KR102151011B1 - Composition for preventing, improving or treating liver disease comprising fermented liquor of aged sprout ginseng extract as effective component - Google Patents
Composition for preventing, improving or treating liver disease comprising fermented liquor of aged sprout ginseng extract as effective component Download PDFInfo
- Publication number
- KR102151011B1 KR102151011B1 KR1020170139596A KR20170139596A KR102151011B1 KR 102151011 B1 KR102151011 B1 KR 102151011B1 KR 1020170139596 A KR1020170139596 A KR 1020170139596A KR 20170139596 A KR20170139596 A KR 20170139596A KR 102151011 B1 KR102151011 B1 KR 102151011B1
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- KR
- South Korea
- Prior art keywords
- sprout ginseng
- ginseng extract
- liver
- aged
- liver disease
- Prior art date
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Abstract
본 발명은 숙성새싹삼 추출물의 발효물을 유효성분으로 함유하는 간질환의 예방, 개선 또는 치료용 건강기능식품 및 약학 조성물에 관한 것이다.The present invention relates to a health functional food and a pharmaceutical composition for the prevention, improvement or treatment of liver disease containing a fermented product of mature sprout ginseng extract as an active ingredient.
Description
본 발명은 숙성새싹삼 추출물의 발효물을 유효성분으로 함유하는 간질환의 예방, 개선 또는 치료용 조성물에 관한 것이다.The present invention relates to a composition for the prevention, improvement or treatment of liver disease containing a fermented product of mature sprout ginseng extract as an active ingredient.
간은 인간의 신체장기 중 생체 내 대사가 가장 활발하게 일어나는 장기로, 인체 내 소화계와 전신순환계 사이에 위치하면서 외부에서 들어온 생체 외 물질로부터 전신을 방어하는 기능을 수행하고 있다. 생체 내로 들어온 생체 외 물질은 일단 간을 통과하고, 영양소 이외에 많은 독성물질에 노출될 위험이 다른 장기에 비해 많으므로 그만큼 손상될 확률도 매우 높다. 그러나 간은 재생능력이 우수하여 약간의 손상이 있을 경우에는 충분히 정상으로 회복 가능하지만, 손상이 지속될 경우에는 간 조직의 일부가 완전히 파괴되거나 간 기능이 저하되어 정상적인 간으로의 회복이 어려울 수 있다. 이러한 간 손상이 만성화되면 그 원인에 상관없이 간 섬유화, 간경화 또는 간암 등으로 진행된다.The liver is an organ in which metabolism occurs most actively in the body among human organs, and it is located between the digestive system and the systemic circulation system in the human body and functions to protect the whole body from in vitro substances from outside. The ex vivo material that has entered the living body once passes through the liver, and the risk of exposure to many toxic substances other than nutrients is higher than that of other organs, so the probability of damage is very high. However, the liver has excellent regenerative ability and can recover to normal if there is a slight damage, but if the damage persists, a part of the liver tissue is completely destroyed or the liver function is deteriorated, making it difficult to recover to a normal liver. When such liver damage becomes chronic, it progresses to liver fibrosis, cirrhosis or liver cancer, regardless of the cause.
간 손상을 유발하는 원인으로는 알콜의 과다섭취, 스트레스성 만성 피로, 지방성분이 포함된 음식 섭취, 바이러스의 감염, 각종 약품과 같은 유해물질 또는 영양부족 등이 있다. 알코올 섭취에 의해 알코올이 간으로 들어가게 되면, 간세포의 세포질 내에 존재하는 알코올 탈수소효소(alcohol dehydrogenase; ADH)에 의해 알코올이 아세트알데히드(acetaldehyde)로 산화되고, 아세트알데히드는 알데히드 탈수소효소(aldehyde dehydrogenase; ALDH)에 의해 아세테이트(acetate)로 전환된다. 특히 알코올의 최초 대사산물인 아세트알데히드는 알코올에 비해 반응성이 매우 높고 독성이 강하여 간세포에 손상을 줄 수 있고, 이러한 알코올 대사에 의해 지방산이 많이 생성되어 간에 지방이 축적된다. 이때 간에 축적된 지방간을 알코올성 지방간이라고 한다.Causes of liver damage include excessive intake of alcohol, chronic stressful fatigue, ingestion of fat-containing foods, viral infections, harmful substances such as various drugs, or lack of nutrition. When alcohol enters the liver by ingestion of alcohol, alcohol is oxidized to acetaldehyde by alcohol dehydrogenase (ADH) present in the cytoplasm of hepatocytes, and acetaldehyde is aldehyde dehydrogenase (ALDH). ) To acetate (acetate). In particular, acetaldehyde, the first metabolite of alcohol, is highly reactive and highly toxic than alcohol, which can damage hepatocytes, and fatty acids are produced by this alcohol metabolism to accumulate fat in the liver. At this time, the fatty liver accumulated in the liver is called alcoholic fatty liver.
알코올성 지방간은 만성 간질환으로 진행될 수 있는데, 알코올성 지방간 환자들 중에서 10-35%가 간염으로, 8-20%가 간경변증으로 진행되는 것으로 보고되고 있으며, 비알코올성 지방간 환자에 비하여 알코올성 지방간 환자들이 간경변증을 보다 많이 가지고 있고 간염으로 진행될 가능성도 높은 것으로 보고되고 있다.Alcoholic fatty liver can progress to chronic liver disease.It is reported that 10-35% of alcoholic fatty liver patients progress to hepatitis and 8-20% of liver cirrhosis, and alcoholic fatty liver patients suffer from cirrhosis compared to non-alcoholic fatty liver patients. It has been reported to have more and is likely to develop into hepatitis.
이러한 간 손상에 의한 간 질환을 치료하기 위한 방법으로는 크게 식이요법과 약제요법이 병용되고 있다. 대표적인 간 질환 치료제로는 실리마린(silymarin)과 비페닐디메틸디카르복실레이트(biphenyl dimethyl dicarboxylate, BDD)가 있으나, 이들 약물 역시 근복적인 치료제가 되지 못하고 있으며, 이외에 개발된 화합물에 의한 치료제는 체내 다양한 부작용을 유발할 수 있어 제품화되지 못하고 있는 실정이다. 따라서, 기존에 사용되고 있는 치료제를 대체할 수 있을 만큼 치료효과가 우수하며, 대량 또는 장기간 투여 시에도 부작용이 없는 천연물을 이용한 새로운 간질환 치료제의 개발이 요구되고 있다.Diet and pharmaceutical therapy are largely used in combination as a method for treating liver disease caused by such liver damage. Representative treatments for liver disease include silymarin and biphenyl dimethyl dicarboxylate (BDD), but these drugs also have not been able to provide a near-term treatment, and other treatments with developed compounds have various side effects in the body. It is not possible to commercialize the product because it can cause a problem. Therefore, there is a need to develop a new therapeutic agent for liver disease using natural products that are excellent enough to replace the existing therapeutic agents and have no side effects even when administered in large quantities or for a long period of time.
한편, 새싹삼은 보통 1~2년의 기간 동안 재배된 인삼으로, 뿌리보다 잎과 줄기에 사포닌 성분이 약 7~8배 더 함유되어 있으며 수경재배, 연중생산 가능 및 재배기간의 단축으로 인해 종래에 이용되는 뿌리인삼보다 낮은 가격을 형성하고 있다는 장점이 있어, 인삼가공품의 원료로서 사용이 용이하다. 사포닌의 주요 약리작용은 중추신경계, 내분비계, 면역계, 대사계 등에 영향을 미쳐 신체조절 기능에 다양한 효과를 발휘한다. 그 이외에도 사포닌은 지방분해력이 크고, 영양분흡수와 소화를 촉진시키며, 세포내의 효소활성화로 신진대사촉진, 에너지를 증가시켜 원기회복, 피로, 무력감, 식욕부진 개선, 혈청단백질 합성 촉진 등의 효과가 있다고 알려져 있다.On the other hand, sprout ginseng is usually grown for a period of 1 to 2 years, and it contains about 7 to 8 times more saponin in leaves and stems than the roots, and due to hydroponic cultivation, year-round production, and shortened cultivation period. It has the advantage of forming a lower price than the used root ginseng, so it is easy to use as a raw material for processed ginseng products. The main pharmacological action of saponin affects the central nervous system, endocrine system, immune system, metabolic system, etc., and exerts various effects on body control functions. In addition, saponin is said to have great lipolysis power, promote nutrient absorption and digestion, promote metabolism by activating intracellular enzymes, and increase energy to restore energy, improve fatigue, helplessness, loss of appetite, and promote serum protein synthesis. Is known.
한편, 한국등록특허 제1688002호에는 '초음파 처리된 인삼열매 추출물을 유효성분으로 함유하는 간질환 예방 또는 치료용 조성물'이 개시되어 있고, 한국등록특허 제1179171호에는 '산삼 또는 인삼을 포함한 인삼류의 형성층 유래 식물줄기세포주를 유효성분으로 함유하는 간질환의 예방 또는 치료용 조성물'이 개시되어 있으나, 본 발명의 숙성새싹삼 추출물의 발효물을 유효성분으로 함유하는 간질환의 예방, 개선 또는 치료용 조성물에 대해서는 기재된 바가 없다.On the other hand, Korean Patent No. 168002 discloses'a composition for preventing or treating liver disease containing an ultrasonic-treated ginseng fruit extract as an active ingredient', and Korean Patent No. 11179171 discloses'Ginseng including wild ginseng or ginseng. A composition for preventing or treating liver disease containing the cambium-derived plant stem cell line as an active ingredient is disclosed, but for preventing, improving or treating liver disease containing the fermented product of the mature sprout ginseng extract of the present invention as an active ingredient There is no description of the composition.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명자들은 락토바실러스 플란타룸(Lactobacillus plantarum)과 락토바실러스 브레비스(L. brevis)가 혼합된 유산균 혼합물에 의한 발효과정을 통해 제조된 숙성새싹삼 추출물의 발효물이 발효되지 않은 새싹삼 추출물에 비해, 알코올성 간손상 실험동물의 혈중 ALT 및 AST 함량을 감소시키고 간조직 내 지질축적을 억제시킬 뿐만 아니라, 간염 실험동물의 혈중 ALT 및 AST 함량을 감소시키는 것을 확인함으로써, 본 발명을 완성하였다. The present invention was derived from the above requirements, the inventors of the present inventors matured sprout ginseng prepared through a fermentation process using a mixture of Lactobacillus plantarum and Lactobacillus brevis ( L. brevis ). Compared to the unfermented sprout ginseng extract, the fermented product of the extract not only reduces blood ALT and AST content in alcoholic liver-damaged experimental animals, inhibits lipid accumulation in liver tissue, but also reduces blood ALT and AST content in hepatitis test animals. By confirming what to do, the present invention was completed.
상기 과제를 해결하기 위해, 본 발명은 숙성새싹삼 추출물의 발효물을 유효성분으로 함유하는 간질환의 예방 또는 개선용 건강기능식품 조성물을 제공한다.In order to solve the above problems, the present invention provides a health functional food composition for preventing or improving liver disease containing a fermented product of mature sprout ginseng extract as an active ingredient.
또한, 본 발명은 숙성새싹삼 추출물의 발효물을 유효성분으로 함유하는 간질환의 예방 또는 치료용 약학 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for preventing or treating liver disease, containing a fermented product of mature sprout ginseng extract as an active ingredient.
본 발명의 숙성새싹삼 추출물의 발효물을 유효성분으로 포함하는 조성물은 천연 약재인 새싹삼을 유효성분으로 사용하므로 인체에 부작용이 없으며, 알코올성 간손상 및 감염 동물모델에서 혈중 ALT 및 AST 함량의 감소, 간조직 내 지질축적 또는 간세포 사멸의 억제 효과가 우수하므로, 본 발명의 조성물은 간질환의 예방, 개선 또는 치료용 건강기능식품 및 의약품에 유용하게 사용될 수 있을 것으로 기대된다. The composition containing the fermented product of the aged sprout ginseng extract of the present invention as an active ingredient uses sprout ginseng, a natural medicinal material, so there is no side effect to the human body, and decreases the content of ALT and AST in blood in an animal model of alcoholic liver injury and infection. , Since the inhibitory effect of lipid accumulation or hepatocyte death in liver tissue is excellent, the composition of the present invention is expected to be useful in health functional foods and pharmaceuticals for preventing, improving or treating liver disease.
도 1은 알코올성 간손상 동물모델에서 새싹삼 추출물 및 숙성새싹삼 추출물의 발효물을 투여한 후 혈중 ALT 및 AST 함량의 감소를 확인한 결과이다. A; 10 ml/kg의 에탄올 섭취군, B; 30 ml/kg의 에탄올 섭취군, No treatment; 무처리 대조군(정상 마우스), Water; 물(대조군), SG; 새싹삼 추출물, FSG; 숙성새싹삼 추출물의 발효물.
도 2는 알코올성 간손상 동물모델에서 새싹삼 추출물 및 숙성새싹삼 추출물의 발효물을 투여한 후 간조직 내 지질축적 억제 효과를 헤마톡실린-에오신 염색을 통해 확인한 결과이다. A; 100 배율, B; 200 배율, No treatment; 무처리 대조군(정상 마우스), Water; 물(대조군), SG; 새싹삼 추출물, FSG; 숙성새싹삼 추출물의 발효물.
도 3은 GalN/LPS로 유도된 염증성 간손상 동물모델에서 새싹삼 추출물 및 숙성새싹삼 추출물의 발효물을 투여한 후 혈중 ALT 및 AST 함량의 감소 효과를 확인한 결과이다. control; 무처리 대조군(정상 마우스), Water; 물(대조군), SG; 새싹삼 추출물, FSG; 숙성새싹삼 추출물의 발효물.
도 4는 GalN/LPS로 유도된 염증성 간손상 동물모델에서 새싹삼 추출물 및 숙성새싹삼 추출물의 발효물을 투여한 후 간 조직에서 간 세포 괴사 억제 효과를 헤마톡실린-에오신 염색을 통해 확인한 결과이다. control; 무처리 대조군(정상 마우스), Water; 물(대조군), SG; 새싹삼 추출물, FSG; 숙성새싹삼 추출물의 발효물.
도 5는 GalN/LPS로 유도된 염증성 간손상 동물모델에서 새싹삼 추출물 및 숙성새싹삼 추출물의 발효물을 투여한 후 TUNEL 염색을 통해 간조직 내 세포사멸 억제 효과를 확인한 결과이다. Control; 무처리 대조군(정상 마우스), water; 물(대조군), SG; 새싹삼 추출물, FSG; 숙성새싹삼 추출물의 발효물. 1 is a result of confirming the decrease in blood ALT and AST content after administration of fermented products of sprout ginseng extract and aged sprout ginseng extract in an alcoholic liver injury animal model. A; 10 ml/kg ethanol intake group, B; 30 ml/kg ethanol intake group, No treatment; Untreated control (normal mouse), Water; Water (control), SG; Sprout ginseng extract, FSG; Fermented product of aged sprout ginseng extract.
2 is a result of confirming the effect of inhibiting lipid accumulation in liver tissue through hematoxylin-eosin staining after administration of fermented products of sprout ginseng extract and mature sprout ginseng extract in an alcoholic liver injury animal model. A; 100 magnification, B; 200 magnification, No treatment; Untreated control (normal mouse), Water; Water (control), SG; Sprout ginseng extract, FSG; Fermented product of aged sprout ginseng extract.
3 is a result of confirming the effect of reducing ALT and AST content in blood after administration of fermented products of sprout ginseng extract and aged sprout ginseng extract in an animal model of GalN/LPS-induced inflammatory liver injury. control; Untreated control (normal mouse), Water; Water (control), SG; Sprout ginseng extract, FSG; Fermented product of aged sprout ginseng extract.
Figure 4 is a result of confirming the effect of inhibiting liver cell necrosis in liver tissue through hematoxylin-eosin staining after administration of fermented products of sprout ginseng extract and mature sprout ginseng extract in an animal model of inflammatory liver injury induced by GalN/LPS. . control; Untreated control (normal mouse), Water; Water (control), SG; Sprout ginseng extract, FSG; Fermented product of aged sprout ginseng extract.
5 is a result of confirming the effect of inhibiting apoptosis in liver tissue through TUNEL staining after administering a fermented product of a sprout ginseng extract and a mature sprout ginseng extract in an animal model of GalN/LPS-induced inflammatory liver injury. Control; Untreated control group (normal mouse), water; Water (control), SG; Sprout ginseng extract, FSG; Fermented product of aged sprout ginseng extract.
본 발명의 목적을 달성하기 위하여, 본 발명은 숙성새싹삼 추출물의 발효물을 유효성분으로 함유하는 간질환의 예방 또는 개선용 건강기능식품 조성물을 제공한다. In order to achieve the object of the present invention, the present invention provides a health functional food composition for preventing or improving liver disease containing a fermented product of mature sprout ginseng extract as an active ingredient.
본 발명에 따른 건강기능식품 조성물에 있어서, 상기 숙성새싹삼 추출물의 발효물은 새싹삼 추출물에 유산균 혼합물을 접종한 후 발효시키는 단계를 포함하여 제조될 수 있으며, 상기 유산균 혼합물은 락토바실러스 플란타룸(Lactobacillus plantarum) 균주 및 락토바실러스 브레비스(Lactobacillus brevis) 균주가 혼합된 것일 수 있으며, 더욱 바람직하게는, 각각 1×108~1×1010 CFU/ml의 농도로 준비된 기탁번호 KACC 91848P인 락토바실러스 플란타룸(Lactobacillus plantarum) P1201 균주 및 기탁번호 KACC 92156P인 락토바실러스 브레비스(Lactobacillus brevis) BMK184 균주가 1:1의 비율로 혼합된 것일 수 있으나, 이에 제한되지 않는다. In the health functional food composition according to the present invention, the fermented product of the aged sprout ginseng extract may be prepared by inoculating the sprout ginseng extract with a lactic acid bacteria mixture and fermenting it, and the lactic acid bacteria mixture is Lactobacillus plantarum ( Lactobacillus plantarum ) strain and Lactobacillus brevis ( Lactobacillus brevis ) strain may be a mixture, and more preferably, Lactobacillus with deposit number KACC 91848P prepared at a concentration of 1×10 8 ~ 1×10 10 CFU/ml, respectively Plantarum ( Lactobacillus plantarum ) P1201 strain and the accession number KACC 92156P Lactobacillus brevis ( Lactobacillus brevis ) BMK184 strain may be mixed in a ratio of 1:1, but is not limited thereto.
본 발명의 숙성새싹삼 추출물의 발효물 제조방법은 구체적으로는, The fermented product manufacturing method of the aged sprout ginseng extract of the present invention is specifically,
(1) 새싹삼을 증자한 후 숙성시키는 단계;(1) aging after increasing sprout ginseng;
(2) 상기 (1) 단계의 숙성된 새싹삼을 건조시키고, 건조된 숙성새싹삼에 물을 혼합하여 추출하는 단계;(2) drying the matured sprout ginseng in step (1), and extracting the dried sprout ginseng by mixing water;
(3) 상기 (2) 단계의 제조된 숙성새싹삼 추출물에 당을 혼합한 후 살균하는 단계; 및(3) sterilizing after mixing sugar in the ripened sprout ginseng extract prepared in step (2); And
(4) 상기 (3) 단계의 살균된 숙성새싹삼 추출물에 유산균 혼합물을 접종한 후 발효시키는 단계를 포함하여 제조될 수 있으며, 더욱 구체적으로는, (4) It may be prepared including the step of fermenting after inoculating a mixture of lactic acid bacteria in the sterilized aged sprout ginseng extract of step (3), and more specifically,
(1) 25~30일간 재배된 새싹삼을 흐르는 물에 3회 세척하고 3~5cm 두께로 세절하여 95~120℃에서 25~35분 동안 증자한 후 70~80℃에서 5~10일간 숙성시키는 단계;(1) Wash sprout ginseng grown for 25 to 30 days in running water 3 times, cut it into 3 to 5 cm thickness, increase it at 95 to 120°C for 25 to 35 minutes, and then mature at 70 to 80°C for 5 to 10 days. step;
(2) 상기 (1) 단계의 숙성된 새싹삼을 50~55℃에서 2~3일간 건조시키고, 건조된 숙성새싹삼에 물을 중량 대비 20배 가하여 95~100℃에서 4시간 동안 추출하여 1.5 브릭스(brix)의 숙성새싹삼 추출물을 제조하는 단계;(2) Dry the aged sprout ginseng in step (1) for 2 to 3 days at 50-55°C, add water 20 times by weight to the dried mature sprout ginseng, and extract it at 95-100°C for 4 hours to 1.5 Preparing an aged sprout ginseng extract of brix;
(3) 상기 (2) 단계의 제조된 숙성새싹삼 추출물에 당을 추출물 중량 대비 2% (w/v)로 혼합한 후 121℃에서 15분 동안 살균하는 단계; 및(3) sterilizing at 121° C. for 15 minutes after mixing sugar in 2% (w/v) based on the weight of the extract in the matured sprout ginseng extract prepared in step (2); And
(4) 상기 (3) 단계의 살균된 숙성새싹삼 추출물에 각각 1×109 CFU/ml의 농도로 준비된 기탁번호 KACC 91848P인 락토바실러스 플란타룸(Lactobacillus plantarum) P1201 균주 및 기탁번호 KACC 92156P인 락토바실러스 브레비스(Lactobacillus brevis) BMK184 균주가 1:1의 비율로 혼합된 유산균 혼합물을 추출물 중량 대비 5% (v/v)로 접종한 후, 25~35℃에서 2~5일 동안 발효시키는 단계를 포함하여 제조될 수 있으나, 이에 제한되지 않는다.(4) Lactobacillus plantarum ( Lactobacillus plantarum ) P1201 strain and accession number KACC 91848P prepared at a concentration of 1×10 9 CFU/ml, respectively, in the sterilized aged sprout ginseng extract of step (3) Lactobacillus brevis ( Lactobacillus brevis ) After inoculating a mixture of lactic acid bacteria mixed in a ratio of 1:1 BMK184 strain at 5% (v/v) based on the weight of the extract, fermentation at 25 to 35° C. for 2 to 5 days was performed. It may be manufactured including, but is not limited thereto.
상기 (3)단계의 당은 유산균이 당을 분해하여 영양분을 형성함으로써 발효를 더욱 원활하게 이루도록 하기 위하여 첨가된다. 본 발명에 사용할 수 있는 당류로는 식품으로 사용할 수 있는 것으로 설탕, 포도당, 과당, 엿류, 당시럽류, 덱스트린, 올리고당류 중에서 하나를 사용하거나 이들을 혼합하여 사용할 수 있고, 바람직하게는 설탕일 수 있으나, 이에 제한되지 않는다.The sugar in step (3) is added to facilitate fermentation by forming nutrients by decomposing sugar by lactic acid bacteria. Sugars that can be used in the present invention may be used as food, and one of sugar, glucose, fructose, malt, sugar syrup, dextrin, and oligosaccharides may be used, or a mixture thereof may be used, preferably sugar. It is not limited thereto.
본 발명의 건강기능식품 조성물에 있어서, 상기 간질환은 알코올성 지방간, 지방간, 만성 간염, 급성 간염, 알코올성 간염, 간경변 또는 알코올성 간경변일 수 있고, 바람직하게는 알코올성 지방간 또는 간염일 수 있으나, 이에 제한되지 않는다.In the health functional food composition of the present invention, the liver disease may be alcoholic fatty liver, fatty liver, chronic hepatitis, acute hepatitis, alcoholic hepatitis, cirrhosis or alcoholic cirrhosis, preferably alcoholic fatty liver or hepatitis, but is not limited thereto. Does not.
용어 '지방간(fatty liver)'은 정상 간에서 지방이 차지하는 비율(5%)보다 많은 양의 지방이 간에 축적된 것을 의미한다. 상기 지방간은 알코올성 지방간, 또는 비만, 당뇨병, 고지혈증 또는 약물 등으로 인한 비알코올성 지방간일 수 있다.The term'fatty liver' means that a greater amount of fat is accumulated in the liver than the percentage (5%) of fat in the normal liver. The fatty liver may be alcoholic fatty liver, or non-alcoholic fatty liver due to obesity, diabetes, hyperlipidemia, or drugs.
용어 '간염'은 간세포 및 간 조직의 염증을 의미하며, 만성 간염에 의하여 장기간에 걸쳐 간세포가 파괴되고 재생하는 과정이 반복되면 간에 섬유조직과 재생 결절이 증가하여 간경변 또는 간경화가 진행될 수 있다. 간경변이 일정 수준 이상으로 진행되면, 간성뇌증(Hepatic encephalopathy), 식도정맥류(Esophageal varix) 등의 합병증을 유발할 수 있다.The term'hepatitis' refers to inflammation of hepatocytes and liver tissues, and when hepatocytes are destroyed over a long period of time due to chronic hepatitis and the process of regeneration is repeated, fibrous tissues and regenerated nodules in the liver may increase, leading to cirrhosis or cirrhosis of the liver. If cirrhosis progresses above a certain level, complications such as hepatic encephalopathy and esophageal varix can occur.
상기 건강기능식품 조성물은 분말, 과립, 환, 정제, 캡슐, 캔디, 시럽, 발포정 및 음료 중에서 선택된 어느 하나의 제형으로 제조될 수 있으나, 이에 제한되지 않는다. 상기 건강기능식품 조성물은 간질환을 예방하거나 개선하기 위해 섭취할 수 있는 것이면 특별히 제한되지 않는다.The health functional food composition may be prepared in any one formulation selected from powders, granules, pills, tablets, capsules, candy, syrup, effervescent tablets, and beverages, but is not limited thereto. The health functional food composition is not particularly limited as long as it can be consumed to prevent or improve liver disease.
본 발명의 숙성새싹삼 추출물의 발효물을 식품첨가물로 사용하는 경우, 상기 숙성새싹삼 추출물의 발효물을 그대로 첨가하거나 다른 식품 또는 식품성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효성분은 그의 사용 목적(예방 또는 개선)에 따라 적절하게 사용될 수 있다. 일반적으로, 식품 또는 음료의 제조시에 본 발명의 숙성새싹삼 추출물의 발효물은 원료에 대하여 15 중량부 이하, 바람직하게는 10 중량부 이하의 양의로 첨가된다. 그러나 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 안전성 면에서 아무런 문제가 없는 범위의 양으로 사용될 수 있다.When the fermented product of the aged sprout ginseng extract of the present invention is used as a food additive, the fermented product of the matured sprout ginseng extract may be added as it is, or may be used with other foods or food ingredients, and may be appropriately used according to a conventional method. have. The active ingredient may be appropriately used depending on the purpose of use (prevention or improvement). In general, in the manufacture of food or beverage, the fermented product of the aged sprout ginseng extract of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less based on the raw material. However, in the case of long-term intake for the purpose of health control, it can be used in an amount within a range without any problems in terms of safety.
상기 식품의 종류에는 특별한 제한은 없다. 상기 건강기능식품 조성물을 첨가할 수 있는 식품의 예로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스낵류, 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. 상기 음료는 탄산음료, 기능성이온음료, 쥬스(예를 들어, 사과, 배,포도, 알로에, 감귤, 복숭아, 당근, 토마토쥬스 등), 식혜 등을 포함한다. There is no particular limitation on the type of food. Examples of foods to which the health functional food composition can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea There are drinks, alcoholic beverages, and vitamin complexes, and all health foods in the usual sense are included. The beverage includes carbonated beverages, functional ionic beverages, juices (eg, apples, pears, grapes, aloe, tangerines, peaches, carrots, tomato juice, etc.), sikhye, and the like.
본 발명의 기능성 식품은 식품 제조 시에 통상적으로 첨가되는 성분을 포함하며, 예를 들어, 단백질, 탄수화물, 지방, 영양소 및 조미제를 포함한다. 예컨대, 드링크제로 제조되는 경우에는 유효성분 이외에 천연 탄수화물 또는 향미제를 추가 성분으로서 포함시킬 수 있다. 상기 천연 탄수화물은 모노사카라이드(예컨대, 글루코오스, 프럭토오스 등), 디사카라이드(예컨대, 말토스, 수크로오스 등), 올리고당, 폴리사카라이드(예컨대, 덱스트린, 시클로덱스트린 등) 또는 당알코올(예컨대, 자일리톨, 소르비톨, 에리쓰리톨 등)인 것이 바람직하다. 상기 향미제는 천연 향미제(예컨대, 타우마틴, 스테비아 추출물 등)와 합성 향미제(예컨대, 사카린, 아스파르탐 등)를 이용할 수 있다. 상기 건강기능식품 조성물 외에 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, 탄산음료에 사용되는 탄산화제 등을 더 함유할 수 있다.Functional foods of the present invention include ingredients that are commonly added during food production, and include, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, when prepared as a drink, natural carbohydrates or flavoring agents may be included as additional ingredients in addition to the active ingredient. The natural carbohydrates are monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, etc.), oligosaccharides, polysaccharides (eg, dextrins, cyclodextrins, etc.) or sugar alcohols (eg , Xylitol, sorbitol, erythritol, etc.). The flavoring agent may be a natural flavoring agent (eg, taumatin, stevia extract, etc.) and a synthetic flavoring agent (eg, saccharin, aspartame, etc.). In addition to the health functional food composition, various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonic acid It may further contain a carbonation agent used in beverages.
본 발명의 또 다른 목적을 달성하기 위하여, 본 발명은 숙성새싹삼 추출물의 발효물을 유효성분으로 함유하는 간질환의 예방 또는 치료용 약학 조성물을 제공한다.In order to achieve another object of the present invention, the present invention provides a pharmaceutical composition for preventing or treating liver disease containing a fermented product of mature sprout ginseng extract as an active ingredient.
본 발명의 상기 약학 조성물은, 유산균 혼합물에 의한 발효과정을 통해 간 세포 보호 효과, 혈중 ALT 및 AST의 함량의 감소 효과가 증가된 숙성새싹삼 추출물의 발효물을 유효성분으로 포함하고 있어, 간질환 예방 또는 치료에 효과가 있다. 상기 숙성새싹삼 추출물의 발효물 제조방법은 전술한 것과 같다.The pharmaceutical composition of the present invention contains, as an active ingredient, a fermented product of aged sprout ginseng extract having an increased effect of protecting liver cells and reducing the content of ALT and AST in blood through a fermentation process by a mixture of lactic acid bacteria, It is effective for prevention or treatment. The fermented product manufacturing method of the aged sprout ginseng extract is the same as described above.
본 발명의 상기 숙성새싹삼 추출물의 발효물을 포함하는 조성물은 약학적 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 또는 희석제를 더 포함할 수 있다. The composition containing the fermented product of the aged sprout ginseng extract of the present invention may further include a suitable carrier, excipient, or diluent commonly used in the manufacture of pharmaceutical compositions.
본 발명의 약학 조성물의 약학적 투여 형태는 단독으로 또는 타 약학적 활성 화합물과 결합뿐만 아니라 적당한 조합으로 사용될 수 있다.The pharmaceutical dosage form of the pharmaceutical composition of the present invention may be used alone or in combination with other pharmaceutically active compounds, as well as in an appropriate combination.
본 발명에 따른 약학 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제제, 외용제, 좌제 및 주사제의 형태로 제형화하여 사용될 수 있다. 상기 약학 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로오스, 메틸 셀룰로오스, 미정질 셀룰로오스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유 등을 포함한 다양한 화합물 혹은 혼합물을 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구 투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형 제제는 상기 발효물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트, 수크로오스 또는 락토오스, 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이 외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜, 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔, 마크로골, 트윈 61, 카카오지, 라우린지, 글리세로젤라틴 등이 사용될 수 있다.The pharmaceutical composition according to the present invention can be formulated and used in the form of oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and injections according to a conventional method. . Carriers, excipients, and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , Methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and these solid preparations contain at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. in the fermented product. It is prepared by mixing. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, and other excipients, such as water and liquid paraffin, which are commonly used simple diluents, such as wetting agents, sweeteners, fragrances, and preservatives. have. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc. may be used as the non-aqueous solvent and suspension. As a base for suppositories, Witepsol, Macrogol, Tween 61, cacao butter, laurin paper, glycerogelatin, and the like can be used.
본 발명의 상기 약학 조성물의 바람직한 투여량은 환자의 상태 및 체중,질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 본 발명의 약학 조성물은 쥐, 생쥐, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁 내 경막 또는 뇌혈관 내(intracerebroventricular) 주사에 의해 투여될 수 있다.The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and weight of the patient, the degree of disease, the form of the drug, the route of administration and the duration, but may be appropriately selected by those skilled in the art. The pharmaceutical composition of the present invention can be administered to mammals such as mice, mice, livestock, and humans by various routes. All modes of administration can be expected and may be administered by, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura mater or intracerebroventricular injection.
이하, 본 발명을 실시예에 의해 상세히 설명한다. 단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail by examples. However, the following examples are only illustrative of the present invention, and the contents of the present invention are not limited to the following examples.
제조예 1. 새싹삼 추출물 제조Preparation Example 1. Preparation of sprout ginseng extract
드림팜(유)로부터 공급받은 25일~30일간 재배된 새싹삼을 흐르는 물에 3회 세척하고, 물기를 제거하여 3~5cm 크기로 세절한 후 50~55℃에서 2~3일간 건조시켰다. 상기 건조된 새싹삼에 정제수를 중량 대비 60배 가하여 95~100℃에서 12시간 동안 추출하여 새싹삼 추출물을 제조하였다. The sprout ginseng cultivated for 25 to 30 days supplied from Dream Farm was washed three times with running water, dried, cut into 3 to 5 cm size, and dried at 50 to 55°C for 2 to 3 days. Purified water was added 60 times by weight to the dried sprout ginseng and extracted for 12 hours at 95-100°C to prepare sprout ginseng extract.
제조예 2. 숙성새싹삼 추출물의 발효물 제조Preparation Example 2. Preparation of fermented product of ripened sprout ginseng extract
상기 제조예 1에서 세절된 새싹삼을 95~100℃에서 30분 동안 증자하고 70~80℃에서 5~10일간 숙성시킨 다음 50~55℃에서 2~3일간 건조시켰다. 상기 건조된 숙성새싹삼에 정제수를 중량 대비 20배 첨가하고 95~100℃에서 4시간 동안 추출하여 발효를 위한 1.5 브릭스(brix)의 숙성새싹삼 추출물을 제조하였다.The sprout ginseng chopped in Preparation Example 1 was increased for 30 minutes at 95 to 100°C, aged at 70 to 80°C for 5 to 10 days, and then dried at 50 to 55°C for 2 to 3 days. Purified water was added 20 times by weight to the dried mature sprout ginseng and extracted for 4 hours at 95-100° C. to prepare a 1.5 brix aged sprout ginseng extract for fermentation.
상기 숙성새싹삼 추출물에 설탕을 추출물의 중량 대비 2%(w/v)로 첨가한 후 121℃에서 15분 동안 살균시키고, 살균된 추출물에 1×109 CFU/ml의 락토바실러스 플란타룸(Lactobacillus plantarum) P1201 균주(KACC 91848P) 및 1×109 CFU/ml의 락토바실러스 브레비스(L. brevis) BMK184 균주(KACC 92156P)를 추출물 중량 대비 2.5%(v/v)로 접종한 후 30~35℃에서 2~5일 동안 발효시켜 숙성새싹삼 추출물의 발효물을 제조하였다.Sugar was added to the aged sprout ginseng extract in an amount of 2% (w/v) based on the weight of the extract, and then sterilized at 121° C. for 15 minutes, and 1×10 9 CFU/ml of Lactobacillus plantarum ( Lactobacillus plantarum ) P1201 strain (KACC 91848P) and 1×10 9 CFU/ml of Lactobacillus brevis (L. brevis ) BMK184 strain (KACC 92156P) were inoculated at 2.5% (v/v) of the extract weight and 30-35 Fermented for 2 to 5 days at ℃ to prepare a fermented product of the aged sprout ginseng extract.
실시예 1. 숙성새싹삼 추출물의 발효물이 알코올성 간손상 동물모델에 미치는 영향 분석Example 1. Analysis of the effect of fermented product of ripened sprout ginseng extract on alcoholic liver injury animal model
알코올에 의해 유발된 간손상(지방간) 동물모델에서, 숙성새싹삼 추출물의 발효물이 혈중 AST와 ALT의 함량 및 간조직의 형태적 변화에 미치는 영향을 분석하였다. In an alcohol-induced liver injury (fatty liver) animal model, the effect of fermented products of aged sprout ginseng extract on blood AST and ALT content and morphological changes in liver tissue was analyzed.
1-1. 알코올에 의해 유발된 간손상 동물모델 준비1-1. Preparation of an animal model of alcohol-induced liver damage
4~5주령의 수컷 ICR 마우스는 사료 섭취로 인해 나타날 수 있는 위장관을 통한 알코올의 흡수 방해 현상을 배재하기 위해 12시간 동안 절식시켰다. 먼저 마우스에 물, 10 ml/kg의 새싹삼 추출물(SG), 2.5 ml/kg 또는 10 ml/kg의 숙성새싹삼 추출물의 발효물(FSG)을 각각 경구투여하고, 30분 후에 10 ml/kg 및 30 ml/kg의 18% 에탄올을 각각 복강투여하였다. Male ICR mice aged 4 to 5 weeks were fasted for 12 hours to rule out the obstruction of absorption of alcohol through the gastrointestinal tract that may occur due to feed intake. First, water, 10 ml/kg of sprout ginseng extract (SG), 2.5 ml/kg or 10 ml/kg of fermented sprout ginseng extract (FSG) were each orally administered to mice, and after 30 minutes, 10 ml/kg And 30 ml/kg of 18% ethanol, respectively, were administered intraperitoneally.
1-2. 혈중 AST 및 ALT 함량 변화 분석1-2. Analysis of changes in blood AST and ALT content
아스파라긴산 아미노전이효소(Aspartate aminotransferase, AST)와 알라닌 아미노전이효소(Alanine aminotransferase, ALT)는 간세포 안에서 아미노산 생성에 관여하는 효소로, 간세포가 손상될 경우 혈관 내로 방출되어 혈중 수치가 증가하게 된다. 따라서, 본 발명에서는 알코올 섭취에 따른 간 손상에 대해 숙성새싹삼 추출물의 발효물의 간손상 보호 효과를 분석하기 위해, 상기 실시예 1-1의 마우스에 에탄올 투여하고 6시간 후 혈액을 채취하고 혈장을 분리한 다음 혈중 AST 및 ALT의 함량을 측정하였다. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) are enzymes involved in the production of amino acids in hepatocytes, and when hepatocytes are damaged, they are released into blood vessels and blood levels increase. Therefore, in the present invention, in order to analyze the liver damage protection effect of the fermented product of the fermented sprout ginseng extract against liver damage caused by alcohol intake, ethanol was administered to the mouse of Example 1-1, and blood was collected and plasma was collected. After separation, the content of AST and ALT in blood was measured.
그 결과, 마우스의 혈중 AST 및 ALT 함량은, 에탄올 무처리군(no treatment)에 비해 에탄올 처리구에서 현저히 증가되었으며, 물을 전처리하고 에탄올을 투여한 대조군(water)에 비해 숙성새싹삼 추출물의 발효물을 전처리하고 에탄올을 투여한 실험군(FSG)에서 혈중 AST 및 ALT 함량이 감소하는 것이 확인되었다. 특히 같은 농도의 숙성새싹삼 추출물의 발효물을 섭취한 실험군은 새싹삼 추출물을 섭취한 실험군에 비해 혈중 AST 및 ALT 함량이 보다 감소되었으며, 숙성새싹삼 추출물의 발효물 처리 농도에 비례하여 혈중 AST 및 ALT 함량이 감소하는 것을 확인하였다(도 1). As a result, the blood AST and ALT content of mice was significantly increased in the ethanol-treated group compared to the ethanol-free group (no treatment), and the fermented product of the aged sprout ginseng extract compared to the control group (water) pretreated with water and administered ethanol. It was confirmed that the AST and ALT content in the blood decreased in the experimental group (FSG) that was pretreated and administered ethanol. In particular, the experimental group ingesting the fermented product of the same concentration of the fermented sprout ginseng extract decreased blood AST and ALT content more than the experimental group ingesting the sprout ginseng extract. It was confirmed that the ALT content decreased (FIG. 1).
1-3. 간조직의 형태적 변화 분석1-3. Analysis of morphological changes in liver tissue
알코올에는 영양소가 없지만 칼로리가 높기 때문에 간 조직의 지방축적에 원인이 될 수 있다. 따라서, 알코올로 유발된 간손상(지방간) 동물모델에서 숙성새싹삼 추출물의 발효물이 손상된 간조직에 대한 보호 효과를 분석하기 위해, 상기 실시예 1-1의 마우스에서 에탄올 투여 6시간 후에 간조직을 적출하였다. 적출된 간조직은 10% 중성 포르말린 용액(10% neutral formalin solution)으로 고정시키고 파라핀 포매(paraffin-embedded) 과정을 거친 후 5 ㎛ 두께의 절편으로 잘랐다. 상기 잘라진 간조직을 유리 슬라이드에 고정시키고 헤마톡실린-에오신 염색(hematoxylin and eosin staining)을 실시한 다음 100 및 200 배율에서 관찰하였다.Alcohol has no nutrients, but because it is high in calories, it can cause fat accumulation in liver tissue. Therefore, in order to analyze the protective effect of the fermented sprout ginseng extract on the damaged liver tissue in an alcohol-induced liver injury (fatty liver) animal model, liver tissue 6 hours after ethanol administration in the mouse of Example 1-1 Was extracted. The extracted liver tissue was fixed with 10% neutral formalin solution, paraffin-embedded, and then cut into 5 μm-thick sections. The cut liver tissue was fixed on a glass slide and subjected to hematoxylin and eosin staining, and then observed at 100 and 200 magnification.
그 결과, 무처리 정상 마우스의 간조직과 비교하여, 에탄올만 처리한 군(water + 30 ml/kg EtOH 또는 water + 10 ml/kg EtOH)에서는 지방의 축적이 증가되어 에오신으로 염색되지 않고 하얗게 관찰되는 부분이 증가된 것이 확인되었으나, 이러한 지방의 축적은 새싹삼 추출물을 전처리한 실험군에서 억제된 것이 확인되었고, 숙성새싹삼 추출물의 발효물을 전처리한 실험군에서는 지질 축적이 더욱 억제된 것이 확인되었다(도 2). As a result, compared with the liver tissue of untreated normal mice, in the ethanol-treated group (water + 30 ml/kg EtOH or water + 10 ml/kg EtOH), the accumulation of fat increased, and thus, it was not stained with eosin, but observed as white. It was confirmed that the amount of fat was increased, but it was confirmed that the accumulation of fat was inhibited in the experimental group pretreated with the sprout ginseng extract, and lipid accumulation was further suppressed in the experimental group pretreated with the fermented product of the aged sprout ginseng extract ( Fig. 2).
실시예 2. 숙성새싹삼 추출물의 발효물이 염증성 간손상 동물모델에 미치는 영향 분석Example 2. Analysis of the effect of fermented product of ripened sprout ginseng extract on inflammatory liver injury animal model
GalN/LPS로 유도된 염증성 간손상 동물모델에서, 숙성새싹삼 추출물의 발효물이 혈중 AST와 ALT의 함량, 간조직의 형태적 변화 및 간 세포 사멸에 미치는 영향을 분석하였다. In an animal model of GalN/LPS-induced inflammatory liver injury, the effect of fermented products of aged sprout ginseng extract on blood AST and ALT content, morphological changes in liver tissue, and liver cell death were analyzed.
2-1. 염증성 간손상 동물모델 준비2-1. Preparation of inflammatory liver injury animal model
8주령의 수컷 C57BL/6 마우스에 물, 10 ml/kg의 새싹삼 추출물(SG), 10 ml/kg의 숙성새싹삼 추출물의 발효물(FSG)을 각각 경구투여하고, 1시간 후에 LPS(lipopolysaccharide) 5 ㎍/㎏과 GalN(galactosamine) 700 ㎎/㎏을 복강투여하였다.Water, 10 ml/kg of sprout ginseng extract (SG), and 10 ml/kg of fermented product (FSG) of mature sprout ginseng extract were each orally administered to 8-week-old male C57BL/6 mice, and 1 hour later LPS (lipopolysaccharide ) 5 ㎍ / ㎏ and GalN (galactosamine) 700 ㎎ / ㎏ intraperitoneally administered.
2-2. 혈중 AST 및 ALT 함량 변화 분석2-2. Analysis of changes in blood AST and ALT content
GalN/LPS에 의한 염증성 간손상 동물모델에서 숙성새싹삼 추출물의 발효물의 간 손상 보호 효과를 분석하기 위해, 상기 실시예 2-1의 마우스에 GalN 및 LPS 투여 5시간 후 혈액을 채취하여 혈장을 분리한 다음 혈중 AST 및 ALT의 함량을 측정하였다. In order to analyze the liver damage protection effect of fermented sprout ginseng extract in an animal model of inflammatory liver injury by GalN/LPS, blood was collected 5 hours after the administration of GalN and LPS to the mouse of Example 2-1 to separate plasma. Then, the content of AST and ALT in blood was measured.
그 결과, 정상 마우스(control)에 비해, 물을 전처리하고 GalN/LPS를 처리한 군에서 혈중 AST 및 ALT 함량이 현저히 증가된 것이 확인되었고, 증가된 AST 및 ALT 함량은 새싹삼 추출물 및 숙성새싹삼 추출물의 발효물을 전처리한 실험군에서 현저히 감소된 것을 확인할 수 있었다. 특히, 숙성새싹삼 추출물의 발효물이 동일 농도의 새싹삼 추출물에 비해 혈중 AST 및 ALT 함량의 감소 효과가 더욱 우수함을 확인하였다(도 3). As a result, compared to normal mice (control), it was confirmed that the blood AST and ALT contents were significantly increased in the water-pretreated and GalN/LPS-treated group, and the increased AST and ALT contents were sprout ginseng extract and mature sprout ginseng. It was confirmed that the fermentation product of the extract was significantly reduced in the pretreated experimental group. In particular, it was confirmed that the fermented product of the aged sprout ginseng extract has a more excellent effect of reducing AST and ALT content in blood compared to the sprout ginseng extract of the same concentration (FIG. 3).
2-3. 간조직의 형태적 변화 분석2-3. Analysis of morphological changes in liver tissue
GalN/LPS에 의한 염증성 간손상 동물모델에서 숙성새싹삼 추출물의 발효물의 간 손상 보호 효과를 분석하기 위해, 상기 실시예 2-1의 GalN 및 LPS 투여 5시간 후의 마우스 간조직을 적출하였고, 헤마톡실린-에오신 염색을 수행하였다. 염색 과정은 상기 실시예 1-3과 동일하게 수행하였다. In order to analyze the liver damage protection effect of fermented sprout ginseng extract in an animal model of inflammatory liver injury by GalN/LPS, mouse liver tissues 5 hours after the administration of GalN and LPS of Example 2-1 were excised, and hematoxylin Cylin-eosin staining was performed. The dyeing process was performed in the same manner as in Examples 1-3.
그 결과, 물만 섭취한 대조군에서는 간세포의 괴사와 같은 간 손상 현상이 관찰되었으나, 새싹삼 추출물 및 숙성새싹삼 추출물의 발효물을 섭취한 실험군에서는 간 손상의 현상이 대조군에 비해 현저하게 억제된 것을 확인하였으며, 특히 숙성새싹삼 추출물의 발효물이 동일농도의 새싹삼 추출물에 비해 간손상 억제 효과가 더욱 우수함을 확인하였다(도 4).As a result, liver damage such as necrosis of hepatocytes was observed in the control group that consumed only water, but it was confirmed that the symptoms of liver damage were significantly suppressed in the experimental group ingesting the fermented product of sprout ginseng extract and mature sprout ginseng extract. In particular, it was confirmed that the fermented product of the aged sprout ginseng extract had more excellent effect of inhibiting liver damage than the sprout ginseng extract of the same concentration (FIG. 4).
2-4. 간 조직 내 세포사멸(apoptosis)의 억제 효과2-4. Inhibitory effect of apoptosis in liver tissue
GalN/LPS에 의한 염증성 간손상 동물모델에서 숙성새싹삼 추출물의 발효물이 간세포 사멸을 보호하는 효과가 있는지 분석하기 위해, 상기 실시예 2-1의 GalN 및 LPS 투여 6시간 후의 마우스에서 간 조직을 적출하였고, TUNEL 분석법(terminal deoxynucleotidyl transferase dUTP nick end labeling assay)을 실시하였다. 구체적으로, 적출된 간조직을 10% 중성 포르말린 용액으로 고정시키고 파라핀 포매 과정을 거친 후 5 ㎛ 두께의 절편으로 잘랐으며, 상기 잘라진 간조직은 in situ cell death detection kit(Roche, 스위스)를 사용하여 제조사의 지시에 따라 실험을 수행하였다. 사멸된 세포는 초록색 형광(green fluorescence)을 나타낸다.In order to analyze whether the fermented product of the fermented sprout ginseng extract has the effect of protecting hepatocyte death in an animal model of inflammatory liver injury caused by GalN/LPS, liver tissue in the mouse 6 hours after the administration of GalN and LPS of Example 2-1 was Extraction was performed, and a TUNEL assay (terminal deoxynucleotidyl transferase dUTP nick end labeling assay) was performed. Specifically, the extracted liver tissue was fixed with a 10% neutral formalin solution and then cut into 5 μm-thick sections after paraffin embedding, and the cut liver tissue was used in situ cell death detection kit (Roche, Switzerland). Experiments were performed according to the manufacturer's instructions. Dead cells show green fluorescence.
따라서, 도 5에 나타난 바와 같이, 무처리 정상 마우스에서 관찰되지 않던 사멸된 세포가 GalN/LPS에 의해 급격히 증가되었으나, 새싹삼 추출물 및 숙성새싹삼 추출물의 발효물을 섭취한 실험군에서는 초록색 형광의 사멸된 세포의 수가 감소한 것으로 보아, 본 발명의 숙성새싹삼 추출물의 발효물이 GalN/LPS에 의해 유도된 간세포 사멸을 억제하는 것을 확인하였으며, 특히 새싹삼 추출물에 비해 숙성새싹삼 추출물의 발효물에서 간세포 사멸이 더욱 효과적으로 억제됨을 알 수 있었다.Therefore, as shown in Figure 5, the dead cells that were not observed in the untreated normal mice were rapidly increased by GalN/LPS, but in the experimental group in which the fermented product of the sprout ginseng extract and the mature sprout ginseng extract was consumed, the green fluorescence was killed. It was found that the number of cells was decreased, and it was confirmed that the fermented product of the aged sprout ginseng extract of the present invention inhibits hepatocyte death induced by GalN/LPS. In particular, hepatocytes in the fermented product of the aged sprout ginseng extract compared to sprout ginseng extract It was found that death was more effectively suppressed.
상기의 결과를 통해, 본 발명의 숙성새싹삼 추출물의 발효물은 알코올 또는 염증 유발 인자에 의해 증가된 혈액 내 AST와 ALT의 함량을 효과적으로 감소시키며, 간 조직 내 지질 축적 또는 세포사멸을 억제함으로써, 알코올성 간손상 또는 비알코올성 간염과 같은 간질환을 개선하는 효과가 있음을 유추할 수 있었다.Through the above results, the fermented product of the aged sprout ginseng extract of the present invention effectively reduces the content of AST and ALT in the blood, which is increased by alcohol or inflammatory factors, and inhibits lipid accumulation or apoptosis in liver tissue, It could be inferred that it has the effect of improving liver diseases such as alcoholic liver injury or non-alcoholic hepatitis.
Claims (6)
상기 숙성새싹삼 추출물의 발효물은,
(1) 25~30일간 재배된 새싹삼을 흐르는 물에 세척하고 세절하여 95~120℃에서 25~35분 동안 증자한 후 70~80℃에서 5~10일간 숙성시키는 단계;
(2) 상기 (1) 단계의 숙성된 새싹삼을 50~55℃에서 2~3일간 건조시키고, 건조된 숙성새싹삼에 물을 가하여 95~100℃에서 추출하여 숙성새싹삼 추출물을 제조하는 단계;
(3) 상기 (2) 단계의 제조된 숙성새싹삼 추출물에 당을 혼합한 후 살균하는 단계; 및
(4) 상기 (3) 단계의 살균된 숙성새싹삼 추출물에 기탁번호 KACC 91848P인 락토바실러스 플란타룸(Lactobacillus plantarum) P1201 균주 및 기탁번호 KACC 92156P인 락토바실러스 브레비스(Lactobacillus brevis) BMK184 균주가 혼합된 유산균 혼합물을 접종한 후, 25~35℃에서 2~5일 동안 발효시키는 단계를 포함하여 제조된 것을 특징으로 하는 간질환의 예방 또는 개선용 건강기능식품 조성물.As a health functional food composition for preventing or improving liver disease containing a fermented product of aged sprout ginseng extract as an active ingredient,
The fermented product of the aged sprout ginseng extract,
(1) Washing sprout ginseng grown for 25 to 30 days under running water, mincing, increasing the steam at 95 to 120°C for 25 to 35 minutes, and then aging at 70 to 80°C for 5 to 10 days;
(2) Drying the aged sprout ginseng in step (1) for 2 to 3 days at 50-55°C, adding water to the dried mature sprout ginseng and extracting at 95-100°C to prepare a mature sprout ginseng extract ;
(3) sterilizing after mixing sugar in the ripened sprout ginseng extract prepared in step (2); And
(4) Lactobacillus plantarum ( Lactobacillus plantarum ) P1201 strain with accession number KACC 91848P and Lactobacillus brevis BMK184 strain with accession number KACC 92156P are mixed with the sterilized ripened sprout ginseng extract of step (3) After inoculating a mixture of lactic acid bacteria, a health functional food composition for preventing or improving liver disease, characterized in that it is prepared, including fermenting for 2 to 5 days at 25 to 35°C.
상기 숙성새싹삼 추출물의 발효물은,
(1) 25~30일간 재배된 새싹삼을 흐르는 물에 세척하고 세절하여 95~120℃에서 25~35분 동안 증자한 후 70~80℃에서 5~10일간 숙성시키는 단계;
(2) 상기 (1) 단계의 숙성된 새싹삼을 50~55℃에서 2~3일간 건조시키고, 건조된 숙성새싹삼에 물을 가하여 95~100℃에서 추출하여 숙성새싹삼 추출물을 제조하는 단계;
(3) 상기 (2) 단계의 제조된 숙성새싹삼 추출물에 당을 혼합한 후 살균하는 단계; 및
(4) 상기 (3) 단계의 살균된 숙성새싹삼 추출물에 기탁번호 KACC 91848P인 락토바실러스 플란타룸(Lactobacillus plantarum) P1201 균주 및 기탁번호 KACC 92156P인 락토바실러스 브레비스(Lactobacillus brevis) BMK184 균주가 혼합된 유산균 혼합물을 접종한 후, 25~35℃에서 2~5일 동안 발효시키는 단계를 포함하여 제조된 것을 특징으로 하는 간질환의 예방 또는 치료용 약학 조성물.As a pharmaceutical composition for the prevention or treatment of liver disease containing a fermented product of ripened sprout ginseng extract as an active ingredient,
The fermented product of the aged sprout ginseng extract,
(1) Washing sprout ginseng grown for 25 to 30 days under running water, mincing, increasing the steam at 95 to 120°C for 25 to 35 minutes, and then aging at 70 to 80°C for 5 to 10 days;
(2) Drying the aged sprout ginseng in step (1) for 2 to 3 days at 50-55°C, adding water to the dried mature sprout ginseng and extracting at 95-100°C to prepare a mature sprout ginseng extract ;
(3) sterilizing after mixing sugar in the ripened sprout ginseng extract prepared in step (2); And
(4) Lactobacillus plantarum ( Lactobacillus plantarum ) P1201 strain with accession number KACC 91848P and Lactobacillus brevis BMK184 strain with accession number KACC 92156P are mixed with the sterilized ripened sprout ginseng extract of step (3) After inoculating the mixture of lactic acid bacteria, a pharmaceutical composition for the prevention or treatment of liver disease, characterized in that it is prepared including the step of fermenting for 2 to 5 days at 25 to 35 ℃.
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