KR20150017880A - Composition for detoxification or relieving hangover comprising fermented natural plant as effective component - Google Patents
Composition for detoxification or relieving hangover comprising fermented natural plant as effective component Download PDFInfo
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- KR20150017880A KR20150017880A KR1020130094039A KR20130094039A KR20150017880A KR 20150017880 A KR20150017880 A KR 20150017880A KR 1020130094039 A KR1020130094039 A KR 1020130094039A KR 20130094039 A KR20130094039 A KR 20130094039A KR 20150017880 A KR20150017880 A KR 20150017880A
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- 230000001225 therapeutic effect Effects 0.000 description 1
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- 208000037816 tissue injury Diseases 0.000 description 1
- 235000015193 tomato juice Nutrition 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 229940116269 uric acid Drugs 0.000 description 1
- 235000015041 whisky Nutrition 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
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- A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
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Abstract
Description
본 발명은 천연 식물소재 발효물을 유효성분으로 함유하는 간 해독 또는 숙취해소용 조성물에 관한 것으로, 더욱 상세하게는 대두 분말, 녹두 분말, 미나리 분말, 다시마 분말, 갈근 분말, 무 분말 및 오이 분말의 혼합 발효물; 상기 혼합 발효물의 저온 숙성물; 또는 상기 혼합 발효물 또는 저온 숙성물의 추출물을 유효성분으로 함유하는 간 해독 또는 숙취해소용 조성물 및 건강식품에 관한 것이다.The present invention relates to a composition for liver detoxification or hangover detoxification comprising a fermented product of natural plant material as an active ingredient, and more particularly to a composition for liver detoxification or hangover decomposition which contains a fermented product of natural plant material as an active ingredient. Mixed fermentation products; Low temperature aging of the mixed fermented product; Or an extract of the mixed fermented product or low-temperature aged product as an active ingredient, and a health food.
급성·만성 알코올의 섭취는 간, 신장 및 뇌 등 여러 장기에 손상을 유발하며 특히 알코올성 간염과 간경화를 유도해 사망을 초래하게 된다. 정상적인 상태에서 소량의 알코올을 섭취할 경우 간 장내로 들어온 에탄올은 세포 기질 내의 알코올탈수소효소(alcohol dehydrogenase, ADH)와 알데하이드탈수소효소(aldehyde dehydrogenase, ALDH)의 작용에 의해 아세테이트(acetate)로 전환되고, 이는 순환계를 통해 간세포 밖으로 배설하게 된다. 이중 특히 에탄올의 최초 대사산물인 아세트알데하이드(acetaldehyde)는 에탄올에 비해 반응성이 매우 높고 독성이 강해 알코올성 간 장애의 주원인 물질로 숙취를 유발할 뿐만 아니라, 세포 내 에너지 생성기관인 미토콘드리아(mitochondria)의 기능을 저해하여 간경변을 유발하고 심장 및 뇌 기능을 저해하게 된다. 또한 아세트알데하이드는 미오피브로블라스트(myofibroblast)의 콜라겐 합성을 촉진하여 간 섬유화와 간세포의 변성 종대를 일으키며, 생체 내 거대 분자와 반응하여 어덕트(adduct)를 형성하기도 한다.The intake of acute and chronic alcohol causes damage to various organs such as liver, kidney and brain, and induces alcoholic hepatitis and cirrhosis in particular, resulting in death. When a small amount of alcohol is consumed in a normal state, ethanol entering the liver is converted to acetate by the action of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in the cell matrix, It is excreted through the circulation system outside the hepatocytes. In particular, acetaldehyde, the first metabolite of ethanol, is highly reactive and toxic, which is the main cause of alcoholic liver disorder. It causes not only hangover but also inhibits the function of mitochondria, an intracellular energy producing organ. Cirrhosis of the liver and heart and brain function is inhibited. Acetaldehyde also promotes collagen synthesis in myofibroblast, causing hepatic fibrosis and metamorphosis of hepatocytes, and reacting with macromolecules in vivo to form adducts.
간에서 알코올의 분해 경로는 알코올 탈수소효소계(alcohol dehydrogenase; ADH pathway), 마이크로좀의 알코올 산화계(microsomal ethanol oxidizing systemm; MEOS) 및 카탈라제계(catalase pathway)로 3 종류의 효소계로 나누어 진다. 임상적으로 중요한 경로는 ADH계와 MEOS계로, 양자 모두 알코올이 아세트알데하이드를 거쳐 아세테이트로 산화되며, AHD계는 조직의 알코올 농도가 낮을 때, MEOS계는 높을 때 알코올대사에 관여하므로 각각 급성과 만성 알코올 대사라고 한다. 숙취성 간손상을 예방하기 위해서는 섭취된 알코올을 아세테이트로 빠르게 분해해야 하고, 그러기 위해서는 알코올탈수소효소(ADH)와 아세트알데히드탈수소효소(ALDH)의 생합성과 이들 효소의 활성을 증강시켜야 한다. 또한, 급성·만성 알코올 섭취에 의한 독작용을 예방하기 위해서는 섭취된 알코올의 대사과정을 촉진과 조직의 손상에 관여하는 ROS의 생성을 저해 혹은 제거함으로써 숙취해소 및 조직손상을 경감시킬 수 있다. 오래 전부터 알코올에 의한 숙취의 해소와 장기손상을 경감 또는 예방하기 위한 다양한 의약품과 건강기능식품이 국내외적으로 개발 및 판매되고 있으나 알코올 섭취에 의한 장기손상의 기전이 매우 다양하고 복잡하기 때문에 전반적인 효과를 인정하기에는 부족한 상태이다.The degradation pathway of alcohol in the liver is divided into three types of enzymes: alcohol dehydrogenase (ADH pathway), microsomal ethanol oxidizing system (MEOS), and catalase pathway. The clinically important pathway is the oxidation of alcohol to acetal via both acetaldehyde and ADH system and MEOS system. The AHD system is involved in alcohol metabolism when the tissue alcohol concentration is low, and the MEOS system is involved in alcohol metabolism. It is called alcohol metabolism. In order to prevent hangover liver damage, the consumed alcohol should be rapidly degraded to acetate. To do so, the biosynthesis of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) and the activity of these enzymes should be enhanced. In addition, in order to prevent the poisoning due to acute and chronic alcohol ingestion, it is possible to reduce the hangover and tissue damage by promoting the metabolism of the consumed alcohol and inhibiting or eliminating the production of ROS involved in tissue damage. Various medicines and health functional foods have been developed and sold domestically in order to alleviate hangovers caused by alcohol and to prevent or prevent long-term damage. However, since the mechanism of organ damage by alcohol consumption is very diverse and complex, It is not enough to acknowledge.
한편, 한국등록특허 제1150885호에는 숙취해소용 홍삼, 용아초, 미배아대두 추출물 및 혼합조성물이 개시되어 있으나, 본 발명과 같은 천연 식물소재 발효물을 유효성분으로 함유하는 간 해독 또는 숙취해소용 조성물에 대해서는 밝혀진 바가 없다.On the other hand, Korean Patent No. 1150885 discloses red ginseng roots, red ginseng roots, ungerminated soybean extract and mixed compositions of hangover, but it is also possible to use the roots of the present invention as the active ingredients of liver plant detoxification or hangover The composition has not been disclosed.
본 발명은 상기와 같은 요구에 의해 안출된 것으로서, 본 발명에서는 인체에 안전하고 쉽게 구할 수 있는 천연 소재 유래의 간 해독 또는 숙취해소용 조성물을 제공하고자, 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이 분말을 일정비율로 혼합하고 이를 발효 및 저온숙성하여 쥐에 경구투여한 결과, 알코올 섭취로 유발되는 간 조직 손상에 대한 예방 및 치유 효과와 숙취해소에 탁월한 효과를 확인함으로써, 본 발명을 완성하였다.SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned needs, and it is an object of the present invention to provide a composition for liver detoxification or hangover detoxification derived from natural materials which is safe and easily obtainable from human body, And cucumber powder were mixed at a certain ratio and fermented and matured at a low temperature to be orally administered to rats. As a result, it was found that the present invention was completed by confirming the prevention and healing effect on liver tissue damage caused by alcohol consumption and the excellent effect on the elimination of hangover Respectively.
상기 목적을 달성하기 위하여, 본 발명은 대두 분말, 녹두 분말, 미나리 분말, 다시마 분말, 갈근 분말, 무 분말 및 오이 분말의 혼합 발효물; 상기 혼합 발효물의 저온 숙성물; 또는 상기 혼합 발효물 또는 저온 숙성물의 추출물을 유효성분으로 함유하는 간 해독 또는 숙취해소용 조성물을 제공한다.To achieve the above object, the present invention provides a fermented product of soybean powder, mung bean powder, parsley powder, kelp powder, pomegranate powder, powder-free powder and cucumber powder; Low temperature aging of the mixed fermented product; Or an extract of the fermented product or low-temperature aged product as an active ingredient.
또한, 본 발명은 대두 분말, 녹두 분말, 미나리 분말, 다시마 분말, 갈근 분말, 무 분말 및 오이 분말의 혼합 발효물; 상기 혼합 발효물의 저온 숙성물; 또는 상기 혼합 발효물 또는 저온 숙성물의 추출물을 유효성분으로 함유하는 간 해독 또는 숙취해소용 건강식품을 제공한다.The present invention also relates to a fermented product of soybean powder, mung bean powder, parsley powder, kelp powder, powdered pomegranate powder, powder-free powder and cucumber powder; Low temperature aging of the mixed fermented product; Or an extract of the mixed fermented product or low temperature aged product as an active ingredient.
본 발명의 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이의 혼합 발효물 또는 상기 혼합 발효물의 저온 숙성물은 기존의 고가의 약용식물을 이용한 간 해독 및 숙취해소 조성물을 대체할 수 있도록 안전할 뿐만 아니라 쉽게 구할 수 있고 저렴한 원료인 천연 식물소재를 사용하여 제조되었는데, 상기 조성물은 간 조직 보호·치유 및 숙취해소 효과가 탁월하므로, 소비자들의 건강에 유익하고 안전한 간 해독 또는 숙취해소용 조성물로 제공될 수 있을 것이다.The fermented product of the fermented product of soybean, mung bean, parsley, kelp, parsley, radish, and cucumber or the fermented product at low temperature of the present invention is safe to replace the liver detoxification and hangover composition using existing expensive medicinal plants In addition, it is manufactured using a natural plant material which is easily available and inexpensive raw material. The composition is excellent in protection of liver tissue, healing and hangover, so that it is useful for consumers' health and safe for liver detoxification or hangover .
도 1은 본 발명의 조성물을 제조하는 과정을 도시한 흐름도이다.
도 2는 본 발명의 조성물을 경구투여한 쥐의 알코올로 유도된 간 조직의 손상 정도를 평가한 결과를 나타낸 그래프이다. (A) 글루타치온 함량, (B) 과산화지질 함량, (C) 중성지방 함량, (D) 알라닌 아미노전이효소(ALT), (E) 아스파르테이트 아미노전이효소(AST), NC: 정상군, EN: 에탄올 대조군, NFME: 혼합 추출물 5.0% 첨가 식이군, FME: 발효 혼합 추출물 5.0% 첨가 식이군, FLME: 저온숙성 혼합 추출물 5.0% 첨가 식이군.
도 3은 본 발명의 조성물을 경구투여한 쥐의 알코올 대사 관련 효소의 활성 정도를 평가한 결과를 나타낸 그래프이다. (A) 알코올 탈수소효소 활성, (B) 미토콘드리아-알데히드탈수소효소 활성, (C) 세포질-알데히드탈수소효소 활성, (D) 세포질-시토크롬 p450 2E1 활성, NC: 정상군, EN: 에탄올 대조군, NFME: 혼합 추출물 5.0% 첨가 식이군, FME: 발효 혼합 추출물 5.0% 첨가 식이군, FLME: 저온숙성 혼합 추출물 5.0% 첨가 식이군.
도 4는 본 발명의 조성물을 경구투여한 쥐의 혈중 알코올 농도 변화를 측정한 결과를 나타낸 그래프이다. NC: 정상군, EN: 에탄올 대조군, NFME: 혼합 추출물 5.0% 첨가 식이군, FME: 발효 혼합 추출물 5.0% 첨가 식이군, FLME: 저온숙성 혼합 추출물 5.0% 첨가 식이군.BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a flow chart illustrating a process for preparing the composition of the present invention. FIG.
FIG. 2 is a graph showing the results of evaluating the degree of damage of liver-induced liver tissue of rats orally administered with the composition of the present invention. (A) glutathione content, (B) lipid peroxide content, (C) triglyceride content, (D) alanine aminotransferase (ALT), (E) aspartate aminotransferase : Ethanol control, NFME: mixed extracts added 5.0%, FME: fermented mixed extracts added 5.0%, FLME: low temperature aged mixed extracts added 5.0%.
FIG. 3 is a graph showing the results of evaluation of the degree of activity of an alcohol metabolism-related enzyme in rats orally administered with the composition of the present invention. (A) Alcohol dehydrogenase activity, (B) mitochondria-aldehyde dehydrogenase activity, (C) cytoplasmic-aldehyde dehydrogenase activity, (D) cytoplasmic cytochrome p450 2E1 activity, NC: normal group, EN: ethanol control group, NFME: FME: fermented mixed extract of 5.0%, FLME: low temperature fermented mixed extract of 5.0% added dietary group.
4 is a graph showing the results of measurement of changes in blood alcohol concentration in rats orally administered with the composition of the present invention. NC: normal group, EN: ethanol control group, NFME: mixed extract 5.0% added diet group, FME: fermented mixed extract 5.0% added diet group, FLME: low temperature aged mixed extract 5.0% diet group.
본 발명의 목적을 달성하기 위하여, 본 발명은 대두 분말, 녹두 분말, 미나리 분말, 다시마 분말, 갈근 분말, 무 분말 및 오이 분말의 혼합 발효물; 상기 혼합 발효물의 저온 숙성물; 또는 상기 혼합 발효물 또는 저온 숙성물의 추출물을 유효성분으로 함유하는 간 해독 또는 숙취해소용 조성물을 제공한다.In order to achieve the object of the present invention, the present invention provides a fermented product of soybean powder, mung bean powder, parsley powder, kelp powder, pomegranate powder, powder-free powder and cucumber powder; Low temperature aging of the mixed fermented product; Or an extract of the fermented product or low-temperature aged product as an active ingredient.
본 발명의 일 구현 예에 따른 상기 혼합 발효물은 혼합물 100 중량부 기준으로, 대두 분말 68~72 중량부, 녹두 분말 3~7 중량부, 미나리 분말 3~7 중량부, 다시마 분말 3~7 중량부, 갈근 분말 3~7 중량부, 무 분말 3~7 중량부 및 오이 분말 3~7 중량부로 혼합하고 발효미생물을 첨가하여 발효하여 제조될 수 있으며, 바람직하게는 혼합물 100 중량부 기준으로, 대두 분말 70 중량부, 녹두 분말 5 중량부, 미나리 분말 5 중량부, 다시마 분말 5 중량부, 갈근 분말 5 중량부, 무 분말 5 중량부 및 오이 분말 5 중량부로 혼합하고 발효미생물을 첨가하여 발효하여 제조될 수 있으나, 이에 제한되지 않는다.The mixed fermented product according to one embodiment of the present invention may be prepared by mixing 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, 3 to 7 parts by weight of kelp powder, 3 to 7 parts by weight of powdered powder, 3 to 7 parts by weight of powder-free powder and 3 to 7 parts by weight of cucumber powder, fermenting by adding fermenting microorganism, preferably 100 parts by weight of the mixture, Mixed with 70 parts by weight of powder, 5 parts by weight of mungbean powder, 5 parts by weight of powdered mung bean, 5 parts by weight of tallow powder, 5 parts by weight of powdered pomegranate powder, 5 parts by weight of powderless powder and 5 parts by weight of cucumber powder, But is not limited thereto.
상기 혼합 발효물은 보다 구체적으로,The mixed fermentation product is more specifically,
(a) 세척한 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이를 동결건조한 후 100~140 메쉬로 분말화하는 단계;(a) lyophilizing washed soybeans, mung beans, buttercups, kelp, parsley, radishes and cucumbers, followed by pulverization to 100-140 mesh;
(b) 상기 분말화한 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이 분말을 혼합물 100 중량부 기준으로, 대두 분말 68~72 중량부, 녹두 분말 3~7 중량부, 미나리 분말 3~7 중량부, 다시마 분말 3~7 중량부, 갈근 분말 3~7 중량부, 무 분말 3~7 중량부 및 오이 분말 3~7 중량부를 혼합하여 75~85℃에서 30~50분 동안 살균하고 발효미생물과 혼합하는 단계; 및 (b) 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, mung bean, parsley, kelp, parsley, radish and cucumber powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered powder and 3 to 7 parts by weight of cucumber powder are mixed and sterilized at 75 to 85 ° C for 30 to 50 minutes, ; And
(c) 상기 발효미생물과 혼합한 혼합물을 습도 70~90%, 35~40℃에서 2~3일 동안 발효하는 단계에 의해 제조될 수 있으나, 이에 제한되지 않는다.(c) fermenting the mixture with the fermenting microorganism at a temperature of 35 to 40 ° C for 2 to 3 days under a humidity of 70 to 90%, but is not limited thereto.
본 발명의 일 구현 예에 따른 상기 혼합 발효물의 저온 숙성물은 혼합물 100 중량부 기준으로, 대두 분말 68~72 중량부, 녹두 분말 3~7 중량부, 미나리 분말 3~7 중량부, 다시마 분말 3~7 중량부, 갈근 분말 3~7 중량부, 무 분말 3~7 중량부 및 오이 분말 3~7 중량부로 혼합하고 발효미생물을 첨가하여 발효한 후, 저온 숙성하여 제조될 수 있으며, 바람직하게는 혼합물 100 중량부 기준으로, 대두 분말 70 중량부, 녹두 분말 5 중량부, 미나리 분말 5 중량부, 다시마 분말 5 중량부, 갈근 분말 5 중량부, 무 분말 5 중량부 및 오이 분말 5 중량부로 혼합하고 발효미생물을 첨가하여 발효한 후, 저온 숙성하여 제조될 수 있으나, 이에 제한되지 않는다.The low-temperature fermented product according to an embodiment of the present invention may be prepared by mixing 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, , 3 to 7 parts by weight of powdered powder, 3 to 7 parts by weight of powdered powder and 3 to 7 parts by weight of cucumber powder, fermenting the mixture by adding a fermenting microorganism, and then aging at low temperature, The mixture was mixed with 70 parts by weight of soybean powder, 5 parts by weight of mung bean powder, 5 parts by weight of parsley powder, 5 parts by weight of seaweed powder, 5 parts by weight of powdered pear powder, 5 parts by weight of no powder and 5 parts by weight of cucumber powder based on 100 parts by weight of the mixture Fermenting microorganism may be added and fermented, followed by low-temperature aging, but the present invention is not limited thereto.
상기 혼합 발효물의 저온 숙성물은 보다 구체적으로,More specifically, the low-temperature aged product of the above-
(a) 세척한 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이를 동결건조한 후 100~140 메쉬로 분말화하는 단계;(a) lyophilizing washed soybeans, mung beans, buttercups, kelp, parsley, radishes and cucumbers, followed by pulverization to 100-140 mesh;
(b) 상기 분말화한 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이 분말을 혼합물 100 중량부 기준으로, 대두 분말 68~72 중량부, 녹두 분말 3~7 중량부, 미나리 분말 3~7 중량부, 다시마 분말 3~7 중량부, 갈근 분말 3~7 중량부, 무 분말 3~7 중량부 및 오이 분말 3~7 중량부를 혼합하여 75~85℃에서 30~50분 동안 살균하고 발효미생물과 혼합하는 단계; 및(b) 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, mung bean, parsley, kelp, parsley, radish and cucumber powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered powder and 3 to 7 parts by weight of cucumber powder are mixed and sterilized at 75 to 85 ° C for 30 to 50 minutes, ; And
(c) 상기 발효미생물과 혼합한 혼합물을 습도 70~90%, 35~40℃에서 2~3일 동안 발효한 후, 습도 40~60%, 10~20℃에서 4~10일 동안 저온숙성하는 단계에 의해 제조될 수 있으나, 이에 제한되지 않는다.(c) fermenting the mixture obtained by mixing the fermenting microorganism with 70 to 90% of humidity at 35 to 40 ° C for 2 to 3 days and then aging at 40 to 60% of humidity and 10 to 20 ° C for 4 to 10 days at a low temperature But it is not limited thereto.
본 발명의 일 구현 예에 따른 상기 혼합 발효물 또는 저온 숙성물의 추출물은 상기 혼합 발효물 또는 상기 혼합 발효물의 저온 숙성물을 효소추출, 열수추출 또는 주정추출하여 제조될 수 있으나, 이에 제한되지 않는다. 효소추출은 알칼라아제 추출일 수 있으나, 이에 제한되지 않는다. 또한, 상기 혼합 발효물 또는 저온 숙성물의 추출물은 상기 혼합 발효물 또는 상기 혼합 발효물의 저온 숙성물을 효소추출, 열수추출 또는 주정추출한 후 각각의 추출물을 합하여 제조할 수 있다.The mixed fermented product or the low-temperature aged extract according to an embodiment of the present invention may be prepared by enzymatic extraction, hot water extraction or alcoholic extraction of the low-temperature aged product of the mixed fermented product or the mixed fermented product, but is not limited thereto. Enzyme extraction may be, but is not limited to, alkalase extraction. Further, the mixed fermented product or the low-temperature aged product may be prepared by subjecting the low-temperature aged product of the mixed fermented product or the mixed fermented product to enzyme extraction, hot water extraction or alcohol extraction, and then combining the respective extracts.
본 발명의 일 구현 예에 따른 간 해독 또는 숙취해소용 조성물에서, 상기 발효미생물은 바실러스 리체니포르미스(Bacillus licheniformis), 바실러스 푸밀루스(Bacillus pumilus), 바실러스 소노렌시스(Bacillus sonorensis) 및 바실러스 서브틸리스(Bacillus subtilis)로 이루어지는 군으로부터 선택되는 1종 이상일 수 있으며, 바람직하게는 바실러스 리체니포르미스(Bacillus licheniformis)일 수 있으나, 이에 제한되지 않는다.In a composition for liver detoxification or hangover resolution according to an embodiment of the present invention, the fermenting microorganism is selected from the group consisting of Bacillus sp. licheniformis , Bacillus pumilus), Bacillus Sono alkylene sheath (Bacillus sonorensis , and Bacillus subtilis . Preferably, it may be Bacillus licheniformis . However, the present invention is not limited thereto.
임상에서 간 조직 손상 정도는 간 조직의 글루타치온, 과산화지질, 중성지방, 알라닌 아미노전이효소(ALT), 아스파르테이트 아미노전이효소(AST)의 활성을 측정을 통해 알 수 있는데, 본 발명의 간 해독 또는 숙취해소용 조성물을 4주간 식이시킨 쥐를 12시간 절식시킨 후 에탄올을 경구투여하여 상기 활성을 측정한 결과, 본 발명의 조성물을 식이한 쥐는 에탄올을 경구투여하지 않은 정상 쥐에 준하는 간 조직의 상태를 보여 알코올에 의한 간 손상이 개선 및 예방된 것을 확인할 수 있었으며(도 2), 알코올 대사 관련 효소인 미토콘드리아-알데히드탈수소효소 및 세포질-알데히드탈수소효소의 활성이 탁월하였다(도 3). 또한 혈중 에탄올의 경시적 농도변화를 측정한 결과, 본 발명의 조성물을 식이시킨 쥐에서 혈중 에탄올의 농도는 1시간 경과 후부터 확연히 줄어드는 것을 확인할 수 있었다(도 4). 따라서, 본 발명의 상기 조성물은 간염 및 간경변의 예방 및 개선을 위한 용도로 이용될 수 있으나, 이에 제한되지 않는다.In clinical practice, the degree of hepatic tissue damage can be determined by measuring the activities of glutathione, lipid peroxidation, triglyceride, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in liver tissue. Or rats were sacrificed for 4 weeks. After 12 hours of fasting, ethanol was orally administered to measure the activity. As a result, rats fed with the composition of the present invention were found to have liver tissue similar to that of normal rats without oral administration of ethanol (FIG. 2), and the activities of mitochondrial-aldehyde dehydrogenase and cytoplasmic-aldehyde dehydrogenase, which are alcohol metabolism-related enzymes, were excellent (FIG. 3). In addition, it was confirmed that the concentration of ethanol in the blood of the rats fed with the composition of the present invention was markedly decreased after 1 hour (FIG. 4). Therefore, the composition of the present invention can be used for prevention and improvement of hepatitis and cirrhosis, but is not limited thereto.
또한, 본 발명은In addition,
(a) 세척한 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이를 건조한 후 분말화하는 단계;(a) drying and then pulverizing the washed soybean, mung bean, parsley, kelp, parsley, radish and cucumber;
(b) 상기 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이 분말을 살균하여 발효미생물과 혼합하는 단계; 및(b) sterilizing the soybean, mung bean, parsley, kelp, parsley, radish, and cucumber powder and mixing with the fermenting microorganism; And
(c) 상기 혼합물을 발효하는 단계를 포함하는 것을 특징으로 하는 간 해독 또는 숙취해소용 조성물의 제조방법을 제공한다.(c) fermenting the mixture. The present invention also provides a method for preparing a composition for liver detoxification or hangover decomposition.
본 발명의 간 해독 또는 숙취해소용 조성물의 제조방법은 구체적으로는The method of the present invention for preparing liver-decrypting or hangover-
(a) 세척한 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이를 동결건조한 후 100~140 메쉬로 분말화하는 단계;(a) lyophilizing washed soybeans, mung beans, buttercups, kelp, parsley, radishes and cucumbers, followed by pulverization to 100-140 mesh;
(b) 상기 분말화한 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이 분말을 혼합물 100 중량부 기준으로, 대두 분말 68~72 중량부, 녹두 분말 3~7 중량부, 미나리 분말 3~7 중량부, 다시마 분말 3~7 중량부, 갈근 분말 3~7 중량부, 무 분말 3~7 중량부 및 오이 분말 3~7 중량부를 혼합하여 75~85℃에서 30~50분 동안 살균하고 발효미생물과 혼합하는 단계; 및 (b) 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, mung bean, parsley, kelp, parsley, radish and cucumber powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered powder and 3 to 7 parts by weight of cucumber powder are mixed and sterilized at 75 to 85 ° C for 30 to 50 minutes, ; And
(c) 상기 발효미생물과 혼합한 혼합물을 습도 70~90%, 35~40℃에서 2~3일 동안 발효하는 단계를 포함할 수 있다.(c) fermenting the mixture with the fermenting microorganism at a temperature of 35 to 40 ° C for 2 to 3 days under a humidity of 70 to 90%.
또한, 본 발명은In addition,
(a) 세척한 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이를 건조한 후 분말화하는 단계;(a) drying and then pulverizing the washed soybean, mung bean, parsley, kelp, parsley, radish and cucumber;
(b) 상기 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이 분말을 살균하여 발효미생물과 혼합하는 단계; 및(b) sterilizing the soybean, mung bean, parsley, kelp, parsley, radish, and cucumber powder and mixing with the fermenting microorganism; And
(c) 상기 혼합물을 발효한 후, 저온숙성하는 단계를 포함하는 것을 특징으로 하는 간 해독 또는 숙취해소용 조성물의 제조방법을 제공한다.(c) fermenting the mixture, and then aging the mixture at a low temperature. The present invention also provides a method for preparing a composition for liver detoxification or hangover decomposition.
본 발명의 간 해독 또는 숙취해소용 조성물의 제조방법은 구체적으로는The method of the present invention for preparing liver-decrypting or hangover-
(a) 세척한 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이를 동결건조한 후 100~140 메쉬로 분말화하는 단계;(a) lyophilizing washed soybeans, mung beans, buttercups, kelp, parsley, radishes and cucumbers, followed by pulverization to 100-140 mesh;
(b) 상기 분말화한 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이 분말을 혼합물 100 중량부 기준으로, 대두 분말 68~72 중량부, 녹두 분말 3~7 중량부, 미나리 분말 3~7 중량부, 다시마 분말 3~7 중량부, 갈근 분말 3~7 중량부, 무 분말 3~7 중량부 및 오이 분말 3~7 중량부를 혼합하여 75~85℃에서 30~50분 동안 살균하고 발효미생물과 혼합하는 단계; 및(b) 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, mung bean, parsley, kelp, parsley, radish and cucumber powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered powder and 3 to 7 parts by weight of cucumber powder are mixed and sterilized at 75 to 85 ° C for 30 to 50 minutes, ; And
(c) 상기 발효미생물과 혼합한 혼합물을 습도 70~90%, 35~40℃에서 2~3일 동안 발효한 후, 습도 40~60%, 10~20℃에서 4~10일 동안 저온숙성하는 단계를 포함할 수 있다.(c) fermenting the mixture obtained by mixing the fermenting microorganism with 70 to 90% of humidity at 35 to 40 ° C for 2 to 3 days and then aging at 40 to 60% of humidity and 10 to 20 ° C for 4 to 10 days at a low temperature Step < / RTI >
또한, 본 발명은 대두 분말, 녹두 분말, 미나리 분말, 다시마 분말, 갈근 분말, 무 분말 및 오이 분말의 혼합 발효물; 상기 혼합 발효물의 저온 숙성물; 또는 상기 혼합 발효물 또는 저온 숙성물의 추출물을 유효성분으로 함유하는 간 해독 또는 숙취해소용 건강식품을 제공한다.The present invention also relates to a fermented product of soybean powder, mung bean powder, parsley powder, kelp powder, powdered pomegranate powder, powder-free powder and cucumber powder; Low temperature aging of the mixed fermented product; Or an extract of the mixed fermented product or low temperature aged product as an active ingredient.
본 발명의 혼합 발효물 또는 상기 혼합 발효물의 저온 숙성물 또는 상기 혼합 발효물 또는 저온 숙성물의 추출물을 식품첨가물로 사용하는 경우, 상기 혼합 발효물 또는 상기 혼합 발효물의 저온 숙성물 또는 상기 혼합 발효물 또는 저온 숙성물의 추출물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효 성분의 혼합양은 그의 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. 일반적으로, 식품 또는 음료의 제조시에 본 발명의 발효물 또는 저온 숙성물 또는 추출물은 원료에 대하여 15 중량부 이하, 바람직하게는 10 중량부 이하의 양의로 첨가된다. 그러나, 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다. When the fermented product of the present invention or an extract of the fermented product at low temperature or the fermented product or the fermented product at low temperature is used as a food additive, the fermented product or the fermented product or the fermented product of the fermented product The extract of the low-temperature aged product can be directly added or used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, health or therapeutic treatment). Generally, the fermentation product or the low-temperature aged product or extract of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on the raw material. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .
상기 식품의 종류에는 특별한 제한은 없다. 구체적인 예로, 상기 발효물 또는 저온 숙성물 또는 추출물을 이용하여 농산물, 축산물 또는 수산물의 특성을 살려 변형시키는 동시에 저장성을 좋게 한 가공식품을 제조할 수 있다. 이런 가공식품에는 예를 들어, 과자, 음료, 주류, 발효식품, 통조림, 우유가공식품, 육류가공식품, 국수 등을 포함한다. 과자는 비스킷, 파이, 빵, 캔디, 젤리, 껌, 시리얼(곡물푸레이크 등의 식사대용품류 포함) 등을 포함한다. 음료는 탄산음료, 기능성이온음료, 쥬스(예를 들어, 사과, 배,포도, 알로에, 감귤, 복숭아, 당근, 토마토쥬스 등), 식혜 등을 포함한다. 주류는 청주, 위스키, 소주, 맥주,양주, 과실주 등을 포함한다. 발효식품은 간장, 된장, 고추장 등을 포함한다. 통조림은 수산물 통조림(예를 들어, 참치, 고등어, 공치, 소라 통조림 등), 축산물 통조림(쇠고기, 돼지고기, 닭고기, 칠면조 통조림 등), 농산물 통조림(옥수수, 복숭아, 파인애플 통조림 등)을 포함한다. 우유가공식품은 치즈, 버터, 요구르트 등을 포함한다. 육류가공식품은 돈까스, 비프까스, 치킨까스, 소세지, 탕수육, 너겟류, 너비아니 등을 포함한다. 밀봉포장생면 등의 국수를 포함한다. 이 외에도 상기 조성물은 레토르트식품, 스프류 등에 사용될 수 있다.There is no particular limitation on the kind of the food. As a specific example, the fermented product, the low-temperature aged product or the extract may be used to produce a processed food having good storability while being modified by utilizing characteristics of agricultural products, livestock products or aquatic products. Such processed foods include, for example, confectionery, drinks, liquor, fermented foods, canned foods, milk processed foods, meat processed foods, noodles and the like. The sweets include biscuits, pies, breads, candies, jellies, gums, cereals (including dinner utensils such as cereal flakes). Drinks include carbonated beverages, functional ionic beverages, juices (such as apples, pears, grapes, aloes, citrus fruits, peaches, carrots, tomato juices, etc.) and sikhye. The mainstream includes sake, whiskey, shochu, beer, liquor, and fruit wine. Fermented foods include soy sauce, miso, and kochujang. Canned products include canned products (for example, tuna, mackerel, sandwiches, canned fish, etc.), canned products (canned beef, pork, chicken and turkey canned products) and canned products (corn, peach and pineapple canned products). Milk processed foods include cheese, butter, yogurt and the like. Meat processed foods include pork cutlet, beef cutlet, chicken cutlet, sausage, sweet and sour pork, nuggets, nubucki, and the like. And noodles such as sealed packaging raw noodles. In addition, the composition may be used in retort food, soup and the like.
또한, 상기 발효물 또는 저온 숙성물 또는 추출물을 이용하여 기능성식품, 건강식품 또는 건강보조식품을 제조할 수 있다. 기능성식품, 건강식품 또는 건강보조식품은 영양 기능 외에도 생리활성 성분을 포함하여 생체조절 기능을 제공하는 식품을 의미하고, 본 발명의 발효물 또는 저온 숙성물 또는 추출물은 간 기능 회복 및 보호 효과를 가지는 상기 여러 가지의 특정 천연물을 유효 성분으로 포함하므로 기능성식품, 건강식품 또는 건강보조식품 등의 제조에 이용될 수 있다.
In addition, functional foods, health foods or health supplements can be produced by using the above-mentioned fermented products or low-temperature aged products or extracts. A functional food, a health food or a health supplement food means a food which provides a biomodulating function including a physiologically active ingredient in addition to a nutrition function, and the fermented product or the low-temperature aged product or extract of the present invention has a function Since the above-mentioned various natural products are contained as an effective ingredient, they can be used for the production of functional foods, health foods or health supplements.
이하, 본 발명을 실시예 및 제조예에 의해 상세히 설명한다. 단, 하기 실시예 및 제조예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예 및 제조예에 한정되는 것은 아니다.
Hereinafter, the present invention will be described in detail with reference to Examples and Production Examples. However, the following examples and preparative examples are merely illustrative of the present invention, and the present invention is not limited to the following examples and preparative examples.
제조예Manufacturing example 1: 간 해독 또는 숙취해소용 혼합 1: liver detox or hangover mixed for small 발효물Fermentation product 및 혼합 And mixing 발효물의Fermented 저온 숙성물 제조 Production of low temperature aged water
본 발명에 사용한 대두(Glycine max L.)는 강원도 화천에서 재배된 것을 사용하였으며, 녹두(Phaseolus radiatus, Vigna radita)는 경남 김해시에서, 다시마(Laminaria japonica)는 전남 완도군에서 구입하여 사용하였다. 갈근(Puerariae Radix), 미나리(Oenanthe javanica), 오이(Cucumis sativus L.) 및 무(Raphanus sativus L.)는 국내산 유기농 상품을 구입하여 사용하였다. 균주로는 재래청국장으로부터 우수균주로 선발된 바실러스 리체니포르미스(Bacillus licheniformis) 균주를 사용하였다. 알칼라아제(Alcalase, 2.4L(2.4 AU/g))는 Sigma 사로부터 구입하여 사용하였으며, 효능분석에 사용한 시약은 일반 특급시약을 사용하였다.The soybean ( Glycine max L.) was grown in Hwacheon, Gangwon province, and mung bean ( Phaseolus radiatus, Vigna radita) is in Gimhae, Gyeongnam, kelp (Laminaria japonica ) were purchased from Wando County, Chonnam Province. Puerariae Radix , Oenanthe javanica ), cucumber ( Cucumis sativus L.) and radish ( Raphanus sativus L.) purchased and used domestic organic products. As a strain, Bacillus ricinifolium ( Bacillus sp. licheniformis strains were used. Alcalase (2.4 L (2.4 AU / g)) was purchased from Sigma, and the reagents used for the efficacy analysis were standard limited reagents.
본 발명의 간 해독 또는 숙취해소용 혼합 발효물 및 혼합 발효 숙성물의 저온 숙성물은 도 1의 흐름도를 거쳐 제조되었다.The low-temperature aged product of the liver-detoxification or hangover-reducing small-scale fermentation product and the mixed-fermentation aged product of the present invention was produced through the flow chart of FIG.
제1공정은 건조 및 분말화 단계로, 먼저 대두, 다시마, 녹두, 미나리, 갈근, 무 및 오이를 선별하고 세척한 후 영양분의 손실을 최소화하기 위해 각각의 재료를 급속 동결건조하였다. 각각의 건조물은 유용성분 추출 및 발효의 촉진을 증대시키기 위하여 1차적으로 파쇄기로 파쇄를 한 뒤 미세분쇄기를 이용하여 120 메쉬의 분말을 제조하였다.The first step is a drying and pulverizing step. First, soy beans, kelp, mung bean, parsley, parsley, radish, and cucumber were screened and washed, and each material was rapidly lyophilized to minimize loss of nutrients. In order to increase the extraction of useful components and the promotion of fermentation, each of the dried products was first pulverized with a crusher and then a 120 mesh powder was prepared using a fine pulverizer.
제2공정은 배합, 살균 및 발효미생물을 혼합하는 단계로, 각종 생리활성이 뛰어난 대두와 다시마, 녹두, 미나리, 칡, 무 및 오이 분말을 숙취와 소화기능 개선의 효능을 유도하기 위한 적정 비율(7:0.5:0.5:0.5:0.5:0.5:0.5, w/w)로 고르게 혼합한 다음 80℃에서 40분간 살균하였다. 발효는 유용성이 입증된 고초균(B. licheniformis)을 사용하였으며, 이를 37~40℃에서 24시간 동안 배양하여 수세한 균체를 분말에 균일하게 접종하여 사용하였고, 이 때 고체발효가 충분히 이뤄지도록 일정량의 증류수(분말 중량의 5% 함량)를 가한 다음 일정크기로 성형한 후 상기 분말 혼합물과 혼합하였다.The second step is a step of blending the compounding, sterilizing and fermenting microorganisms. It is a step of mixing the soybean, kelp, mung bean, parsley, bean, radish and cucumber powder having various physiological activities with a proper ratio 7: 0.5: 0.5: 0.5: 0.5: 0.5: 0.5, w / w) and sterilized at 80 ° C for 40 minutes. The fermentation was used for Bacillus subtilis (B. licheniformis) demonstrated the usefulness, was used to uniformly inoculated with this cell was washed by incubation at 37 ~ 40 ℃ for 24 hours in a powder, a certain amount of time so that the solid fermentation is fully yirwoji Distilled water (5% of the powder weight) was added and then molded to a predetermined size and mixed with the powder mixture.
제3공정은 발효 및 저온숙성 단계로, 충분한 발효가 이뤄지도록 상기 발효 미생물을 접종한 분말 혼합물을 습도 70~90%, 35~40℃에서 2~3일간 발효하였으며, 본 단계까지 거친 조성물을 '발효 혼합물'로 사용하였다. 상기 발효 혼합물을 습도 40~60%, 10~20℃에서 7일간 저온숙성하였으며, 본 단계를 거친 조성물을 '발효 혼합물의 저온 숙성물'로 사용하였다.In the third step, the powder mixture inoculated with the fermenting microorganisms was fermented at 70 to 90% of humidity and at 35 to 40 ° C for 2 to 3 days so that sufficient fermentation could be achieved. Fermented mixture '. The fermentation mixture was aged at a temperature of 10 to 20 ° C for 7 days under a humidity of 40 to 60%, and the composition thus obtained was used as a 'low-temperature aged product of the fermentation mixture'.
제4공정은 추출 및 건조 단계로, 상기 발효 및 저온숙성시킨 고체발효 산물은 부패의 위험을 감소시키기 위하여 적외선 건조장치에서 수분율이 5% 이하가 되도록 60℃에서 12~24시간 건조하였다. 추출은 조성물의 기능성 유용성분이 충분히 추출되도록 효소추출, 열수추출, 주정추출의 3단계의 추출과정으로 진행하였다. 제3공정의 혼합 발효물 또는 혼합 발효물의 저온숙성물에 10배의 물을 첨가하여 50℃에서 초음파 추출한 후, 알칼라아제를 첨가하여 60분 동안 효소추출하여 95℃에서 10분 동안 효소를 불활성화시켜 1차 추출액을 제조하고, 이의 잔사에 10배량의 물을 첨가하여 80℃에서 9시간 동안 열수 추출하여 2차 추출액을 제조하고, 이의 잔사를 10배량의 70% 주정을 첨가하여 60℃에서 3차 추출액을 제조하였다.The fourth step is an extraction and drying step. In order to reduce the risk of corruption, the solid fermentation product fermented and fermented at low temperature was dried at 60 ° C for 12 to 24 hours so as to have a moisture content of 5% or less in an infrared drying apparatus. The extraction was carried out in three steps of extraction, hot water extraction and alcohol extraction so that the functional oil component of the composition could be sufficiently extracted. 10 times water was added to the low-temperature fermented product or mixed fermented product of the third step, and the mixture was ultrasonically extracted at 50 ° C. Then, the enzyme was extracted for 60 minutes by the addition of alkalase, and enzyme was removed for 10 minutes at 95 ° C And a 10-fold amount of water was added to the residue, and the mixture was subjected to hot extraction at 80 ° C for 9 hours to prepare a second extract. The residue was added with 10-fold amount of 70% alcohol at 60 ° C A third extract was prepared.
제5공정은 혼합, 농축 및 제형화하는 단계로, 제4공정의 1차, 2차 및 3차의 추출액을 각각 원심분리하여 얻은 상등액을 효소 불활성화를 시킨 다음 혼합하고 여과한 후 13~15 브릭스가 되도록 감압농축하여 발효 혼합조성물 추출액을 획득하였다. 제4공정의 열풍 건조물과 상기 발효 혼합조성물 추출 농축액을 95:5(w/v)의 비율로 혼합한 후 60℃에서 12~24시간 동안 수분함량 6% 이하가 되도록 건조하고 이를 환 제형으로 가공하였다.
The fifth step is a step of mixing, concentrating and formulating. The supernatant obtained by centrifuging the first, second and third extracts of the fourth step is inactivated with enzyme, mixed, filtered, The mixture was concentrated under reduced pressure to obtain a brick, thereby obtaining a fermented mixed composition extract. The hot-air dried product of the fourth step and the fermented mixed composition extract concentrate were mixed at a ratio of 95: 5 (w / v) and dried at 60 ° C for 12 to 24 hours to have a water content of 6% or less. Respectively.
실험방법Experimental Method
1. 실험동물 및 1. Experimental animals and 식이의Dietary 조제 pharmacy
실험동물은 6주령의 평균체중이 150±10g인 흰쥐(Sprague-Dawly SPF/VAF outbred rats, 오리엔트)를 사용하였고, 실험 사료 조성은 표 1과 같이 제조하였으며, 사료에 첨가된 각각의 추출물은 13 브릭스인 것을 사용하였다. 실험동물을 1주일간 환경에 적응시킨 후 정상군(NC), 에탄올 대조군(EN), 대두분말 : 분말(다시마, 녹두, 미나리, 칡, 무, 오이; 1:1:1:1:1:1:1)을 7:3의 비율로 고르게 배합한 혼합 추출물(발효미생물로 발효하지 않은 혼합물의 추출물) 5.0% 첨가 식이군, 발효혼합분말 추출물 5.0% 첨가 식이군 및 발효 숙성혼합분말 추출물 5.0% 첨가 식이군으로 나누어(총 5군) 각각 7마리씩으로 하여 4주간 사육하였다. 4주간 사육한 실험동물은 공복 상태에서 2일간 일일 1회 50% 에탄올 용액을 체중 kg당 5g씩 경구투여 하였으며, 투여하고 12시간이 경과한 다음 처치하였다. 사육장은 스테인레스 스틸 케이지를 사용하고, 온도 및 습도는 60±5% 및 23±2℃로 조정하고, 명암주기는 12시간 간격으로 설정하였으며, 물과 사료의 섭취는 자유 섭취시켰다.Experimental animals were fed with Sprague-Dawly SPF / VAF outbred rats (Orient) with an average body weight of 150 ± 10 g at 6 weeks of age. The experimental diets were prepared as shown in Table 1, Bricks. 1: 1: 1: 1: 1: 1: 1) was added to the test animals (NC), the ethanol control (EN) and the soybean flour powder (kelp, mung bean, : 5.0% addition of fermented mixed powder extract, 5.0% fermented mixed powder extract and 5.0% addition of fermented aged mixed powder extract (mixture of fermented microbial extract and non fermented mixture) (
1)NC: 정상군, EN: 에탄올 대조군, NFME: 혼합 추출물 5.0% 첨가 식이군, FME: 발효 혼합 추출물 5.0% 첨가 식이군, FLME: 발효, 저온숙성 혼합 추출물 5.0% 첨가 식이군. 1) NC: Normal group, EN: Ethanol control group, NFME: Mixed extract 5.0% added diet group, FME: Fermented mixed extract 5.0% added diet group, FLME: Fermentation, Low temperature aged mixed extract 5.0%
2)5L79 diet of PMI Nutrition(LLC, PO Box 19798, Brentwood MO 63144, USA). 성분함량: 조단백 18%, 조지방 5%, 조섬유 5%, 회분 8%.
2) 5 L79 diet of PMI Nutrition (LLC, PO Box 19798, Brentwood MO 63144, USA). Ingredients: crude protein 18%,
간 손상 정도를 알아보기 위하여, 간 조직 글루타치온(glutathione: GSH)의 함량은 적출한 동물의 간 조직 균질액에 2-니트로벤조산(2-nitrobenzoic acid)를 가해 생성되는 티오페놀(thiophenol)의 양을 측정하였다.To determine the degree of liver damage, the content of glutathione (GSH) in the liver was calculated by subtracting the amount of thiophenol produced by adding 2-nitrobenzoic acid to the liver homogenate of the extracted animal Respectively.
과산화지질(Lipid peroxide, LPO)의 함량은 간 조직 균질액에 티오바비투릭산(thiobarbituric acid, TBA) 용액을 가하여 반응시킨 후 부탄올(n-butanol)을 가해 이행되는 티오바비투릭산 반응물질(TBA-reactive substance)의 흡광도를 532 nm에서 측정하였으며, 분자흡광계수(ε=1.5×105 M-1 cm-1)를 이용하여 함량을 산출하였다.The content of lipid peroxide (LPO) was determined by adding thiobarbituric acid (TBA) solution to the liver tissue homogenate and adding thiobarbituric acid reactant (TBA -active substance was measured at 532 nm and the content was calculated using molecular extinction coefficient (ε = 1.5 × 10 5 M -1 cm -1 ).
간 조직의 지질은 Folch의 방법(Folch J et al., J Biol Chem 226: 497-509, 1957)으로 추출하여 지질측정용 시료로 사용하였으며 총지질 함량은 Frings와 Dunn의 방법(Frings CS and Dumm RT, Am J Clin Path 53: 89-91, 1970)으로 측정하였다.The lipid content of liver tissues was measured by Folch's method (Folch J et al., J Biol Chem 226: 497-509, 1957) and used as a lipid measurement sample. The total lipid content was determined by the method of Frings and Dunn RT, Am J Clin Path 53: 89-91, 1970).
혈청 알라닌 아미노전이효소(alanine aminotransferase, ALT) 활성도는 키트 시약(아산제약, 대한민국)을 사용하여 측정하였으며 활성도는 혈청 1㎖당 칼멘 유닛(Karmen unit)로 나타내었다. 간 조직의 단백질 함량은 소의 혈청 알부민(bovine serum albumin, BSA)을 표준용액으로 하여 측정하였다.The activity of alanine aminotransferase (ALT) was measured using a kit reagent (ASAN PHARMACEUTICAL, Korea). The activity was expressed as Karmen unit per 1 mL of serum. Protein content of liver tissue was measured by using bovine serum albumin (BSA) as a standard solution.
간 조직의 효소 활성 정도는 잔틴 산화환원효소(xanthine oxidoreductase; XOD), 슈퍼옥사이드 디스뮤타아제(superoxide dismutase: SOD), 카탈라아제(catalase: CAT), 글루타치온 퍼옥시다아제(glutathione peroxidase: GPX), 글루타치온 에스-트랜스퍼라제(glutathione S-transferase: GST), 알코올탈수소효소(alcohol dehydrogenase: ADH), 알데히드탈수소효소(aldehyde dehydrogenase: ALDH) 및 시토크롬 P450 2E1(Cytochrome P450 2E1)의 활성을 측정하여 확인하였다.측정용 효소원은 적출한 동물의 간 조직 일정량에 4배의 빙냉의 0.25M 설탕 용액을 가하여 균질화한 다음 10,000rpm에서 20분간 원심분리하여 나온 상징액(postmitochondrial fraction, PMF) 및 미토콘드리아 분획을 사용하였다. The degree of enzymatic activity of hepatic tissues is determined by xanthine oxidoreductase (XOD), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione S- The activity of glutathione S-transferase (GST), alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH) and cytochrome P450 2E1 (Cytochrome P450 2E1) The circles were homogenized by adding 4 times ice cold 0.25 M sugar solution to a fixed amount of hepatic tissues of the extracted animals and then using a postmitochondrial fraction (PMF) and a mitochondrial fraction centrifuged at 10,000 rpm for 20 minutes.
잔틴 산화환원효소의 활성 측정에서 토탈 타입(total type) 활성은 니코틴산아마이드의 존재 하에서, O 타입(O type) 활성은 니코틴산아마이드를 첨가하지 않은 상태에서 생성되는 요산의 함량을 292nm에서 흡광도를 측정하였다.In the measurement of the activity of xanthine oxidoreductase, the total type activity was measured in the presence of nicotinic acid amide, and the content of uric acid produced in the absence of nicotinic acid amide (O type) activity was measured at 292 nm .
슈퍼옥사이드 디스뮤타아제 활성은 흡광도 560nm에서 측정하였으며, 활성도는 효소 부재 하에서 헤마토자일렌의 자동산화를 50% 억제하는 단백의 함량(Unit)으로 나타내었다. 카탈라아제의 활성은 반응시키는 동안 분해되는 과산화수소의 함량을 240nm에서 측정하였다. The superoxide dismutase activity was measured at an absorbance of 560 nm, and the activity was expressed as the protein content (unit) of 50% inhibition of autoxidation of hematoxylin in the absence of the enzyme. The catalase activity was measured at 240 nm in the content of hydrogen peroxide which was decomposed during the reaction.
글루타치온 퍼옥시다아제의 활성은 기질인 과산화수소를 환원시키는데 소모되는 환원된 글루타치온(reduced glutathione)을 글루타치온 환원효소(glutathione reductase)가 글루타치온로 재환원시키는데 이용하는 니코틴아미드 아데닌 디뉴클레오티드 인산(NADPH)의 함량을 340nm에서 측정하였고, 글루타치온 에스 트랜스퍼라제의 활성은 기질인 1-클로로-2,4-디니트로벤젠(1-chloro-2,4-dinitroezene, CDNB)과 환원된 글루타치온이 포합하여 생성되는 티오에테르(thioether)를 340nm에서 흡광도를 측정하였다.The activity of glutathione peroxidase decreases the content of nicotinamide adenine dinucleotide phosphate (NADPH), which is used to convert glutathione reductase to glutathione by reducing reduced glutathione, which is consumed to reduce the substrate hydrogen peroxide, The activity of glutathione S-transferase was determined by measuring the activity of thioether produced by incorporating 1-chloro-2,4-dinitroezene (CDNB) and reduced glutathione, Absorbance at 340 nm.
알데히드탈수소효소의 활성은 기질인 아세트알데하이드와 니코틴산아마이드를 첨가하여 반응시키는 동안 생성된 니코틴아미드 아데닌 디뉴클레오티드의 함량을 340nm에서 흡광도를 측정하였으며, 활성도는 분당 단백 1 mg이 생성시킨 니코틴아미드 아데닌 디뉴클레오티드의 양을 nmole로 나타내었다.The activity of the aldehyde dehydrogenase was determined by measuring the absorbance at 340 nm of the content of nicotinamide adenine dinucleotide produced during the reaction of acetaldehyde and nicotinic acid amide as substrates, and the activity was determined by measuring the activity of nicotinamide adenine dinucleotide In terms of nmole.
시토크롬 P450 2E1(CYP2E1) 활성은 효소원에 아닐린(aniline, Sigma aldrich, USA)과 니코틴아미드 아데닌 디뉴클레오티드를 첨가하여 생성된 파라-아미노페놀(p-aminophenol)의 함량을 측정하였으며, 활성도는 단백 1mg/hr당 생성된 파라-아미노페놀을 nmole로 나타내었다.The activity of cytochrome P450 2E1 (CYP2E1) was determined by the addition of aniline (Sigma Aldrich, USA) and nicotinamide adenine dinucleotide to the enzyme source, and the activity of p-aminophenol / hr < / RTI > in nmole.
체중, 식이 및 음용수 섭취량은 전 실험 기간을 통하여 매일 일정한 시간에 측정하였다. 식이 효율은 일일 증체량을 일일 식이 섭취량으로 나눈 값으로 하였다.Weight, dietary, and drinking water intake were measured at regular intervals throughout the day. Dietary efficiency was calculated by dividing daily gain by daily dietary intake.
실험 식이로 4주간 사육한 흰쥐를 물은 자유롭게 먹게 하고, 12시간 동안 절식시킨 후 에테르 마취 하에서 복부 대동맥으로부터 채혈한 다음, 빙냉의 생리식염수로 간을 관류하고 장기를 적출하고 습기를 제거하여 무게를 측정하였다. 채취한 혈액은 실온에서 응고시킨 다음 4℃에서 2,500rpm으로 20분간 원심분리하여 혈청을 분리한 후 -70℃에 보관하면서 분석용 시료로 사용하였다.
The rats fed the diet for 4 weeks were allowed to drink water freely. After fasting for 12 hours, blood was drawn from the abdominal aorta under ether anesthesia. The liver was perfused with ice-cold physiological saline, the organs were removed, Respectively. The collected blood was coagulated at room temperature, centrifuged at 2500 rpm for 20 minutes at 4 ° C, serum was separated, and stored at -70 ° C for analysis.
실시예Example 1: 본 발명의 간 해독 또는 숙취해소용 혼합 발효물 및 혼합 발효 숙성물 1: Mixed fermented product for liver detoxification or hangover treatment of the present invention and mixed fermented aged product 의of 알코올성 Alcoholic 간손상Liver damage 예방 및 치유 효과 Prevention and healing effects
4주간 실험 식이로 성장시킨 실험동물에 체중 kg당 5g에 해당하는 50% 에탄올을 2회 경구 투여하여 알코올성 간손상 예방 및 치유 효과를 분석한 결과는 도2와 같다. FIG. 2 shows the result of the oral administration of 50% ethanol equivalent to 5 g / kg of body weight to the experimental animals grown for 4 weeks on the prevention of alcoholic liver damage and the healing effect.
체내의 과산화지질을 무독화시키는 것으로 알려진 글루타치온 함량을 분석한 결과, 50% 에탄올을 급성적으로 투여한 에탄올 대조군은 정상군과 혼합 추출물, 혼합 발효물 및 혼합 발효 숙성물 첨가 식이군에 비해 증가하였으며, 혼합 발효물의 경우 정상군에 비해 다소 낮은 글루타치온의 함량을 나타내었고, 혼합 발효 숙성물은 정상군 수준을 유지하였다(도 2A).As a result of the analysis of glutathione content, which is known to detoxify lipid peroxidation in the body, the ethanol control group treated with 50% ethanol was increased compared to the normal group and the mixed extract, mixed fermented product and mixed fermented product added diet, In the case of mixed fermented product, the content of glutathione was lower than that of the normal group, and the fermented fermented product remained in the normal group level (FIG. 2A).
활성산소종에 의해 생성되는 세포막 손상의 지표로 알려져 있는 과산화지질의 함량의 경우 에탄올의 급성적 처치에 의해 에탄올 대조군이 정상군에 비해 급격하게 증가한 반면, 천연 식물소재 혼합 추출물, 혼합 발효물 및 혼합 발효 숙성물 첨가 식이군에서는 에탄올에 의해 현저히 증가되었던 과산화지질의 함량이 정상군 수준으로 회복되었으며, 특히 혼합 발효 숙성물 첨가 식이군은 정상군 및 그 외 모든 추출물 첨가 식이군에 비해 감소하였다. 이로써, 천연 식물소재 혼합 추출물, 혼합 발효물 및 혼합 발효 숙성물의 경우 이들에 함유되어 있는 다양한 유용성분들은 에탄올 급성 독성을 예방 혹은 완화시키는 작용이 있어 간 조직 과산화지질의 함량이 낮게 나타난 것으로 판단된다. 특히 혼합 발효 숙성물 식이군의 경우 글루타치온은 정상적인 수치를 유지하면서 과산화지질과 중성지방의 함량을 현저하게 감소되었는데, 이는 발효 및 숙성에 의하여 생성되는 다양한 유용물질이 에탄올성 간 독성의 예방 및 회복에 도움을 주는 것으로 판단된다(도 2B).The content of lipid peroxidation, which is known to be an indicator of cell membrane damage caused by reactive oxygen species, was rapidly increased by the acute treatment of ethanol compared to that of the normal group, whereas the mixed extract of natural plant material, In the fermented aged group, the amount of lipid peroxidation, which was significantly increased by ethanol, recovered to the normal level. Especially, the mixed fermented aged diet - supplemented group was decreased compared with the normal group and all other extract supplemented diet group. Therefore, it was concluded that the various nutrients contained in natural plant material mixed extract, mixed fermented product and mixed fermented aged product have a function of preventing or alleviating acute toxicity of ethanol, resulting in a low content of liver lipid peroxidation. Particularly, in the case of the mixed fermented water diet group, the content of lipid peroxidation and triglyceride was remarkably decreased while the normal value of glutathione was maintained. This indicates that various useful substances produced by fermentation and aging can prevent and recover ethanolic liver toxicity (Fig. 2B).
에탄올 섭취시 간 조직에 중성지질이 배출되지 못하고 축적하여 지방간을 야기하게 된다. 이에 간 조직의 중성지방 함량을 분석한 결과, 에탄올 대조군(EN)에서는 정상군에 비해 간 조직 중성지방의 함량이 현저히 증가한 반면, 천연 식물소재 혼합 추출물(NFME), 혼합 발효물(FME) 및 혼합 발효 숙성물(FLME) 식이군의 경우 에탄올 대조군에 비해 각각 22.8%, 37.2%, 38.1%가 감소하였다. 이러한 결과로, 에탄올 대조군에서 간 조직 중성지방의 함량이 증가된 것은 에탄올의 급성 독성에 의해 나타난 것으로 사료되며 이상의 실험 결과들을 종합해 볼 때, 천연 식물소재 혼합물의 혼합 발효 숙성물은 에탄올의 급성 독성을 예방 혹은 경감시켜 줄 것으로 판단된다(도 2C).When ethanol is consumed, neutral lipids are not released into the liver tissue and accumulate and cause fatty liver. Analysis of triglyceride content of liver tissues showed that the content of triglyceride of liver tissue was significantly increased in the ethanol control group (EN) compared to that of the normal group, while that of natural plant material mixed extract (NFME), mixed fermented product (FME) In the fermented (FLME) diet group, 22.8%, 37.2% and 38.1% of the ethanol group were decreased, respectively. These results suggest that the increase in liver triglyceride content in the ethanol control group is due to the acute toxicity of ethanol. Based on the above experimental results, the mixed fermentation ages of the natural plant material mixture showed the acute toxicity of ethanol (Fig. 2C).
혈청 알라닌 아미노전이효소(ALT)와 아스파르테이트 아미노전이효소(AST)의 활성은 간 조직 손상에 의해 증가하는 것으로, 간 조직 손상의 지표로 널리 이용되고 있다. 혈청 알라닌 아미노전이효소(ALT)와 아스파르테이트 아미노전이효소(AST)를 분석한 결과, 음성 대조군인 에탄올 대조군은 정상군에 비해 증가한 반면, 각종 시료 추출물을 투여한 모든 실험군에서는 유의하게 정상군 수준으로 회복되었다(도 2D 및 도 2E).The activity of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) is increased by hepatic tissue injury and is widely used as an index of liver tissue damage. Analysis of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) showed that the ethanol control group, which was negative control group, showed a significant increase compared to the normal group, (Figs. 2D and 2E).
이러한 결과로, 에탄올 대조군에서 혈청 알라닌 아미노전이효소 및 아스파르테이트 아미노전이효소의 활성과 간 조직 과산화지질과 중성지질의 함량이 현저히 증가된 것은 급성 에탄올의 독성에 의해 나타난 것으로 생각되며, 각 시료 추출물 첨가 식이에 의해 급성 에탄올의 독성이 예방 혹은 경감되어 간 조직의 손상이 발생하지 않은 것을 확인할 수 있었다.
These results suggest that the activity of serum alanine aminotransferase and aspartate aminotransferase and the content of hepatic lipid peroxidation and triglyceride in the ethanol control group were significantly increased by the toxicity of acute ethanol. It was confirmed that the toxicity of acute ethanol was prevented or reduced by the diet, and no damage of the liver tissue occurred.
실시예Example 2: 본 발명의 간 해독 또는 숙취해소용 혼합 2: liver detoxification of the present invention or mixture of hangover 발효물Fermentation product 및 혼합 발효 And mixed fermentation 숙성물의Aged 간 조직의 효소활성 평가 Evaluation of enzymatic activity of liver tissue
본 발명의 간 해독 또는 숙취해소용 혼합 발효물 및 혼합 발효 숙성물이 간 조직의 잔틴 산화환원효소의 활성에 미치는 영향을 알아보기 위하여, 잔틴 산화환원효소, 슈퍼옥사이드 디스뮤타아제, 글루타치온 퍼옥시다아제, 카탈라아제 및 글루타치온 에스 트랜스퍼라제, 알코올 탈수소효소, 미토콘드리아-알데히드탈수소효소, 세포질-알데히드탈수소효소 및 세포질-시토크롬 p450 2E1의 활성을 측정하였다.In order to investigate the effect of the mixed fermented product and the mixed fermentation aged product of the present invention on the activity of xanthine oxidoreductase in liver tissues, xanthine oxidase, superoxide dismutase, glutathione peroxidase, The activities of catalase and glutathione estersparation, alcohol dehydrogenase, mitochondria-aldehyde dehydrogenase, cytoplasmic-aldehyde dehydrogenase and cytoplasmic cytochrome p450 2E1 were measured.
잔틴 산화효소는 정상 생리 상태 하에서는 니코틴산아마이드를 전자수용체로 이용하여 잔틴탈수소효소(xanthine dehydrogenase: XOR type D)로 존재하지만, 간 조직 손상시 니코틴산아마이드 대신 분자상 산소(O2)를 전자수용체로 이용하는 잔틴옥시다제(xanthine oxidase: XOR type O)로 전환되어 과산화물(superoxide) 및 과산화수소(hydrogen peroxide) 등과 같은 활성산소종(ROS)를 생성하게 된다. 이에 4주간 실험 식이로 성장시킨 실험동물에 체중 kg당 5g에 해당하는 50% 에탄올을 2회 경구 투여한 다음 처치하여 간 조직의 잔틴 산화환원효소 활성을 측정하였다. 잔틴 산화환원효소의 토탈 타입(XOR total type)의 경우 에탄올 급성 투여에 의해 에탄올 대조군은 정상군에 비해 감소한 반면, 천연 식물소재 혼합 추출물, 혼합 발효물 및 혼합 발효 숙성물은 에탄올 대조군에 비해 현저히 증가하였고 특히, 혼합 발효 숙성물 식이군은 정상군보다도 높은 활성을 나타내었다. 한편 잔틴 산화환원효소의 O 타입(XOR O type)의 활성은 에탄올 급성 투여에 의해 정상군에 비해 증가한 반면, 천연 식물소재 혼합 추출물, 혼합 발효물 및 혼합 발효 숙성물 식이군에서는 정상군 이하로 감소하는 것을 확인할 수 있었다(표 2).Xanthine oxidase is the normal physiological condition under the nicotinic acid amide xanthine dehydrogenase, using as an electron acceptor: using the present as (xanthine dehydrogenase XOR type D), but between textural molecule instead of nicotinic acid amide damage the oxygen (O 2) as an electron acceptor And converted to xanthine oxidase (XOR type O) to produce reactive oxygen species (ROS) such as superoxide and hydrogen peroxide. After 4 weeks, 50% ethanol equivalent to 5g / kg of body weight was orally administered to experimental animals, and the activity of xanthine oxidase activity of liver tissue was measured. In the case of total type of xanthine oxidase (XOR total type), the ethanol control group was decreased by acute administration compared with the normal group, whereas the mixed extract of natural plant material, mixed fermented product and mixed fermented product were significantly increased Especially, the fermented fermented diet group showed higher activity than the normal group. On the other hand, the activity of xanthine oxidase (XOR O type) of xanthine oxidase was increased by the acute administration of ethanol compared with that of the normal group, but decreased to below the normal group in the mixed extract of natural plant material, mixed fermented product and mixed fermented water (Table 2).
실험군
Experimental group
(Total type) Total Type
(Total type)
(O type)Oh type
(O type)
(O/T, %)Rate 1 )
(O / T,%)
1)(오타입/토탈타입)×100=비율(%). 1) (O type / total type)
2)7마리 측정치의 평균값 및 표준편차.
2) Mean value and standard deviation of 7 measurements.
본 발명의 간 해독 또는 숙취해소용 혼합 발효물 및 혼합 발효 숙성물이 간 조직의 활성산소 소거계 효소류의 활성에 미치는 영향을 알아보기 위하여, 간 조직의 슈퍼옥사이드 디스뮤타아제, 글루타치온 퍼옥시다아제, 카탈라아제 및 글루타치온 에스 트랜스퍼라제 활성을 측정하였고, 그 결과는 표 3과 같다. 천연 식물소재 혼합 추출물, 혼합 발효물 및 혼합 발효 숙성물 식이군에서 초과산화물(O2 -)을 독성이 약한 과산화수소로 전환시키고, 다시 과산화수소를 물로 전화하여 무독화시키는 슈퍼옥사이드 디스뮤타아제, 글루타치온 퍼옥시다아제, 카탈라아제의 활성은 음성 대조구인 에탄올 대조군에 비해 전반적으로 증가하였으며, 특히 혼합 발효 숙성물 식이군에서 현저히 증가한 것을 확인할 수 있었다. 체내에 미쳐 제거하지 못한 활성산소종이 과산화 지질을 만들 때 글루타치온을 이용하여 이를 독성이 약한 에탄올성 지방으로 전환하여 무독화시키는 것이 글루타치온 에스 트랜스퍼라제인데, 본 발명의 간 해독 또는 숙취해소용 혼합 발효물 및 혼합 발효 숙성물의 글루타치온 에스 트랜스퍼라제 활성에서는 에탄올 대조군에서 다소 감소하였으나 모든 실험군에서 유의한 변동은 관찰되지 않았다(표 3).In order to investigate the effect of the mixed fermented product and the mixed fermentation aged product of the present invention on the activity of the active oxygen scavenging enzymes of hepatic tissues, superoxide dismutase, glutathione peroxidase, Catalase and glutathione S-transferase activities were measured, and the results are shown in Table 3. (O 2 - ) in the group of natural plant material mixed extracts, mixed fermented products and mixed fermented aged diets were converted into hydrogen peroxide which was weakly toxic, and again hydrogen peroxide was added to water to make a detoxified superoxide dismutase, glutathione peroxidase , Catalase activity was generally increased in comparison with the ethanol control group, which was a negative control, and it was confirmed that the activity was significantly increased in the mixed fermented water diet group. It is glutathione S-transferase that converts glutathione into a weakly toxic ethanolic fat and makes it non-toxic when making active lipid peroxidation lipid peroxidation. Glutathione S-transferase activity of the mixed fermented product was slightly decreased in the ethanol control group, but no significant change was observed in all experimental groups (Table 3).
이러한 결과로, 천연 식물소재 혼합물의 혼합 발효 숙성물은 간의 항산화계 효소류의 활성증가에 도움을 주며, 급성에탄올의 투여에도 이러한 현상이 나타남에 따라 에탄올에 의한 간 손상을 예방 및 완화하는 것을 확인할 수 있었다.As a result, the mixed fermentation aged mixture of the natural plant material helps increase the activity of the antioxidant enzymes in the liver, and this phenomenon also appears in the administration of acute ethanol, so that it is confirmed that the liver damage caused by ethanol is prevented and alleviated I could.
1-4)수퍼옥시드 디스뮤타아제 단위: 유닛(헤마토자일렌의 자동산화를 50% 억제하는 단백질의 함량), 카탈라아제 단위: nmole/단백질 mg, 글루타치온 퍼옥시다아제 단위: nmole/단백질 mg, 글루타치온 에스 트랜스퍼라제: nmole/단백질 mg. 1-4) Superoxide dismutase unit: unit (content of protein that inhibits 50% of autoxidation of hematoxylin), catalase unit: nmole / protein mg, glutathione peroxidase unit: nmole / protein mg, glutathione S-transferase: nmole / protein mg.
5)7마리 측정치의 평균값 및 표준편차.
5) Mean value and standard deviation of 7 measurements.
본 발명의 간 해독 또는 숙취해소용 혼합 발효물 및 혼합 발효 숙성물이 간 조직의 알코올 대사 관련 효소류의 활성에 미치는 영향을 알아보기 위하여, 알코올 탈수소효소, 미토콘드리아-알데히드탈수소효소, 세포질-알데히드탈수소효소 및 세포질-시토크롬 p450 2E1의 활성을 평가한 결과는 도 3과 같다. In order to investigate the effect of the mixed fermented product and the mixed fermentation aged product of the present invention on the activity of the alcohol metabolism-related enzymes in liver tissues, the alcohol dehydrogenase, mitochondrial-aldehyde dehydrogenase, cytoplasmic-aldehyde dehydrogenase The results of evaluating the activity of the enzyme and cytoplasmic cytochrome p450 2E1 are shown in FIG.
아세트알데히드로 산화되며, 아세트알데히드탈수소에 의해 다시 아세트산(acetic acid)으로 산화되어 최종적으로 아세틸 조효소(acetyl CoA)로 전환되어 에너지를 생산하는 것으로 알려진 알코올탈수소효소의 활성은 정상군 및 모든 실험군간에 유의한 변동은 관찰되지 않은 반면(도 3A), 아세트알데히드의 주 대사효소인 간 조직 미토콘드리아(mitochondria) 분획 및 세포질의 알데히드탈수소효소는 에탄올 대조군에 비해 현저히 증가하였다(도 3B 및 도 3C). 반면 시토크롬 P450 2E1의 활성은 천연 식물소재 혼합 추출물, 혼합 발효물 및 혼합 발효 숙성물 식이군에서 현저히 감소되는 것을 확인할 수 있었다(도 3D).The activity of alcohol dehydrogenase, which is oxidized to acetaldehyde and converted to acetic acid by acetaldehyde dehydrogenation and finally converted to acetyl CoA to produce energy, (Fig. 3A), whereas the mitochondrial fraction of hepatic tissue, the major metabolic enzyme of acetaldehyde, and cytoplasmic aldehyde dehydrogenase were significantly increased compared to the ethanol control (Fig. 3B and Fig. 3C). On the other hand, the activity of cytochrome P450 2E1 was significantly reduced in the natural plant material mixed extract, mixed fermented product, and mixed fermented aged diet (FIG. 3D).
이러한 결과로, 천연 식물소재 혼합물의 혼합 발효 숙성물은 높은 아세트알데히드탈수소효소의 활성을 나타냄으로써 다른 추출물에 비하여 강한 독성물질인 아세트알데히드를 신속하게 대사시키므로, 급성적인 음주로 인한 간 손상과 숙취를 예방 혹은 경감 효과가 탁월한 것을 확인할 수 있었다.
As a result, the mixed fermentation aging of the mixture of natural plant material shows the activity of high acetaldehyde dehydrogenase, which rapidly metabolizes acetaldehyde, which is a strong toxic substance, compared with other extracts. Therefore, It was confirmed that the prevention or alleviation effect was excellent.
실시예Example 3: 본 발명의 간 해독 또는 숙취해소용 혼합 발효물 및 혼합 발효 숙성물의 식이 효율 및 혈중 알코올 농도 변화 평가 3: Evaluation of Dietary Efficacy and Blood Alcohol Concentration Change of Mixed Fermented Products and Mixed Fermentation Agents for Liver Detoxification or Hanging Habits of the Present Invention
4주간 실험 식이로 성장시키는 동안 증체량, 식이 섭취량 및 식이 효율을 관찰한 결과는 표 4와 같다. 체중과 초기체중의 차이로부터 산출한 1일 평균 체중 증가량은 정상군이 4.54 g인데 비하여 에탄올 대조군의 경우 3.96 g으로 11.89%가 감소하였다. 반면, 천연 식물소재 혼합 추출물, 혼합 발효물 및 혼합 발효 숙성물의 경우 에탄올만 투여한 대조군 보다 높은 식이 섭취량을 나타내었다. 음용수 섭취량은 에탄올을 투여한 실험군에서 다소 높아지는 경향을 보였다. 체중 증가량 및 식이 효율은 에탄올 대조군에서 감소하는 반면 발효숙성 혼합 추출군의 경우 그 외 실험대조군에 비해 높은 값을 나타내어 알코올 섭취에 의한 생리변동의 현상이 낮은 것을 확인할 수 있었다.Table 4 shows the results of observing weight gain, dietary intake and dietary efficiency during the 4 weeks experimental diet. The daily average weight gain from the differences in body weight and initial body weight was 4.54 g in the normal group and 3.96 g in the ethanol control group, which was decreased by 11.89%. On the other hand, mixed extracts of natural plant material, mixed fermented product and mixed fermented product showed higher dietary intake than ethanol - only control. The intake of drinking water tended to be somewhat higher in the ethanol - treated group. Weight gain and dietary efficiency were decreased in the ethanol control group, but the fermented mixed extract group had a higher value than the other experimental control groups, indicating that the phenomenon of physiological fluctuation due to alcohol consumption was low.
(g/일)Dietary intake
(g / day)
(g/일)Drinking water intake
(g / day)
(g/일)Weight gain
(g / day)
1)식이 효율: 일일 증체량/일일 식이 섭취량 1) Dietary efficiency: daily gain / daily dietary intake
2)7마리 측정치의 평균값 및 표준편차
2) Mean value and standard deviation of 7 measurements
본 발명의 간 해독 또는 숙취해소용 혼합 발효물 및 혼합 발효 숙성물이 혈중 에탄올의 경시적 농도변화에 미치는 영향을 알아보기 위하여, 4주간 사육한 실험동물에게 50% 에탄올을 체중 kg당 2g의 농도로 경구투여한 후 혈중 에탄올의 경시적 농도변화를 조사하였다. 에탄올 대조군은 최고 농도에 도달하였다가 이후 서서히 감소하여 4시간 경과 후에는 40% 정도로 감소하였다. 반면 천연 식물소재 혼합 추출물의 경우 30분 이후부터 혈중 에탄올 농도가 서서히 감소하였고, 본 발명의 혼합 발효물 및 혼합 발효 숙성물 식이군의 경우 에탄올 농도는 현저하게 감소하였다. 특히 혼합 발효 숙성물에서 경시적 혈중 알코올 농도의 변화가 큰 것으로 나타났는데 이와 같은 결과는 알코올대사의 보조적인자인 CAT 활성이 높게 나타난 상기의 실험결과에 따른 것으로 판단된다(도 4).To investigate the effect of the mixed fermented product and the mixed fermentation aged product of the present invention on the change of the ethanol concentration with time, 50% ethanol was added to 2 g per kg of body weight And the changes of ethanol concentration in blood were examined. The ethanol control group reached its peak concentration, then gradually decreased and decreased to about 40% after 4 hours. On the other hand, in the case of the mixed extract of natural plant material, the concentration of ethanol in the blood gradually decreased after 30 minutes, and the ethanol concentration of the mixed fermented product and mixed fermented aged food group of the present invention was remarkably decreased. In particular, the change in the blood alcohol concentration with time was found to be large in the mixed fermentation aged product, and this result is considered to be the result of the above experiment in which the CAT activity, which is an auxiliary agent of alcohol metabolism, is high (FIG.
이와 같이, 각종 시료 추출물 첨가 식이군에서 간 조직 알코올탈수소효소의 활성은 음성 대조군인 에탄올 대조군보다 혈중 에탄올 농도가 낮아진 것은 에탄올을 아세트알데히드로 전환하는데 보조적으로 관여하는 효소인 카탈라아제의 활성과 체내에 아세트알데히드를 아세트산으로 전환시키는 알데히드탈수소효소의 활성을 증가시킴과 동시에 간 조직손상에 관여하는 활성산소 생성계 효소는 저해하고 활성산소 소거계 효소의 활성이 증가하여 나타난 것으로 해석할 수 있다.Thus, the activity of liver tissue alcohol dehydrogenase in the dietary group with various sample extracts was lower than that of the ethanol control group, which is a negative control group, because the activity of catalase, which is an enzyme involved in the conversion of ethanol into acetaldehyde, It can be interpreted that the activity of the aldehyde dehydrogenase which converts the aldehyde to acetic acid is increased and the active oxygen generating enzyme which is involved in the liver tissue damage is inhibited and the activity of the active oxygen scavenging enzyme is increased.
Claims (8)
(a) 세척한 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이를 동결건조한 후 100~140 메쉬로 분말화하는 단계;
(b) 상기 분말화한 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이 분말을 혼합물 100 중량부 기준으로, 대두 분말 68~72 중량부, 녹두 분말 3~7 중량부, 미나리 분말 3~7 중량부, 다시마 분말 3~7 중량부, 갈근 분말 3~7 중량부, 무 분말 3~7 중량부 및 오이 분말 3~7 중량부를 혼합하여 75~85℃에서 30~50분 동안 살균하고 발효미생물과 혼합하는 단계; 및
(c) 상기 발효미생물과 혼합한 혼합물을 습도 70~90%, 35~40℃에서 2~3일 동안 발효하는 단계.2. The composition for liver detoxification or hangover as claimed in claim 1, wherein the mixed fermented product is produced by the following steps:
(a) lyophilizing washed soybeans, mung beans, buttercups, kelp, parsley, radishes and cucumbers, followed by pulverization to 100-140 mesh;
(b) 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, mung bean, parsley, kelp, parsley, radish and cucumber powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered powder and 3 to 7 parts by weight of cucumber powder are mixed and sterilized at 75 to 85 ° C for 30 to 50 minutes, ; And
(c) fermenting the mixture mixed with the fermenting microorganism at a humidity of 70 to 90% at 35 to 40 DEG C for 2 to 3 days.
(a) 세척한 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이를 동결건조한 후 100~140 메쉬로 분말화하는 단계;
(b) 상기 분말화한 대두, 녹두, 미나리, 다시마, 갈근, 무 및 오이 분말을 혼합물 100 중량부 기준으로, 대두 분말 68~72 중량부, 녹두 분말 3~7 중량부, 미나리 분말 3~7 중량부, 다시마 분말 3~7 중량부, 갈근 분말 3~7 중량부, 무 분말 3~7 중량부 및 오이 분말 3~7 중량부를 혼합하여 75~85℃에서 30~50분 동안 살균하고 발효미생물과 혼합하는 단계; 및
(c) 상기 발효미생물과 혼합한 혼합물을 습도 70~90%, 35~40℃에서 2~3일 동안 발효한 후, 습도 40~60%, 10~20℃에서 4~10일 동안 저온숙성하는 단계.The composition according to claim 1, wherein the low-temperature aged product of the mixed fermented product is prepared by the following steps:
(a) lyophilizing washed soybeans, mung beans, buttercups, kelp, parsley, radishes and cucumbers, followed by pulverization to 100-140 mesh;
(b) 68 to 72 parts by weight of soybean powder, 3 to 7 parts by weight of mung bean powder, 3 to 7 parts by weight of powdered mung bean, mung bean, parsley, kelp, parsley, radish and cucumber powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered pear powder, 3 to 7 parts by weight of powdered powder and 3 to 7 parts by weight of cucumber powder are mixed and sterilized at 75 to 85 ° C for 30 to 50 minutes, ; And
(c) fermenting the mixture obtained by mixing the fermenting microorganism with 70 to 90% of humidity at 35 to 40 ° C for 2 to 3 days and then aging at 40 to 60% of humidity and 10 to 20 ° C for 4 to 10 days at a low temperature step.
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KR20190044177A (en) * | 2017-10-20 | 2019-04-30 | 대구한의대학교산학협력단 | Composition for preventing and protecting liver injury comprising the extract of sea tangle or fucoxanthin |
KR20200092532A (en) * | 2019-01-24 | 2020-08-04 | 동신대학교산학협력단 | Anti-inflammatory composition using an extract of radish root, etc. |
WO2020231127A1 (en) * | 2019-05-14 | 2020-11-19 | 한국식품연구원 | Strain showing liver function improving activity, and use thereof |
KR102195980B1 (en) * | 2020-05-21 | 2020-12-29 | 김일 | Fermentation pills and its manufacturing method |
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KR950010794A (en) * | 1993-10-14 | 1995-05-15 | 김정순 | Hangover beverage composition |
KR0140057B1 (en) * | 1994-10-25 | 1998-06-01 | 박대규 | Alcohol metabolism drinks including plant extract & its method |
KR100968109B1 (en) * | 2008-01-15 | 2010-07-06 | 주식회사농심 | Calcium?compound containing natural product composition for curing a hangover and preparation method thereof |
KR101291388B1 (en) * | 2011-03-14 | 2013-07-30 | 이현구 | Crude Saponin Mixture Extracted from Fermentation Mixture of Soybean and Ginseng Having a Functionality for Hangover and It's Preparation Methods |
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KR20190044177A (en) * | 2017-10-20 | 2019-04-30 | 대구한의대학교산학협력단 | Composition for preventing and protecting liver injury comprising the extract of sea tangle or fucoxanthin |
KR20200092532A (en) * | 2019-01-24 | 2020-08-04 | 동신대학교산학협력단 | Anti-inflammatory composition using an extract of radish root, etc. |
WO2020231127A1 (en) * | 2019-05-14 | 2020-11-19 | 한국식품연구원 | Strain showing liver function improving activity, and use thereof |
KR102195980B1 (en) * | 2020-05-21 | 2020-12-29 | 김일 | Fermentation pills and its manufacturing method |
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