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KR20000006848A - Anticancerous functional Danmuji and process for preparation thereof - Google Patents

Anticancerous functional Danmuji and process for preparation thereof Download PDF

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Publication number
KR20000006848A
KR20000006848A KR1019990049257A KR19990049257A KR20000006848A KR 20000006848 A KR20000006848 A KR 20000006848A KR 1019990049257 A KR1019990049257 A KR 1019990049257A KR 19990049257 A KR19990049257 A KR 19990049257A KR 20000006848 A KR20000006848 A KR 20000006848A
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South Korea
Prior art keywords
radish
salt
functional
anticancer
weight
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KR1019990049257A
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Korean (ko)
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KR100396841B1 (en
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박건영
유종현
안미영
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박건영
유종현
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Priority to KR10-1999-0049257A priority Critical patent/KR100396841B1/en
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L19/00Products from fruits or vegetables; Preparation or treatment thereof
    • A23L19/20Products from fruits or vegetables; Preparation or treatment thereof by pickling, e.g. sauerkraut or pickles
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00Function of food ingredients
    • A23V2200/30Foods, ingredients or supplements having a functional effect on health
    • A23V2200/308Foods, ingredients or supplements having a functional effect on health having an effect on cancer prevention
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/90Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Mycology (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

PURPOSE: A pickled radish and a preparation method thereof are provided, which pickled radish blocks the growth of solid tumor, strengthen the immunity of spleen, and increases the activity of glutathione-S-transferase which removes toxic materials in a liver. CONSTITUTION: The pickled radish composition comprises 0.2-0.6 wt% of a natural dye, 1-3 wt% of salt, 0.1-0.4 wt% of sugar, 0.3-0.7 wt% of an active material and the balance of water. The natural dye is a mixture of gardenia yellow and safflower yellow in a ratio of 7:3. The salt is a mixture of the broiled salt and KCl substitute salt in a ratio of 7:3-3:7. The sager is stevioside and the active material is sodium alginate. The pickled radish is prepared by soaking a washed radish into a 15% brine containing the broiled salt and KCl substitute salt (3:7-7:3); desalting the radish; seasoning the radish with the composition; and ripening the radish for 4 days in a refrigerator.

Description

항암 기능성 단무지 및 그 제조방법{Anticancerous functional Danmuji and process for preparation thereof}Anti-cancer functional radish and method for preparing the same {Anticancerous functional Danmuji and process for preparation etc}

본 발명은 항암 기능성 단무지 및 그 제조방법에 관한 것으로 더욱 상세하게는 본 발명은 단무지 제조시에 천연색소, 구운소금, KCl 대체염, 스테비오사이드 (stevioside) 및 알긴산나트륨(sodium alginate)을 사용하여 항암효과를 증진시킨 단무지 및 그 제조방법에 관한 것이다.The present invention relates to anticancer functional radish and a method for preparing the same. More specifically, the present invention relates to anticancer using natural pigment, roasted salt, KCl substitute salt, stevioside and sodium alginate in the production of radish. It is related with the pickled radish and the manufacturing method which enhanced the effect.

단무지는 주로 가정에서 담그는 것은 드물고 거의 공장에서 담그어 시판되고 있다. 단무지의 제조방법은 주로 염적 침지법을 사용하여 단순히 무우를 소금에 절여 일정기간(2-3개월) 경과한 후 적정 수준의 염도가 유지되도록 탈염한 다음 조미하는 제조 공정을 거쳐 식품으로 유통되고 있는데 유통과정에서 효소적인 분해와 산화에 의해 제품의 변질과는 관계없이 색상이 조금씩 갈변화되는 문제점이 있다.Pickled radish is rarely immersed at home, and is almost commercially immersed in factories. The production method of pickled radish is mainly distributed by using salt immersion method, simply salted radish, and then desalted to maintain proper salinity after a certain period of time (2-3 months). There is a problem that the color changes little by little regardless of product deterioration by enzymatic decomposition and oxidation during distribution.

한편, 단무지의 맛은 주로 짠맛과 단맛으로 느껴지기 때문에 이 두 맛을 내는 원료의 선택은 매우 중요하다. 그러나 현재 시판되는 단무지는 짠맛은 천일염으로, 단맛은 사카린나트륨(sodium saccharin)을 사용하고 있다. 천일염은 요즘 바다의 오염으로 인하여 중금속이 많이 함유되어 있으며, 사카린 나트륨은 과다 복용시 암을 유발할 수 있으므로 식품첨가물로서 바람직하지 않다.On the other hand, since the taste of pickled radish is mainly felt as salty and sweet, the choice of raw materials for these two flavors is very important. However, commercially available pickled radish uses the salt of the sun salt and the salt of sodium saccharin (sodium saccharin). Natural salts are heavy metals due to sea pollution these days, and sodium saccharin is not preferable as a food additive because it can cause cancer when overdose.

본 발명자들은 상기와 같은 문제점을 해결하기 위해 단무지가 어느정도 갈변이 일어나더라도 제품의 질을 떨어뜨리지 않도록 천연색소 치자(gardenia yellow)와 홍화(safflower yellow)를, 소금은 천일염의 대용으로 고열에서 정제하여 불순물을 제거한 구운소금과 KCl 대체염 등을, 당으로는, 사카린나트륨의 대용으로는 천연감미료인 스테비오사이드를, 활성물질은 소금 흡착 기능이 있다고 알려진 알긴산 나트륨(sodium alginate)을 이용하여 단무지를 제조하였으며 제조한 단무지가 항암효과가 있음을 확인하므로써 본 발명을 완성하였다.In order to solve the above problems, the present inventors refined the natural pigments of gardenia and safflower yellow so that the radish does not degrade the product even if browning occurs to some extent, and salt is purified at high temperature as a substitute for sun salt. Roasted salt and KCl alternative salts free of impurities are used, sugar, stevioside, a natural sweetener, and sodium alginate, which are known to have salt adsorption, are used as a substitute for sodium saccharin. The present invention was completed by confirming that the prepared radish had anti-cancer effects.

따라서, 본 발명의 목적은 천연색소, 구운소금, KCl 대체염, 스테비오사이드 (Stevioside), 알긴산나트륨(sodium alginate)을 사용하여 제조한 항암 기능성 단무지를 제공함에 있다. 본 발명의 다른 목적은 상기 항암 기능성 단무지의 제조방법을 제공함에 있다.Therefore, an object of the present invention is to provide an anticancer functional radish prepared using natural pigments, baked salt, KCl substitute salt, stevioside, sodium alginate. Another object of the present invention to provide a method for producing the anticancer functional radish.

본 발명의 상기 목적은 천연색소, 구운소금과 KCl 혼합염, 스테비오사이드 (stevioside)및 알긴산나트륨(sodium alginate)을 사용하여 본 발명 항암 기능성 단무지를 제조하고 같은 방법으로 재료를 달리하여 시판단무지를 제조한 후 본 발명 항암 기능성 단무지와 시판 단무지의 메탄올 추출물을 종양세포를 이식한 마우스에 투여하고 고형암의 무게와 자연살해세포의 활성 및 글루타치온-S-트랜스퍼라아제(glutathione-S-transferase)의 활성을 각각 조사하여 본 발명 항암 기능성 단무지가 항암성이 있음을 확인하고 상기 마우스 장기의 중량변화를 조사하여 단무지 메탄올 추출물이 항암성을 증진시키는 효과외에 다른 장기에 미치는 영향을 조사하며 또 본 발명 항암 기능성 단무지의 메탄올 추출물을 투여한 마우스의 간조직내 글라이코겐 분포와 면역조직학적 반응을 조사하므로써 달성하였다.The object of the present invention is to prepare the anticancer functional radish of the present invention using natural pigments, roasted salt and KCl mixed salt, stevioside and sodium alginate and to produce commercially available radish by different materials in the same manner. The anticancer functional radish and commercial radish of the present invention were then administered to mice transplanted with tumor cells, and the weight of solid cancer, the activity of natural killer cells, and the activity of glutathione-S-transferase. Investigate each of the anticancer functional radish of the present invention has anticancer properties and examine the weight change of the mouse organs to investigate the effect of the radish extract of radish on other organs besides the effect of enhancing anticancer activity and the anticancer functional radish of the present invention Distribution of Glycogen and Immunohistologic Responses in Mice Treated with Methanol Extract It was achieved by irradiation.

이하, 본 발명의 구성 및 작용을 설명한다.Hereinafter, the configuration and operation of the present invention.

도 1은 본 발명 항암 기능성 단무지의 제조과정을 나타낸 순서도이다.1 is a flow chart showing the manufacturing process of the anticancer functional radish of the present invention.

도 2는 본 발명 항암 기능성 단무지와 시판 단무지가 마우스 비장 임파구의 자연살해세포 활성에 미치는 영향을 나타낸 그래프이다.Figure 2 is a graph showing the effect of the anticancer functional radish and commercial radishes of the present invention on natural killer cell activity of mouse spleen lymphocytes.

도 3은 본 발명 항암 기능성 단무지와 시판 단무지가 마우스 간내 글루타치온-S-트랜스퍼라아제 활성에 미치는 영향을 나타낸 그래프이다.Figure 3 is a graph showing the effect of the anticancer functional radish and commercial radish of the present invention on glutathione-S-transferase activity in mouse liver.

도 4a∼4d는 정상 간조직(a)과 종양세포 sarcoma-180을 이식한 마우스에 PBS만을 투여한 군(b), 시판 단무지(c)와 본 발명 항암 기능성 단무지(d)의 메탄올 추출물을 각각 투여한 군의 간조직내 글라이코겐 분포를 헤마토실린-에오신 (hematoxylin eosin)염색으로 확인하는 결과 사진도이다.4A to 4D show methanol extracts of group (b), commercial radish (c), and anticancer functional radish (d) of the present invention, in which only PBS was administered to mice transplanted with normal liver tissue (a) and tumor cells sarcoma-180, respectively. The result of confirming the glycogen distribution in the liver tissue of the group administered by hematoxylin eosin staining.

도 5a∼5d는 정상 간조직(a)과 종양세포 sarcoma-180을 이식한 마우스에 PBS만을 투여한 군(b), 시판 단무지(c)와 본 발명 항암 기능성 단무지(d)의 메탄올 추출물을 각각 투여한 군의 간조직내 글라이코겐 분포를 피리오딕 산 시프(periodic acid Schiff;PAS) 반응으로 실시한 결과 사진도이다.5A to 5D show methanol extracts of group (b), commercial radish (c), and anticancer functional radish (d) of the present invention, which were administered with PBS only to mice transplanted with normal liver tissue (a) and tumor cells sarcoma-180, respectively. Glycogen distribution in the hepatic tissue of the group administered is a photographic view of the result of the pyiodic acid Schiff (PAS) reaction.

도 6a∼6d는 정상 간조직(a)과 종양세포 sarcoma-180을 이식한 마우스에 PBS만을 투여한 군(b), 시판 단무지(c)와 본 발명 항암 기능성 단무지(d)의 메탄올 추출물을 각각 투여한 군의 간조직에 대한 종양관련 단백질중 항체 Bax에 나타낸 반응 결과 사진도이다.6a to 6d are methanol extracts of the group (b), commercially available radish (c), and anticancer functional radish (d) of the present invention, which were administered with PBS only to mice transplanted with normal liver tissue (a) and tumor cells sarcoma-180, respectively. It is a photograph of the response result shown to antibody Bax among tumor-related proteins with respect to liver tissue of administration group.

도 7a∼7d는 정상 간조직(a)과 종양세포 sarcoma-180을 이식한 마우스에 PBS만을 투여한 군(b), 시판 단무지(c)와 본 발명 항암 기능성 단무지(d)의 메탄올 추출물을 각각 투여한 군의 간조직에 대한 종양관련 단백질중 항체 Rb(sc-050)에 나타낸 반응 결과 사진도이다.7A to 7D are methanol extracts of the group (b), commercial radish (c), and anticancer functional radish (d) of the present invention, which were administered with only PBS to mice transplanted with normal liver tissue (a) and tumor cells sarcoma-180, respectively. The photograph shows the result of the reaction shown in antibody Rb (sc-050) among tumor-associated proteins to the liver tissues of the administered group.

도 8a∼8d는 정상 간조직(a)과 종양세포 sarcoma-180을 이식한 마우스에 PBS만을 투여한 군(b), 시판 단무지(c)와 본 발명 항암 기능성 단무지(d)의 메탄올 추출물을 각각 투여한 군의 간조직에 대한 종양관련 단백질중 항체 c-fos(sc-253)에 나타낸 반응 결과 사진도이다.8a to 8d are methanol extracts of the group (b), commercial radish (c), and anticancer functional radish (d) of the present invention, which were administered with PBS only to mice transplanted with normal liver tissue (a) and tumor cells sarcoma-180, respectively. The photograph shows the result of the reaction shown in the antibody c-fos (sc-253) among the tumor-related proteins to the liver tissue of the group to which the administration was performed.

본 발명은 천연색소, 구운소금과 KCl 혼합염, 스테비오사이드 및 알긴산나트륨을 사용하여 본 발명 항암 기능성 단무지를 제조하고 색소를 사용치 않고 천일염과 사카린을 사용하여 상기 본 발명 항암 기능성 단무지와 동일한 방법으로 시판 단무지를 제조하는 단계; 시판 단무지와 본 발명 항암 기능성 단무지를 동결건조시켜 분말화한 후 10배의 메탄올을 첨가하여 메탄올 추출물을 얻은 후 진공농축하는 단계; 마우스에 종양세포를 이식하고 시판 단무지와 본 발명 항암 기능성 단무지 추출물 시료를 투여한 후 종양의 무게를 측정하여 본 발명 항암 기능성 단무지의 고형암 성장 저지 효과를 조사하는 단계; 종양세포를 이식한 마우스에 PBS를 투여한 경우와 시판 단무지 및 본 발명 항암 기능성 단무지 추출물 시료를 투여한 경우 각 장기의 중량변화를 측정하여 시료가 다른 장기에 미치는 영향을 조사하는 단계; 종양세포를 이식한 마우스의 비장에서 자연살해(natural killer;NK) 세포의 활성을 조사하여 본 발명 항암 기능성 단무지의 면역활성 증가 효과를 조사하는 단계; 종양세포를 이식한 마우스 간에서 체내 독성물질과 과산화물질을 전이, 배설함으로서 무독화에 관여하는 효소 글루타치온-S-트랜스퍼라아제(glutathione-S-transferase) 활성을 측정하는 단계; 종양세포를 이식한 마우스의 간조직내 글라이코겐 분포를 조사하여 본 발명 항암 기능성 단무지의 간세포 손상정도를 조사하는 단계 및; 종양세포를 이식한 마우스의 간조직에 종양 관련 단백질에 대한 면역조직학적 반응을 조사하는 단계로 구성된다.The present invention prepares the anticancer functional radish of the present invention using natural pigments, roasted salt and KCl mixed salt, stevioside and sodium alginate, and the same method as the anticancer functional radish of the present invention using sun salt and saccharin without using a pigment. Preparing commercial radish; Lyophilizing commercially available radish and the anticancer functional radish of the present invention, followed by pulverization to obtain methanol extract by adding 10-fold methanol; Implanting tumor cells into the mouse and administering a commercial radish extract and an anticancer functional radish extract sample of the present invention, and then measuring the tumor weight to investigate the effect of inhibiting solid cancer growth of the anticancer functional radish of the present invention; Investigating the effect of the sample on other organs by measuring the weight change of each organ when PBS was administered to a mouse transplanted with tumor cells and when the commercial radish extract and the anticancer functional radish extract sample of the present invention were administered; Investigating the activity of natural killer (NK) cells in the spleen of mice transplanted with tumor cells to investigate the effect of increasing the immune activity of the anticancer functional radish of the present invention; Measuring the enzyme glutathione-S-transferase activity involved in detoxification by transferring and excreting toxic substances and peroxides in the liver of mice transplanted with tumor cells; Investigating glycogen distribution in liver tissue of a mouse transplanted with tumor cells to examine the degree of hepatocellular damage of the anticancer functional radish of the present invention; Examining the immunohistochemical response to tumor-associated proteins in the liver tissue of the mouse transplanted tumor cells.

본 발명에서 사용한 천연색소는 단무지가 어느 정도 갈변이 일어나더라도 제품의 질이 떨어지지 않도록 치자(gardenia yellow)와 홍화(safflower yellow)를 3: 7∼7:3의 비율로 혼합하여 사용하였으며 항암 기능성 단무지용 조미 조성물 총 중량에 대해 0.2∼0.6중량%를 사용하였다.Natural pigment used in the present invention was mixed with Gardenia yellow and safflower yellow in a ratio of 3: 7 to 7: 3 so that the quality of the product does not deteriorate even if the radish is browned to some extent. 0.2 to 0.6 wt% was used relative to the total weight of the seasoning composition.

본 발명에서 사용한 소금은 천일염의 대용으로 고열 정제하여 불순물을 제거한 구운소금과 KCl 대체염(KCl과 NaCl이 3:7로 함유)을 7:3∼3:7의 비율로 하고 항암 기능성 단무지용 조미 조성물 총 중량에 대해 0.5∼0.8중량%를 사용하였다.Salt used in the present invention is a high-temperature refined as a substitute for sun salt and baked salt and KCl substitute salt (containing KCl and NaCl as 3: 7) in a ratio of 7: 3 to 3: 7, and the anti-cancer functional radish seasoning 0.5 to 0.8 weight percent was used relative to the total weight of the composition.

본 발명에서 사용한 당은 사카린 나트륨 대용으로 천연감미료인 스테비오사이드(stevioside)를 사용하였으며 항암 기능성 단무지용 조미 조성물 총 중량에 대해 0.08∼0.10중량%를 사용하였다.The sugar used in the present invention used stevioside, a natural sweetener, as a substitute for saccharin sodium, and 0.08 to 0.10% by weight based on the total weight of the seasoning composition for anticancer functional radish.

본 발명에서 사용한 활성물질은 소금 흡착기능이 있다고 알려진 알긴산나트륨(sodium alginate)을 사용하였으며 항암 기능성 단무지용 조미 조성물 총 중량에 대해 0.10∼30중량%를 사용하였다.The active material used in the present invention used sodium alginate known to have a salt adsorption function and 0.10 to 30% by weight based on the total weight of the seasoning composition for anticancer functional radish.

본 발명 항암 기능성 단무지는 탈염공정이 끝나고 조미공정에서 유익한 천연색소를 이용하여 단무지의 갈변화를 방지하였고 활성물질과 구운 소금, 대체염 등을 이용하여 깔끔한 맛을 더하였으며 항암성 증진에도 중요하게 작용하였다.The anti-cancer functional radish of the present invention prevents browning of the radish by using natural pigments beneficial in seasoning process after the desalting process, and adds a refreshing taste by using active substances, roasted salts, alternative salts, etc. It was.

이하, 본 발명의 구체적인 방법을 실시예를 들어 상세히 설명하고자 하지만 본 발명의 권리범위는 이들 실시예에만 한정되는 것은 아니다.Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

실시예 1: 본 발명 항암 기능성 단무지 및 시판 단무지 제조Example 1: Anticancer functional radish and commercial radish preparation of the present invention

가을 무를 깨끗하게 세척하여 15% 농도의 구운소금과 KCl 염을 3:7의 비율로 혼합한 혼합염 소금물에 침지시킨 후 완전히 탈염하고 색소, 활성물질, 당 및 소금으로 조미한 후 숙성시켰다. 이때, 조미 조성액은 표 1에 나타낸 바와 같이 사용한 색소로는 치자황색소와 홍화황색소를 7:3비율로 하고, 소금은 KCl 대체염(KCl:NaCl=3:7)과 구운소금을 3:7의 비율로 하였으며 당으로는 스테비오사이드(stevioside100s)를, 활성물질로 알긴산 나트륨(sodium alginate 0.5%)으로 조미하였다. 이렇게 조미한 후 4일간 냉장고에서 침적, 숙성시켰다(도 1). 시판 단무지도 상기 본 발명 항암 기능성 단무지와 동일한 방법으로 제조하되 사용하는 재료는 역시 표 1에 나타낸 바와 같다.Autumn radish was washed clean and immersed in mixed salt brine mixed with 15% of roasted salt and KCl salt in a ratio of 3: 7, then completely desalted, seasoned with pigments, active substances, sugars and salts, and aged. At this time, the seasoning composition is shown in Table 1 as a pigment used in the gardening yellow and safflower yellow (7: 3) ratio, salt is KCl alternative salt (KCl: NaCl = 3: 7) and baked salt 3: The ratio was 7 and stevioside (stevioside 100s) was used as sugar and sodium alginate (0.5% sodium alginate) as an active substance. After seasoning was soaked and aged in the refrigerator for 4 days (Fig. 1). Commercial radish also produced in the same manner as the anticancer functional radish of the present invention, but the materials used are also shown in Table 1.

본 발명 항암 기능성 단무지와 시판 단무지의 재료배합비Material mixing ratio of anticancer functional radish and commercial radish of the present invention 시판 단무지Commercial pickled radish 항암 기능성 단무지Anticancer functional radish 탈염한 무Desalted radish 1kg1 kg 1kg1 kg water 1kg1 kg 1kg1 kg 시트르산Citric acid 0.6kg0.6kg 0.6g0.6g 아세트산 결정Acetic acid crystals 2g2 g 2g2 g Party 사카린 0.6gSaccharin 0.6g 스테비오사이드 1.8gStevioside 1.8g 글라이신Glycine 0.26g0.26 g 0.26g0.26 g 비타민 CVitamin c 0.166g0.166 g 0.166g0.166 g 사과산Malic acid 0.266g0.266 g 0.266g0.266 g 1 Salt 1 20g20 g 구운소금 14g: KCl + NaCl(KCl:1.8g, NaCl: 4.2g)Baked salt 14g: KCl + NaCl (KCl: 1.8g, NaCl: 4.2g) 인산염(STPP)Phosphate (STPP) 0.2g0.2 g 0.2g0.2 g 천연색소2 Natural colors 2 -- 치차황색소(1.4∼4.2g) + 홍화황색소(0.6∼1.8g)Yellow tea (1.4-4.2g) + yellow safflower (0.6-1.8g) 활성물질Active substance -- 알긴산염(0.5%) 5gAlginate (0.5%) 5g 1 소금: 구운소금 : 대체염 = 7:32 천연색소: 치자황색소 : 홍화황색소 = 7 : 3(gardenia yellow) (safflower yellow)1 Salt: Baked salt: Alternative salt = 7:32 Natural color: Gardenia yellow: Safflower yellow = 7: 3 (gardenia yellow) (safflower yellow)

실시예 2: 조미 단무지의 메탄올 추출물 제조Example 2: Preparation of Methanol Extract of Seasoned Radish

상기 실시예 1에서 4일간 냉장고에서 침적, 숙성한 각각의 단무지를 동결건조시켜 분말화한 후 시료 중량의 10배의 메탄올을 추가하고, 60 ~ 70℃의 수욕조에서 3 ~ 4시간 동안 시료를 추출하는데 이 과정을 3회 반복하였다. 여기서 얻은 메탄올 추출물을 회전식 진공 농축기로 농축하여 메탄올을 완전히 제거하였다. 실험에 사용된 시료는 DMSO(dimethyl sulfoxide)에 10% 용액으로 녹인 다음 냉동실에서 냉동시킨 뒤 필요시 해동하여 적당한 농도로 희석하여 사용하였다.In Example 1, four days of immersion and maturation of the dried radish were aged by lyophilization and pulverization, and then methanol was added 10 times the weight of the sample, and the sample for 3 to 4 hours in a 60 ~ 70 ℃ water bath. This process was repeated three times for extraction. The methanol extract obtained here was concentrated in a rotary vacuum concentrator to completely remove the methanol. The sample used in the experiment was dissolved in 10% solution in DMSO (dimethyl sulfoxide), frozen in a freezer, thawed if necessary, and diluted to an appropriate concentration.

실시예 3: 본 발명 항암 기능성 단무지의 고형암 성장저지 효과Example 3 Solid Cancer Growth Inhibition Effect of Anticancer Functional Radish of the Present Invention

본 실시예에서는 상기 실시예 2에서 얻은 본 발명 항암 기능성 단무지와 시판 단무지로부터 얻은 메탄올 추출물 시료에 의한 항종양효과를 보기 위하여 Balb/c 마우스의 왼쪽 서혜부 피하에 sarcoma-180 종양세포를 이식시킨 후 20일 동안 시료를 투여하고 이로부터 32일이 지난 다음 마우스를 해부하여 종양을 분리하고 그 무게를 측정하였다. 즉, In vitro에서 생존력 시험(viablility test)를 바탕으로 기능성단무지와 시판단무지 모두 0.25mg/mL의 농도로 결정하고 0.25mg/kg의 단무지 메탄올 추출물을 시료로 투여하였다. 실험결과, 표 2에 나타낸 바와 같이 종양세포만 이식한 대조군(S-180+PBS, 시료대신 동량의 PBS만 투여)은 종양의 무게가 6.08g인 반면 기능성 단무지를 주사한 군(S-180+FD)은 4.38g으로 감소하여 27.96%의 종양생성 억제효과를 보였으며 시판단무지를 주사한 군(S-180+CD)은 5.93g으로 대조군과 큰 차이를 보이지 않았다. 따라서 기능성 단무지가 고형암 성장저지 효과가 있음을 알 수 있었다.In this example, in order to see the anti-tumor effect of the anticancer functional radish obtained from Example 2 and the methanol extract obtained from commercial radish obtained in Example 2, sarcoma-180 tumor cells were implanted 20 subcutaneously under the left groin of Balb / c mice. Samples were administered for one day and 32 days later, mice were dissected to isolate tumors and weighed. That is, based on the viability test (viblility test) in vitro, both the functional and commercial radish was determined at a concentration of 0.25mg / mL and 0.25mg / kg of radish methanol extract was administered as a sample. As a result, as shown in Table 2, the control group implanted with only tumor cells (S-180 + PBS, administered with the same amount of PBS instead of the sample) had a tumor weight of 6.08g while the group was injected with functional mussels (S-180 + FD) decreased to 4.38 g, showing 27.96% of tumor suppression effect. The group injected with commercially available radish (S-180 + CD) was 5.93 g, which was not significantly different from the control group. Therefore, the functional radish was found to have a solid growth inhibitory effect.

sarcoma-180 세포을 이식한 Balb/c 마우스에 단무지 메탄올 추출물을 투여한 후 종양무게 측정Tumor weight measurement after administration of radish methanol extract to Balb / c mice transplanted with sarcoma-180 cells 샘플Sample Dose(mg/kg)Dose (mg / kg) 종양 무게(g)Tumor Weight (g) 저해비율(%)Inhibition Ratio (%) S-180+PBSS-180 + PBS 0.250.25 6.08±0.046.08 ± 0.04 -- S-180+CD1 S-180 + CD 1 0.250.25 5.93±0.845.93 ± 0.84 2.472.47 S-180+FD2 S-180 + FD 2 0.250.25 4.38±1.224.38 ± 1.22 27.9627.96 [주] 1: 시판 단무지2: 항암 기능성 단무지[Note] 1: commercially available radish 2: anti-cancer functional radish

실시예 4: 단무지 메탄올 추출물를 식이로 급여한 마우스 각 장기 중량변화Example 4 Weight Changes of Organs of Rats Fed Methanol Extract by Diet

본 발명 항암 기능성 단무지 메탄올 추출물과 시판 단무지 메탄올 추출물이 종양외에 다른 기타 기관에서의 작용을 측정하기 위하여, 대조군(Control)은 자유식이만을 시켰고, 종양세포대조군(S-180+PBS)에는 자유식이를 시키고 주사로 PBS를 투여하였고, 처리군은 자유식이를 주고 주사로 종양세포대조군과 동량인 단무지 메탄올 추출물을 종양세포(Sarcoma-180 cell)이식 후 상기 실시예 3과 동일하게 시료를 20일 동안 투여하고 각 장기의 중량을 측정하였다. 실험결과, 표 3에 나타낸 바와 같이, 먼저 대사과정 중에 생기는 여러 유해한 물질들의 해독작용을 담당하는 간의 중량비는 종양세포만 이식한 대조군(S-180+PBS)과 시판단무지를 주사한 군(S-180+CD)이 기능성단무지를 주사한 군(S-180+FD)에 비해 다소 높았고 비장의 경우도 기능성단무지를 주사한 군이 종양세포만 이식한 대조군에 비해 낮았으나 시판단무지를 주사한 군과는 큰 차이는 없었다.In order to measure the action of the anticancer functional radish methanol extract and commercial radish methanol extract of the present invention in other organs besides the tumor, the control was only free diet, and the tumor cell control group (S-180 + PBS) was free diet. PBS was administered by injection, and the treated group was given a free diet and injected the sample for 20 days after transplanting tumor cells (Sarcoma-180 cell) with the same extract of radish as the tumor cell control group. And the weight of each organ was measured. As a result of the experiment, as shown in Table 3, the weight ratio of liver, which is responsible for the detoxification of various harmful substances generated during metabolic processes, was the control group (S-180 + PBS) transplanted with tumor cells only and the group injected with commercially available anchovy (S- 180 + CD) was slightly higher than the group injected with functional monoclonal (S-180 + FD) and the spleen was significantly lower than the control group implanted with only tumor cells. There was no big difference.

본 발명 항암 기능성 단무지의 메탄올 추출물과 시판 단무지의 메탄올 추출물을 식이로 공급한 마우스 각 장기의 무게Weight of each organ of mice fed with methanol extract of anticancer functional radish and commercial radish methanol extract of the present invention as a diet 샘플Sample 체중(g)Weight (g) 비장/몸무게(%)Spleen / Weight (%) 간/몸무게(%)Liver / Weight (%) 심장/몸무게(%)Heart / Weight (%) 신장/몸무게(%)Height / weight (%) 대조군Control 28.95±1.928.95 ± 1.9 0.33±0.020.33 ± 0.02 5.74±0.65.74 ± 0.6 0.50±0.150.50 ± 0.15 1.85±0.051.85 ± 0.05 S-180+PBSS-180 + PBS 29.5±1.329.5 ± 1.3 1.55±0.121.55 ± 0.12 7.72±0.97.72 ± 0.9 0.34±0.010.34 ± 0.01 1.58±0.041.58 ± 0.04 S-180+CD1 S-180 + CD 1 29.9±1.429.9 ± 1.4 1.47±0.091.47 ± 0.09 7.57±0.77.57 ± 0.7 0.34±0.010.34 ± 0.01 1.70±0.181.70 ± 0.18 S-180+FD2 S-180 + FD 2 28.6±2.128.6 ± 2.1 1.40±0.011.40 ± 0.01 6.68±0.46.68 ± 0.4 0.35±0.030.35 ± 0.03 1.66±0.071.66 ± 0.07 [주] 1: 시판 단무지2: 항암 기능성 단무지[Note] 1: commercially available radish 2: anti-cancer functional radish

실시예 5: 본 발명 항암 기능성 단무지와 시판 단무지의 면역활성 증가 효과Example 5 Increasing Immune Activity of Anticancer Functional Radishes and Commercial Radishes of the Present Invention

본 실시예에서는 실시예 4와 동일한 방법으로 종양세포를 이식하고 20일 동안 본 발명 항암 기능성 단무지 추출물과 시판 단무지 추출물 시료를 투여한 후 각 마우스의 비장을 무균적으로 적출하여 비장의 자연살해(natural killer;NK) 세포 활성을 살펴보았다. 자연살해세포의 활성을 분석하기 위해 먼저 이펙터 세포(Effector cell)인 NK세포를 준비하였다. 마우스의 비장을 클린 벤치(clean bench) 내에서 무균적으로 적출하여 페니실린(100U/mL), 스트렙토마이신(100㎍/mL)를 함유한 5mL의 RPMI 1640 배지로 3회 세척한 후 곱게 마쇄하였다. 이 세포 부유액을 70㎛ 나일론 메쉬로 여과시켜서 원심분리하여 림프구를 모은 다음 이것을 같은 배지에 부유시켰다. 이것을 다시 히스토파쿠에-1077(histopaque-1077)을 이용하여 원심분리(500×g, 30 min, 18℃)를 통해서 림프구를 분리해냈다. 준비한 세포를 10% FCS, 2mM L-글루타민, 페니실린(100U/mL), 스트렙토마이신(100㎍/mL)을 함유한 RPMI 1640 배지에 재현탁시켰다. 이것을 37℃, CO2배양기에서 1 ~ 2 시간 배양시켜서 세포가 배양 플라스크(6cm 페트리디쉬)에 부착되도록 하였다. 비부착성 자연살해 세포를 원심분리하여 수집한 후 배양배지에 재현탁시킨 다음 적정 세포수를 계수하여 사용하였다. 이어서, 자연살해 세포의 세포파괴활성은 MTT 방법으로 분석하였다. Yac-1 마우스 림포마 세포(Yac-1 mouse lymphoma cell)는 타겟 세포(target cell)로 사용하였다. Yac-1 세포는 NK 세포 활성을 측정하는데 가장 편리한 타겟 세포주이다. Yac-1 세포는 10% FCS, 2mM L-글루타민 페니실린(100U/mL), 스트렙토마이신(100㎍/mL)를 함유한 RPMI 1640 배지에 5×104cells/mL의 밀도가 되도록 희석한 후 각 웰에 50㎕씩 가하였다. 그리고 NK cell은 1×107cells/mL의 NK 세포 활성 분석을 위해서 이펙터 세포와 Yac-1 cell 각각 50㎕씩을 96웰 플레이트에 첨가하였다. 이를 37℃, CO2배양기 안에서 3일간 배양시킨 후 MTT(5mg/mL) 용액을 10㎕씩 각 웰에 가한 다음 37℃에서 4시간 배양하였다. 여기에 SDS(10% in 0.02N HCl)을 25㎕ 첨가하여 실온에서 30분간 발색시킨 후 540nm에서 OD를 측정하였다. 세포 독성 퍼센트는 하기 공식에 의해 계산하였다.In this embodiment, tumor cells were transplanted in the same manner as in Example 4, and after administration of the anticancer functional radish extract and commercial radish extract sample of the present invention for 20 days, spleens of each mouse were sterilely extracted and natural killing of spleen was performed. killer (NK) cell activity was examined. In order to analyze the activity of natural killer cells, first, effector cells (NK cells) were prepared. The spleens of the mice were aseptically extracted in a clean bench, washed three times with 5 mL of RPMI 1640 medium containing penicillin (100 U / mL) and streptomycin (100 μg / mL) and then finely ground. The cell suspension was filtered through a 70 μm nylon mesh, centrifuged to collect lymphocytes, and then suspended in the same medium. Lymphocytes were isolated by centrifugation (500 × g, 30 min, 18 ° C.) using histopacque-1077 (histopaque-1077). The prepared cells were resuspended in RPMI 1640 medium containing 10% FCS, 2 mM L-glutamine, penicillin (100 U / mL), streptomycin (100 μg / mL). This was incubated for 1 to 2 hours at 37 ° C. in a CO 2 incubator to allow cells to adhere to the culture flask (6 cm Petri dish). Non-adhesive natural killer cells were collected by centrifugation, resuspended in culture medium, and the appropriate number of cells was counted. Subsequently, cell destruction activity of natural killer cells was analyzed by MTT method. Yac-1 mouse lymphoma cells were used as target cells. Yac-1 cells are the most convenient target cell line for measuring NK cell activity. Yac-1 cells were diluted to a density of 5 × 10 4 cells / mL in RPMI 1640 medium containing 10% FCS, 2 mM L-glutamine penicillin (100 U / mL) and streptomycin (100 μg / mL). 50 μl was added to the wells. And NK cells were added to the 96 well plate 50μL of effector cells and Yac-1 cells for the analysis of NK cell activity of 1 × 10 7 cells / mL. After incubating for 3 days at 37 ° C. in a CO 2 incubator, 10 μl of MTT (5 mg / mL) solution was added to each well and incubated at 37 ° C. for 4 hours. 25 μl of SDS (10% in 0.02N HCl) was added thereto, followed by color development at room temperature for 30 minutes, and OD was measured at 540 nm. Percent cytotoxicity was calculated by the following formula.

실험결과, 도 2에 나타낸 바와 같이 항암 기능성 단무지투여군(S-180+FD)이 58.39%의 저해효과를 나타내 가장 높았으며 대조군은 50.37%, 종양세포만을 이식한 경우는 32.01%, 종양세포를 이식하고 시판 단무지 메탄올 추출물을 투여한 군은 32.41%를 나타냈다. 상기 결과에서 시판 단무지를 투여한 군은 NK세포 면역활성이 낮아 종양세포만 이식한 군과 비슷하게 나타났다.As a result, as shown in Figure 2, the anti-cancer functional group (S-180 + FD) showed the highest inhibitory effect of 58.39%, the control group was 50.37%, only the tumor cells were transplanted 32.01%, tumor cells were transplanted The group receiving the commercial radish methanol extract showed 32.41%. In the above results, the group administered with commercial radish showed low NK cell immune activity, similar to the group transplanted with only tumor cells.

실시예 6: 본 발명 항암 기능성 단무지와 시판 단무지에 의한 마우스 간내의 글루타치온-S-트랜스퍼라아제 활성 변화Example 6: Glutathione-S-transferase Activity Changes in Mouse Liver by Anticancer Functional Radish and Commercial Radish of the Present Invention

간에서 일어나는 대사효소계의 phaseⅡ 단계는 내인성 물질이나 외부에서 투여되어지는 독성물질을 포함하거나 수용성 물질로 전환시켜 체외로 배출 시킴으로서 이물질을 제거하는 작용을 하는데 글루타치온-S-트랜스퍼라아제(glutathione-S-transferase)는 환원 글루타치온(reduced glutathione)을 이용하여 체내 독성물질과 과산화물질을 전이, 배설함으로서 무독화에 관여하는 효소이다.Phase II of the metabolic enzyme system in the liver removes foreign substances by including endogenous substances or toxic substances administered externally or by converting them into water-soluble substances to be discharged to the body. Glutathione-S-transferase (glutathione-S- Transferase is an enzyme that is involved in detoxification by transferring and excreting toxic substances and peroxides in the body using reduced glutathione.

본 실시예에서는 상기 실시예 5와 같이 시료와 식이를 공급한 쥐의 간내 글루타치온-S-트랜스퍼라아제 활성을 측정하였다. 먼저 글루타치온- S-트랜스퍼라아제(GST) 활성을 측정하기 위해 마우스를 치사시킨 후 4℃ 이하의 생리 식염수로 간을 관류하여 간내에 남아 있는 혈액을 제거한 후 간장을 적출하였다. 간조직 1g 당 0.1M 인산칼륨 버퍼(potassium phospate buffer, pH 7.4)를 가하여 빙냉하에서 글라스 테플론 호모게나이저(glass teflon omogenizer)로 마쇄하였다. 이 마쇄액을 호모게네이트(homogenate) 분획으로 하였으며 이것을 13,000 rpm에서 10분간 원심분리하여 핵 및 미마쇄세포부분을 제거하고 다시 105,000×g에서 1시간 동안 초원심분리하여 얻은 상등액을 사이토졸 분획으로 하였다. Habig 등의 방법에 준하여 0.1M 인산칼륨버퍼(pH 6.5) 중에 0.04M 환원 글루타치온 75㎕을 가한 후 효소액을 0.1mL 넣고 블랭크에는 20% 트리크로아세트산 0.5mL를 가해 효소를 실활시키고 시료는 25℃에서 5분간 반응시킨 후 블랭크와 시료 각각에 0.12M 1-클로로-2,4-디니트로벤젠 25㎕를 가하여 25℃에서 2분간 반응시킨 다음 시료에 20% 트리클로로아세트산을 가해 반응을 완결시킨 후 원심분리하여 얻은 상등액을 340nm에서 흡광도를 측정한 다음 1-클로로-2,4-디니트로벤젠의 몰 흡광계수 9.6mM1cm-1을 이용하여 활성도를 산정하였다. 효소 활성의 단위는 1분간 mg 단백질이 생성한 2,4-디니트로벤젠-글루타치온의 nmole 수로 표시하였다. 실험결과, 도 3에 나타낸 바와 같이 글루타치온-S-트랜스퍼라아제 활성은 종양세포를 이식하지 않은 대조군이 392.2nmol/mg protein/min이었으며 종양세포를 이식한 대조군(S-180 + PBS)에서는 281.4nmol/mg protein/min으로 떨어졌다. 시판단무지를 주사한 군(S-180 + CD)에서는 277.5nmol/mg protein/min으로 종양세포를 이식한 대조군보다 오히려 낮은 수치를 보였으나, 기능성단무지를 주사한 군(S-180 + FD)에서는 510.4nmol/mg protein/min으로 종양세포를 이식한 대조군에 비해 Glutathione-S-transferase의 활성이 현저히 높은 것으로 나타났다.In the present Example, glutathione-S-transferase activity in the rats fed the sample and the diet was measured as in Example 5. First, mice were killed to measure glutathione-S-transferase (GST) activity, and then liver was perfused with physiological saline of 4 ° C. or lower to remove blood remaining in the liver, followed by extraction of the liver. 0.1 M potassium phosphate buffer (pH 7.4) was added per 1 g of liver tissue, and the mixture was ground with a glass teflon omogenizer under ice cooling. This grinding solution was used as a homogenate fraction, which was centrifuged at 13,000 rpm for 10 minutes to remove the nuclei and unbranched cell sections, and the supernatant obtained by ultracentrifugation for 1 hour at 105,000 × g was used as the cytosol fraction. It was. According to the method of Habig et al., 75 μl of 0.04 M reduced glutathione in 0.1 M potassium phosphate buffer (pH 6.5) was added, 0.1 mL of enzyme solution was added, 0.5 mL of 20% trichloroacetic acid was added to the blank, and the sample was inactivated at 25 ° C. After reacting for 5 minutes, add 25 µl of 0.12M 1-chloro-2,4-dinitrobenzene to the blank and each sample, and react for 2 minutes at 25 ° C. Then, add 20% trichloroacetic acid to the sample to complete the reaction. The supernatant obtained from the separation was measured for absorbance at 340 nm, and then the activity was calculated using the molar extinction coefficient of 1-chloro-2,4-dinitrobenzene at 9.6 mM 1 cm -1 . Units of enzymatic activity were expressed as nmole number of 2,4-dinitrobenzene-glutathione produced by mg protein for 1 minute. As shown in FIG. 3, glutathione-S-transferase activity was 392.2 nmol / mg protein / min in the control group without transplanting tumor cells and 281.4 nmol in the control group (S-180 + PBS) without transplantation of tumor cells. dropped to / mg protein / min. In the group injected with commercially available radish (S-180 + CD), the level was lower than the control group implanted with tumor cells at 277.5 nmol / mg protein / min. The activity of Glutathione-S-transferase was significantly higher than that of the control group transplanted with 510.4 nmol / mg protein / min.

실시예 7: 본 발명 항암 기능성 단무지와 시판 단무지에 의한 마우스 간조직내의 글라이코겐 분포조사Example 7 Investigation of Glycogen Distribution in Mouse Liver Tissues by Anticancer Functional Radish and Commercial Radish

본 실시예에서는 상기 실시예 5와 같이 시료와 식이를 공급한 쥐의 간조직내 글라이코겐 분포를 조사하였다. 쥐의 간을 절취하여 4% 파라포름알데하이드 인 포스페이트-버퍼 사린(paraformaldehyde in phosphate-buffered saline;PBS)에 4℃, 12시간 고정하여 순차적인 탈수와 투명화를 거쳐 파라핀(paraffin)에 포매한 후 6㎛ 연속절편을 제작하였다. 간의 조직학적 변화와 글라이코겐 분포관찰을 위해 탈 파라핀한 후 헤마토실린-에오신(hematoxylin-eosin:H-E) 염색과 PAS(periodic acid Schiff)반응을 실시하였다. 실험결과, 도 4a와 도 5a는 정상적인 간조직을 나타내는 것이고 도 4b와 도 5b에는 종양세포 sarcoma-180를 이식하고 PBS를 투여한 군(S-180 + PBS)으로 간 특유의 간소엽 형태를 갖추고 있으나 약한 혼탁종창, 수포성 변성, 지방변성, Kupffer 세포변화, 국소적 괴사 등의 변화와 함께 호산성 세포도 일부 관찰되는 등 간세포의 변화를 보이고 있다. 대조군에서 위와 같은 조직학적 변화가 현저히 증가할 뿐만 아니라 호산성세포와 핵이 농염되는 세포가 증가하는 등 많은 변화를 보이는데 특히 간소엽 주변구역에서 심하였다. 시판 단무지군을 식이한 경우(S-180 + CD)는 도 4c와 도 5c에서 보는 바와 같이 조직학적 변화가 S-180 + PBS와 유사하였으며 항암 기능성 단무지군을 식이한 경우(S-180 + FD)는 도 4d와 도 5d에서 보는 바와 같이 지방변성, Kupffer 세포변화, 국소적 괴사에서 다소 적은 변화를 보였다. 이를 표 4에 정리하였다. 글라이코겐 분포는 정상적인 간조직에 비해 S-180 + PBS의 간소엽 주변구역과 중간구역에서 감소하며 S-180 + CD와 S-180 + FD도 정상군에 비해 간소엽 주변구역에서 다소 감소하였다. 이는 시판단무지보다 본 발명 항암 기능성 단무지가 간세포를 덜 손상시킴을 나타낸다. 이는 표 5에 정리하였다.In this example, the glycogen distribution in the liver tissues of the rats fed the sample and the diet was examined as in Example 5. The rat liver was excised and fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS) at 4 ° C for 12 hours, followed by sequential dehydration and clarification and embedded in paraffin. A micrometer continuous section was produced. Deparaffinization, hematoxylin-eosin (H-E) staining and periodic acid Schiff (PAS) reaction were performed for histological changes and glycogen distribution. As a result, Figures 4a and 5a shows normal liver tissue and Figures 4b and 5b are tumor cells sarcoma-180 transplanted and administered with PBS (S-180 + PBS) has a liver-specific hepatic lobe morphology However, changes in hepatocytes have been observed, such as weak eosinophils, bullous degeneration, adipose degeneration, Kupffer cell change, local necrosis, and some eosinophilic cells. In the control group, not only did the histological changes significantly increase, but also increased the number of eosinophilic cells and the cells of the nucleus-concentrating cells, especially in the periphery of the hepatic lobe. In the case of commercially available radish group (S-180 + CD), histological changes were similar to S-180 + PBS as shown in FIGS. As shown in FIGS. 4D and 5D, a little change was observed in fat degeneration, Kupffer cell change, and local necrosis. This is summarized in Table 4. Glycogen distribution was decreased in the hepatic and peripheral zones of S-180 + PBS compared to normal liver tissue, and S-180 + CD and S-180 + FD were slightly decreased in the hepatic lobe surrounding areas compared to normal groups. . This indicates that the anticancer functional radish of the present invention is less damaging to liver cells than commercially available radish. This is summarized in Table 5.

마우스의 간내 조직변화Intrahepatic Tissue Changes in Mice 그룹/HCGroup / HC CSCS HDHD FCFC KCKC FNFN 정상군Normal ++ 0-+0- + +-+++-++ ++ 0-+0- + S-180 + PBSS-180 + PBS ++++++ ++++ ++++++ ++++++ ++++ S-180 + CDS-180 + CD ++++++ ++++ ++++++ ++++++ ++++ S-180 + FDS-180 + FD ++++++ ++++ ++++ ++++ ++ [주] HC: 조직적 변화, CS: 종기형성, HD: 수증 변질, FC: 지방질 변화,KC: Kupffer 세포 반응, FN: 병소 회저0:변화가 없거나 무시해도 좋을 정도+: 약하게 나타남++: 적당한 수준으로 나타남+++: 심하게 나타남++++: 매우 심하게 나타남Note: HC: histologic change, CS: boil formation, HD: hydrophilic alteration, FC: lipid change, KC: Kupffer cell response, FN: lesion regression 0: no change or negligible +: appears weak ++: moderate Shown as a level +++: Shows bad ++++: Shows very bad

마우스 간내의 글라이코겐 분포Glycogen Distribution in Mouse Liver 그룹group PZHLPZHL IZHLIZHL CZHLCZHL 정상군Normal ++++++++ ++-+++++-+++ ++-+++++-+++ S-180 + PBSS-180 + PBS ++++++ ++++ ++-+++++-+++ S-180 + CDS-180 + CD ++-+++++-+++ ++-+++++-+++ ++-+++++-+++ S-180 + FDS-180 + FD ++++++ ++-+++++-+++ ++-+++++-+++ [주] PZHL: 간소엽 말초지역, IZHL: 간소엽 중간지역,CZHL: 간소엽 중앙지역,0:변화가 없거나 무시해도 좋을 정도+: 약하게 나타남++: 적당한 수준으로 나타남+++: 심하게 나타남++++: 매우 심하게 나타남Note: PZHL: Peripheral section of hepatic lobe, IZHL: Intermediate section of mesenchymal lobe, CZHL: Central section of mesenchymal lobe, 0: No change or negligible +: Appear weak +++: Appear moderate ++++: Very bad

실시예 8: 본 발명 항암 기능성 단무지와 시판 단무지에 의한 마우스 간조직의 면역조직학적 반응Example 8: Immunohistologic Responses of Mouse Liver Tissues by Anticancer Functional Muds and Commercial Muds of the Invention

본 실시예에서는 상기 실시예 5와 같이 시료와 식이를 공급한 쥐의 간조직에 대한 종양관련 단백질의 면역조직화학적 반응을 보기 위해 각 종양과 관련된 단백질에 대한 면역조직학적 관찰을 위해 탈파라핀하여 10mM 소듐 시트레이트 버퍼(pH 6.0)에서 95℃ 5분간 처리하였고 다시 실온에서 동일 버퍼에 20분 두었다. 이를 3% 메탄올 하이드로전 퍼옥사이드에 30분간 실온에서 처리하고 PBS(0.01M, pH 7.5)로 5분간 3회 세척한 후 비특이성 반응을 없애기 위하여 goat normal serum(Vector Lab., PK-6101)으로 실온에서 30분간 처리하였다. 이를 제거하고 Santa Cruz Biotechnology Inc 제품의 Bax(sc-493), Bcl-2(sc-783), Rb(sc-050), c-fos (sc-253), c-jun(sc-1694)을 500:1로 희석하여 4℃ 습실에 16시간 동안 반응시켰다. PBS로 세척하고 biotinylated anti-rabbit lg G(Vector Lab., PK-6101)를 실온에서 30분 동안 반응시켰으며 PBS로 세척하고 ABC 키트(avidin-biotin-peroxidae complex, vector Lab., PK-6101)에 실온에서 60분간 반응시켰다. 다시 PBS로 세척한 후 DAB substract 키트(vector Lab., SK-4100)로 실온에서 5분간 발색시켰다. 증류수로 세척하여 Mayer's hematoxylin으로 대조 염색한 후 광학현미경하에서 반응정도를 판정하였다. 실험결과, 간세포에 대한 종양관련 단백질의 면역조직화학적 반응을 보면 6가지 항체중 Bax, Rb, c-fos에만 반응을 나타내며 세포질에 비해 핵에서 강한 반응을 보였으며 일부 간세포사이의 염증세포에서도 반응을 나타내었다. 도 6a ~ 6d에 나타낸 바와 같이 Bax는 핵에서만 반응을 나타내며 정상군에 비해 S-180 + PBS와 S-180 + CD는 감소하였으며 기능성 단무지군인 S-180 + FD에서는 감소정도가 적었다. 도 7a ~ 7d에 나타낸 바와 같이 Rb는 핵과 세포질 모두에서 정상군에 비해 S-180 + PBS와 S-180 + CD의 경우에 증가하여 특히 중심정맥주위 간세포의 세포질의 변화가 심하였으나 기능성 단무지군인 S-180 + FD의 경우는 세포질의 변화는 적었다. c-fos는 도 8a ~ 8d에 나타낸 바와 같이 정상군의 핵과 세포질에 비해 S-180 + PBS에서 증가하는데 시판 단무지군 S-180 + CD과 기능성 단무지군S-180 + FD에서도 대조군과 유사하게 증가하였다. 이를 표 6에 정리하였다.In this example, to see the immunohistochemical response of the tumor-associated protein to the liver tissue of the rat fed the sample and diet as in Example 5, deparaffinizing 10 mM for immunohistochemical observation of the protein associated with each tumor Treated at 95 ° C. for 5 minutes in sodium citrate buffer (pH 6.0) and again placed in the same buffer at room temperature for 20 minutes. This was treated with 3% methanol hydroperoxide at room temperature for 30 minutes, washed three times with PBS (0.01M, pH 7.5) three times for 5 minutes, followed by goat normal serum (Vector Lab., PK-6101). Treatment was at room temperature for 30 minutes. Remove it and replace Bax (sc-493), Bcl-2 (sc-783), Rb (sc-050), c-fos (sc-253) and c-jun (sc-1694) from Santa Cruz Biotechnology Inc. Diluted to 500: 1 and reacted for 4 hours in 4 ℃ wet. Washed with PBS and reacted with biotinylated anti-rabbit lg G (Vector Lab., PK-6101) for 30 minutes at room temperature, washed with PBS and ABC kit (avidin-biotin-peroxidae complex, vector Lab., PK-6101) Was reacted at room temperature for 60 minutes. After washing with PBS again, it was developed for 5 minutes at room temperature with a DAB substract kit (vector Lab., SK-4100). After washing with distilled water and control dyeing with Mayer's hematoxylin, the reaction degree was determined under an optical microscope. As a result, immunohistochemical reactions of tumor-associated proteins against hepatocytes showed only Bax, Rb, and c-fos among six antibodies, and showed stronger responses in the nucleus than cytoplasm, and also in inflammatory cells between some hepatocytes. Indicated. As shown in FIGS. 6a to 6d, Bax showed a reaction only in the nucleus, and S-180 + PBS and S-180 + CD were decreased compared with the normal group, and the decrease was small in the functional group of S-180 + FD. As shown in Figure 7a to 7d Rb increased in the case of S-180 + PBS and S-180 + CD in the nucleus and cytoplasm compared to the normal group, especially the change in the cytoplasm of the peripheral venous hepatocytes, but S functional group In the case of -180 + FD, cytoplasmic changes were small. c-fos is increased in S-180 + PBS compared to the nucleus and cytoplasm of the normal group as shown in Figures 8a to 8d, similar to the control group in the commercial radish group S-180 + CD and S-180 + FD It was. This is summarized in Table 6.

쥐의 간조직내 종양관련 단백질의 면역조직화학적 반응Immunohistochemical Responses of Tumor-associated Proteins in Rat Liver 그룹group 지역/항체Region / Antibody BaxBax Bcl-2Bcl-2 RbRb c-junc-jun c-fosc-fos 정상군Normal 핵세포질Nuclear cytoplasm ++++0++++ 0 0000 +++0-++++ 0- + 0000 +++0-++++ 0- + S-180 + PBSS-180 + PBS 핵세포질Nuclear cytoplasm ++0++ 0 0000 ++++++++++++ 0000 ++++++++++++ S-180 + CDS-180 + CD 핵세포질Nuclear cytoplasm ++-+++0++-+++ 0 0000 ++++++++++ 0000 ++++++++ S-180 + FDS-180 + FD 핵세포질Nuclear cytoplasm +++0+++ 0 0000 ++++++++ 0000 ++++-++++++-++ [주] 0:변화가 없거나 무시해도 좋을 정도+: 약하게 나타남++: 적당한 수준으로 나타남+++: 심하게 나타남++++: 매우 심하게 나타남Note: 0: No change or negligible +: Appears weak +++: Appropriate

이상, 상기 실시예를 통하여 설명한 바와 같이 천연색소로 치자(gardenia yellow)와 홍화(safflower yellow), 소금으로 구운소금과 KCl 대체염, 당으로 스테비오사이드(stevioside) 및 활성물질로 소금흡착기능이 있는 알긴산나트륨(sodium alginate)을 사용하여 조미 및 숙성시킨 본 발명 항암 기능성 단무지는 고형암 성장을 저지하고 비장의 면역기능을 강화하며 간에서 독성물질을 제거하는 글루타치온-S-트랜스퍼라아제의 활성을 증가시키고 우수한 항암성을 나타내는 뛰어난 효과가 있으므로 식품산업상 매우 유용한 발명인 것이다.As described above, as described in the above examples, gardenia (gardenia yellow) and safflower (safflower yellow), salt-to-salt salt and KCl substitute salt, stevioside as sugar, and a salt adsorbing function as an active substance as described in the above embodiment The anticancer functional radish of the present invention seasoned and aged using sodium alginate increases the activity of glutathione-S-transferase which inhibits solid cancer growth, enhances the spleen's immune function, and removes toxic substances from the liver. It is a very useful invention in the food industry because it has an excellent effect of showing excellent anticancer properties.

Claims (4)

천연색소, 소금, 당, 활성물질 및 물로 구성된 것을 특징으로 하는 항암 기능성 단무지용 조미 조성물.Seasoning composition for anticancer functional radish consisting of natural pigments, salts, sugars, active substances and water. 제 1 항에 있어서, 천연색소는 치자(gardenia yellow)와 홍화(safflower yellow)를 7:3으로 혼합한 혼합색소 0.2 ~ 0.6중량%, 소금은 구운소금과 KCl 대체염을 7:3 ~ 3:7의 비율로 혼합한 염 1 ~ 3중량%, 당은 스테비오사이드(stevioside) 0.1 ~ 0.4중량%, 활성물질은 알긴산 나트륨(sodium alginate) 0.3 ~ 0.7중량% 및 나머지는 물로 구성된 것을 특징으로 하는 항암 기능성 단무지용 조미 조성물.The method of claim 1, wherein the natural pigment is 0.2 to 0.6% by weight mixed with gardenia yellow (safflower yellow) and safflower yellow (7: 3), the salt is 7: 3 to 3: 1 to 3% by weight of the salt mixed in a ratio of 7, sugar is stevioside (stevioside) 0.1 to 0.4% by weight, active substance is sodium alginate (sodium alginate) 0.3 to 0.7% by weight of the anti-cancer Seasoning composition for functional radish. 제 2 항 기재의 항암 기능성 단무지용 조미 조성물로 탈염한 무를 조미, 숙성시킨 것을 특징으로 하는 항암 기능성 단무지.An anticancer functional pickled radish characterized by seasoning and ripening radish desalted by the seasoning composition for anticancer functional radish described in claim 2. 세척한 무를 구운소금과 KCl을 3:7 ~ 7:3 범위로 혼합한 15% 농도 소금물에 침지시킨 후 완전히 탈염하고 제 1 항 내지 2 항 기재중 선택된 어느 한 항의 항암 기능성 단무지용 조미 조성물로 조미하여 4일간 냉장고에서 침적, 숙성시키는 것을 특징으로 하는 제 3 항 기재의 항암 기능성 단무지 제조방법.Washed radish is immersed in 15% brine mixed with roasted salt and KCl in the range of 3: 7 to 7: 3, then completely desalted and seasoned with the anti-cancer functional radish seasoning composition of any one of claims 1 to 2. A method for producing anticancer functional radish according to claim 3, wherein the product is deposited and aged in a refrigerator for 4 days.
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US7003976B2 (en) 2002-12-10 2006-02-28 Lg Electronics Inc. Air conditioner
US7165416B2 (en) 2002-12-10 2007-01-23 Lg Electronics Inc. Air conditioner
KR100916518B1 (en) * 2009-07-09 2009-09-08 한광웅 The manufacturing method of picked radish containing natural-pigment and fruit-flavored, and the picked radish thereof

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KR19980024025A (en) * 1996-09-12 1998-07-06 최원배 Manufacturing method of ginseng radish
KR19990080233A (en) * 1998-04-14 1999-11-05 김기영 Manufacturing Method of Gardenia Compressed Monochrome

Cited By (5)

* Cited by examiner, † Cited by third party
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US7003976B2 (en) 2002-12-10 2006-02-28 Lg Electronics Inc. Air conditioner
US7162889B2 (en) 2002-12-10 2007-01-16 Lg Electronics Inc. Air conditioner
US7165416B2 (en) 2002-12-10 2007-01-23 Lg Electronics Inc. Air conditioner
US7174741B2 (en) 2002-12-10 2007-02-13 Lg Electronics Inc. Air conditioner
KR100916518B1 (en) * 2009-07-09 2009-09-08 한광웅 The manufacturing method of picked radish containing natural-pigment and fruit-flavored, and the picked radish thereof

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