27 | 2020-06-02 | 건국대학교 글로컬산학협력단 | Composition comprising Extract of Crataegus pinnatifida Bunge for Antithrombosis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR101471048B1 (en) * | 2013-03-05 | 2014-12-08 | 동의대학교 산학협력단 | A composition comprising Siegesbeckia pubescens extracts having anti-obesity activity |
| KR20200025254A (en) * | 2018-08-29 | 2020-03-10 | 윤화순 | Preparation for improving blood circulation comprising mixed oriental medicinal extracts |
| KR102127726B1 (en) | 2018-08-29 | 2020-06-29 | 윤화순 | Preparation for improving blood circulation comprising mixed oriental medicinal extracts |
| KR20200047010A (en) | 2018-10-26 | 2020-05-07 | 대전대학교 산학협력단 | Compositions comprising extract of Crataegi fructus and Lycopus lucidus for prevention or treatment of lipid-related cardiovascular diseases or obesity |
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