KR20100108869A - Composition containing extract of herbal mixture for prevention or treatment of diabetes - Google Patents
Composition containing extract of herbal mixture for prevention or treatment of diabetes Download PDFInfo
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- KR20100108869A KR20100108869A KR1020090027138A KR20090027138A KR20100108869A KR 20100108869 A KR20100108869 A KR 20100108869A KR 1020090027138 A KR1020090027138 A KR 1020090027138A KR 20090027138 A KR20090027138 A KR 20090027138A KR 20100108869 A KR20100108869 A KR 20100108869A
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- KR
- South Korea
- Prior art keywords
- extract
- diabetes
- insulin
- glucose
- gardenia
- Prior art date
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Abstract
Description
본 발명은 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물을 포함하는 당뇨병 예방 및 치료용 조성물에 관한 것이다.The present invention relates to a composition for preventing and treating diabetes comprising a mixed extract of golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade.
당뇨병(diabetes)은 비전염성 만성질환으로, 췌장의 베타세포(β-cell)에서 분비되는 인슐린이 부족하거나 그 기능을 제대로 발휘하지 못하여 체내에서 포도당이 에너지원으로 이용되지 못하고 혈액 내에 고농도로 남아 있다가 소변으로 배설되는 질환이다.Diabetes is a non-communicable chronic disease. Insulin secreted by the pancreatic beta cells (β-cell) is insufficient or does not function properly, and glucose is not used as an energy source in the body and remains high in blood. Is a disease that is excreted in the urine.
당뇨병은 전세계적으로 중요한 성인병 중의 하나로서, 최근 우리나라에서도 급속한 경제 성장과 더불어 당뇨병 유병율이 7 ~ 8%에 달하며, 60 ~ 70대의 경우 중요한 사망원인이 되고 있다.Diabetes is one of the important adult diseases all over the world. In Korea, the prevalence rate of diabetes mellitus reaches 7-8% with rapid economic growth, and it is an important cause of death for people in their 60s and 70s.
당뇨병은 그 발병원인 및 증상의 치료방법에 따라 크게 인슐린 의존성인 제1 형 당뇨병(Insulin dependent diabetes mellitus: IDDM)과 인슐린 비의존성인 제2형 당뇨병(Non-insulin dependent diabetes mellitus: NIDDM)으로 분류된다.Diabetes is classified into insulin dependent diabetes mellitus (IDDM) and non-insulin dependent diabetes mellitus (NIDDM), which are largely insulin dependent, depending on the cause of the disease and the method of treating the symptoms.
제1형 당뇨병은 유전적 원인, 바이러스 감염 등에 의해 췌장 베타세포의 기능이 저하되어 인슐린이 거의 분비되지 않은 상태로서, 주로 10 ~ 20대에 갑자기 발병하며, 소아형 당뇨병이라고도 한다.
제2형 당뇨병은 가족력, 비만, 스트레스, 식사의 잘못 또는 인슐린과 길항(拮抗)하는 약물 등에 의해 발병되는 것으로, 40대 이후에 잘 나타나며, 성인형 당뇨병이라고도 한다. 실제로는 전체 당뇨병 환자의 대부분이 제2형 당뇨병 환자이다.
제2형 당뇨병은 간, 근육, 지방세포 등의 말초조직에서 인슐린 작용이 저하하여 포도당의 이용이 감소하고 특히 간에서는 포도당의 생성이 증가하여 혈당이 상승할 때 췌장의 베타세포에서 혈당이 상승하는 것을 억제할 수 있을 정도로 인슐린 분비가 충분하지 못할 때 나타나는 질병이다. 즉 제2형 당뇨병은 인슐린 작용이 저하한 인슐린 저항성을 인슐린 분비가 따르지 못해서 발병한다.
제2형 당뇨병에서는 인슐린이 분비되지만 인슐린이 작용하는 간, 근육, 지방세포 등의 말초조직에서 인슐린의 작용력이 저하하여 포도당의 이용이 감소되므로 혈당이 점점 높아지고 이에 대응하여 췌장의 베타세포에서 지속적으로 인슐린이 분비되는 고인슐린혈증을 동반한다[Kadowaki T, Hara K, Yamauchi T, Terauchi Y, Tobe K, Nagai R. 2003. Molecular mechanism of insulin resistance and obesity. Exp Biol Med(Maywood) 228: 1111-1117].In
그러나 제2형 당뇨병에서도 혈당이 상승할 때 췌장의 베타세포에서 혈당이 상승하는 것을 억제할 수 있을 정도로 충분히 인슐린 분비가 이루어지지 않는 경우도 있다. 우리나라의 제2형 당뇨병이 이에 해당하며, 이러한 경우에는 인슐린 작용력과 인슐린 분비가 모두 낮아서 서구에 비해 당뇨병 증세도 심하고 비만이 되지않는 특징을 가지고 있다[Kim J, Choi S, Kong B, Oh Y, Shinn S. 2001. Insulin secretion and sensitivity during oral glucose tolerance test in Korean lean elderly women. J Korean Med Sci 16: 592-597].However, even in
그러므로 우리나라의 제2형 당뇨병의 예방 및 치료을 위해서는, 인슐린 작용력을 향상시킴과 동시에 포도당 자극에 의한 인슐린 분비능도 증가시키는 것이 중요한 것으로 알려져 있다.Therefore, for the prevention and treatment of
제2형 당뇨병 치료제로서 가장 먼저 사용되어 온 것이 인슐린 분비를 촉진시키는 약물들이다. 설포닐우레아(sulfonylurea) 계통의 약물이 이에 해당하며, 이 약물은 췌장의 베타세포의 세포막에 존재하는 KATP(ATP-regulated potassium) 채널에 직접 작용하여 이 채널을 닫아 줌으로써, 세포막을 분극화(depolarization)시키고 Ca2+을 세포 내로 유입시킴으로 인슐린의 분비를 촉진시키는 인슐린 분비 촉진제(insulin secretagogues)이다[Krentz AJ, Bailey CJ. 2005. Oral antidiabetic agents: current role in type 2 diabetes mellitus. Drugs 65: 385-411; Quesada I, Soria B. 2004. Intracellular location of KATP channels and sulphonylurea receptors in the pancreatic beta-cell: new targets for oral antidiabetic agents. Curr Med Chem. 11: 2707-2716]. 그러나, 이 약물은 혈당 농도에 상관없이 KATP 채널을 닫아서 혈당이 낮아도 인슐린을 분비시켜 저혈당을 유발시킬 수 있다는 문제점이 있다.The first drugs used to treat
최근에 제2형 당뇨병의 새로운 치료제로 등장한 것이 인슐린 작용을 향상시킴으로 소량의 인슐린으로 혈당을 정상화할 수 있도록 도와주는 인슐린 민감성 물질들이다. 인슐린 민감성 물질은 제2형 당뇨병의 근본적인 문제인 인슐린 저항성을 완화시켜 소량의 인슐린으로 혈당을 정상화시키고, 궁극적으로 혈당에 필요한 인슐린 양을 저하시켜 고인슐린혈증을 없애 줄 수 있는 것으로 알려지고 있다 [Ciaraldi TP, Kong AP, Chu NV, Kim DD, Baxi S, Loviscach M, Plodkowski R, Reitz R, Caulfield M, Mudaliar S, Henry RR. 2002. Regulation of glucose transport and insulin signaling by troglitazone or metformin in adipose tissue of type 2 diabetic subjects. Diabetes 51: 30-36]. 즉, 인슐린 민감성 물질은 당뇨병뿐 아니라 인슐린 저항성 증후군의 증세를 완화시킬 수 있을 것이며, 이는 인슐린이 인슐린 수용체와 결합한 후 세포 내에서 일어나는 신호전달 체계가 원활하게 일어날 수 있도록 도와주어 세포에서 포도당의 이용을 향상시키는 것이다.Recently, a new treatment for
췌장의 베타세포에서 인슐린이 분비되는 과정은 간접적으로 인슐린/IGF-1(Insulin like Growth Factor-1) 신호전달과 관련이 있고, 이러한 신호전달의 저하는 베타세포의 생성 저하 및 세포사멸(apoptosis) 증가를 거쳐 베타세포의 양을 감소시키고, 결과적으로 인슐린 분비의 필요를 충당하지 못하여 당뇨병으로 진전된 다.The secretion of insulin from pancreatic beta cells is indirectly related to insulin / IGF-1 (Insulin like Growth Factor-1) signaling, and the degradation of beta cells is associated with decreased beta cell production and apoptosis. This increases the amount of beta cells, and consequently leads to diabetes by not meeting the need for insulin secretion.
또한, GLP-1은 생체내 소장 세포의 L-cell에서 분비되는 펩타이드로서, 베타세포에서 포도당 자극에 의한 인슐린의 분비를 촉진하고, 베타세포의 성장 촉진 및 사멸 억제를 통해서 베타세포의 양을 증가시켜 베타세포의 기능을 향상시키는 것으로 알려져 있다.In addition, GLP-1 is a peptide secreted by L-cells of small intestine cells in vivo, and promotes the secretion of insulin by glucose stimulation in beta cells, and increases the amount of beta cells by promoting the growth and suppression of beta cells. It is known to improve the function of beta cells.
췌장 베타세포의 생존 및 기능을 향상시키기 위한 방법으로서 분리된 랑게르한스섬 내의 인슐린 수용체 기질-2(insulin receptor substrate-2: IRS2), 췌장의 호메오박스-1(pancreas duodenum homeobox-1: PDX-1) 및 글루코키나제(glucokinase)의 mRNA 양을 증가시키는 방법이 있다. IRS2 발현의 유도는 Min6 세포와 랑게르한스섬의 크렙(cAMP response element-binding protein: CREB)의 활성화에 의해 일어나는 것으로 알려져 있다(Hennige, A. M. et al., J. Clin.Invest. 112: 1521-1532, 2003; Jhala, U. S. et al., Genes Dev. 17: 1575-1580, 2003; Park, S. et al. J. Biol. Chem. 281: 1159-1168, 2006).Insulin receptor substrate-2 (IRS2), pancreas duodenum homeobox-1 (PDX-1) in isolated Langerhans as a method for improving pancreatic beta cell survival and function And a method of increasing the amount of mRNA of glucokinase. Induction of IRS2 expression is known to be caused by the activation of Min6 cells and the cAMP response element-binding protein (CREB) of Langerhans Island (Hennige, AM et al., J. Clin. Invest. 112: 1521-1532, 2003 Jhala, US et al., Genes Dev. 17: 1575-1580, 2003; Park, S. et al. J. Biol. Chem. 281: 1159-1168, 2006).
최근에 개발된 포도당-자극 인슐린 분비 촉진제(glucose-stimulated insulin secretagogue)로는 글루카곤 유사 펩타이드-1 수용체 작용제(glucagon like peptide-1(GLP-1) receptor agonist)인 엑센딘-4(exendin-4, exenatide)가 있다.A recently developed glucose-stimulated insulin secretagogue is the glucagon like peptide-1 (GLP-1) receptor agonist, exendin-4 (exendin-4, exenatide). There is).
엑센딘-4는 설포닐우레아 계통의 약물과는 달리 KATP 채널에 직접적으로 작용하지 않고, 세포막에 존재하는 GLP-1 수용체에 결합하여 세포 내의 cAMP 농도를 상승시킴으로써 단백질 키나제 A(protein kinase A; PKA)를 활성화시킨다. 활성된 PKA만으로는 세포 내로의 Ca2+ 유입을 상승시키지 못하고, 혈당이 높아져 베타세포 내로 포도당이 흡수되어 해당작용(glycolysis)을 거쳐 ATP/ADP의 농도가 높아지면서 PKA가 활성화될 때, Ca2+의 유입이 증가하여 인슐린 분비를 촉진시킨다[Drucker DJ. 2001. Development of glucagon-like peptide-1-based pharmaceuticals as therapeutic agents for the treatment of diabetes.Curr Pharm Res 7: 1399-1412]. 그러므로 엑센딘-4는 혈당이 정상일 때는 인슐린 분비를 촉진시키지 않고, 혈당이 상승될 때만 인슐린 분비를 촉진시키는 포도당-자극 인슐린 분비 촉진제로 작용한다. 또한, 인슐린 분비를 향상시킬 뿐 아니라 인슐린 작용도 향상시키는 것으로 알려져 있다.Exendin-4, unlike drugs of the sulfonylurea family, does not act directly on the KATP channel, but binds to the GLP-1 receptor present in the cell membrane and raises the cAMP concentration in the cell, thereby enhancing protein kinase A (PKA). ) Is activated. Not only the active PKA not increase the Ca 2+ influx into the cell, when glucose is increased glucose is absorbed into the beta-cell As via glycolysis (glycolysis) due to high concentrations of ATP / ADP is PKA is activated, Ca 2+ Increased influx to promote insulin secretion [Drucker DJ. 2001. Development of glucagon-like peptide-1-based pharmaceuticals as therapeutic agents for the treatment of diabetes. Curr Pharm Res 7: 1399-1412]. Therefore, exendin-4 does not promote insulin secretion when blood sugar is normal, but acts as a glucose-stimulating insulin secretagogue to promote insulin secretion only when blood glucose is elevated. It is also known to improve insulin secretion as well as to improve insulin action.
또 다른 약물로서, 탄수화물의 소화흡수를 방해하여 식후 혈당의 상승을 방지하는 아카보스(acarbose) 계통의 약물이 있다. 아카보스는 가역적으로 α-글루코아밀라제 활성을 억제시키는 약물로, 주로 소장 세포의 점막에 존재하는 말타제(maltase)의 활성을 억제시키는 것으로 알려져 있다[Carrascosa JM, Molero JC, Fermin Y, Martinez C, Andres A, Satrustegui, J. 2001. Effects of chronic treatment with acarbose on glucose and lipid metabolism in obese diabetic Wistar rats. Diabetes Obes Metab 3: 240-248].Another drug is the acarbose family of drugs that interferes with the digestion and absorption of carbohydrates and prevents elevated blood sugar after meals. Acarbose is a drug that reversibly inhibits α -glucoamylase activity and is known to inhibit the activity of maltase in the mucosa of small intestine cells [Carrascosa JM, Molero JC, Fermin Y, Martinez C, Andres] A, Satrustegui, J. 2001. Effects of chronic treatment with acarbose on glucose and lipid metabolism in obese diabetic Wistar rats. Diabetes Obes Metab 3: 240-248].
그 외에, 인슐린 작용력을 증진시키는 피오글리타존(pioglitazone) 계통의 약물이 있으며, 간에서 당신생 합성을 감소시키는 메트포민(metformin) 계통의 물질이 있다.In addition, there are drugs of the pioglitazone line that enhance insulin action, and there are metformin lines that reduce your biosynthesis in the liver.
또한, 국내를 비롯한 아시아에서는 민간요법으로 여러 가지 약초들을 당뇨병 및 여러 질병의 치료제로 사용하여 왔다. 크레니스키(Krenisky) 등은 페루 전통의 약식물 Otholobium pubescens에서 바쿠치올(bakuchiol)을 분리하여 인슐린 민감성에 관여하고 있는 물질이라고 보고하였고[Krenisky JM, Luo J, Carney JR. 1999. Isolation and antihyperglycemic activity of bakuchiol from Otholobium pubsecens (Fabaceae), a peruvian medicinal plant used for the treatment of diabetes. Biol Pharm Bull 22: 1137-1140], Hong 등은 인삼, 천문동, 황금, 지골피, 황백 및 맥문동으로 구성된 혼합처방이 3T3-L1 지방세포에서 포도당 흡수를 증가시켰다고 보고하고 있다[Hong SJ, Fong JC. Hwang JH. 2000. Effect of crude drugs on glucose uptake in 3T3-L1 adipocyte. Gaxiong Yi Xue Ke Xue Za Zhi 16: 445-451].In addition, in Korea and in Asia, various medicinal herbs have been used for the treatment of diabetes and various diseases as folk remedies. Krenisky et al. Reported the separation of bakuchiol from the Peruvian traditional plant, Otholobium pubescens , as a substance involved in insulin sensitivity [Krenisky JM, Luo J, Carney JR. Isolation and antihyperglycemic activity of bakuchiol from Otholobium pubsecens (Fabaceae), a peruvian medicinal plant used for the treatment of diabetes. Biol Pharm Bull 22: 1137-1140], Hong et al. Reported that a mixed prescription consisting of ginseng, astronomical dong, gold, phalanges, baekbaek and munmundong increased glucose uptake in 3T3-L1 adipocytes [Hong SJ, Fong JC. Hwang JH. 2000. Effect of crude drugs on glucose uptake in 3T3-L1 adipocyte. Gaxiong Yi Xue Ke Xue Za Zhi 16: 445-451].
현재까지 당뇨에 효과가 있는 것으로 알려져 있는 것으로는 후박(대한민국 공개특허 제 2003-0039241호), 인삼 잎(대한민국 공개특허 제 2005-0054905호), 황련(대한민국 공개특허 제 2004-0014556호) 등이 있다. 또한, 인슐린의 작용을 향상시키는 인슐린 민감성 효과, 인슐린처럼 작용하는 인슐린성 효과, 탄수화물의 소화 흡수를 방해하는 α-글루코아밀라제 억제 효과를 가지고 있는 한약재로는 율무, 상백피, 둥굴레 뿌리, 오미자(대한민국 공개특허 제 2004-0016145호), 목단피(대한민국 공개특허 제 2003-0100558호)의 분획물에는 인슐린 민감성 물질이 함유되어 있는 것을 이미 발견하였다. 특히, 둥굴레 뿌리 추출물은 90% 췌장을 제거한 백서에서 인슐린 민감성을 호전시킴으로 체내 포도당 이용을 증가시켜 혈당을 강하시키는 효과를 나타내었다[Choi SB, Park S. 2002. The effects of water extract of Polygonatum Odoratum (Mill) Druce on insulin resistance in 90% pancreatectomized rats. J Food Sci 67: 2375-2379]. 또한, 둥굴레 뿌리에 함유되어 있는 스테로이드성 글리코시드(steroidal glycosides)가 90% 췌장 제거 백서에서 인슐린 민감성 물질로 작용한다는 것이 확인되었다[Choi BS, Park S. 2002. A Steroidal Glycoside from Polygonatum odoratum (Mill.) Druce. Improves Insulin Resistance but does not Alter Insulin Secretion in 90% Pancreatectomized Rats. Biosci Biotech Biochem 66: 2036-2043].To date, known to be effective in diabetes, Hubak (Korea Patent Publication No. 2003-0039241), Ginseng leaves (Korea Patent Publication No. 2005-0054905), Hwangyeon (Korea Patent Publication No. 2004-0014556) have. In addition, herbal medicines that have an insulin-sensitive effect that enhances the action of insulin, an insulin-like effect that acts like insulin, and an α -glucoamylase inhibitory effect that interferes with the digestion and absorption of carbohydrates include Yulmu, Morus bark, Roots, and Schisandra chinensis. Patent 2004-0016145) and fractions of Mokdanpi (Korean Patent Publication No. 2003-0100558) have already been found to contain insulin sensitive substances. In particular, the extract of Rhubarb Root showed the effect of lowering blood glucose by increasing glucose utilization in the body by improving insulin sensitivity in 90% of the pancreas from which the pancreas was removed [Choi SB, Park S. 2002. The effects of water extract of Polygonatum Odoratum ( Mill) Druce on insulin resistance in 90% pancreatectomized rats. J Food Sci 67: 2375-2379. It has also been shown that steroidal glycosides contained in the roots act as insulin-sensitive substances in 90% of pancreas-removed white papers [Choi BS, Park S. 2002. A Steroidal Glycoside from Polygonatum odoratum (Mill. ) Druce. Improves Insulin Resistance but does not Alter Insulin Secretion in 90% Pancreatectomized Rats. Biosci Biotech Biochem 66: 2036-2043.
그러나 민간요법으로 사용되고 있는 많은 약초에 대해서도 아직까지 그 효과가 밝혀진 것은 많지 않다.However, for many herbs that are used as folk remedies, the effect is still not clear.
이에, 본 발명자들은 다양한 생약재 가운데에서 베타세포에 있어서 포도당 자극에 의한 인슐린 분비를 향상시키고 대장암세포에서 GLP-1 분비를 증가시키는 생약재를 찾아내고, 이러한 생약재를 포함한 혼합 추출물이 생체내에서 혈당 강하 효과 및 인슐린 분비 증가 효과를 나타내고, 생체내 인슐린의 작용력 증가 효과 및 인슐린 저항성 감소 효과를 나타내며, 생체내에서 인슐린 분비량을 증가시키고, 랑게르한스섬에서 IRS2의 발현을 증가시켜 당뇨병의 예방 및 치료 효과가 있음을 확인하고, 본 발명을 완성하였다.Accordingly, the present inventors have found a herbal medicine that improves insulin secretion by glucose stimulation in beta cells and increases GLP-1 secretion in colorectal cancer cells among various herbal medicines, and the mixed extract including such herbal medicines has a hypoglycemic effect in vivo. And increase insulin secretion, increase insulin action and decrease insulin resistance in vivo, increase insulin secretion in vivo, and increase expression of IRS2 in island of Langerhans, thus preventing and treating diabetes. The present invention was completed.
본 발명은 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물을 포함하는 당뇨병 예방 및 치료용 조성물과 당뇨병 예방 및 개선용 건강식품을 제공하고자 한다.The present invention is to provide a composition for preventing and treating diabetes and health food for preventing and improving diabetes comprising a mixed extract of golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade porridge.
상기 목적을 달성하기 위하여, 본 발명은 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물을 포함하는 당뇨병 예방 및 치료용 조성물을 제공한다.In order to achieve the above object, the present invention provides a composition for preventing and treating diabetes comprising a mixed extract of gold, yellow white, gardenia, yellow lotus, Schisandra chinensis and jade.
또한, 본 발명은 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물을 포함하는 당뇨병 예방 및 개선용 건강식품을 제공한다.In addition, the present invention provides a health food for preventing and improving diabetes comprising a mixed extract of gold, yellow white, gardenia, yellow lotus, Schisandra chinensis and jade porridge.
본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물은 생체내에서 혈당 강하 효과 및 인슐린 분비 증가 효과를 나타내고, 생체내 인슐린의 작용력 증가 효과 및 인슐린 저항성 감소 효과를 나타냈다. 특히, 생체내에서 인슐린 분비량을 증가시키고, 랑게르한스섬에서 IRS2의 발현을 증가시키며, 세포 및 동물에서 급성 독성을 나타내지 않았다. 따라서, 본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물을 포함하는 본 발명의 조성물은 당뇨병 예방 및 치료용 조성물로 유용하게 사용될 수 있다.The mixed extracts of golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade porridge according to the present invention showed a hypoglycemic effect and an insulin secretion increasing effect in vivo, and an effect of increasing insulin action and decreasing insulin resistance in vivo. In particular, it increased insulin secretion in vivo, increased expression of IRS2 in the island of Langerhans, and showed no acute toxicity in cells and animals. Therefore, the composition of the present invention comprising a mixed extract of gold, yellow white, gardenia, yellow lotus, Schisandra chinensis and jade according to the present invention can be usefully used as a composition for preventing and treating diabetes.
이하, 본 발명에 관하여 상세히 설명한다.Hereinafter, the present invention will be described in detail.
황금(Scutellaria baicalensis)은 다년생 초본으로서 황금의 뿌리를 말려서 사용한다. 황금의 뿌리에는 바이칼레인(baicalein), 바이칼린 (baicalin), 워고닌(wogonin), 네오바이칼레인 (neobaicalein) 등이 함유되어 있다. 황금의 생리활성 및 약리작용으로 해열, 소염, 항알레르기 작용을 나타내며, 혈관을 확장시키고 더욱이 혈압을 강하시키는 작용을 하며 죽상 동맥경화를 억제한다. 특히, 황금에 함유된 바이칼린은 진정작용을 하며 모세혈관의 투과성을 저하시키므로 지혈 (止血)하게 된다. 또한, 비만 세포막을 강화하여 화학 전달 물질의 유리를 억제하여 항알러지 작용을 하며, 특히 알러지로 인한 간염을 개선하고 혈관투과성 항진을 억제하며 (정 보섭 및 신 민교; 향약대사전, 영림사, pp864-865, 1998), 항염증, 고지혈증 개선작용이 있는 것으로 알려져 있다 (Uchiyama et al., J. Med. Pharm. Soc. Wakan-Yaku, 6, 158-164, 1989; Kubo et al., Chem. Pharm. Bull., 32, 2724-2729, 1984).Golden ( Scutellaria baicalensis ) is a perennial herb used to dry golden roots. The roots of gold contain baicalein, baicalin, wogonin and neobaiicalein. Physiological activity and pharmacological action of gold, antipyretic, anti-inflammatory, anti-allergic action, and expands blood vessels, and further lowers blood pressure and inhibits atherosclerosis. In particular, Baikalin contained in gold is sedating and hemostasis (止血) because it reduces the permeability of capillaries. In addition, by strengthening the mast cell membrane to suppress the release of chemical transport material, it has anti-allergic effect, in particular to improve hepatitis caused by allergies and inhibit vascular permeability (Jung Bo-seop and Shin, Min-Kyo; Hyangjedae, Yeonglimsa, pp864- 865, 1998), anti-inflammatory and hyperlipidemic effects (Uchiyama et al., J. Med. Pharm. Soc. Wakan-Yaku, 6, 158-164, 1989; Kubo et al., Chem. Pharm Bull., 32, 2724-2729, 1984).
황련(Coptis chinensis)은 미나리제비과 (Ranuculaceae)에 속하는 다년생 초본식물로서, 그 근경을 햇볕에 말려서 사용한다. 황련의 뿌리에는 베르베린(berberine), 콥티신(coptisine), 워레닌(worenine), 팔마틴 (palmatine) 등의 알칼로이드가 유효성분으로 함유되어 있다. 황련은 항미생물 작용 및 항병원충 작 용이 있으며, 특히 베르베린은 아세틸콜린의 작용을 강화시키고, 혈관 평활근의 이완작용(정 보섭 및 신 민교; 향약대사전, 영림사, pp490-493, 1998), 항염증 작용(Ko et al., Eur. J. Pharmacol., 399, 187-196, 2000), 항산화 억제작용 (Yokozawa et al., J. Pharm. Pharmacol., 56, 547-556, 2004)및 혈압 강하 효과가 있는 것으로 알려져 있다. Coptis chinensis is a perennial herbaceous plant belonging to the family Ranuculaceae , and its roots are dried in the sun. The roots of nasturtium contain alkaloids such as berberine, coptisine, warenine, and palmatine as active ingredients. Lactobacillus has antimicrobial action and anti-pathogenic action, especially berberine enhances the action of acetylcholine, relaxes vascular smooth muscle (Jung Bo-seop and Shin Min-kyo; Yaksaimsa, Yeonglimsa, pp490-493, 1998), anti-inflammatory Action (Ko et al., Eur. J. Pharmacol., 399, 187-196, 2000), antioxidant inhibitory activity (Yokozawa et al., J. Pharm. Pharmacol., 56, 547-556, 2004) and lowering blood pressure It is known to be effective.
황백(Phellodendron amurensis)은 황벽나무의 껍질을 약재로 이르는 말로, 황경피라고도 한다. 낙엽활엽 교목으로 높이 10m 정도로 자라는데, 껍질에는 코르크가 발달했으며 속껍질은 황색이다. 잎은 마주나며, 깃꼴겹잎으로 작은 잎사귀는 5~13장이며, 달걀모양의 피침형(披針形) 또는 긴 원형이다. 잎 가장자리에는 작은 톱니 및 연한 털이 있으며, 중간 맥(脈)의 기부(基部)에는 긴 털이 약간 있다. 5~6월에 황색의 꽃이 피는데, 자웅이주(雌雄異株)의 단성(單性)이다. 원추 모양의 수술은 긴 원형으로, 수꽃에는 5개의 수술이 있으며 암꽃에는 퇴화된 수술이 비늘 조각 모양을 하고 있다. 자방은 5실로 되어 있으며 암술대는 짧고 암술머리는 5갈래로 갈라진다. 열매는 장과상(漿果狀)의 육질이 많고 중심부에 단단한 핵을 갖고 있으며, 동그란 모양으로 익었을 때에는 자흑색을 띤다. 우리나라에서는 제주ㅇ전남 지역을 제외한 전지역에서 분포하고, 일본, 만주, 중국, 아무르 및 우수리 등지에 분포하며, 잡목 숲 또는 산속의 계곡에서 자란다. 황백나무줄기에서 껍질을 벗겨내어 조피(粗皮:거친 껍질)를 제거하거나 썰어서 햇볕에 말린 후, 이것 즉 황백은 약으로 쓰이며 혈당저하 작용을 한다. 또한, 폐렴쌍구균, 인형결핵균, 포도상구균 등에 대하여 발육저지 작용을 함과 동시에 종양세포의 번식을 저지시키고, 살 균작용을 한다. 복용하는 경우에는 미각 반사의 항진에 의하여 위액의 분비를 촉진시키고, 식욕의 항진도 가져오게 한다. 또한, 일반 알칼로이드가 지니는 전신작용을 하지 않기 때문에 다량으로 투여해도 부작용이 없으므로 정장제뿐 아니라 건위제로 사용할 수 있다. 또한, 이 약재에 대하여 여러 세균의 내성(耐性)도 생기지 않으므로 유행성 눈병이 유행할 때 세안 소독약으로도 사용할 수 있다. 그 외에도 혈압강하, 중추신경계 억제, 항염증 등의 효과도 보고되어 있으며, 동양의학에서는 황련해독탕, 시호청간탕, 형개연교탕 등에 사용되고 있다. Hwangbaek ( Phellodendron amurensis ) is a term that refers to the bark of the yellow bark as a medicinal herb, also known as hwanggyeongpi . It is a deciduous broad-leaved arboreous tree, growing up to 10m in height. The leaves are opposite each other. The pinnate leaves are 5 ~ 13 sheets long, egg-shaped lanceolate or long round. The leaf edge has small jagged and soft hairs, and the base of the middle vein has some long hairs. Yellow flowers bloom in May-June, which is a monolith of the male sex family. The cone-shaped stamen is a long circle, with five stamens in the male flower, and the degenerated stamens in the female flower are scaly. It has 5 rooms, with short pistils and 5 pistils. The fruit has a lot of berry and has a hard core in the center, and when rounded, it is purple-black. In Korea, it is distributed in all regions except Jeju and Jeonnam, and is distributed in Japan, Manchuria, China, Amur, and Ussuri, and grows in scrub forests or valleys in the mountains. Peel off the bark and remove the skin (조: rough bark) or cut and dried in the sun, this is the white pill is used as a medicine to lower blood sugar. In addition, pneumococcal pneumoniae, pneumococcal tuberculosis, staphylococcus, etc., while inhibiting the development of tumor cells, and inhibits the growth and sterilization. When taken, palpation of gastric juice is promoted by palpation of taste reflexes, which also leads to an increase in appetite. In addition, since the general alkaloids do not have a systemic effect, even if administered in large amounts, there are no side effects, so it can be used as a formal agent as well as a stomach agent. In addition, since the resistance of various bacteria to this medicine does not occur, it can be used as a face wash disinfectant when a pandemic eye disease is epidemic. In addition, blood pressure lowering, central nervous system inhibition, anti-inflammatory effects have been reported, and is used in oriental medicine such as Hwangnyeonhaedoktang, Shihocheonggantang, Hyeonggaeyeongyotang.
치자(Gardeniae Fructus)는 꼭두서니과(Rubiaceae)에 속하는 치자나무(Gardenia jasmoides Ellis) 또는 그 밖의 동속식물의 열매로 예로부터 식품의 착색제로 사용되어왔다. 한방에서는 소염, 이뇨, 지혈, 해열, 진정, 항균, 담즙분비, 간장염 치료약으로 황달, 토혈 등에 이용하고 있으며, 진정약으로 응용되고 있다. 치자의 약리 작용으로는 담즙분비촉진작용과 약물성 간장해에 대한 보호 작용 등이 보고되었으며 (Yamauchi 등 Plant Med. 1974, 25, 219, 285; 김강원 등 생약학회지 1994, 25, 368), 혈청 트리글리세라이드 (triglyceride), 인지질, 지질과산화물, 혈당, 유리 지방산을 감소시키며 (Kimura 등 Chem. Pharm. Bull. 1982, 30, 4444), 위운동 억제, 긴장감소를 나타나고 (Aburada 등 J. Pharm. Dyn. 1980, 3, 423), 혈중 빌리루빈 (bilirubin)양의 상승을 억제하며, 동맥경화 예방등의 효과가 있다고 보고 되어있다 (Gainer 등 Experimentia 1975, 31, 548; Nishizawa 등 Chem. Pharm. Bull. 1987, 35, 2133).Gardenia ( Fructus ) is the fruit of Gardenia jasmoides Ellis or other cohorts belonging to the Rubiaceae family, and has been used as a food coloring agent since ancient times. In oriental medicine, it is used for anti-inflammatory, diuretic, hemostatic, antipyretic, soothing, antibacterial, bile secretion, hepatitis, jaundice, hemostasis, and is applied as a sedative. The pharmacological action of gardenia was reported to promote bile secretion and protect against drug-induced hepatic disorders (Yamauchi et al. Plant ed. 1974, 25, 219, 285; Journal of Pharmacognosy et al. 1994, 25, 368), serum trigly Decreases triglycerides, phospholipids, lipid peroxides, blood sugar and free fatty acids (Kimura et al. Chem. Pharm. Bull. 1982, 30, 4444), suppresses gastric motility and decreases tension (Aburada et al. J. Pharm. Dyn. 1980, 3, 423), suppresses the increase in the amount of bilirubin in the blood, and has been reported to be effective in preventing atherosclerosis (Gainer et al. Experimentia 1975, 31, 548; Nishizawa et al. Chem. Pharm. Bull. 1987, 35, 2133).
오미자(Schizandrae Fructus)는 낙엽 활엽 덩굴나무로 잎은 어긋나고 난형 (卵形) 또는 도란형(倒卵形)이며 끝은 급히 뾰족하고 톱니가 있으며 뒷면 맥상에 다소 털이 있다. 꽃은 홍백색으로 6~7월에 피며 자웅이주이다. 과실은 식용으로 쓰이고 가지 끝에 이삭모양으로 늘어지며 8~9월에 붉게 익는다. 과실을 오미자(오매자), 북오미자(Schizandrae Fructus)라고 한다. 표고 200~1,600m 지리산, 전북, 강원도가 주산지이며 충남, 충북을 제외한 전국에 야생하고, 지리적으로 일본, 사할린, 만주, 중국에 분포한다. 과실에는 시잔드롤(Schizandrol), 시잔드린 (Schizandrin) A, B, C, 안질로일고미신(angeloylgomisin) O, P, 에피고미신 (epigomisin), 프레고모이신(pregomoisin), 데옥시시잔드린(Deoxyschizandrin) 등을 함유한다. 오미자는 오래전부터 수렴, 자양, 강장, 진해약, 해주독, 목마름, 수렴고삽, 익기생진, 보신염심 등의 약효를 가져 생약원료로 한방에서 사용해오던 재료이며, 오미자주의 원료로 오래전부터 사용하여 체내 섭취에 따른 인체 안정성이 이미 확인되어 있다. Schizandrae Fructus is a deciduous broad-leaved vine, leaves alternate, ovate or obovate, with acutely pointed, jagged, and somewhat hairy on the back vein. Flower is reddish white and blooms in June ~ July. Fruits are used for food and hang on the ends of branches. They ripen red in August-September. Fruits are called Schisandra (Fructus) and Schizandrae Fructus. 200 ~ 1,600m above sea level Jirisan, Jeonbuk, and Gangwon-do are the main production areas, and are wild throughout the country except Chungnam and Chungbuk, and geographically distributed in Japan, Sakhalin, Manchuria, and China. Fruits include chizandrol, chizandrin A, B, C, angeloylgomisin O, P, epigomisin, pregomoisin, deoxyschizandrin ), And the like. Schisandra chinensis has long been used in herbal medicine as a raw material of medicinal herbs and has the effects of astringent, nourishment, tonic, ginhae medicine, haejudok, thirsty, convergent shovel, ripe ginjin, bosin dyed heart. Human stability with ingestion has already been confirmed.
옥죽(Polygonatum Officinale)은 둥글레의 근경으로 약용하며, 생약의 이명(異名)은 지절(地節), 황지(黃地), 여위(女委) 등으로 불리운다. 옥죽의 기원식물인 둥글레는 다년생의 초본으로 이 식물의 근경인 옥죽은 가을 또는 이른 봄에 채취하여 줄기와 수염뿌리를 제거하고 응달에서 말리거나 아니면 수증기로 쪄서 햇볕에 말려서 약용한다. 이 옥죽에 들어 있는 성분으로는 콘발라마린(convallamarin), 콘발라린(convallarin)등의 스테로이성 사포닌(steroidal saponin)과 다수의 플라보노이드(flavonoids)를 함유하여 강심작용, 부신피질호르몬 유사작용, 갈증완화작용, 항산화작용, 피부노화작용 등이 있는 것으로 알려져 있다. 동양의학에서 이 약의 성질은 평(平)하고 감(甘)하며, 자양, 윤폐, 지갈(止渴) 등을 효능으로 하며, 허약체질, 번갈(煩渴), 당뇨병, 협심통 등에 탕제, 환제(丸劑) 또는 산제(散劑)로 하여 복용한다.Jadeok ( Polygonatum Officinale ) is medicinal as the root of the round, medicinal tinnitus (異 名) is called as Jijeol (地 節), Hwangji (黄 地), skinny (女 委) and so on. Dungle, the origin of jade, is a perennial herb. The root of this plant, jade, is harvested in the fall or early spring to remove stems and beard roots, dried in the shade, or steamed and sun-dried in the sun. This jade contains components such as steroidal saponin (convallamarin) and convallarin (steroidal saponin) and a number of flavonoids (flavonoids), cardiovascular action, corticosteroid-like action, It is known to have a thirst quenching action, antioxidant action, and skin aging action. In oriental medicine, the properties of this medicine are flat and sensation, nourishment, lubrication, and jigal, and it is effective for weak constitution, bungalow, diabetes, angina pain. Take it as a pill or powder.
본 발명에 따른 추출물의 제조과정은 하기와 같다.The preparation process of the extract according to the present invention is as follows.
황금, 황백, 치자, 황련, 오미자 및 옥죽를 세척하여 이물질을 제거한 후, 그늘에서 건조한다. 건조된 황금, 황백, 치자, 황련, 오미자, 옥죽 또는 이들을 적합한 비율, 바람직하게는 1:1:1:1:2:2의 중량비로 섞어 분쇄한다. 분쇄된 황금, 황백, 치자, 황련, 오미자, 옥죽 또는 이들의 혼합물에 적당한 양의 용매를 첨가하여 완전히 침지되도록 한다. 추출 용매로는 물, 탄소수 1 내지 4의 저급 알콜 또는 이들의 혼합 용매로부터 선택된 용매가 바람직하며, 더욱 바람직하게는 물이다. 상기 추출 용매의 양은 바람직하게 황금, 황백, 치자, 황련, 오미자, 옥죽 또는 이들의 혼합물 건중량의 5 ~ 20배이며, 바람직하게는 10배이다. 추출 방법은 실온에서 함침하거나, 가온할 수 있으며, 바람직하게는 열수 추출한다. 제조된 황금, 황백, 치자, 황련, 오미자, 옥죽 또는 이들의 혼합 추출물은 여과, 농축 및/또는 동결건조 등의 과정을 거쳐 사용할 수 있다.Golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade porridge are washed to remove foreign substances, and then dried in the shade. Dried golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis, Jade porridge or these are mixed and ground in a suitable ratio, preferably in a weight ratio of 1: 1: 1: 1: 2: 2. Appropriate amount of solvent is added to the pulverized golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis, jade porridge or mixtures thereof so as to be completely immersed. The extraction solvent is preferably a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, more preferably water. The amount of the extraction solvent is preferably 5 to 20 times, preferably 10 times, the dry weight of golden, yellowish white, gardenia, yellow lotus, schizandra, jade porridge or mixtures thereof. The extraction method can be impregnated at room temperature or warmed, preferably hot water extraction. The prepared golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis, jade porridge or a mixed extract thereof may be used through filtration, concentration and / or lyophilization.
본 발명에 따른 추출물인 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물(추출물B)을 2개월간 투여한 제2형 당뇨병 증세를 나타내는 90% 췌장 제거 랫트군에서는 대조군에 비하여 포도당의 경구 투여 후의 혈청 포도당이 현저히 낮게 나타나서 혈당강하 효과가 우수함을 알 수 있었으며, 인슐린 분비량도 증가되는 것을 확인할 수 있었다(3a 및 3b 참조). 이 때, 추출물A는 황금, 황백, 치자 및 황련의 혼합 추출물이고, 추출물B는 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물이다. Oral administration of glucose compared to the control group in the 90% pancreas-removed rat
또한, 생체내 인슐린 저항성을 측정하기 위한 고인슐린혈증 정상혈당 클램프(hyperinsulinemic euglycemic clamp) 실험에서 추출물B를 투여한 군에서는 포도당 주입률이 현저히 증가되어 인슐린 민감성이 증가된 것을 확인할 수 있었고(도 4a 참조), 고인슐린 상태에서의 간에서의 포도당 생성속도가 효과적으로 감소되어 간의 인슐린 저항성이 감소하는 것을 알 수 있었으며(도 4b 참조), 고혈당 상태에서의 혈청 인슐린 농도도 1차 단계 및 2차 단계 모두에서 대조군 및 기타 군보다 높게 나타남을 확인할 수 있었다(도 5a 및 5b 참조).In addition, in the hyperinsulinemic euglycemic clamp experiment for measuring insulin resistance in vivo, the group administered with extract B showed a significant increase in glucose infusion rate and increased insulin sensitivity (see FIG. 4A). It was found that the glucose production rate in the liver in the high insulin state was effectively decreased (see FIG. 4B), and the serum insulin concentration in the hyperglycemic state was also increased in the first and second stages. It can be seen that the control group and other groups appear higher (see FIGS. 5A and 5B).
또한, 추출물B는 인슐린/IGF-1 신호전달의 첫 번째 신호전달자인 랑게르한스섬 내의 IRS2의 발현을 유도함으로써(도 6 참조), 베타세포의 기능과 생존력을 증가시켜 혈당 강하 작용을 증진시킴을 확인할 수 있었다.In addition, extract B induces the expression of IRS2 in the Langerhans islet, the first signal of insulin / IGF-1 signaling (see FIG. 6), thereby increasing the function and viability of beta cells to enhance blood glucose lowering action. there was.
상기의 효과들은 황금, 황백, 치자, 황련, 오미자, 옥죽 추출물을 각각 단독으로 사용할 때보다 우수하였으므로, 본 발명에 따른 추출물은 상기 생약재의 혼합으로 인해 예측되는 결과와는 다른 예상하지 못한 현저한 효과가 나타남을 확인할 수 있었다. 즉, 도 3b 및 도 5a에서 나타난 바와 같이 오미자 추출물은 인슐린 분비량을 증가시키지 못하였고, 도 6에서 나타난 바와 같이 IRS2의 발현을 증가시키지 못하였음에도 불구하고, 오미자 및 옥죽을 황금, 황백, 치자 및 황련과 혼합하여 본 발명에 따른 새로운 생약재 혼합 추출물을 얻었고, 상기 추출물B는 인슐린 분비 증진 및 인슐린 작용 향상 활성에서 기존의 오미자 추출물 또는 황금, 황백, 치자 및 황련으로 이루어진 생약재 혼합 추출물보다 우수한 효과를 나타냄을 확인하였다.Since the above effects were superior to the use of gold, yellowish white, gardenia, yellow lotus, Schisandra chinensis, and jade extract alone, the extract according to the present invention had an unexpected and unexpected effect different from the results expected due to the mixing of the herbal medicines. Appeared. That is, the Schizandra chinensis extract did not increase insulin secretion as shown in FIGS. 3b and 5a, and did not increase the expression of IRS2 as shown in FIG. 6. When mixed with and obtained a new herbal mixed extract according to the present invention, the extract B shows an excellent effect than the conventional extract of Schizandra chinensis or the herbal extracts consisting of golden, yellow, white, gardenia and yellow lotus in the insulin secretion and insulin action enhancement activity Confirmed.
본 발명에 따른 추출물은 예전부터 사용되어온 생약재의 혼합 추출물로서 안전한 물질로 판단된다.Extract according to the present invention is considered to be a safe substance as a mixed extract of herbal medicines that have been used in the past.
본 발명의 조성물은 당뇨병의 치료 및 예방에 유용하게 이용될 수 있으며, 상기 당뇨병은 제2형 당뇨병을 특징으로 한다.The composition of the present invention can be usefully used for the treatment and prevention of diabetes, the diabetes is characterized by
따라서, 본 발명의 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물은 효과적인 혈당 강하 작용 및 인슐린 분비 작용을 나타내고, 생체내 인슐린 저항성을 감소시키며, IRS2의 발현을 증가시키므로, 상기 추출물을 함유하는 본 발명의 조성물은 당뇨병 예방 및 치료용 조성물로 유용하게 이용될 수 있다.Therefore, the mixed extracts of the golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade porridge of the present invention exhibits an effective hypoglycemic action and insulin secretion action, decreases insulin resistance in vivo and increases the expression of IRS2, thus containing the extract. The composition of the present invention can be usefully used as a composition for preventing and treating diabetes.
본 발명의 조성물은 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물에 추가로 동일 또는 유사한 기능을 나타내는 유효성분을 1종 이상 함유할 수 있다.The composition of the present invention may contain one or more active ingredients exhibiting the same or similar functions in addition to the mixed extract of gold, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade.
본 발명의 조성물은, 투여를 위해서 상기 기재한 유효성분 이외에 추가로 약제학적으로 허용 가능한 담체를 1종 이상 포함하여 제조할 수 있다. 약제학적으로 허용 가능한 담체는 식염수, 멸균수, 링거액, 완충 식염수, 덱스트로즈 용액, 말토 덱스트린 용액, 글리세롤, 에탄올 및 이들 성분 중 1 성분 이상을 혼합하여 사용할 수 있으며, 필요에 따라 항산화제, 완충액, 정균제 등 다른 통상의 첨가제를 첨가 할 수 있다. 또한 희석제, 분산제, 계면활성제, 결합제 및 윤활제를 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립 또는 정제로 제제화할 수 있다. 더 나아가 당분야의 적정한 방법으로 또는 Remington's Pharmaceutical Science(Mack Publishing Company, Easton PA, 18th, 1990)에 개시되어 있는 방법을 이용하여 각 질환에 따라 또는 성분에 따라 바람직하게 제제화할 수 있다.The composition of the present invention can be prepared by including one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients for administration. Pharmaceutically acceptable carriers may be used in combination with saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components, if necessary, as antioxidants, buffers Other conventional additives such as bacteriostatic agents can be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, it may be preferably formulated according to each disease or component by a suitable method in the art or using a method disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA, 18th, 1990).
본 발명의 조성물은 목적하는 방법에 따라 비경구 투여(예를 들어 정맥 내, 피하, 복강 내 또는 국소에 적용)하거나 경구 투여할 수 있으며, 투여량은 환자의 체중, 연령, 성별, 건강상태, 식이, 투여시간, 투여방법, 배설율 및 질환의 중증도 등에 따라 그 범위가 다양하다. 1일 투여량은 본 발명에 따른 추출물을 동결건조하였을 때의 양으로 10 ~ 500 ㎎/㎏, 바람직하게는 100 ㎎/㎏로, 하루 1회 ~ 수회에 나누어 투여하는 것이 바람직하다.The compositions of the present invention may be administered parenterally (eg, intravenously, subcutaneously, intraperitoneally or topically) or orally, depending on the desired method, and the dosage may be based on the weight, age, sex, health status, The range varies depending on the diet, the time of administration, the method of administration, the rate of excretion and the severity of the disease. The daily dose is 10 to 500 mg / kg, preferably 100 mg / kg, in an amount of lyophilized extract according to the present invention, preferably administered once to several times a day.
본 발명의 조성물은 당뇨의 예방 및 치료를 위하여 단독으로, 또는 수술, 방사선 치료, 호르몬 치료, 화학 치료 및 생물학적 반응 조절제를 사용하는 방법들과 병용하여 사용할 수 있다.The composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the prevention and treatment of diabetes.
본 발명의 조성물은 당뇨의 예방 및 개선을 목적으로 건강식품에 첨가될 수 있다. 본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물을 식품 첨가물로 사용할 경우, 상기 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통 상적인 방법에 따라 적절하게 사용될 수 있다. 유효 성분의 혼합양은 그의 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. 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 있음은 확실하다.The composition of the present invention can be added to health food for the purpose of preventing and improving diabetes. When using the mixed extract of the golden, yellow white, gardenia, yellow lotus, Schisandra chinensis and jade porridge according to the present invention as a food additive, the mixed extract of the golden, yellow white, gardenia, yellow lotus, schisandra chinensis and jade porridge is added as it is or with other food or food ingredients It can be used together and can be appropriately used according to a conventional method. The blending amount of the active ingredient can be suitably determined according to the purpose of its use (prevention, health or therapeutic treatment). 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. Is sure.
상기 식품의 종류에는 특별한 제한은 없다. 상기 물질을 첨가할 수 있는 식품의 예로는 육류, 소세지, 빵, 쵸코렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다.There is no particular limitation on the kind of food. Examples of the food to which the substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, drinks, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the conventional sense.
본 발명의 건강음료 조성물은 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 ㎖당 일반적으로 약 0.01∼0.04 g, 바람직하게는 약 0.02∼0.03 g 이다.The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The proportion of said natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.
상기 외에 본 발명의 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 조성물은 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 혼합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부 당 0.01∼0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, And a carbonation agent used for the carbonated beverage. In addition, the composition of the present invention may contain a flesh for preparing natural fruit juice, fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not critical but is usually selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
이하, 본 발명의 이해를 돕기 위하여 바람직한 실시예 및 실험예를 제시한다. 그러나 하기의 실시예 및 실험예는 본 발명을 보다 쉽게 이해하기 위하여 제공되는 것일 뿐, 실시예에 의해 본 발명의 내용이 한정되는 것은 아니다.Hereinafter, preferred examples and experimental examples are presented to help understand the present invention. However, the following Examples and Experimental Examples are provided only to more easily understand the present invention, and the contents of the present invention are not limited by the Examples.
<실시예> 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물의 제조EXAMPLES Preparation of Mixed Extracts of Gold, Yellowish White, Gardenia, Rhododendron, Schisandra chinensis and Jade Rice
황금, 황백, 치자, 황련, 오미자 및 옥죽을 세척하고 그늘에서 건조시켰다. 건조된 혼합물A(황금:황백:치자:황련=1:1:1:1 중량비), 혼합물B(황금:황백:치자:황련:오미자:옥죽=1:1:1:1:2:2 중량비), 오미자 및 황금을 각각 1.2 kg 씩 분쇄하였다. 분쇄된 재료에 증류수 8 ℓ을 넣고, 90℃에서 70분 동안 초음파탕전기(소니메디, 한국)를 이용해 추출하고, 12시간 동안 냉장침전 후에 2번 필터(No 2. filter)로 여과하였다. 여과액을 450 × g로 원심분리 (Megafuge 1.0R, USA)하여 상등액을 회수하고, 이를 회전진공 농축기 (Buchi R-114, USA)로 35℃에서 감압농축한 후, 동결건조 (Ilshin Freeze dryer, Korea)하여 분말 형태의 각 추출물을 제조하였다. 이 때 , 수율은 8%이었다.Golden, yellow white, gardenia, yellow lotus, schizandra and jade porridge were washed and dried in the shade. Dried Mixture A (Golden: Yellow White: Gardenia: Ranum = 1: 1: 1: 1 Weight Ratio), Mixture B (Golden: Yellow White: Gardenia: Russium: Schisandra: Jeju = 1: 1: 1: 1: 2: 2 Weight Ratio ), Schisandra chinensis and Gold were ground each 1.2 kg. 8 L of distilled water was added to the pulverized material, and extracted using an ultrasonic mixer (Sony Medi, Korea) for 70 minutes at 90 ° C., and filtered through a filter twice (
추출물을 농축하여 생성된 분말은 멸균된 디메틸술폭시화물(Dimethylsulfoxide; DMSO; Sigma Co., St. Louis, USA)에 용해 후 배양액으로 재희석하여 최종 농도를 50ug/mL로 하여 사용하였다.The powder produced by concentrating the extract was dissolved in sterile dimethylsulfoxide (DMSO; Sigma Co., St. Louis, USA) and re-diluted with a culture solution to obtain a final concentration of 50ug / mL.
<실험예 1> 대장암 세포주에서의 GLP-1 분비 측정Experimental Example 1 Measurement of GLP-1 Secretion in Colorectal Cancer Cell Lines
1. 세포 배양1. Cell Culture
실험에 사용한 NCI-H716 세포주는 미국세포주은행(ATCC)에서 구입하였고, 10% FBS(fetal bovine serum), 2 mM의 엘-글루타민(L-glutamine), 100 IU/㎖의 페니실린(penicillin), 100 ㎍/㎖의 스트렙토마이신(streptomycin) 등을 첨가한 RPMI(Roswell Park Memorial Institulete) 배지에서 현탁세포배양하였다. 배양된 세포를 Matrigel(Becton Dickinson, Bedford, MA, USA)을 입힌 디쉬(dish)로 옮겨서 GLP-1 분비 실험에 사용하였다.The NCI-H716 cell line used for the experiment was purchased from the American Cell Line Bank (ATCC), 10% fetal bovine serum (FBS), 2 mM L-glutamine, 100 IU / ml penicillin, 100 Suspension cells were cultured in Roswell Park Memorial Institulete (RPMI) medium containing μg / ml of streptomycin and the like. Cultured cells were transferred to dishes coated with Matrigel (Becton Dickinson, Bedford, Mass., USA) and used for GLP-1 secretion experiments.
실험 2일 전에 1 x 106 세포/㎖의 분화된 세포를 Matrigel을 입힌 12웰 플레이트(12-well culture plates)로 옮긴 후 10% FBS, 2 mM의 엘-글루타민, 100 IU/㎖의 페니실린, 100 ㎍/㎖의 스트렙토마이신 등을 첨가한 RPMI 배지에서 배양하였다.Two days before the experiment, 1 x 10 6 cells / ml of differentiated cells were transferred to Matrigel coated 12-well culture plates, followed by 10% FBS, 2 mM el-glutamine, 100 IU / ml penicillin, The cells were cultured in RPMI medium to which 100 µg / ml of streptomycin and the like were added.
2. GLP-1 분비 측정 실험2. GLP-1 Secretion Measurement Experiment
분화된 세포가 배양된 플레이트의 배양액을 0.2% (w/v) 소혈청 알부민(Bovine Serum Albumin, BSA)와 황금, 황백, 치자, 황련, 오미자 및 옥죽의 각 추출물을 넣은 KRB 완충용액(Krebs Ringer bicarbonate, pH 7.2)으로 바꾸어 주고 2시간 배양하였다. 각각의 웰로부터 KRB 용액을 모아서 여기에 50 ㎍/㎖의 PMSF(phenylmethylsulphonyl fluoride)와 34 ㎍/㎖의 디프로틴-A(diprotin-A)를 첨가하여 GLP-1 농도를 측정할 때까지 -80℃에 보관하였다.The culture medium of the plate in which the differentiated cells were cultured was prepared with KRB buffer solution containing 0.2% (w / v) bovine serum albumin (BSA) and extracts of each of golden, yellow, white, gardenia, barberry, schisandra chinensis and jade. bicarbonate, pH 7.2) and incubated for 2 hours. Collect KRB solution from each well and add it to -80 ° C until 50 μg / ml of phenylmethylsulphonyl fluoride (PMSF) and 34 μg / ml of diprotin-A were added to measure GLP-1 concentration. Stored in.
웰에 있는 세포는 긁어서 분쇄 완충용액(homogenization buffer, 5% (v/v)의 포름산(HCOOH)을 포함한 1 M의 염산(HCl), 1% (v/v)의 트리플루오로 아세트산(trifluoroacetic acid) 및 1% (w/v)의 염화나트륨)으로 분해시키고, 분해된 세포내의 펩타이드를 알코올 추출법 (alcohol extraction method)으로 추출하였다.The cells in the wells were scraped and scraped with 1 M hydrochloric acid (HCl), with 1% (v / v) trifluoroacetic acid, including homogenization buffer (5% (v / v) formic acid (HCOOH). ) And 1% (w / v) sodium chloride) and the digested intracellular peptides were extracted by alcohol extraction method.
린코 GLP-1 키트(GLP-1(736) Active RIA Kit, Linco Research Inc., St. Charles)를 사용한 방사선면역측정법(radioimmunoassay)으로 KRB 용액의 GLP-1 분비량과 세포에 있는 GLP-1의 양을 정량하였고, 세포의 양을 보정하기 위해서 세포 분쇄물(homogenate)의 단백질 양을 브래드포드 단백질 정량법(Bradford protein assay, Bio-Rad, Inc.)으로 정량하였다.Radioimmunoassay using the Linco GLP-1 Kit (GLP-1 (736) Active RIA Kit, Linco Research Inc., St. Charles) for the release of GLP-1 in KRB solution and the amount of GLP-1 in cells In order to correct the amount of cells, the protein amount of the cell homogenate was quantified by the Bradford protein assay (Bio-Rad, Inc.).
3. 생약재의 각 추출물이 NCI-H716 대장암세포의 GLP-1 분비에 미치는 영향3. Effect of each extract of herbal medicine on GLP-1 secretion of NCI-H716 colorectal cancer cells
NCI-H716 세포주에서의 GLP-1의 분비 측정 결과, 황금, 황백, 치자, 황련, 오미자 및 옥죽의 각 추출물 중 황금과 황백이 GLP-1 분비를 증가시켰다(도 1a 참조).As a result of measuring the secretion of GLP-1 in the NCI-H716 cell line, among the extracts of gold, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade, the gold and yellow white increased GLP-1 secretion (see FIG. 1A).
모든 결과는 평균± 표준편차(SD)로 표현하였고, 도면 중 *는 One-way analyses of variance (ANOVA)를 이용하여 P<0.05 수준으로 다른 군과 현저한 차이 를 보이는 것을 표시하였다. a,b 값은 Tukey 검정법에 의한 P<0.05 수준으로 같은 항의 값과 현저한 차이를 보이는 것을 나타내었다. 하기의 모든 실험예에서의 통계분석은 상기와 같이 하였다.All results were expressed as mean ± standard deviation (SD), and * in the figure indicated that the one-way analyses of variance (ANOVA) showed a remarkable difference from other groups at P <0.05 level. The a and b values are P <0.05 by Tukey's test, and the same terms It showed a difference. Statistical analysis in all the following Experimental Examples were as described above.
<실험예 2> 췌장 베타세포주에서의 인슐린 분비 측정Experimental Example 2 Insulin Secretion Measurement in Pancreatic Beta Cell Line
1. 세포 배양1. Cell Culture
실험에 사용한 Min6 세포주는 미국세포주은행에서 구입하였고, 15% FBS, 2 mM의 엘-글루타민, 100 IU/㎖의 페니실린, 100 ㎍/㎖의 스트렙토마이신 등을 첨가한 고농도 포도당을 함유한 DMEM(Dulbecco's modifiedEagle's medium)에서 배양하였다.The Min6 cell line used in the experiment was purchased from the American Cell Line Bank and contained high concentrations of DMEM (Dulbecco's) containing 15% FBS, 2 mM el-glutamine, 100 IU / ml penicillin, and 100 μg / ml streptomycin. cultured in modifiedEagle's medium).
실험 2일 전에 1 x 106 세포/㎖의 세포를 12웰 플레이트로 옮긴 후 10% FBS, 2 mM의 엘-글루타민, 100 IU/㎖의 페니실린, 100 ㎍/㎖의 스트렙토마이신 등을 첨가한 고농도 포도당 DMEM 배지에서 배양하고, 다음날 0.2% BSA를 포함하는 저농도 포도당 DMEM 배지로 바꾸어 배양하였다.Two days before the experiment, 1 x 10 6 cells / ml of cells were transferred to a 12-well plate, followed by high concentration of 10% FBS, 2 mM el-glutamine, 100 IU / ml of penicillin, and 100 μg / ml of streptomycin. Cultured in glucose DMEM medium, the next day was incubated with a low concentration of glucose DMEM medium containing 0.2% BSA.
2. 인슐린 분비 측정 실험2. Insulin Secretion Measurement Experiment
세포가 배양된 플레이트의 배양액을 0.2% (w/v) BSA와 황금, 황백, 치자, 황련, 오미자 및 옥죽의 각 추출물을 넣은 KRB 용액(pH 7.2)으로 바꾸어 주고 2시간 배양하였다. 각각의 웰로부터 KRB 용액을 모아서 인슐린 농도를 측정할 때까지 -80℃에 보관하였다.The culture medium of the plate in which the cells were cultured was changed to KRB solution (pH 7.2) containing 0.2% (w / v) BSA and extracts of golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade, and incubated for 2 hours. KRB solutions from each well were collected and stored at -80 ° C until insulin concentration was measured.
웰에 있는 세포는 긁어서 분쇄 완충용액으로 분해시키고, 분해된 세포내의 펩타이드를 알코올 추출법으로 추출하였다.The cells in the wells were scraped and digested with the grinding buffer, and the peptides in the digested cells were extracted by alcohol extraction.
린코 인슐린 키트(Linco Research Inc., St. Charles)를 사용한 방사선면역측정법으로 KRB 용액의 인슐린 분비량과 세포에 있는 인슐린의 양을 정량하였고, 세포의 양을 보정하기 위해서 세포 분쇄물의 단백질 양을 브래드포드 단백질 정량법으로 정량하였다.Radioimmunoassay using Linco Research Inc. (St. Charles) was used to quantify the amount of insulin secreted from the KRB solution and the amount of insulin present in the cells. Quantification by protein quantification.
3. 생약재의 각 추출물이 Min6 세포의 인슐린 분비에 미치는 영향3. Effect of each extract of herbal medicine on insulin secretion of Min6 cells
Min6 세포주에서의 인슐린의 분비 측정 결과, 황금, 황백, 치자, 황련, 오미자 및 옥죽의 각 추출물 중 황금과 황백이 인슐린 분비를 증가시켰다(도 1b 참조).As a result of measuring insulin secretion in the Min6 cell line, among the extracts of gold, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade, the gold and yellow white increased insulin secretion (see FIG. 1B).
<실험예 3> 실험 동물의 제작과 생약재 추출물의 투여Experimental Example 3 Preparation of Experimental Animals and Administration of Herbal Extracts
1. 당뇨 증상을 나타내는 실험 동물의 제작1. Production of experimental animals showing diabetes symptoms
Hosokawa 등의 방법에 의해 생후 8주령의 수컷 SD 쥐(Sprague Dawley rat)의 췌장의 90%를 제거하여 경미한 당뇨 증세를 가진 실험동물(90% pancreatectomized rats)을 제작하였다. 실험동물 대조군(sham operated rats)은 췌장 절제 수술과 동일한 수술을 하여 췌장을 절제하지 않고 손가락으로 만져 준 후 봉합하였다.90% of pancreas of 8-week-old male SD rats (Sprague Dawley rats) were removed by Hosokawa et al. To produce experimental animals with mild diabetes (90% pancreatectomized rats). Experimental animal controls (sham operated rats) were sutured after touching the fingers with the pancreas without excision by the same operation as the surgery for pancreas resection.
수술 후 2주 동안 고형사료로 사육하였고, 수술 경과가 좋지 않은 개체와 혈당이 9.4 mmol/l 미만으로 나타나는 개체는 실험 대상에서 제외하였으며, 이러한 당뇨모델 실험동물 제작 방법으로 약 40-50%가 당뇨 증상을 나타내는 것으로 판명되었다.Two weeks after the operation, the animals were fed with solid feed, and those with poor surgical procedures and those with less than 9.4 mmol / l of blood sugar were excluded from the test subjects. About 40-50% of the diabetic model animals were manufactured. It turned out to be symptomatic.
실험동물 대조군은 당뇨 증세를 나타내지 않았다.The experimental animal control group did not show diabetes.
2. 공복시와 식후의 혈당 및 혈청 인슐린 농도 측정2. Determination of Blood Glucose and Serum Insulin Levels on Fasting and After Meal
공복시의 혈당을 측정하기 위해서 1주일에 1회 오후 6시부터 다음날 오전 10시까지 실험동물을 절식시킨 후에 혈액을 대퇴부 정맥(femoral vein)으로부터 채취하였다. 채취한 혈액의 혈장을 분리한 후 혈당 분석기 (Glucose Analyzer, Beckman Instruments, Fullerton, CA)로 혈당을 측정하였다.In order to measure fasting blood glucose, the animals were fasted from 6 pm to 10 am the following day once a week, and blood was collected from the femoral vein. Plasma of the collected blood was separated and blood glucose was measured by a glucose analyzer (Glucose Analyzer, Beckman Instruments, Fullerton, CA).
식후 혈당은 1주일에 1회 오전 10시에 정상적으로 식이가 주어진 실험동물로부터 혈액을 채취하여 측정하였다.Post-prandial blood glucose was measured at 10 am once a week by taking blood from experimental animals given normal diet.
혈청내 인슐린 농도는 린코 키트(Rat Insulin Specific RIA kit, Linco Research Inc., St. Charles, USA)를 사용하여 방사선면역측정법으로 정량하였다.Serum insulin concentrations were quantified by radioimmunoassay using a Rat Insulin Specific RIA kit, Linco Research Inc., St. Charles, USA.
3. 당뇨 증상을 나타내는 실험 동물에 대한 생약재 추출물의 투여3. Administration of Herbal Extracts to Experimental Animals with Diabetic Symptoms
상기에서 제작한 당뇨 증상을 나타내는 실험동물에게 대조군(DMSO), 혼합물A로부터 얻은 추출물A, 혼합물B로부터 얻은 추출물B, 황금 추출물 및 오미자 추출물을 2개월 동안 경구 투여한 후에 이하의 실험을 수행하였다.The following experiment was performed after oral administration of the control group (DMSO), the extract A obtained from the mixture A, the extract B obtained from the mixture B, the golden extract and the Schizandra chinensis extract for 2 months.
각 추출물의 투여량은 추출물A에서는 황금, 황백, 치자 및 황련이 각각 600mg/kg체중/일, 추출물B에서는 황금, 황백, 치자 및 황련은 각각 300mg/kg체중/ 일, 오미자와 옥죽은 600 mg/kg체중/일이 되도록 투여하였다. 황금 추출물은 2,400mg/kg체중/일, 오미자 추출물은 2,400mg/kg체중/일이 되도록 실험동물에게 경구 투여하였다.The dosage of each extract was 600 mg / kg body weight / day of golden, yellow white, gardenia and yellow lotus in extract A, and 300 mg / kg body weight / day of golden, yellow white, gardenia and yellow lotus in extract B, respectively. It was administered to / kg body weight / day. The gold extract was orally administered to the experimental animals such that 2,400 mg / kg body weight / day and Schisandra chinensis extract was 2,400 mg / kg body weight / day.
4. 실험 동물의 식이 섭취량, 체중 및 내장 지방 함량의 변화 여부 4. Changes in dietary intake, body weight and visceral fat content of experimental animals
상기와 같이 각 조성물을 8주간 투여하면서, 일정 간격으로 1일 식이 섭취량을 기록하였고, 투여 완료 후의 체중 및 내장지방 정도를 측정하였다.While administering each composition as described above for 8 weeks, the daily dietary intake was recorded at regular intervals, and the body weight and visceral fat level after completion of the administration were measured.
1일 식이 섭취량, 체중 및 내장지방 정도는 대조군, 추출물A 처리군, 추출물B 처리군, 황금 처리군, 오미자 처리군에서 큰 차이는 없었다. 이것은 1일 식이 섭취량의 결과가 체중과 내장 지방 정도(visceral fat, epidydimal fat pads)에 반영된 것으로 보인다(도 2 참조).Daily dietary intake, body weight and visceral fat were not significantly different in the control group, extract A treatment group, extract B treatment group, golden treatment group and Schizandra treatment group. It seems that the results of the daily dietary intake were reflected in the body weight and visceral fat (epidydimal fat pads) (see Figure 2).
<실험예 4> 생약재 추출물의 생체내에서의 혈당 강하 효과 및 인슐린 분비량에 미치는 효과<
상기에서 제작한 제2형 당뇨병 증세를 나타내는 실험 동물에 대조군(DMSO), 추출물A, 추출물B, 황금 추출물, 오미자 추출물을 2개월 동안 투여하였다.The control animals (DMSO), extract A, extract B, golden extract, Schisandra chinensis extract were administered to experimental
이러한 실험동물에 대하여 경구 포도당 내성 시험법(oral glucose tolerance test: OGTT)을 실시하기 위해서 체중 1 kg 당 2 g의 포도당을 경구 투여한 후 혈청내 포도당 및 인슐린 분비량을 측정하였다. 이 후, 0, 10, 20, 30, 45, 60, 90, 120 분이 지난 후에 각각 꼬리로부터 혈액을 채취하여 혈당량을 측정하였으며, 그 곡선의 아랫부분 면적을 계산하였다. *는 P<0.05로 다른 군과 현저한 차이를 보였으며, Tukey 검정법에 의한 막대기의 a,b,c 값은 P<0.05로 같은 군의 값과 현저한 차이를 보였다.In order to perform the oral glucose tolerance test (OGTT) on these test animals, glucose and insulin secretion were measured after oral administration of 2 g of glucose per kg of body weight. Thereafter, after 0, 10, 20, 30, 45, 60, 90 and 120 minutes, blood was collected from the tail to measure blood glucose levels, and the area of the lower portion of the curve was calculated. * Was significantly different from other groups with P <0.05, and a, b, c of the rods by Tukey test The value is P <0.05, which is remarkable Showed a difference.
포도당의 혈청 중 농도를 도 3a에 나타냈고, 혈청 중 포도당과 인슐린의 AU(Artificial Unit)를 계산하여 도 3b에 나타냈다. 이 때, AU는 혈청 중 최고 농도를 나타내는 시간으로부터 1분 동안의 평균 농도를 말한다. 도 3b의 AU를 혈청 중 포도당의 감소 정도 및 인슐린 분비량의 증가 정도를 나타내는 지표로 사용하였다.The concentration of glucose in serum is shown in FIG. 3A, and the AU (Artificial Unit) of glucose and insulin in serum is calculated and shown in FIG. 3B. At this time, AU means the average concentration for 1 minute from the time which shows the highest concentration in serum. AU of FIG. 3b was used as an indicator of the degree of decrease in glucose and increase of insulin secretion in serum.
결과에서 보는 바와 같이, 추출물A와 오미자 추출물을 처리한 군에서 혈당이 낮게 나타났으나 통계적으로 유의하지 않았고, 추출물B 처리군만이 대조군과 비교하여 통계적으로 유의하게 혈당을 낮추는 효과를 보였다(도 3a 참조).As shown in the results, the blood glucose levels were low in the group treated with Extract A and Schisandra chinensis extract, but not statistically significant, and only the extract B treated group showed a statistically significant effect of lowering blood glucose compared to the control group (Fig. 3a).
인슐린 분비량은 추출물A, 추출물B, 황금 추출물 처리군에서 대조군에 비하여 통계적으로 유의하게 높은 값을 보였다(도 3b 참조). 특히, 본원 발명인 추출물B를 처리한 군에서 가장 높은 인슐린 분비량을 나타내어, 추출물B를 처리한 군에서 가장 낮은 혈당값을 나타내는 것과 관련이 있음을 시사하였다.Insulin secretion was significantly higher than the control group in the extract A, extract B, golden extract treatment group (see Figure 3b). In particular, it showed the highest insulin secretion in the group treated with the extract B of the present invention, suggesting that it is related to the lowest blood glucose value in the group treated with the extract B.
<실험예 5> 생약재 추출물이 인슐린의 작용력 및 인슐린 저항성에 미치는 효과Experimental Example 5 Effect of Medicinal Herb Extracts on Insulin's Action and Insulin Resistance
1. 실험 동물에 대한 추출물 투여 및 수술1. Extract Administration and Surgery for Experimental Animals
인슐린 감수성을 측정하기 위해, 고인슐린혈증 정상혈당 클램프 실 험(hyperinsulinemic euglycemic clamp study)를 시행하였다. 상기 <실험예 3>의 3에서와 같은 방식으로, 추출물A, 추출물B, 황금 추출물 및 오미자 추출물을 각 군의 랫트에 2개월 동안 경구 투여한 후에 이하의 실험을 수행하였다.To measure insulin sensitivity, a hyperinsulinemic euglycemic clamp study was performed. In the same manner as in <Experiment 3>, the following experiment was performed after oral administration of extract A, extract B, golden extract and Schizandra extract for 2 months to rats of each group.
물질 투여와 혈액 채취를 위해 랫트의 오른쪽의 경정맥(jugular vein)과 왼쪽의 경동맥(carotid artery)에 카테터(catheter, PE-10, Intramedic, Clay Adams, Parsippany, NJ)를 삽입하는 수술을 하였고, 수술 후 1주일 동안 회복시켰다.A catheter (PE-10, Intramedic, Clay Adams, Parsippany, NJ) was inserted into the jugular vein on the right side and the carotid artery on the left side for substance administration and blood collection. After 1 week of recovery.
2. 인슐린의 저항성 측정을 위한 고인슐린혈증 정상혈당 클램프 실험2. Hyperinsulinemia Normal Glucose Clamp Experiment to Measure Insulin Resistance
회복된 랫트에 표지된 포도당([3-3H]glucose)을 0.15mCi/분의 속도로 1시간 동안 주입한 후, 표지된 포도당을 계속 주입하면서 인슐린과 25% 포도당용액을 12 mU/kg체중/분의 속도로 정맥 주입하여, 정상혈당(euglycemia)이 유지되는 정상 상태(steady-state)를 유지하였다.The recovered rats were infused with labeled glucose ([3- 3 H] glucose) at a rate of 0.15 mCi / min for 1 hour, followed by continued infusion of labeled glucose with 12 mU / kg of insulin and 25% glucose solution. Intravenously injected at a rate of / min to maintain a steady-state in which normal blood glucose (euglycemia) is maintained.
상기 기간 동안 5분 간격으로 동맥으로부터 혈액을 채취하여 혈당이 90-100 mg/dl로 유지하도록 하여 포도당 주입률(glucose infusion rate, GIR)을 결정하였다. 실험 중 혈액 손실에 대하여는 다른 랫트로부터 혈액을 채취하여 헤파린 처리된 생리식염수(10 U/ml)로 1:1 (v:v)로 희석하여 정맥으로 주입하여 보충하였다.Glucose infusion rate (GIR) was determined by collecting blood from the arteries at 5 minute intervals during this period to maintain blood glucose at 90-100 mg / dl. For blood loss during the experiment, blood was collected from other rats and supplemented by intravenous dilution with 1: 1 (v: v) in heparinized saline (10 U / ml).
포도당 주입률은 혈당을 90-100 mg/dl로 유지하는데 필요한 포도당 주입속도에 주입하는 용액의 포도당 농도를 곱하고 랫트의 체중으로 나누어서 분 당, 단위 체중 당, 주입되는 포도당 함량으로 표시하였다(도 4a 참조).The glucose infusion rate was expressed as the glucose content injected per minute, per unit weight, and injected per minute by multiplying the glucose concentration of the solution to be injected to the glucose infusion rate required to maintain blood sugar at 90-100 mg / dl (Figure 4a). Reference).
결과에서 보는 바와 같이, 대조군과 비교하여 추출물B와 오미자 추출물을 처 리한 군의 포도당 주입률이 통계적으로 유의하게 증가하였고, 특히 본원 발명인 추출물B를 처리한 군에서 가장 높은 포도당 주입률을 나타냈다.As shown in the results, compared with the control group, the glucose injection rate of the group treated with the extract B and Schisandra chinensis extract statistically significantly increased, especially the group treated with extract B of the present invention showed the highest glucose injection rate.
3. 포도당의 특이활성(specific activity) 측정3. Measurement of glucose specific activity
혈청내 표지된 포도당의 특이활성은 Somogi 방법을 이용하여 측정하였다. 혈청 단백질을 수산화바륨(Ba(OH)2)과 황산납(ZnSO4)으로 침전시키고, 상등액을 분리한 후 55oC에서 건조시켜 3H2O를 제거하였다. 얻어진 건조물을 0.1 ml의 증류수로 녹인 후 3ml의 방사선 측정용액(scintiverse)을 넣어 이것을 액체 방사능 측정기(liquid scintillation counter)로 방사능 정도(dpm)를 측정하였다. 혈청내 3H2O의 방사능(dpm)은 혈청 단백질 제거 후의 건조시키지 않은 상등액의 방사능 측정값과 상등액을 건조시킨 후의 방사능 측정값의 차이로 계산하였다.The specific activity of labeled glucose in serum was measured using Somogi method. The serum protein was precipitated with barium hydroxide (Ba (OH) 2 ) and lead sulfate (ZnSO 4 ), and the supernatant was separated and dried at 55 ° C. to remove 3 H 2 O. The obtained dried product was dissolved in 0.1 ml of distilled water, and then 3 ml of a radiation solution (scintiverse) was added thereto, and the radioactivity (dpm) was measured by a liquid scintillation counter. The radioactivity (dpm) of 3 H 2 O in serum was calculated as the difference between the radioactivity measurement value of the undried supernatant after serum protein removal and the radioactivity measurement value after drying the supernatant.
이 때, 포도당 섭취율(uptake)은 혈관계로 포도당이 유입되는 정도를 나타내는 값이고, 포도당 소실률(glucose disposal rate)은 인슐린의 작용에 의해 혈관계로부터 포도당이 소실되는 정도를 나타내는 값이며, 정상 상태에서는 포도당 섭취율과 포도당 소실률이 같다. 포도당 소실률은 표지된 포도당의 주입률(dpm/분)을 정상 상태에서의 표지된 포도당의 특이활성(dpm/mg)으로 나눈 값으로 계산하였고, 이 값은 포도당 섭취율을 의미한다(도 4a 참조).In this case, the glucose uptake rate is a value representing the degree of glucose inflow into the vascular system, and the glucose disposal rate is a value indicating the degree of glucose loss from the vascular system due to the action of insulin. Intake rate and glucose loss rate are the same. Glucose loss rate was calculated as the infusion rate of labeled glucose (dpm / min) divided by the specific activity of labeled glucose (dpm / mg) at steady state, which means the glucose intake rate (see FIG. 4A). .
상기 기재한 바와 같이 포도당 주입률은 추출물B를 처리한 군에서 가장 높은 값을 나타냈으나, 포도당 섭취율은 도 4a에서 보는 바와 같이 추출물A, 추출물B, 황금 추출물, 오미자 추출물 처리군에서 차이가 없었음을 알 수 있다. 즉, 본원발명의 추출물인 추출물B를 처리한 군에서는 실험동물에 주입되는 포도당의 절대량이 많음에도 불구하고 혈관으로 유입되는 포도당의 양은 일정하여, 결과적으로 생체내의 혈관이 아닌 다른 기관에서 포도당 소비가 크다는 것을 시사하고 있다.As described above, the glucose infusion rate showed the highest value in the group treated with Extract B, but the glucose intake rate was not different in the Extract A, Extract B, Golden Extract, and Schisandra extract treated groups as shown in FIG. I can see that. That is, in the group treated with the extract B, the extract of the present invention, the amount of glucose introduced into the blood vessel is constant despite the absolute amount of glucose injected into the experimental animal, and consequently, glucose consumption is increased in organs other than the blood vessel in vivo. It's big.
4. 간에서의 인슐린 저항성(Hepatic glucose production)에 미치는 효과4. Effect on Hepatic Insulin Resistance
상기의 수술을 한 실험 동물에 표지된 포도당을 일정한 속도로 40분간 주입하여 혈청내 표지된 포도당의 특이활성이 정상 상태를 유지하도록 하였다.Labeled glucose was injected into the experimental animals subjected to the above surgery at a constant rate for 40 minutes to maintain the specific state of the labeled glucose in serum.
고인슐린혈증(고인슐린혈증 정상혈당 클램프 실험 상태) 에서의 간에서의 포도당 생성속도는 혈관계로의 포도당 섭취율에서 외부로부터의 포도당 주입률을 뺀 값으로 계산하였고, 간에서의 포도당의 기저 생성속도와 고인슐린혈증에서의 생성속도를 도 4b에 나타냈다.Glucose production rate in the liver in hyperinsulinemia (hyperglycemic normal glucose clamp test) was calculated by subtracting the glucose intake from the outside from the glucose intake into the vasculature and basal glucose production rate in the liver. The production rate in hyperinsulinemia is shown in Figure 4b.
도 4b에 나타난 바와 같이, 혈액내 인슐린 농도가 높을 때에는 추출물B와 오미자 추출물을 처리한 군의 포도당 생성속도가 통계적으로 유의하게 감소하였고, 특히 본원 발명인 추출물B를 처리한 군에서 가장 낮은 포도당 생성속도를 나타냈다. 이러한 결과로부터 본원발명의 추출물B가 간에서의 인슐린 저항성을 효과적으로 감소시킴을 알 수 있었다. 공복상태에서는 본원 발명인 추출물B를 처리한 군에서 가장 낮은 포도당 생성속도를 나타냈지만 통계적으로 유의하지는 않았다.As shown in Figure 4b, when the blood insulin concentration is high, the glucose production rate of the group treated with the extract B and Schizandra extract statistically significantly decreased, especially the lowest glucose production rate in the group treated with the extract B of the present invention Indicated. From these results, it can be seen that the extract B of the present invention effectively reduces insulin resistance in the liver. The fasting state showed the lowest glucose production rate in the group treated with the extract B of the present invention, but was not statistically significant.
<실험예 6> 생약재 추출물이 췌장 베타세포의 인슐린 분비능에 미치는 효과Experimental Example 6 Effect of Medicinal Herb Extracts on Insulin Secretion of Pancreatic Beta Cells
인슐린 분비능을 측정하기 위해, 고혈당증 클램프 실험(Hyperglycemic clamp)를 시행하였다. 물질 투여와 혈액 채취를 위해 랫트의 오른쪽의 경정맥과 왼쪽의 경동맥에 카테터를 삽입하는 수술을 하였고, 수술 후 1주일 동안 회복시켰다. 회복 후 체중이 정상으로 돌아오면 12시간 금식시킨 후 자유롭게 움직이는 상태에서 고혈당증 클램프 실험을 시행하였다.To measure insulin secretion, a hyperglycemic clamp test was performed. The catheter was inserted into the jugular vein on the right side and the carotid artery on the left side for the administration of the substance and blood collection, and recovered for 1 week after the surgery. After recovery, the body returned to normal fasting for 12 hours, and then the hyperglycemic clamp test was performed while moving freely.
실험 동물에 25% 포도당을 주입하면서 10분에 한번씩 혈액을 채취하여 혈당이 230mg/dL로 유지하도록 포도당 주입량을 조절하였다. 채취한 혈액의 0, 2, 5, 10, 60, 90, 120분의 혈청은 -70oC에서 보관하였다가 혈청 인슐린 농도를 측정하였다.Blood was collected every 10 minutes while injecting 25% glucose into the experimental animals to adjust the glucose infusion to maintain blood glucose at 230 mg / dL. Serum of 0, 2, 5, 10, 60, 90, 120 minutes of the collected blood was stored at -70 o C and serum insulin concentration was measured.
시간에 따른 혈청 인슐린 농도를 도 5a에 나타냈고, 1차 단계와 2차 단계에서의 인슐린의 AU를 도 5b에 나타냈다. 이 때, 1차 단계 및 2차 단계의 AU란 혈청 인슐린 농도 곡선에서 각각 첫번째 봉우리의 최고 농도와 두번째 봉우리의 최고 농도를 나타내는 시간으로부터 1분 동안의 평균 농도를 말한다.The serum insulin concentration over time is shown in FIG. 5A and the AU of insulin in the first and second stages is shown in FIG. 5B. In this case, the AU of the first stage and the second stage refers to the average concentration for one minute from the time indicating the highest concentration of the first peak and the highest concentration of the second peak in the serum insulin concentration curve, respectively.
도 5a에 나타난 바와 같이, 혈청 중 인슐린의 농도는 본원 발명인 추출물B를 처리한 군에서 높게 나타났다.As shown in Figure 5a, the concentration of insulin in the serum was high in the group treated with extract B of the present invention.
또한, 도 5b에 나타난 바와 같이 1차 단계에서는 추출물B를 처리한 군에서 통계적으로 유의하게 증가되었고, 2차 단계에서는 추출물A와 추출물B를 처리한 군에서 통계적으로 유의하게 증가되었다. 특히, 본원 발명인 추출물B를 처리한 군에서는 1차 단계 및 2차 단계에서 가장 높은 인슐린 농도값을 나타냈다.In addition, as shown in Figure 5b in the first step was statistically significantly increased in the group treated with extract B, and in the second step was significantly increased in the group treated with extract A and extract B. In particular, the group treated with the extract B of the present invention showed the highest insulin concentration value in the first and second stages.
<실험예 7> 생약재 추출물이 췌장 베타세포의 IRS2의 발현에 미치는 효과Experimental Example 7 Effects of Medicinal Herb Extracts on the Expression of IRS2 in Pancreatic Beta Cells
베타세포에서 IRS2 발현의 증가 여부 및 베타세포의 기능과 밀접한 관련이 있는 글루코키나제(glucokinase) 및 GLUT2의 발현 여부를 측정하여 IRS2의 발현 정도를 알아 보았다.The expression level of IRS2 was examined by measuring the expression of glucokinase and GLUT2, which are closely related to beta cell function and whether IRS2 expression was increased in beta cells.
분리된 랑게르한스섬에 상기의 각 추출물을 처리하여 8시간 동안 배양한 후, 대조군 또는 트라이졸(GIBCO BRL, Life Technologies, Inc., Grand Island, New York)로 처리된 랑게르한스섬에서 총 RNA를 분리하였다. 분리된 RNA에 대하여 올리고 dT를 사용하는 RETRO script kit(Ambion Inc., Austin, Texas)으로 cDNA를 합성하였다. 그리고 iCycler PCR 기구(Bio-Rad Laboratories, Hercules, California)와 QuantiTect SYBR Green PCR kit(Qiagen Inc., Valencia, California)를 이용하여 정량 Realtime PCR[94℃에서 4 분, (94℃에서 30초, 55℃에서 30초, 72℃에서 30초) ×n 및 72℃에서 5 분]을 수행하였다.After treatment with each of the extracts on the isolated island of Langerhans and incubated for 8 hours, total RNA was isolated from the island of Langerhans treated with a control or trizol (GIBCO BRL, Life Technologies, Inc., Grand Island, New York). CDNA was synthesized with the RETRO script kit (Ambion Inc., Austin, Texas) using oligo dT on the isolated RNA. And quantitative Realtime PCR [4 minutes at 94 ° C., 30 minutes at 94 ° C., 55 minutes] using iCycler PCR instrument (Bio-Rad Laboratories, Hercules, California) and QuantiTect SYBR Green PCR kit (Qiagen Inc., Valencia, California). 30 seconds at 0 ° C., 30 seconds at 72 ° C.) × n and 5 minutes at 72 ° C.].
내부의 대조가 되는 싸이클로필린과 관심있는 유전자 PCR 산물의 상대적인 비를 계산함으로써 정량 분석을 수행하였다.Quantitative analysis was performed by calculating the relative ratios of the internal control cyclophilin and the gene PCR product of interest.
결과는 도 6에 나타냈다.The results are shown in FIG.
도 6에서 보는 바와 같이, 추출물A, 추출물B 및 황금 추출물을 처리한 군에서 IRS2의 발현이 통계적으로 유의하게 증가되었다.As shown in Figure 6, the expression of IRS2 was significantly increased in the group treated with Extract A, Extract B and golden extract.
<제제예 1> 약학적 제제의 제조Preparation Example 1 Preparation of Pharmaceutical Formulation
1-1. 산제의 제조1-1. Manufacture of powder
본 발명에 따른 생약재 혼합 추출물 2g2g herbal medicine mixed extract according to the present invention
유당 1g1g lactose
상기의 성분을 혼합하고 기밀포에 충진하여 산제를 제조하였다.The above components were mixed and packed in airtight bags to prepare powders.
1-2. 정제의 제조1-2. Manufacture of tablets
본 발명에 따른 생약재 혼합 추출물 100㎎100 mg herbal extract extract according to the present invention
옥수수전분 100㎎Corn Starch 100mg
유 당 100㎎Lactose 100mg
스테아린산 마그네슘 2㎎2 mg magnesium stearate
상기의 성분을 혼합한 후, 통상의 정제의 제조방법에 따라서 타정하여 정제를 제조하였다.After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.
1-3. 캡슐제의 제조1-3. Preparation of Capsules
본 발명에 따른 생약재 혼합 추출물 100㎎100 mg herbal extract extract according to the present invention
옥수수전분 100㎎Corn Starch 100mg
유 당 100㎎Lactose 100mg
스테아린산 마그네슘 2㎎2 mg magnesium stearate
상기의 성분을 혼합한 후, 통상의 캡슐제의 제조방법에 따라서 젤라틴 캡슐에 충전하여 캡슐제를 제조하였다.After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.
1-4. 주사액제의 제조1-4. Preparation of Injection
생약재 혼합 추출물 10 ㎍/㎖10 ㎍ / ㎖ herbal medicine extract
묽은 염산 BP pH 3.5로 될 때까지Dilute hydrochloric acid BP until pH 3.5
주사용 염화나트륨 BP 최대 1 ㎖Injectable sodium chloride BP up to 1 ml
적당한 용적의 주사용 염화나트륨 BP 중에 본 발명에 따른 생약재 혼합 추출물을 용해시키고, 생성된 용액의 pH를 묽은 염산 BP를 사용하여 pH 3.5로 조절하고, 주사용 염화나트륨 BP를 사용하여 용적을 조절하고 충분히 혼합하였다. 용액을 투명 유리로 된 5 ㎖ 타입 I 앰플 중에 충전시키고, 유리를 용해시킴으로써 공기의 상부 격자하에 봉입시키고, 120℃에서 15 분 이상 오토클래이브시켜 살균하여 주사액제를 제조하였다.Dissolve the herbal mixture extract according to the present invention in a suitable volume of injectable sodium chloride BP, adjust the pH of the resulting solution to pH 3.5 with dilute hydrochloric acid BP, adjust the volume with injectable sodium chloride BP and mix well It was. The solution was filled into a 5 ml Type I ampoule made of clear glass, encapsulated under an upper grid of air by dissolving the glass, and sterilized by autoclaving at 120 ° C. for at least 15 minutes to prepare an injection solution.
<제제예 2> 식품의 제조Preparation Example 2 Preparation of Food
본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물을 포함하는 식품들을 다음과 같이 제조하였다.Food products containing a mixed extract of golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade porridge according to the present invention were prepared as follows.
2-1. 밀가루 식품의 제조2-1. Manufacture of flour food products
본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물 0.5 ~ 5.0 중량%를 밀가루에 첨가하고, 이 혼합물을 이용하여 빵, 케이크, 쿠키, 크래커 및 면류를 제조하여 건강 증진용 식품을 제조하였다.Add 0.5 ~ 5.0% by weight of the mixed extract of golden, yellow white, gardenia, yellow lotus, Schisandra chinensis and jade porridge according to the present invention to flour, and prepare bread, cake, cookies, crackers and noodles using this mixture to improve health Was prepared.
2-2. 스프 및 육즙(gravies)의 제조2-2. Preparation of soups and gravy
본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물을 각각 0.1 ~ 1.0 중량부를 스프 및 육즙에 첨가하여 통상의 방법으로 건강 증진용 육가공 제품, 면류의 수프 및 육즙을 제조하였다.0.1 to 1.0 parts by weight of the mixed extracts of golden, yellow and white, gardenia, yellow lotus, Schisandra chinensis and jade porridge according to the present invention were added to soups and broth, respectively, to prepare meat products for health promotion, soups and broths in a conventional manner.
2-3. 그라운드 비프(ground beef)의 제조2-3. Preparation of Ground Beef
본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물을 각각 10 중량부를 그라운드 비프에 첨가하여 통상의 방법으로 건강 증진용 그라운드 비프를 제조하였다.10 parts by weight of the mixed extracts of golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade in accordance with the present invention were added to ground beef to prepare ground beef for health promotion in a conventional manner.
2-4. 유제품(dairy products)의 제조2-4. Manufacture of dairy products
본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물 5 ~ 10 중량%를 우유에 첨가하고, 상기 우유를 이용하여 버터 및 아이스크림과 같은 다양한 유제품을 제조하였다.5-10% by weight of the mixed extract of golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade porridge according to the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.
2-5. 선식의 제조2-5. Manufacture of wire
현미, 보리, 찹쌀, 율무를 공지의 방법으로 알파화시켜 건조시킨 것을 배전한 후 분쇄기로 입도 60메쉬의 분말로 제조하였다.Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted to prepare a powder having a particle size of 60 mesh.
검정콩, 검정깨, 들깨도 공지의 방법으로 쪄서 건조시킨 것을 배전한 후 분쇄기로 입도 60 메쉬의 분말로 제조하였다.Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer.
본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물을 진공 농축기에서 감압농축하고, 분무, 열풍건조기로 건조하여 얻은 건조물을 분쇄기로 입도 60 메쉬로 분쇄하여 건조분말을 얻었다.The extract of golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade porridge according to the present invention was concentrated under reduced pressure in a vacuum concentrator, and dried by spraying and drying with a hot air dryer to pulverize the dried product to a particle size of 60 mesh to obtain a dry powder.
상기에서 제조한 곡물류, 종실류 및 본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물의 건조분말을 다음의 비율로 배합하여 제조하였다.The dry powders of the grains, seeds and the extracts of the golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade porcine according to the present invention were prepared by blending in the following ratios.
곡물류(현미 30중량%, 율무 15중량%, 보리 20중량%),Cereals (30% by weight brown rice, 15% by weight radish, 20% by weight barley),
종실류(들깨 7중량%, 검정콩 8중량%, 검정깨 7중량%),Seeds (7% by weight perilla, 8% by weight black beans, 7% by weight black sesame),
본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물의 건조분말(3 중량%),Dry powder (3% by weight) of the mixed extract of golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade porridge according to the present invention,
영지(0.5중량%),Ganoderma lucidum (0.5% by weight),
지황(0.5중량%)Foxglove (0.5 wt%)
<제제예 3> 음료의 제조Preparation Example 3 Preparation of Beverage
3-1. 탄산음료의 제조3-1. Preparation of Carbonated Drinks
설탕 5~10%, 구연산 0.05~0.3%, 카라멜 0.005~0.02%, 비타민 C 0.1~1%의 첨가물을 혼합하고, 여기에 79~94%의 정제수를 섞어서 시럽을 만들고, 상기 시럽을 85~98℃에서 20~180초간 살균하여 냉각수와 1:4의 비율로 혼합한 다음 탄산가스를 0.5~0.82%를 주입하여 본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물을 포함하는 탄산음료를 제조하였다.5-10% of sugar, 0.05-0.3% citric acid, 0.005-0.02% caramel, 0.1-1% of vitamin C are mixed, and 79-94% purified water is mixed to make syrup, and the syrup is 85-98 Sterilizing at 20 ℃ for 180 seconds and mixed with a cooling water at a ratio of 1: 4, and then inject 0.5 ~ 0.82% of carbon dioxide gas, containing a mixed extract of gold, yellow white, gardenia, yellow lotus, Schisandra chinensis and jade according to the present invention Carbonated drinks were prepared.
3-2. 건강음료의 제조3-2. Manufacture of health drinks
액상과당(0.5%), 올리고당(2%), 설탕(2%), 식염(0.5%), 물(75%)과 같은 부재료와 본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물을 균질하게 배합하여 순간 살균을 한 후 이를 유리병, 패트병 등 소포장 용기에 포장하여 건강음료를 제조하였다.Of subsidiary materials such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%), water (75%), and gold, yellowish white, gardenia, yellow lotus, schizandra and jade porridge according to the present invention. The mixed extract was homogeneously blended and sterilized immediately and then packaged in a small packaging container such as a glass bottle or a plastic bottle to prepare a healthy beverage.
3-3. 야채쥬스의 제조3-3. Preparation of Vegetable Juice
본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물 5g을 토마토 또는 당근 쥬스 1,000㎖에 가하여 건강 증진용 야채쥬스를 제조하였다.5 g of a mixed extract of golden, yellowish white, gardenia, yellow lotus, Schisandra chinensis and jade porridge according to the present invention was added to 1,000 ml of tomato or carrot juice to prepare vegetable juice for health promotion.
3-4. 과일쥬스의 제조3-4. Preparation of Fruit Juice
본 발명에 따른 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물 1g을 사과 또는 포도 쥬스 1,000㎖에 가하여 건강 증진용 과일쥬스를 제조하였다.1 g of a mixed extract of golden, yellow and white, gardenia, yellow lotus, Schisandra chinensis and jade in accordance with the present invention was added to 1,000 ml of apple or grape juice to prepare a fruit juice for health promotion.
도 1a는 황금, 황백, 치자, 황련, 오미자 및 옥죽의 각 추출물이 NCI-H716 대장암세포의 GLP-1 분비에 미치는 영향을 나타낸 도이다. Figure 1a is a diagram showing the effect of GLP-1 secretion of NCI-H716 colorectal cancer cells extract of gold, yellow white, gardenia, yellow lotus, Schisandra chinensis and jade porridge.
도 1b는 황금, 황백, 치자, 황련, 오미자 및 옥죽의 각 추출물이 Min6 베타세포의 인슐린 분비에 미치는 영향을 나타낸 도이다. Figure 1b is a diagram showing the effect of each extract of gold, yellow white, gardenia, yellow lotus, Schisandra chinensis and jade porridge on insulin secretion of Min6 beta cells.
도 2는 대조군, 추출물A, 추출물B, 황금 추출물 및 오미자 추출물을 투여한 실험 동물군의 1일 식이 섭취량, 체중 및 내장 지방 정도(epidydimal fat pad)를 나타낸 도이다. Figure 2 is a diagram showing the daily dietary intake, body weight and degree of epididymal fat pad of the control animals, extract A, extract B, golden extract and Schizandra extract.
도 3a는 대조군, 추출물A, 추출물B, 황금 추출물 및 오미자 추출물을 투여한 실험 동물군에 포도당을 투여한 후의 시간에 따른 혈청 중 포도당 농도를 나타낸 도이다. Figure 3a is a diagram showing the concentration of glucose in serum with time after administration of glucose to the experimental animal groups to which the control group, extract A, extract B, golden extract and Schizandra extract.
도 3b는 대조군, 추출물A, 추출물B, 황금 추출물 및 오미자 추출물을 투여한 군의 혈청 중 포도당 농도와 인슐린 농도의 AU를 나타낸 도이다(이 때, AU(Artificial Unit)는 혈청 중 최고 농도를 나타내는 시간으로부터 1분 동안의 평균 농도를 말한다. 또한, 추출물A는 황금, 황백, 치자 및 황련의 혼합 추출물이고, 추출물B는 황금, 황백, 치자, 황련, 오미자 및 옥죽의 혼합 추출물이다.) Figure 3b is a diagram showing the AU of glucose and insulin concentrations in the serum group of the control group, extract A, extract B, golden extract and Schizandra extract (wherein AU (Artificial Unit) represents the highest concentration in the serum) Mean concentration for 1 minute from time, and extract A is a mixed extract of golden, yellow white, gardenia and yellow lotus, and extract B is a mixed extract of golden, yellow white, gardenia, yellow lotus, Schisandra chinensis and jade.)
도 4a는 대조군, 추출물A, 추출물B, 황금 추출물 및 오미자 추출물을 투여한 군의 포도당 주입률 및 포도당 섭취율을 나타낸 도이다. Figure 4a is a diagram showing the glucose infusion rate and glucose intake rate of the group administered the control group, extract A, extract B, golden extract and Schizandra extract.
도 4b는 대조군, 추출물A, 추출물B, 황금 추출물 및 오미자 추출물을 투여한 군의 간에서의 기저 포도당 생성속도와 고농도 인슐린 상태에서의 포도당 생성 속도를 나타낸 도이다. Figure 4b is a diagram showing the baseline glucose production rate in the liver of the control group, extract A, extract B, golden extract and Schisandra chinensis extract and glucose production rate in high insulin state.
도 5a는 대조군, 추출물A, 추출물B, 황금 추출물 및 오미자 추출물을 투여한 군에 대하여 포도당을 투여한 후의 혈청 중 인슐린 농도을 나타낸 도이다. Figure 5a is a diagram showing the concentration of insulin in the serum after administration of glucose for the group administered the control group, extract A, extract B, golden extract and Schizandra extract.
도 5b는 대조군, 추출물A, 추출물B, 황금 추출물 및 오미자 추출물을 투여한 군의 1차 단계와 2차 단계의 AU를 나타낸 도이다. Figure 5b is a diagram showing the AU of the first step and the second step of the group administered the control group, extract A, extract B, golden extract and Schizandra extract.
도 6은 대조군, 추출물A, 추출물B, 황금 추출물 및 오미자 추출물을 투여한 군의 IRS2의 발현 정도를 나타낸 도이다. Figure 6 is a diagram showing the expression level of IRS2 in the group administered with the control, extract A, extract B, golden extract and Schizandra extract.
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Cited By (2)
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CN102935136A (en) * | 2012-11-28 | 2013-02-20 | 重庆开洲九鼎牧业科技开发有限公司 | Heat-clearing and toxicity-removing composition and feed for livestock as well as preparation method and application thereof |
WO2014109587A1 (en) * | 2013-01-10 | 2014-07-17 | 한국한의학연구원 | Pharmaceutical composition and functional food comprising natural extracts for preventing or treating diabetic complications or angiodema |
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Cited By (3)
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CN102935136A (en) * | 2012-11-28 | 2013-02-20 | 重庆开洲九鼎牧业科技开发有限公司 | Heat-clearing and toxicity-removing composition and feed for livestock as well as preparation method and application thereof |
WO2014109587A1 (en) * | 2013-01-10 | 2014-07-17 | 한국한의학연구원 | Pharmaceutical composition and functional food comprising natural extracts for preventing or treating diabetic complications or angiodema |
US9844575B2 (en) | 2013-01-10 | 2017-12-19 | Korea Institute Of Oriental Medicine | Pharmaceutical composition and functional food comprising natural extracts for preventing or treating diabetic complications or angiodema |
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