KR101939192B1 - Composition for treating acute inflammatory and immuine enhancement - Google Patents
Composition for treating acute inflammatory and immuine enhancement Download PDFInfo
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- KR101939192B1 KR101939192B1 KR1020170101554A KR20170101554A KR101939192B1 KR 101939192 B1 KR101939192 B1 KR 101939192B1 KR 1020170101554 A KR1020170101554 A KR 1020170101554A KR 20170101554 A KR20170101554 A KR 20170101554A KR 101939192 B1 KR101939192 B1 KR 101939192B1
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- A23L—FOODS, 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
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Abstract
Description
본 발명은 급성 염증 개선 및 면역 증진용 조성물에 관한 것으로, 더욱 상세하게는 녹용 분말 및 천연 약재 분말을 함유하여 급성 염증의 개선 및 면역 증진의 효과를 나타내는 조성물에 관한 것이다.The present invention relates to a composition for improving acute inflammation and immunity, and more particularly, to a composition for improving acute inflammation and improving immunity by containing an antler powder and a natural pharmaceutical powder.
최근 생약 성분을 이용한 염증의 치료, 예방, 개선에 대한 다양한 기술이 개발되고 있다.Recently, various techniques for the treatment, prevention, and improvement of inflammation using herbal ingredients have been developed.
염증 피부의 치료, 개선을 위한 기술로는 대한민국 등록특허공보 10-0532633호에서 당귀, 숙지황, 천궁, 작약을 일정 혼합비로 추출하여 얻은 혼합 식물 추출물을 통해 항염증 효과를 나타내는 화장료 조성물을 들 수 있다. 이러한 화장료 조성물은 피부에 도포함으로써 염증 피부의 치료뿐만 아니라 세포막 보호나 세포증식의 효과도 얻을 수 있는 것으로 보고되고 있다.Examples of techniques for treating and improving inflammatory skin include cosmetic compositions that exhibit an anti-inflammatory effect through a mixed plant extract obtained by extracting Angelicae radix, Rhododendron japonica, Rhodiola japonica, Angelica japonica, and Angelica keiskei at a certain mixing ratio in Korean Patent Publication No. 10-0532633 . Such a cosmetic composition has been reported to be able to obtain not only treatment of inflammatory skin but also protection of cell membrane and cell proliferation by applying to the skin.
한편, 대한민국 공개특허공보 10-2017-0047555호에서 백선피, 천궁 및 참당귀로 이루어진 군에 선택된 1종 이상을 초임계 추출법으로 추출한 추출물을 유효성분으로 함유함으로써 피부염 등의 염증 또는 부종을 예방 및 치료할 수 있는 조성물이 개시되어 있고, 대한민국 공개특허공보 10-2017-0033590호에서는 목과, 홍화), 당귀, 방풍, 오가피 및 천궁을 필수적으로 포함하는 복합 생약 추출물 또는 상기 추출물의 분획물을 유효성분으로 함유하여 염증성 질환의 예방 또는 치료의 효과를 나타내는 조성물이 개시되어 있다. 상기 선행기술들에서는 염증성 사이토카인을 농도 의존적으로 감소시키고 염증반응의 주요 전사인자들을 전좌 및 인산화 저해시킴에 따라 염증반응을 억제하는 효과를 얻고 있다.Korean Patent Laid-Open Publication No. 10-2017-0047555 discloses a method for preventing or treating inflammation or swelling such as dermatitis by containing, as an active ingredient, an extract obtained by extracting at least one selected from the group consisting of white radish, And in Korean Patent Laid-Open Publication No. 10-2017-0033590, a herbal extract or a herbal extract which essentially contains essential oils such as ginseng, safflower, ginseng, persimmon, Thereby showing the effect of preventing or treating an inflammatory disease. In the prior arts, inflammatory cytokines are reduced in a concentration-dependent manner, and translocation and phosphorylation inhibition of major transcription factors of the inflammatory response have resulted in the suppression of the inflammatory response.
따라서 생약 성분의 적절한 조성을 통해 염증성 사이토카인을 저감시키는 효과를 얻을 수 있다면 급성염증의 예방이나 개선에 적합한 조성물을 얻을 수 있을 것으로 기대된다.Therefore, if an effect of reducing the inflammatory cytokine through the proper composition of the herbal medicine component can be obtained, it is expected that a composition suitable for prevention or improvement of acute inflammation can be obtained.
본 발명은 상기와 같은 종래기술을 감안하여 안출된 것으로, 천연 약재 성분을 함유함으로써 급성 염증 개선뿐만 아니라 면역 증진의 효과를 나타내는 조성물을 제공하는 것을 그 목적으로 한다.Disclosure of the Invention The present invention has been made in view of the above-described prior art, and it is an object of the present invention to provide a composition showing the effect of improving immunity as well as acute inflammation by containing a natural medicinal ingredient.
상기와 같은 과제를 해결하기 위한 본 발명의 급성 염증 개선 및 면역 증진용 조성물은 녹용 분말 및 길경, 맥문동, 오미자, 인삼으로 이루어진 천연 약재 분말을 함유하는 것을 특징으로 한다.In order to solve the above-mentioned problems, the composition for acute inflammation improvement and immunity enhancement of the present invention is characterized by containing antler powder and natural medicinal powders composed of Gakyung, McMundong, Omija and Ginseng.
이때, 상기 천연 약재 분말은 길경, 맥문동, 오미자, 인삼을 환류 추출하고 감압 농축하여 얻어진 용액을 동결 건조하여 얻어진 분말인 것을 특징으로 한다.At this time, the natural medicinal ingredient powder is a powder obtained by refluxing and extracting Gakyung, McMundong, Omija and Ginseng, and concentrating under reduced pressure, followed by lyophilization.
또한, 상기 녹용 분말 및 천연 약재 분말은 2:1 내지 1:2의 중량비로 혼합될 수 있다.The antler seed powder and the natural medicinal powder may be mixed at a weight ratio of 2: 1 to 1: 2.
본 발명의 조성물은 경구투여용일 수 있으며, 식품 조성물일 수도 있다.The composition of the present invention may be for oral administration or may be a food composition.
본 발명에 따른 조성물은 천연 약재 성분을 함유함으로써 급성 염증 개선뿐만 아니라 면역 증진의 효과를 나타낸다.The composition according to the present invention exhibits the effect of improving immunity as well as improving acute inflammation by containing a natural medicinal ingredient.
도 1은 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 간 무게 변화를 측정한 결과이다.
도 2는 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 비장 무게 변화를 측정한 결과이다.
도 3은 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 AST를 측정한 결과이다.
도 4는 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 ALT를 측정한 결과이다.
도 5는 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 BUN을 측정한 결과이다.
도 6은 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 Creatinine을 측정한 결과이다.
도 7은 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈액 내 백혈구를 측정한 결과이다.
도 8은 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈액 내 단핵구를 측정한 결과이다.
도 9는 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈액 내 호중구를 측정한 결과이다.
도 10은 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈액 내 호산구를 측정한 결과이다.
도 11은 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈액 내 호염기구를 측정한 결과이다.
도 12는 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈액 내 림프구를 측정한 결과이다.
도 13은 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈청 내 IL-1β를 측정한 결과이다.
도 14는 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈청 내 IL-5 생성량을 측정한 결과이다.
도 15는 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈청 내 IL-6 생성량을 측정한 결과이다.
도 16은 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈청 내 IL-10 생성량을 측정한 결과이다.
도 17은 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈청 내 IL-12 생성량을 측정한 결과이다.
도 18은 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈청 내 TNF-α 생성량을 측정한 결과이다.
도 19는 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈청 내 INF-γ 생성량을 측정한 결과이다.
도 20은 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈청 내 ROS 생성량을 측정한 결과이다.
도 21은 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈청 내 PGE2 생성량을 측정한 결과이다.
도 22는 본 발명의 조성물을 적용했을 때 급성 염증 유발 쥐의 혈청 내 LTB4 생성량을 측정한 결과이다.
도 23은 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 간 무게 변화를 측정한 결과이다.
도 24는 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 비장 무게 변화를 측정한 결과이다.
도 25는 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 AST를 측정한 결과이다.
도 26은 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 ALT를 측정한 결과이다.
도 27은 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 BUN을 측정한 결과이다.
도 28은 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 Creatinine을 측정한 결과이다.
도 29는 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 혈액 내 백혈구를 측정한 결과이다.
도 30은 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 혈액 내 단핵구를 측정한 결과이다.
도 31은 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 혈액 내 호중구를 측정한 결과이다.
도 32는 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 혈액 내 호산구를 측정한 결과이다.
도 33은 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 혈액 내 호염기구를 측정한 결과이다.
도 34는 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 혈액 내 림프구를 측정한 결과이다.
도 35는 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 혈청 내 IgA를 측정한 결과이다.
도 36은 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 혈청 내 IgG를 측정한 결과이다.
도 37은 본 발명의 조성물을 적용했을 때 면역 억제 유도 쥐의 혈청 내 IgM을 측정한 결과이다.FIG. 1 shows the result of measurement of liver weight change in acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 2 shows the results of measurement of spleen weight change in acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 3 shows the results of measurement of AST of acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 4 shows the results of measurement of ALT in acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 5 shows the results of measurement of BUN of acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 6 shows the result of measurement of creatinine in acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 7 shows the results of measurement of leukocytes in the blood of acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 8 shows the results of measurement of mononuclear cells in the blood of acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 9 shows the results of measurement of neutrophils in the blood of acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 10 shows the results of measurement of eosinophils in the blood of acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 11 shows the results of measurement of basophils in blood of acute inflammation-induced rats when the composition of the present invention was applied.
Fig. 12 shows the result of measurement of lymphocytes in blood of acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 13 shows the results of measurement of serum IL-1 beta in acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 14 shows the results of measurement of IL-5 production in serum of acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 15 shows the results of measurement of IL-6 production in serum of acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 16 shows the results of measurement of IL-10 production in serum of acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 17 shows the results of measurement of IL-12 production in serum of acute inflammation-induced rats when the composition of the present invention was applied.
18 shows the results of measurement of TNF-α production in serum of acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 19 shows the results of measurement of INF-γ production in serum of acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 20 shows the results of measurement of ROS production in serum of acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 21 shows the results of measurement of PGE 2 production in serum of acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 22 shows the results of measurement of serum LTB 4 production in acute inflammation-induced rats when the composition of the present invention was applied.
FIG. 23 shows the results of measurement of liver weight change in immunosuppressed rats when the composition of the present invention was applied.
FIG. 24 shows the results of measurement of spleen weight change in immunosuppressed rats when the composition of the present invention was applied.
25 shows the results of measurement of AST of the immunosuppressed rats when the composition of the present invention was applied.
FIG. 26 shows the results of measurement of ALT of immunosuppressed rats when the composition of the present invention was applied.
FIG. 27 shows the results of measurement of the BUN of the immunosuppressed rats when the composition of the present invention was applied.
FIG. 28 shows the result of measuring the creatinine of the immunosuppressed rats when the composition of the present invention was applied.
FIG. 29 shows the results of measurement of leukocytes in the blood of the immunosuppression-induced rats when the composition of the present invention was applied.
FIG. 30 shows the results of measurement of mononuclear cells in the blood of the immunosuppressed rats when the composition of the present invention was applied.
FIG. 31 shows the results of measurement of neutrophils in the blood of immunosuppressed rats when the composition of the present invention was applied.
Figure 32 shows the result of measurement of eosinophils in the blood of immunosuppressed rats when the composition of the present invention was applied.
FIG. 33 shows the result of measuring the basophil count in the blood of the immunosuppressed rats when the composition of the present invention was applied.
FIG. 34 shows the results of measurement of lymphocytes in the blood of the immunosuppression-induced rats when the composition of the present invention was applied.
FIG. 35 shows the results of measurement of IgA in the serum of the immunosuppression-induced rats when the composition of the present invention was applied.
FIG. 36 shows the results of measurement of IgG in the serum of immunosuppression-induced rats when the composition of the present invention was applied.
FIG. 37 shows the results of measurement of serum IgM in immunosuppression-induced rats when the composition of the present invention was applied.
이하 도면을 참조하여 본 발명을 보다 상세히 설명한다. 본 명세서 및 청구범위에 사용된 용어나 단어는 통상적이거나 사전적인 의미로 한정해서 해석되어서는 아니되며, 발명자는 그 자신의 발명을 가장 최선의 방법으로 설명하기 위해 용어의 개념을 적절하게 정의할 수 있다는 원칙에 입각하여 본 발명의 기술적 사상에 부합하는 의미와 개념으로 해석되어야만 한다.Hereinafter, the present invention will be described in more detail with reference to the drawings. The terms and words used in the present specification and claims should not be construed as limited to ordinary or dictionary terms and the inventor may appropriately define the concept of the term in order to best describe its invention It should be construed as meaning and concept consistent with the technical idea of the present invention.
본 발명에 따른 급성 염증 개선 및 면역 증진용 조성물은 녹용 분말, 및 길경, 맥문동, 오미자, 인삼로 이루어진 천연 약재 분말을 함유하며, 특히 바람직하게는 녹용 분말과 길경, 맥문동, 오미자, 인삼으로 이루어진 천연 약재 분말로 이루어지는 것을 특징으로 한다.The composition for improving acute inflammation and improving immunity according to the present invention comprises a natural anticorrosive powder consisting of antler powder, gakgyeong, moonmundong, omija and ginseng, and more preferably natural anticorrosive powder composed of antler powder, Characterized in that it is made of a pharmaceutical powder.
녹용 성분의 항염증 효과에 대해서는 대한민국 공개특허공보 10-2009-0065575호에서 녹용으로부터 얻어진 글라이코사민 - 단백질 추출물 및 그 분획물이 항염증 효과를 나타내는 것으로 보고되어 있는 등 염증의 예방이나 개선에 효과가 있는 성분이다. 본 발명에서는 이러한 녹용을 주성분으로 하여 천연 약재 분말과 혼합하여 조성물을 형성함으로써 급성 염증의 예방이나 개선에 효과를 얻고 있다.Regarding the anti-inflammatory effect of the antler ingredient, Korean Patent Laid-Open Publication No. 10-2009-0065575 reports that the glycosamine-protein extract and its fractions obtained from antler exhibit anti-inflammatory effects, and are effective for preventing or improving inflammation It is an ingredient. In the present invention, it is effective to prevent or ameliorate acute inflammation by forming a composition by mixing such antler antler as a main ingredient and mixing with a natural medicinal ingredient powder.
본 발명에서는 녹용 분말로 뉴질랜드산 녹용을 추출한 후 여기에 L. acidophilus, L. casci, S. thermophilus, L. delbruekil 중 어느 하나의 균주를 투여하여 24시간 배양하고 이를 동결건조하여 분말화한 발효 녹용 분말을 사용하고 있다.In the present invention, a New Zealand antler extract is extracted with an antler granule, followed by culturing for 24 hours with a strain of L. acidophilus, L. casci, S. thermophilus, or L. delbruek, which is then lyophilized, Powder is used.
상기 녹용 분말과 혼합되는 상기 천연 약재 분말은 당귀, 천궁으로 이루어진다.The natural medicinal powders to be mixed with the antler granules are composed of Angelica gigas and Angelica gigas.
상기 천연 약재 분말은 세절된 약재를 환류 추출하여 여과액을 얻은 후 이를 회전감압농축기(rotary vacuum evaporator) 등을 사용하여 감압 농축하고 상기 감압 농축된 용액을 동결 건조기 등을 사용하여 동결 건조하여 제조될 수 있다.The natural medicinal ingredient powder is prepared by refluxing the crude medicinal material to obtain a filtrate, concentrating it under reduced pressure using a rotary vacuum evaporator or the like, and lyophilizing the reduced-pressure concentrated solution using a freeze dryer or the like .
상기 길경, 맥문동, 오미자, 인삼으로 이루어진 천연 약재 분말은 The natural medicinal powders composed of Gil-gyeong, McMundong, Omija, and Ginseng
길경 10 내지 20 중량부, 맥문동 10 내지 20 중량부, 오미자 10 내지 20 중량부, 인삼 10 내지 20 중량부를 혼합하여 환류 추출하고 감압 농축하여 얻어진 용액을 동결 건조함으로써 제조될 수 있다.10 to 20 parts by weight of gum arabic, 10 to 20 parts by weight of gum arabic, 10 to 20 parts by weight of omiza and 10 to 20 parts by weight of ginseng, refluxing the mixture, and concentrating under reduced pressure.
이와 같이 얻어진 천연 약재 분말과 상기 녹용 분말을 혼합하여 본 발명의 조성물을 제조하게 되는데, 상기 녹용 분말 및 천연 약재 분말을 2:1 내지 1:2의 중량비로 조성하는 것이 바람직하다. 상기 녹용 분말이나 천연 약재 분말 중 1 성분의 함량이 상기 범위를 벗어나 지나치게 많아지면 본 발명에서 요구하는 급성 염증에 대한 개선 효과가 감소하는 것으로 나타났다.The composition of the present invention is prepared by mixing the thus obtained natural medicine powder and the deer antler powder. Preferably, the antler powder and the natural medicine powder are mixed at a weight ratio of 2: 1 to 1: 2. When the content of one component in the antler seed powder or the natural medicinal ingredient powder is out of the above range, the improvement effect on the acute inflammation required by the present invention is decreased.
본 발명의 조성물은 급성 염증 개선 및 면역 증진의 효과가 있으므로 경구투여용 또는 식품 조성물에 적용할 수 있다.The composition of the present invention is effective for improving acute inflammation and improving immunity, and thus can be applied to oral administration or food composition.
경구투여용 조성물로 적용할 경우, 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형으로 제형화하여 사용할 수 있다. 상기 조성물의 제형화를 위하여 담체, 부형제 및 희석제를 포함할 수 있는데, 구체적으로는 락토오스, 덱스트로스, 수크로오스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로오스, 메틸 셀룰로오스, 미정질 셀룰로오스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 포함할 수 있다. 제제화 할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 분획물에 적어도 하나 이상의 부형제 예를 들면, 전분, 탄산칼슘, 수크로스 또는 락토오스, 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. When it is applied as a composition for oral administration, it may be formulated into oral form such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like according to a conventional method. Examples of the carrier include excipients such as lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, Calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin, . In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .
상기 조성물의 투여량은 치료 받을 대상의 연령, 성별, 체중과, 치료할 특정 질환 또는 병리 상태, 질환 또는 병리 상태의 심각도, 투여경로 및 처방자의 판단에 따라 달라질 것이다. 이러한 인자에 기초한 투여량 결정은 당업자의 수준 내에 있으며, 일반적으로 투여량은 0.01 내지 2000㎎/㎏/일의 범위이다. 더 바람직한 투여량은 0.1 내지 500㎎/㎏/일이다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다. The dosage of the composition will vary depending on the age, sex, body weight of the subject to be treated, the particular disease or condition to be treated, the severity of the disease or condition, the route of administration and the judgment of the prescriber. Dosage determinations based on these factors are within the level of ordinary skill in the art and generally the dosage ranges from 0.01 to 2000 mg / kg / day. A more preferable dosage is 0.1 to 500 mg / kg / day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.
또한, 상기 조성물은 쥐, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있으므로 경구 투여 외에도 직장 또는 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관내 주사에 의해 투여될 수도 있다.In addition, the composition may be administered to mammals such as rats, livestock, humans, and the like in various routes. Thus, the composition may be administered by rectal or intravenous, muscular, subcutaneous, intrauterine or intracerebral injection or intravenous injection.
또한, 상기 조성물은 식품학적으로 허용 가능한 식품보조 첨가제를 포함함으로써 식품 조성물로서 적용될 수도 있다. 이러한 식품 조성물은 분말, 정제, 캡슐제, 환제 또는 액제 등의 형태를 포함하며, 상기 건강기능식품으로는 드링크제, 육류, 소세지, 빵, 캔디류, 스넥류, 면류, 아이스크림을 포함한 낙농제품, 스프, 이온음료 등을 포함한 음료수, 알코올 음료 및 비타민 복합제를 포함한 영양 공급용 제품 등이 포함될 수 있다.In addition, the composition may be applied as a food composition by including a food-acceptable food-aid additive. Such food compositions may be in the form of powders, tablets, capsules, pills or liquids, and the health functional foods include dairy products including soups, meats, sausages, breads, candies, snacks, noodles, Beverages including beverages, nutritional products including alcoholic beverages and vitamin complexes.
예를 들어, 조리용 양념에 적용할 경우 본 발명의 조성물을 0.2~10 중량%로 하여 건강 증진용 조리용 양념을 제조할 수 있다.For example, when applied to a cooking season, the composition of the present invention may be used in an amount of 0.2 to 10% by weight to prepare health-enhancing cooking seasonings.
또한, 밀가루 식품에 적용할 경우 본 발명의 조성물을 0.1~5.0 중량%로 하여 밀가루에 첨가하고, 이 혼합물을 이용하여 빵, 케이크, 쿠키, 크래커 및 면류를 제조하여 건강 증진용 식품을 제조할 수 있다.In addition, when applied to flour food, the composition of the present invention may be added to flour at 0.1 to 5.0 wt%, and bread, cake, cookies, crackers and noodles may be prepared by using the mixture to prepare a food for health promotion have.
또한, 스프 및 육즙(gravies)에 적용할 경우, 본 발명의 조성물을 0.1~1.0 중량%로 하여 스프 및 육즙에 첨가하여 건강 증진용 육가공 제품, 면류의 수프 및 육즙을 제조할 수 있다.In addition, when applied to soups and gravies, the composition of the present invention may be added to soups and gravies at a concentration of 0.1 to 1.0 wt% to prepare health-enhancing meat products, soups and juices of noodles.
또한, 유제품(dairy products)에 적용할 경우, 본 발명의 조성물을 0.1~1.0 중량%로 하여 우유에 첨가하고, 상기 우유를 이용하여 버터 및 아이스크림과 같은 다양한 유제품을 제조할 수 있다.In addition, when applied to dairy products, the composition of the present invention may be added to milk in an amount of 0.1 to 1.0% by weight, and various dairy products such as butter and ice cream may be prepared using the milk.
또한, 주스에 적용하는 경우, 본 발명의 조성물 0.5g을 토마토 또는 당근 쥬스 1,000㎖에 가하여 건강 증진용 야채주스를 제조하거나, 본 발명의 조성물 0.1g을 사과 또는 포도 쥬스 1,000㎖에 가하여 건강 증진용 과일주스를 제조할 수도 있다.When applied to juice, 0.5 g of the composition of the present invention is added to 1,000 ml of tomato or carrot juice to prepare vegetable juice for health promotion, or 0.1 g of the composition of the present invention is added to 1,000 ml of apple or grape juice, Fruit juices may also be prepared.
이하, 본 발명의 조성물에 대한 급성 염증 및 면역 증진의 효과를 탐색하기 위하여 다음과 같이 실험을 실시하였다.Hereinafter, the following experiment was conducted to investigate the effect of acute inflammation and immunity enhancement on the composition of the present invention.
천연 약재 분말을 제조하기 위하여 사용된 약재 및 녹용 분말은 옴니허브에서 구입하였고, 대전대학교 TBRC-RIC에서 정선 후 사용하였다. 분량은 1첩을 기준으로 길경(Platycodi Radix) 14g, 맥문동(Liriopis Tuber) 14g, 오미자(Schisandrae Fructus) 14g, 인삼(Ginseng Radix Alba) 14g으로 이루어진다. The medicinal materials and antler seeds used for the preparation of natural medicinal powders were purchased from OMNI HUB and used after selection in TBRC-RIC, Daejeon University. Platycodi Radix 14 g, Liriopis Tuber 14 g, Schisandrae Fructus 14 g, and Ginseng Radix Alba 14 g.
사용된 시약은 Lipopolysaccharide (LPS : Sigma, U.S.A.), PBS (Welgene Inc., Korea), Ethyl ether (Samchun, Korea), Microtainer® tube with Dipotassium EDTA (BD Co., U.S.A.), Mouse cytokine milliplex map immunoassay kit (Millipore Co., U.S.A.), Mouse reactive oxygen species ELISA kit (MyBioSource, U.S.A.), Prostaglandin E2 parameter assay kit (R&D system, U.S.A.), LTB4 parameter assay kit (R&D system, U.S.A.)등을 사용하였다.The reagents used were Lipopolysaccharide (LPS: Sigma, USA), PBS (Welgene Inc., Korea), Ethyl ether (Samchun, Korea), Microtainer tube with Dipotassium EDTA (BD Co., USA), Mouse cytokine milliplex map immunoassay kit (Millipore Co., USA), Mouse reactive oxygen species ELISA kit (MyBioSource, USA), Prostaglandin E2 parameter assay kit (R & D system, USA) and LTB4 parameter assay kit (R & D system, USA).
또한, 사용된 기기는 환류 추출기 (Mtops, Korea), 회전감압농축기(rotary vacuum evaporator) (Buchi B-480, Switzerland), 동결 건조기(freeze dryer) (IlShinBioBase, Korea), 디프 프리저(deep freezer) (Sanyo, Japan), 오토클레이브(autoclave) (Sanyo, Japan), 볼텍스 믹서(vortex mixer) (Vision scientific, Korea), 원심분리기(Sigma, U.S.A.), 아이스메이커(ice-maker) (Vision scientific, Korea), plate shaker (Lab-Line, U.S.A.), Luminex (Millipore, U.S.A.), ELISA reader (Molecular Devices Co., U.S.A.)를 사용하였다.The apparatus used was a rotary vacuum evaporator (Buchi B-480, Switzerland), a freeze dryer (IlShinBioBase, Korea), a deep freezer Sanyo, Japan), autoclave (Sanyo, Japan), vortex mixer (Vision scientific, Korea), centrifuge (Sigma, USA), ice-maker (Vision scientific, , Luminex (Millipore, USA) and ELISA reader (Molecular Devices Co., USA) were used.
시료의 추출 방법은 다음과 같다.The extraction method of the sample is as follows.
2첩의 천연 약재 혼합물(112 g)에 증류수 1000㎖를 넣고 3시간 동안 환류 추출 후 여과액을 얻어 회전감압농축기에서 감압 농축하였다. 농축된 용액을 동결 건조기로 동결 건조하여 분말 12.8g을 얻었으며, 얻어진 분말은 초저온 냉동고 (-80℃)에서 보관하면서 실험에 따라 필요한 농도로 증류수에 희석하여 사용하였다.1000 ml of distilled water was added to the mixture (112 g) of two concentrates, followed by reflux extraction for 3 hours, followed by filtration and concentration under reduced pressure using a rotary evaporator. The concentrated solution was lyophilized in a freeze dryer to obtain 12.8 g of powder. The obtained powder was diluted with distilled water to the required concentration while being stored in an ultra-low temperature freezer (-80 ° C).
급성 염증 관련 실험을 위해 사용된 ICR mouse (5주령, 수컷, 23∼28 ｇ)는 ㈜샘타코 (Korea)에서 구입하여 사용하였다. 실험동물은 2주간의 안정기를 가지면서 순화를 시켰으며, 안정기 및 실험기간에 모든 실험군에는 일반 사료 (Altromin, Germany)를 자유식이 하며 물을 충분히 공급하였다. 2주간의 안정기 이후 7주령부터 동물 실험을 진행하였다. 동물 사육실의 조건은 conventional system으로 22±2℃, 1일 중 12시간은 200-300 Lux로 조명하고, 12시간은 모든 빛을 차단하였다. 본 실험은 대전대 동물실험윤리 위원회의 승인 (승인번호 DJUARB2017-005)을 받아 동물윤리준칙에 의거하여 실험하였다.ICR mice (5-week-old, male, 23-28 g) used for acute inflammation-related experiments were purchased from Samtaco, Korea. Experimental animals were stabilized for 2 weeks with stabilization period. During the stabilization period and experiment period, all feeds were fed free of general diet (Altromin, Germany) and water. After 2 weeks of stabilization, animals were tested from 7 weeks of age. The conditions of the animal room were 22 ± 2 ℃ for conventional system, 200-300 Lux for 12 hours in a day, and all lights were blocked for 12 hours. This experiment was conducted according to the animal ethics code of the Daejeon National Laboratory Ethics Committee (approval No. DJUARB2017-005).
급성 염증 유발 및 시료 처리를 위하여, 아무것도 처지하지 않는 정상군과 증류수를 투여하는 대조군, 천연 약재 분말에 각각 녹용분말을 1:1의 중량비로 혼합한 조성물을 200, 400㎎/㎏/day로 투여하는 실험군 등 총 8개의 그룹으로 나누어 매일 1회, 오후 2시에 200㎕씩 oral zonde를 이용하여 7일간 경구 투여하였다. (이하, 투여량에 따라 SC2 200, SC2 400으로 부른다.) In order to induce acute inflammation and sample treatment, a composition prepared by mixing a non-aged normal group, a control group to which distilled water was administered, and a natural medicinal powder at a ratio of 1: 1 by weight of antler seeds was administered at 200, 400 mg / kg / day And 2 groups were divided into 8 groups. Each group was orally administered by oral zonde at 2 o'clock at 2 o'clock for 7 days. (Hereinafter referred to as
7일 후 LPS 1㎎/㎏을 복강에 주사하고 3시간 후에 ethyl ether로 마취하여 심장 천자법으로 채혈하였다. 시료 투여량은 성인 체중 1kg당 200, 400㎎을 기준으로 하고 마우스 체중 30g으로 계산하여 산출하였다. 또한, 체중 및 식이섭취량은 매일 오전 10시에 ｇ단위의 저울을 이용하여 측정을 실시하였다. After 7 days, 1 mg / kg of LPS was injected into the peritoneal cavity, and after 3 hours, it was anesthetized with ethyl ether and blood was collected by cardiac puncture. The dose of the sample was calculated on the basis of 200 and 400 mg per 1 kg of adult body weight and 30 g of mouse body weight. The body weight and dietary intake were measured daily at 10 am using a scale of g unit.
또한, 면역 증진 실험을 위하여 7주령이 된 ICR mouse (5주령, 수컷, 23∼28ｇ)는 ㈜샘타코 (Korea)에서 구입하여 사용하였다. 실험동물은 2주간의 안정기를 가지면서 순화를 시켰으며, 안정기 및 실험기간에 모든 실험군에는 일반 사료 (Altromin, Germany)를 자유식이 하며 물을 충분히 공급하였다. 2주간의 안정기 이후 7주령부터 동물 실험을 진행하였다. 동물 사육실의 조건은 conventional system으로 22±2℃, 1일 중 12시간은 200-300 Lux로 조명하고, 12시간은 모든 빛을 차단하였다. 본 실험은 대전대 동물실험윤리 위원회의 승인 (승인번호 DJUARB2017-007)을 받아 동물윤리준칙에 의거하여 실험하였다. ICR mouse (5 weeks old, male, 23 ~ 28g), which was 7 weeks old, was purchased from Korea. Experimental animals were stabilized for 2 weeks with stabilization period. During the stabilization period and experiment period, all feeds were fed free of general diet (Altromin, Germany) and water. After 2 weeks of stabilization, animals were tested from 7 weeks of age. The conditions of the animal room were 22 ± 2 ℃ for conventional system, 200-300 Lux for 12 hours in a day, and all lights were blocked for 12 hours. This experiment was conducted in accordance with the animal ethics code with the approval of Daejeon National Laboratory Ethics Committee (approval number DJUARB2017-007).
면역 억제 유도 및 시료 처리를 위하여, 7주령이 된 ICR mouse 복강에 면역억제제 (Cyclophosphamide)를 100㎎/㎏ 농도로 100㎕ 주사하여 면역 억제를 유도하였다. 아무것도 처지하지 않는 정상군과 면역 억제제 처리 후 증류수를 투여하는 대조군, 면역 억제제 처리 후 천연 약재 분말과 녹용분말을 1:1 비율로 혼합하여 제조한 조성물을 200, 400 ㎎/㎏/day로 투여하는 실험군 등 총 4개의 그룹으로 나누어 매일 1회, 오후 2시에 200㎕씩 oral zonde를 이용하여 7일간 경구 투여하였다. 면역 억제제 및 시료 투여량은 성인 체중 1kg당 200, 400㎎을 기준으로 하고 마우스 체중 30g으로 계산하여 산출하였다. 또한, 체중 및 식이섭취량은 매일 오전 10시에 ｇ단위의 저울을 이용하여 측정을 실시하였다. Immunosuppression was induced by injecting 100 μl of immunosuppressant (Cyclophosphamide) at a concentration of 100 mg / kg into ICR mouse peritoneal cavity at 7 weeks of age for induction of immunosuppression and sample treatment. After administration of the immunosuppressant, the composition prepared by mixing the natural pharmaceutical powder and the deer antler powder at a ratio of 1: 1 was administered at 200, 400 mg / kg / day. The test group was divided into 4 groups. The test group was orally administered once daily for 1 day and 200 μl at 2:00 pm for 7 days using an oral zonde. Immunosuppressants and sample doses were calculated on the basis of 200 and 400 mg per 1 kg of adult body weight and 30 g of mouse body weight. The body weight and dietary intake were measured daily at 10 am using a scale of g unit.
간 및 비장 무게 측정은 실험 종료 후 적출한 장기를 ｇ단위의 저울을 이용하여 측정을 실시하였다. The liver and spleen weights were measured using the scales of g units after excision at the end of the experiment.
또한, 생화학적 검사는 실험 종료 후 심장 천자법을 이용하여 혈액을 채취하여 30분간 상온에서 굳힌 뒤 3,000rpm에서 15분간 원심분리 후 혈청을 분리하여 간 기능 지표 (AST, ALT)와 신 기능 지표 (BUN, Creatinine)를 KPNT (Korea)에 분석 의뢰하였다.In addition, biochemical tests were performed by cardiopulmonary resuscitation and blood samples were collected for 30 minutes at room temperature. After centrifugation at 3,000 rpm for 15 minutes, serum samples were collected and analyzed for liver function index (AST, ALT) BUN, Creatinine) to KPNT (Korea) for analysis.
또한, 혈액 내 면역세포 수 측정은 실험 종료 후 심장 천자법을 이용하여 혈액을 채취하여 EDTA 튜브에 담아 백혈구 및 호중구, 호산구, 호염구, 림프구, 단핵구 수를 KPNT (Korea)에 분석 의뢰하였다.After the end of the experiment, blood samples were collected from the blood using an EDTA tube and analyzed for leukocytes, neutrophils, eosinophils, basophils, lymphocytes, and mononuclear cells in KPNT (Korea).
또한, 혈청 내 사이토카인 측정은 실험 종료 후 분리한 혈청으로 사이토카인 Il-1β, IL-5, IL-6, IL-10, IL-12, TNF-α, IFN-γ는 Mouse cytokine milliplex map immunoassay kit를 이용하여 다음과 같이 측정하였다. 각 well에 standard, control, 2배 희석한 혈청 25㎕씩 분주하고 assay buffer 및 matrix buffer, antibody-immobilized beads를 각 25㎕씩 가하여 혼합한 후 2시간 동안 실온에서 반응시키고 washing 완충 용액을 이용하여 2회 세척하였다. 이를 다시 25㎕의 detection antibody을 가하여 1시간 동안 실온에서 빛을 차단한 채 반응시키고 추가로 25㎕의 Streptavidin-Phycoerythrin을 가하여 30분 동안 실온에서 반응시킨 후 washing 완충 용액을 이용하여 2회 세척하였다. 세척 후 PBS를 150㎕ 넣고 5분 간 shaking한 후 Luminex를 이용하여 측정하였다.The serum cytokine IL-1β, IL-5, IL-6, IL-10, IL-12, TNF-α and IFN-γ were measured by mouse cytokine milliplex map immunoassay kit as follows. 25 μl of standard, control and 2-fold diluted serum were added to each well, and 25 μl of assay buffer, matrix buffer and antibody-immobilized beads were added to each well. The mixture was reacted at room temperature for 2 hours, Lt; / RTI > Then 25 μl of detection antibody was added and reacted at room temperature for 1 hour with blocking the light. An additional 25 μl of Streptavidin-Phycoerythrin was added and reacted at room temperature for 30 minutes, followed by washing twice with washing buffer solution. After washing, 150 μl of PBS was added and shaking was carried out for 5 minutes, and then measured using Luminex.
또한, 혈청 내 염증 유관인자 측정을 위하여 ROS, PGE2, LTB4의 생성량을 측정하였다.In addition, the amount of ROS, PGE 2 , and LTB 4 was measured for the measurement of inflammatory factor in serum.
실험 종료 후 분리한 혈청으로 ROS 생성량은 Mouse reactive oxygen species ELISA kit를 이용하여 다음과 같이 측정하였다. 96 well plate에 standard와 혈청을 50㎕를 넣고 추가로 HRP-conjugate reagent 100㎕를 넣어 37℃에서 1시간 동안 반응시켰다. 이후, wash buffer를 이용하여 넣고 washing 작업을 실행한 후, chromogen solution A 50㎕와 chromogen solution B 50㎕를 넣고 37 ℃에서 30분 동안 빛을 차단한 채 반응시켰다. 마지막으로 stop solution 50 ㎕를 넣고 ELISA reader 450㎚ 파장에서 측정하였다.After the end of the experiment, the amount of ROS produced by the separated serum was measured using Mouse reactive oxygen species ELISA kit as follows. 50 μl of standard and serum was added to a 96-well plate, and 100 μl of HRP-conjugate reagent was added thereto, followed by reaction at 37 ° C for 1 hour. After washing with washing buffer, 50 μl of chromogen solution A and 50 μl of chromogen solution B were added and incubated at 37 ° C for 30 minutes. Finally, 50 μl of the stop solution was added and the ELISA reader was measured at a wavelength of 450 nm.
실험 종료 후 분리한 혈청으로 PGE2 생성량은 Prostaglandin E2 parameter assay kit를 이용하여 다음과 같이 측정하였다. 96 well plate에 standard와 Control, 5배 희석한 혈청을 150 ㎕를 넣고 추가로 primary antibody 50㎕를 넣어 1시간 동안 plate shaker를 이용하여 혼합하였다. 이후, PGE2 conjugate 50㎕를 넣고 다시 2시간 동안 plate shaker를 이용하여 혼합하였다. 그 다음 wash buffer 400㎕씩 넣고 washing 작업을 실행한 후 substrate solution 200㎕를 첨가하고 상온에서 30분 동안 빛을 차단한 채 반응시켰다. 마지막으로 stop solution 100㎕를 넣고 ELISA reader 450㎚ 파장에서 측정하였다.After the end of the experiment, the amount of PGE 2 produced by the separated serum was measured using the Prostaglandin E2 parameter assay kit as follows. 150 μl of standard, control and 5-fold dilutions of serum were added to a 96-well plate, and 50 μl of a primary antibody was added to the plate. The plate was shaken for 1 hour using a plate shaker. Then, 50 μl of PGE 2 conjugate was added and mixed again using a plate shaker for 2 hours. After washing with 400 μl of wash buffer, 200 μl of substrate solution was added, and the mixture was incubated at room temperature for 30 minutes while blocking light. Finally, 100 μl of the stop solution was added and the ELISA reader was measured at a wavelength of 450 nm.
실험 종료 후 분리한 혈청으로 LTB4 생성량은 Prostaglandin E2 parameter assay kit를 이용하여 다음과 같이 측정하였다. 96 well plate에 standard와 3배 희석한 혈청을 50㎕를 넣고 추가로 primary antibody solution 50㎕를 넣어 1시간 동안 plate shaker를 이용하여 혼합하였다. 이후, LTB4 conjugate 50㎕를 넣고 다시 3시간 동안 plate shaker를 이용하여 혼합하였다. 그 다음 wash buffer 400㎕씩 넣고 washing 작업을 실행한 후 substrate solution 200㎕를 첨가하고 상온에서 30분 동안 빛을 차단한 채 반응시켰다. 마지막으로 stop solution 100㎕를 넣고 ELISA reader 450㎚ 파장에서 측정하였다.After the end of the experiment, the amount of LTB 4 produced in the separated serum was measured using the Prostaglandin E2 parameter assay kit as follows. 50 μl of standard and 3-fold dilutions of serum were added to 96-well plates, and 50 μl of a primary antibody solution was added thereto. The mixture was mixed using a plate shaker for 1 hour. Then, 50 μl of LTB 4 conjugate was added and mixed again using a plate shaker for 3 hours. After washing with 400 μl of wash buffer, 200 μl of substrate solution was added, and the mixture was incubated at room temperature for 30 minutes while blocking light. Finally, 100 μl of the stop solution was added and the ELISA reader was measured at a wavelength of 450 nm.
혈청 내 면역글로불린 측정은 실험 종료 후 분리한 혈청으로 면역글로불린 IgA, IgG, IgM은 Mouse immunoglobulin isotyping multiplex assay kit를 이용하여 다음과 같이 측정하였다. 각 well에 standard, control, 25,000배 희석한 혈청을 50 ㎕씩 분주하고 anti-mouse multi-immunoglobulin beads를 각 25 ㎕씩 가하여 혼합하여 15분 동안 실온에서 빛을 차단한 채 반응시키고 washing 완충 용액을 이용하여 2회 세척하였다. 이 후 25㎕의 anti-mouse κLight Chain, PE를 가하여 15분 동안 실온에서 빛을 차단한 채 반응시키고 PBS를 150㎕ 넣고 5분 간 shaking한 후 Luminex를 이용하여 측정하였다.Serum immunoglobulin IgA, IgG, and IgM were measured using the immunoglobulin isotyping multiplex assay kit as follows. Add 50 μl of standard, control, and 25,000-fold diluted serum to each well, add 25 μl of anti-mouse multi-immunoglobulin beads, and incubate for 15 minutes at room temperature with blocking light. And washed twice. After that, 25 μl of anti-mouse κLight Chain, PE was added and reacted with blocking light at room temperature for 15 minutes, and then 150 μl of PBS was added and shaking was performed for 5 minutes and then measured using Luminex.
실험 결과는 SPSS 18.0의 unpaired student's T-test와 ANOVA를 사용하여 통계처리 하였고 p<0.05, p<0.01 및 p<0.001 수준에서 그 유의성을 검정하였다.The results were statistically analyzed using the unpaired Student's T-test and ANOVA of SPSS 18.0, and the significance was tested at p <0.05, p <0.01 and p <0.001.
<급성 염증 개선 효과>≪ Effect of improving acute inflammation &
1) 체중 변화 측정1) Weight change measurement
체중 변화를 측정한 결과는 표 1과 같다. 표 1에서 모든 단위는 g이다. 또한, Normal은 무처리 ICR 쥐이며, Control은 급성 염증 유발 쥐로서 증류수만 경구투여한 경우이며, 이하 표 및 도면에서 동일한 의미로 사용된다.The results of measurement of weight change are shown in Table 1. In Table 1, all units are g. In addition, Normal is an untreated ICR rat, and Control is an acute inflammation-induced mouse in which only distilled water is orally administered, and is used in the following tables and figures in the same sense.
body weightChange of
body weight
표 1의 결과를 살펴보면, 실험 시작일 부터 6일차까지 정상군은 34.50±1.98, 35.60±2.55, 35.85±3.18, 35.45±3.18, 35.37±2.88, 35.35±2.90, 35.60±3.11ｇ으로 나타났으며, 대조군은 34.16±1.59, 34.46±1.23, 34.82±1.16, 34.72±1.60, 34.56±1.82, 35.62±1.43, 35.46±1.06ｇ으로 나타났다. SC2 200 투여군은 34.08±1.45, 34.18±1.59, 34.56±1.58, 34.36±1.36, 34.58±1.52, 35.78±1.33, 35.36±1.13ｇ으로 나타났으며, SC2 400 투여군은 34.36±2.26, 35.18±3.04, 35.36±3.18, 35.40±2.84, 34.96±2.87, 35.54±3.11, 35.76±2.94ｇ으로 나타나, 모든 실험군은 차이가 나타나지 않았다.As shown in Table 1, from the beginning of the experiment to the 6th day, 34.50 ± 1.98, 35.60 ± 2.55, 35.85 ± 3.18, 35.45 ± 3.18, 35.37 ± 2.88, 35.35 ± 2.90 and 35.60 ± 3.11g in the normal group, 34.16 ± 1.59, 34.46 ± 1.23, 34.82 ± 1.16, 34.72 ± 1.60, 34.56 ± 1.82, 35.62 ± 1.43, and 35.46 ± 1.06g, respectively. In the
2) 식이 섭취량 변화 측정2) Measurement of dietary intake change
식이섭취량 변화를 측정한 결과는 표 2와 같다. 표 2에서 모든 단위는 g이다.Table 2 shows the results of the changes in dietary intake. In Table 2, all units are g.
food intakeChange of
food intake
표 2를 살펴보면, 1일차부터 6일차까지 정상군은 4.41, 4.65, 4.06, 5.12, 4.94, 4.65ｇ으로 나타났으며, 대조군은 4.02, 4.48, 4.40, 4.42, 4.56, 4.84ｇ으로 나타났다. SC2 200 투여군은 4.26, 4.78, 4.84, 5.02, 5.30, 4.68ｇ으로 나타났으며, SC2 400 투여군은 4.46, 4.72, 4.86, 4.94, 4.32, 4.74ｇ으로 나타나, 모든 실험군은 차이가 나타나지 않았다.The results are shown in Table 2. As shown in Table 2, 4.41, 4.65, 4.06, 5.12, 4.94 and 4.65g were observed in the normal group from the 1st day to the 6th day, and 4.02, 4.48, 4.40, 4.42, 4.56 and 4.84g in the control group. 4.42, 4.72, 4.86, 4.94, 4.32, and 4.74g in the
3) 장기 무게 변화 측정3) Measurement of long-term weight change
간, 비장의 무게 변화 측정을 수행했으며, 간 무게 변화 측정 결과는 도 1과 같다. Liver, and spleen. The results of the liver weight change measurement are shown in Fig.
간 무게 변화를 측정한 결과, 정상군은 1.76±0.20 ｇ, 대조군은 1.61±0.15ｇ으로 나타났으며, SC2 200 투여군은 1.62±0.22ｇ, SC2 400 투여군은 1.73±0.20ｇ으로 나타나, SC2 투여군은 대조군에 비해 증가가 나타났다.The SC2-treated group showed 1.62 ± 0.22 g and the SC2 400-treated group showed 1.73 ± 0.20 g. In the SC2-treated group, Compared to the control group.
또한, 비장 무게 변화를 측정한 결과, 정상군은 0.15±0.02ｇ, 대조군은 0.15±0.02ｇ으로 나타났으며, SC2 200 투여군은 0.15±0.03ｇ, SC2 400 투여군은 0.15±0.01ｇ으로 나타나, 모든 실험군은 차이가 나타나지 않았다.As a result of the measurement of spleen weight, 0.15 ± 0.02 g was observed in the normal group and 0.15 ± 0.02 g in the control group, 0.15 ± 0.03 g in the
4) 간 기능 측정4) Liver function measurement
간 기능 측정을 위하여 AST 및 ALT 레벨을 측정하였다.AST and ALT levels were measured for liver function measurements.
간 기능 지표인 AST를 측정한 결과, 도 3에서와 같이, 정상군은 102.05±16.97U/ℓ, 대조군은 201.40±15.10U/ℓ로 나타났으며, SC2 200 투여군은 200.55±7.14U/ℓ, SC2 400 투여군은 204.37±13.40U/ℓ로 나타나, SC2 투여군은 대조군에 비해 차이가 나타나지 않았다.As shown in FIG. 3, the AST of the liver function index was 102.05 ± 16.97 U / ℓ in the normal group and 201.40 ± 15.10 U / ℓ in the control group, and 200.55 ± 7.14 U / The
또한, ALT를 측정한 결과, 도 4에서와 같이, 정상군은 23.30±2.31U/ℓ, 대조군은 68.03±5.15U/ℓ로 나타났으며, SC2 200 투여군은 68.69±6.90U/ℓ, SC2 400 투여군은 65.91±4.08U/ℓ로 나타나, SC2 투여군은 대조군에 비해 차이가 나타나지 않았다.As shown in FIG. 4, ALT was measured to be 23.30 ± 2.31 U / ℓ in the normal group and 68.03 ± 5.15 U / ℓ in the control group, 68.69 ± 6.90 U / ℓ in the
5) 신 기능 측정5) Measurement of new functions
신 기능 측정을 위하여 BUN, creatinine 레벨을 측정하였다.BUN and creatinine levels were measured to measure renal function.
신 기능 지표인 BUN을 측정한 결과, 도 5에서와 같이, 신 기능 지표인 BUN을 측정한 결과, 정상군은 23.45±1.50㎎/㎗, 대조군은 32.03±2.70㎎/㎗로 나타났으며, SC2 200 투여군은 32.90±2.17㎎/㎗, SC2 400 투여군은 31.88±4.09㎎/㎗로 나타나, SC2 투여군은 대조군에 비해 차이가 나타나지 않았다.As shown in FIG. 5, the BUN, a new functional index, was measured as 23.45 ± 1.50 mg / dl in the normal group and 32.03 ± 2.70 mg / dl in the control group, 200 administration group showed 32.90 ± 2.17 mg / dl and
또한, creatinine을 측정한 결과, 정상군은 0.28±0.03㎎/㎗, 대조군은 0.31±0.01㎎/㎗로 나타났으며, SC2 200 투여군은 0.29±0.02㎎/㎗, SC2 400 투여군은 0.28±0.02㎎/㎗로 나타나, 모든 실험군은 차이가 나타나지 않았다.The creatinine level was 0.28 ± 0.03 mg / dl in the normal group and 0.31 ± 0.01 mg / dl in the control group, 0.29 ± 0.02 mg / dl in the
6) 혈액 내 면역세포 수 측정6) Measurement of immune cell count in blood
혈액 내의 백혈구, 단핵구, 호중구, 호산구, 호염구, 및 림프구의 수를 각각 측정하였다.The numbers of leukocytes, mononuclear cells, neutrophils, eosinophils, basophils, and lymphocytes in blood were measured respectively.
혈액 내 백혈구 수를 측정한 결과, 도 7에서와 같이, 정상군은 1.30±0.29×103cells/㎕, 대조군은 3.03±0.30×103cells/㎕로 나타났으며, SC2 200 투여군은 1.87±0.31×103cells/㎕, SC2 400 투여군은 1.42±0.20×103cells/㎕로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (** : p<0.01, *** : p<0.001) 감소가 나타났다.As shown in FIG. 7, the number of leukocytes in blood was 1.30 ± 0.29 × 10 3 cells / μl in the normal group and 3.03 ± 0.30 × 10 3 cells / μl in the control group. 0.31 × 10 3 cells / μl, and SC2 400-treated group showed 1.42 ± 0.20 × 10 3 cells / μl. The SC2-treated group showed a significant decrease (**: p <0.01, ***: p <0.001) appear.
또한, 혈액 내 단핵구 수를 측정한 결과, 도 8에서와 같이, 정상군은 0.30±0.05%, 대조군은 0.63±0.09%로 나타났으며, SC2 200 투여군은 0.47±0.03%, SC2 400 투여군은 0.34±0.09%로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (* : p<0.05, ** : p<0.01) 감소가 나타났다.As shown in FIG. 8, the number of mononuclear cells in the blood was 0.30 ± 0.05% in the normal group and 0.63 ± 0.09% in the control group, 0.47 ± 0.03% in the
또한, 혈액 내 호중구 수를 측정한 결과, 도 9에서와 같이, 정상군은 28.05±3.75%, 대조군은 15.72±2.13%로 나타났으며, SC2 200 투여군은 30.70±1.70%, SC2 400 투여군은 31.65±3.04%로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (*** : p<0.001) 증가가 나타났다.As shown in FIG. 9, the number of neutrophils in the blood was 28.05 ± 3.75% in the normal group, 15.72 ± 2.13% in the control group, 30.70 ± 1.70% in the
또한, 혈액 내 호산구 수를 측정한 결과, 도 10에서와 같이, 정상군은 11.30±1.45%, 대조군은 23.40±2.06%로 나타났으며, SC2 200 투여군은 14.10±1.62%, SC2 400 투여군은 13.93±1.50%로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (** : p<0.01) 감소가 나타났다.The eosinophil count in the blood was measured as 11.30 ± 1.45% in the normal group and 23.40 ± 2.06% in the control group as shown in FIG. 10, 14.10 ± 1.62% in the
또한, 혈액 내 호염기구 수를 측정한 결과, 도 11에서와 같이, 정상군은 1.10±0.10%, 대조군은 0.30±0.06%로 나타났으며, SC2 200 투여군은 0.58±0.08%, SC2 400 투여군은 0.94±0.07%로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (* : p<0.05, ** : p<0.01) 증가가 나타났다.As shown in FIG. 11, the number of basophils in the blood was 1.10 ± 0.10% in the normal group, 0.30 ± 0.06% in the control group, 0.58 ± 0.08% in the
또한, 혈액 내 림프구 수를 측정한 결과, 도 12에서와 같이, 정상군은 41.15±4.40%, 대조군은 60.00±3.15%로 나타났으며, SC2 200 투여군은 45.48±3.57%, SC2 400 투여군은 44.15±2.33%로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (** : p<0.01) 감소가 나타났다.As shown in FIG. 12, the number of lymphocytes in the blood was 41.15 + 4.40% in the normal group and 60.00 + 3.15% in the control group, 45.48 + 3.57% in the
7) 혈청 내 사이토카인 측정7) Measurement of serum cytokines
혈청 내 사이토카인으로 IL-1β, IL-5, IL-6, IL-10, IL-12, TNF-α, INF-γ의 생성량을 각각 측정했다.The amounts of IL-1β, IL-5, IL-6, IL-10, IL-12, TNF-α and INF-γ were measured by serum cytokines.
혈청 내 IL-1β 생성량을 측정한 결과, 도 13에서와 같이, 혈청 내 IL-1β 생성량을 측정한 결과, 정상군은 11.08±5.72pg/㎖, 대조군은 193.75±4.58pg/㎖로 나타났으며, SC2 200 투여군은 77.99±6.90pg/㎖, SC2 400 투여군은 59.33±3.40pg/㎖로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (*** : p<0.001) 감소가 나타났다.As shown in FIG. 13, the amount of IL-1β produced in the serum was measured to be 11.08 ± 5.72 pg / ml in the normal group and 193.75 ± 4.58 pg / ml in the control group, , 77.99 ± 6.90 pg / ㎖ for the
또한, 혈청 내 IL-5 생성량을 측정한 결과, 도 14와 같이, 정상군은 0.79±0.10pg/㎖, 대조군은 19.65±1.55pg/㎖로 나타났으며, SC2 200 투여군은 13.76±1.61pg/㎖, SC2 400 투여군은 6.35±0.66pg/㎖로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (* : p<0.05, *** : p<0.001) 감소가 나타났다.As shown in FIG. 14, the amount of IL-5 produced in the serum was 0.79 ± 0.10 pg / ml in the normal group and 19.65 ± 1.55 pg / ml in the control group, and 13.76 ± 1.61 pg / ㎖ and
또한, 혈청 내 IL-6 생성량을 측정한 결과, 도 15에서와 같이, 정상군은 9.10±2.05pg/㎖, 대조군은 5907.41±274.13pg/㎖로 나타났으며, SC2 200 투여군은 4952.91±344.10pg/㎖, SC2 400 투여군은 4024.60±347.38pg/㎖로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (** : p<0.01, *** : p<0.001) 감소가 나타났다.As shown in FIG. 15, the amount of IL-6 produced in the serum was 9.10 ± 2.05 pg / ml in the normal group and 5907.41 ± 274.13 pg / ml in the control group, and 4952.91 ± 344.10 pg / ㎖, and
또한, 혈청 내 IL-10 생성량을 측정한 결과, 도 16에서와 같이, 정상군은 9.19±1.55pg/㎖, 대조군은 869.49±76.91pg/㎖로 나타났으며, SC2 200 투여군은 643.59±22.62pg/㎖, SC2 400 투여군은 344.64±20.48pg/㎖로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (** : p<0.01, *** : p<0.001) 감소가 나타났다.As shown in FIG. 16, the amount of IL-10 produced in the serum was 9.19 ± 1.55 pg / ml in the normal group and 869.49 ± 76.91 pg / ml in the control group, and 643.59 ± 22.62 pg / ㎖, and SC2 400 - treated group was 344.64 ± 20.48 pg / ㎖, and the SC2 treated group showed a significant decrease (**: p <0.01, ***: p <0.001)
또한, 혈청 내 IL-12 생성량을 측정한 결과, 도 17에서와 같이, 정상군은 3.48±0.37pg/㎖, 대조군은 202.07±33.56pg/㎖로 나타났으며, SC2 200 투여군은 308.30±16.47pg/㎖, SC2 400 투여군은 424.11±20.95pg/㎖로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (** : p<0.01, *** : p<0.001) 증가가 나타났다.As shown in FIG. 17, the amount of IL-12 produced in the serum was 3.48 ± 0.37 pg / ml in the normal group and 202.07 ± 33.56 pg / ml in the control group, and 308.30 ± 16.47 pg / / ㎖, and
또한, 혈청 내 TNF-α 생성량을 측정한 결과, 도 18에서와 같이, 정상군은 4.06±0.63pg/㎖, 대조군은 208.28±4.50pg/㎖로 나타났으며, SC2 200 투여군은 125.59±2.53pg/㎖, SC2 400 투여군은 111.48±2.90pg/㎖로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (*** : p<0.001) 감소가 나타났다.As shown in FIG. 18, the amount of TNF-α produced in the serum was 4.06 ± 0.63 pg / ml in the normal group and 208.28 ± 4.50 pg / ml in the control group, and 125.59 ± 2.53 pg / / ㎖, and
또한, 혈청 내 IFN-γ 생성량을 측정한 결과, 도 19에서와 같이, 정상군은 7.33±2.21pg/㎖, 대조군은 192.16±12.66pg/㎖로 나타났으며, SC2 200 투여군은 80.14±11.38pg/㎖, SC2 400 투여군은 50.13±6.70pg/㎖로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (*** : p<0.001) 감소가 나타났다.As shown in FIG. 19, the amount of IFN-γ produced in the serum was 7.33 ± 2.21 pg / ml in the normal group and 192.16 ± 12.66 pg / ml in the control group, and 80.14 ± 11.38 pg / / ㎖, and
8) 혈청 내 염증 유관인자 측정8) Measurement of inflammatory oil factor in serum
혈청 내 염증 유관인자로서 ROS, PGE2, 및 LTB4 생성량을 측정하였다.Serum levels of ROS, PGE 2 and LTB 4 were measured as inflammatory factors.
혈청 내 ROS 생성량을 측정한 결과, 도 20에서와 같이, 정상군은 113.09±2.60pg/㎖, 대조군은 14.90±4.56pg/㎖로 나타났으며, SC2 200 투여군은 21.23±2.30pg/㎖, SC2 400 투여군은 24.81±3.30pg/㎖로 나타나, SC2 투여군은 대조군에 비해 증가가 나타났다.As shown in FIG. 20, the amount of ROS produced in the serum was 113.09 ± 2.60 pg / ml in the normal group and 14.90 ± 4.56 pg / ml in the control group, 21.23 ± 2.30 pg / 400 administration group showed 24.81 ± 3.30 pg / ㎖, and the SC2 treated group showed an increase compared to the control group.
또한, 혈청 내 PGE2 생성량을 측정한 결과, 도 21에서와 같이, 정상군은 89.18±27.54pg/㎖, 대조군은 2348.04±117.45pg/㎖로 나타났으며, SC2 200 투여군은 1443.07±86.69pg/㎖, SC2 400 투여군은 1030.52±78.50pg/㎖로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (*** : p<0.001) 감소가 나타났다.As shown in FIG. 21, the amount of PGE 2 produced in the serum was 89.18 ± 27.54 pg / ml in the normal group and 2348.04 ± 117.45 pg / ml in the control group, and 1443.07 ± 86.69 pg / ㎖, and
또한, 혈청 내 LTB4 생성량을 측정한 결과, 도 22에서와 같이, 정상군은 986.75±123.89pg/㎖, 대조군은 2586.29±205.91pg/㎖로 나타났으며, SC2 200 투여군은 1793.65±211.55pg/㎖, SC2 400 투여군은 1445.40±217.90pg/㎖로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (** : p<0.01, *** : p<0.001) 감소가 나타났다.As shown in FIG. 22, the amount of LTB 4 produced in the serum was 986.75 ± 123.89 pg / ml in the normal group, 2586.29 ± 205.91 pg / ml in the control group, and 1793.65 ± 211.55 pg / (P <0.01, ***: p <0.001) in the SC2 treated group compared to the control group, and 1445.40 ± 217.90 pg /
LPS를 통해 급성염증을 유발한 ICR mouse를 이용하여 급성 염증 개선 효과에 대한 실험 결과를 요약하면 다음과 같다. Experimental results on the acute inflammation improvement effect of ICR mice induced acute inflammation through LPS were summarized as follows.
우선, 체중 및 식이섭취량 변화를 측정한 결과, 체중 및 식이섭취량은 SC2 200, 400 투여군에서 정상군과 대조군에 비해 차이가 나타나지 않았다.First, weight and dietary intakes were not significantly different between the control and SC2 groups.
또한, 간 및 비장 무게 변화를 측정한 결과, 간 무게는 SC2 200, 400 투여군에서 대조군에 비해 증가가 나타났으며, 비장 무게는 SC1, SC2 투여군에서 정상군과 대조군에 비해 차이가 나타나지 않았다.In liver and spleen weights, liver weight was increased in
또한, 간 기능을 측정한 결과, AST, ALT는 SC2 200, 400 투여군에서 대조군에 비해 차이가 나타나지 않았고, 신 기능을 측정한 결과, BUN, Creatinine은 SC2 200, 400 투여군에서 대조군에 비해 차이가 나타나지 않았다.As a result of measuring liver function, there was no difference between AST and ALT in the
또한, 혈액 내 면역세포 수를 측정한 결과, 백혈구, 단핵구, 호산구, 림프구는 SC2 200, 400 투여군에서 대조군에 비해 유의성 있는 감소가 나타났으며, 호중구, 호염기구는 SC2 200, 400 투여군에서 대조군에 비해 유의성 있는 증가가 나타났다.In addition, leukocyte, monocyte, eosinophil, and lymphocyte were significantly decreased in the
또한, 혈청 내 사이토카인 생성량을 측정한 결과 IL-1β, IL-5, IL-6, IL-10, TNF-α, IFN-γ는 SC2 200, 400 투여군에서 대조군에 비해 유의성 있는 감소가 나타났으며, IL-12는 SC2 200, 400 투여군에서 대조군에 비해 유의성 있는 증가가 나타났다.In addition, the amount of IL-1β, IL-5, IL-6, IL-10, TNF-α and IFN-γ was significantly decreased in the
또한, 혈청 내 ROS 생성량을 측정한 결과, SC2 200, 400 투여군에서 대조군에 비해 증가가 나타났으나, 유의성이 나타나지 않았고, 혈청 내 PGE2 생성량을 측정한 결과, SC2 200, 400 투여군에서 대조군에 비해 유의성 있는 감소가 나타났으며, 혈청 내 LTB4 생성량을 측정한 결과, SC2 200, 400 투여군에서 대조군에 비해 유의성 있는 감소가 나타났다.In addition, the amount of ROS produced in the serum was increased in the
따라서 본 발명의 조성물은 LPS를 통해 급성염증을 유발한 ICR mouse에서 체중 변화량의 증가와 식이섭취량의 감소가 나타났으며, 간 및 신 기능이 대조군에 비해 차이가 나타나지 않아 안전성이 확보되었다. 또한 면역세포 수, 사이토카인, 염증 유관인자들의 감소 효능이 실험적으로 규명되었다. Therefore, the composition of the present invention showed an increase in body weight change and a decrease in dietary intake in ICR mice which caused acute inflammation through LPS, and safety was secured because liver and renal function were not different from those in the control group. In addition, the effect of reducing the number of immune cells, cytokines, and inflammatory factors was experimentally determined.
다만, 염증부위에 가장 먼저 도달하는 과립성 백혈구인 호중구와 과립 내 약리활성 물질을 유리시키는 역할을 하는 호염구의 증가는 염증 개선을 위해 증가된 것으로 판단되며, 또한 염증성 사이토카인 중 TH1을 촉진하는 IL-12의 증가는 급성염증 반응에서 TH1을 촉진하기 위해 증가된 것으로 사료된다. However, increased neutrophils, which are the first granulocyte leukocytes to reach the inflammation site, and basophils, which serve to release the pharmacologically active substances in the granules, are thought to be increased for improving inflammation, and IL -12 is thought to be increased to promote TH1 in the acute inflammatory response.
따라서 본 발명에 따른 조성물을 적용함으로써 급성 염증의 개선에 효과를 나타낼 수 있는 것으로 파악되었다.Therefore, it has been found that application of the composition according to the present invention is effective for improving acute inflammation.
<면역 증진 효과><Immune enhancement effect>
1) 체중 변화 측정1) Weight change measurement
체중 변화를 측정한 결과는 표 3과 같다. 표 3에서 단위는 g이다.The results of measurement of weight change are shown in Table 3. In Table 3, the unit is g.
body weightChange of
body weight
표 3의 결과를 살펴보면, 실험 시작일 부터 4주차까지 정상군은 33.65±2.62, 34.55±2.75, 35.87±2.20, 37.35±1.87, 38.50±2.15ｇ으로 나타났으며, 대조군은 32.35±2.61, 32.77±2.31, 33.48±2.74, 34.90±3.00, 36.73±2.57ｇ으로 나타났다. SC2 200 투여군은 34.87±2.98, 35.08±2.94, 36.32±2.97, 37.50±2.75, 38.88±2.91ｇ으로 나타났으며, SC2 400 투여군은 33.43±1.80, 33.67±1.73, 34.50±1.53, 35.33±1.86, 36.57±2.05ｇ으로 나타나, 모든 실험군은 차이가 나타나지 않았다.In the results of Table 3, 33.65 ± 2.62, 34.55 ± 2.75, 35.87 ± 2.20, 37.35 ± 1.87, and 38.50 ± 2.15g were normal groups from the beginning of the experiment to the 4th week of the experiment. The control group was 32.35 ± 2.61, 32.77 ± 2.31 , 33.48 ± 2.74, 34.90 ± 3.00, and 36.73 ± 2.57g, respectively. The
2) 식이섭취량 변화 측정2) Measurement of dietary intake change
식이섭취량 변화를 측정한 결과는 표 4와 같다. 표 4에서 단위는 g이다.The results of measurement of dietary intake changes are shown in Table 4. In Table 4, the unit is g.
food intakeChange of
food intake
표 4의 결과를 살펴보면, 식이섭취량 변화를 측정한 결과, 1주차부터 4주차까지 정상군은 35.00, 34.97, 36.15, 38.20 ｇ으로 나타났으며, 대조군은 30.20, 34.13, 35.63, 33.70 ｇ으로 나타났다. SC2 200 투여군은 30.40, 33.80, 36.43, 34.50 ｇ으로 나타났으며, SC2 400 투여군은 30.60, 31.42, 32.53, 35.90 ｇ으로 나타나, 모든 실험군은 차이가 나타나지 않았다.In the results of Table 4, 35.00, 34.97, 36.15, and 38.20 g of the normal group were observed from the 1st to 4th weeks of the measurement, and 30.20, 34.13, 35.63, and 33.70 g of the control group were measured. The
3) 장기 무게 변화 측정3) Measurement of long-term weight change
간, 비장의 무게 변화 측정을 수행했으며, 간 무게 변화 측정 결과는 도 1과 같다. Liver, and spleen. The results of the liver weight change measurement are shown in Fig.
간 무게 변화를 측정한 결과, 도 23에서와 같이, 정상군은 2.06±0.07ｇ, 대조군은 2.08±0.17ｇ으로 나타났으며, SC2 200 투여군은 2.02±0.18ｇ, SC2 400 투여군은 2.03±0.20ｇ으로 나타나, 모든 실험군은 차이가 나타나지 않았다.As shown in FIG. 23, the liver weight change was 2.06 ± 0.07 g in the normal group and 2.08 ± 0.17 g in the control group, 2.02 ± 0.18 g in the
또한, 비장 무게 변화를 측정한 결과, 도 24에서와 같이, 정상군은 0.13±0.01ｇ, 대조군은 0.13±0.01ｇ으로 나타났으며, SC2 200 투여군은 0.13±0.02ｇ, SC2 400 투여군은 0.13±0.01ｇ으로 나타나, 모든 실험군은 차이가 나타나지 않았다.As a result of measurement of the spleen weight change, as shown in FIG. 24, 0.13 ± 0.01 g of the normal group and 0.13 ± 0.01 g of the control group were observed, and 0.13 ± 0.02 g of the
4) 간 기능 측정4) Liver function measurement
간 기능 측정을 위하여 AST 및 ALT 레벨을 측정하였다.AST and ALT levels were measured for liver function measurements.
간 기능 지표인 AST를 측정한 결과, 도 25에서와 같이, 정상군은 92.19±9.70U/ℓ, 대조군은 124.80±11.11U/ℓ로 나타났으며, SC2 200 투여군은 125.32±8.40U/ℓ, SC2 400 투여군은 124.50±12.04U/ℓ로 나타나, SC2 투여군은 대조군에 비해 차이가 나타나지 않았다. As shown in FIG. 25, the AST of liver function index was 92.19 ± 9.70 U / ℓ in the normal group and 124.80 ± 11.11 U / ℓ in the control group, 125.32 ± 8.40 U / ℓ in the
또한, ALT를 측정한 결과, 도 26에서와 같이, 정상군은 31.06±2.63U/ℓ, 대조군은 38.16±3.78U/ℓ로 나타났으며, SC2 200 투여군은 38.36±3.60U/ℓ, SC2 400 투여군은 36.54±3.20U/ℓ로 나타나, SC2 투여군은 대조군에 비해 차이가 나타나지 않았다.As shown in FIG. 26, ALT was measured to be 31.06 ± 2.63 U / ℓ in the normal group and 38.16 ± 3.78 U / ℓ in the control group, 38.36 ± 3.60 U / The dose of SC2 treated group was 36.54 ± 3.20 U / ℓ.
5) 신 기능 측정5) Measurement of new functions
신 기능 측정을 위하여 BUN, creatinine 레벨을 측정하였다.BUN and creatinine levels were measured to measure renal function.
신 기능 지표인 BUN을 측정한 결과, 도 27에서와 같이, 정상군은 26.56±1.69㎎/㎗, 대조군은 27.40±2.51㎎/㎗로 나타났으며, SC2 200 투여군은 27.73±3.07㎎/㎗, SC2 400 투여군은 26.47±3.24㎎/㎗로 나타나, 모든 실험군은 차이가 나타나지 않았다.As shown in FIG. 27, the BUN of the new functional index was 26.56 ± 1.69 mg / dl in the normal group and 27.40 ± 2.51 mg / dl in the control group, 27.73 ± 3.07 mg / dl in the
또한, creatinine을 측정한 결과, 도 28에서와 같이, 정상군은 0.35±0.04㎎/㎗, 대조군은 0.36±0.05㎎/㎗로 나타났으며, SC2 200 투여군은 0.36±0.05㎎/㎗, SC2 400 투여군은 0.36±0.03㎎/㎗로 나타나, 모든 실험군은 차이가 나타나지 않았다. As shown in FIG. 28, creatinine was 0.35 ± 0.04 mg / dl in the normal group and 0.36 ± 0.05 mg / dl in the control group, 0.36 ± 0.05 mg / dl in the
6) 혈액 내 면역세포 수 측정6) Measurement of immune cell count in blood
혈액 내의 백혈구, 단핵구, 호중구, 호산구, 호염구, 및 림프구의 수를 각각 측정하였다.The numbers of leukocytes, mononuclear cells, neutrophils, eosinophils, basophils, and lymphocytes in blood were measured respectively.
혈액 내 백혈구 수를 측정한 결과, 도 29에서와 같이, 정상군은 6.67±0.85 ×103cells/㎕, 대조군은 4.13±0.61 ×103cells/㎕로 나타났으며, SC2 200 투여군은 4.07±0.72 ×103cells/㎕, SC2 400 투여군은 4.92±0.64 ×103cells/㎕로 나타나, SC2 400 투여군은 대조군에 비해 유의성 있는 (* : p<0.05) 증가가 나타났다.As a result of measurement of white blood cell count in blood, as shown in FIG. 29, 6.67 ± 0.85 × 10 3 cells / μl in the normal group and 4.13 ± 0.61 × 10 3 cells / μl in the control group and 4.07 ± 0.72 × 10 3 cells / μl in
또한, 혈액 내 단핵구 수를 측정한 결과, 도 30에서와 같이, 정상군은 1.60±0.08%, 대조군은 1.95±0.03%로 나타났으며, SC2 200 투여군은 1.65±0.06%, SC2 400 투여군은 1.56±0.05%로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (* : p<0.05) 감소가 나타났다.As shown in FIG. 30, the number of mononuclear cells in the blood was 1.60 ± 0.08% in the normal group and 1.95 ± 0.03% in the control group, 1.65 ± 0.06% in the
또한, 혈액 내 호중구 수를 측정한 결과, 도 31에서와 같이, 정상군은 21.80±1.25%, 대조군은 24.48±1.56%로 나타났으며, SC2 200 투여군은 29.97±2.29%, SC2 400 투여군은 35.12±3.44%로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (** : p<0.01, *** : p<0.001) 증가가 나타났다.As shown in FIG. 31, the number of neutrophils in the blood was 21.80 ± 1.25% in the normal group, 24.48 ± 1.56% in the control group, 29.97 ± 2.29% in the
혈액 내 호산구 수를 측정한 결과, 도 32에서와 같이, 정상군은 15.28±1.41%, 대조군은 15.80±1.30%로 나타났으며, SC2 200 투여군은 20.52±1.95%, SC2 400 투여군은 24.93±1.40%로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (** : p<0.01, *** : p<0.001) 증가가 나타났다.The eosinophil count in the blood was measured as 15.28 ± 1.41% in the normal group and 15.80 ± 1.30% in the control group as shown in FIG. 32, 20.52 ± 1.95% in the
또한, 혈액 내 호염기구 수를 측정한 결과, 도 33에서와 같이, 정상군은 0.34±0.03%, 대조군은 0.35±0.02%로 나타났으며, SC2 200 투여군은 0.36±0.01%, SC2 400 투여군은 0.40±0.03%로 나타나, SC2 400 투여군은 대조군에 비해 유의성 있는 (* : p<0.05) 증가가 나타났다. As shown in FIG. 33, the number of basophils in the blood was 0.34 ± 0.03% in the normal group and 0.35 ± 0.02% in the control group, 0.36 ± 0.01% in the
또한, 혈액 내 림프구 수를 측정한 결과, 도 34에서와 같이, 정상군은 79.66±5.25%, 대조군은 60.47±4.10%로 나타났으며, SC2 200 투여군은 60.87±8.97%, SC2 400 투여군은 68.45±5.30%로 나타나, SC2 400 투여군은 대조군에 비해 유의성 있는 (* : p<0.05) 증가가 나타났다.As shown in FIG. 34, the number of lymphocytes in the blood was 79.66 ± 5.25% in the normal group and 60.47 ± 4.10% in the control group, 60.87 ± 8.97% in the
7) 혈청 내 면역글로불린 생성량 측정7) Measurement of serum immunoglobulin production
혈청 내 면역글로불린 생상량의 지표로서 IgA, IgG, IgM 생성량을 측정하였다.IgA, IgG, and IgM production were measured as an indicator of serum immunoglobulin production.
혈청 내 IgA 생성량을 측정한 결과, 도 35에서와 같이, 정상군은 2294.33±141.35pg/㎖, 대조군은 1480.06±177.61pg/㎖로 나타났으며, SC2 200 투여군은 1618.00±125.39pg/㎖, SC2 400 투여군은 1872.31±143.63pg/㎖로 나타나, SC2 400 투여군은 대조군에 비해 유의성 있는 (* : p<0.05) 증가가 나타났다.As shown in FIG. 35, serum IgA production was 2294.33 ± 141.35 pg / ml in the normal group and 1480.06 ± 177.61 pg / ml in the control group. In the
또한, 혈청 내 IgG 생성량을 측정한 결과, 도 36에서와 같이, 정상군은 3705.88±256.75pg/㎖, 대조군은 2752.39±257.17pg/㎖로 나타났으며, SC2 200 투여군은 4741.19±192.38pg/㎖, SC2 400 투여군은 5141.12±285.21pg/㎖로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (*** : p<0.001) 증가가 나타났다.As shown in FIG. 36, the amount of IgG produced in the serum was 3705.88 ± 256.75 pg / ml in the normal group, 2752.39 ± 257.17 pg / ml in the control group, and 4741.19 ± 192.38 pg / ,
또한, 혈청 내 IgM 생성량을 측정한 결과, 도 37에서와 같이, 정상군은 627.06±56.32 pg/㎖, 대조군은 492.37±62.50 pg/㎖로 나타났으며, SC2 200 투여군은 630.37±43.31 pg/㎖, SC2 400 투여군은 667.70±23.19 pg/㎖로 나타나, SC2 투여군은 대조군에 비해 유의성 있는 (* : p<0.05, ** : p<0.01) 증가가 나타났다.As shown in FIG. 37, the amount of IgM produced in the serum was 627.06 ± 56.32 pg / ml in the normal group and 492.37 ± 62.50 pg / ml in the control group, and 630.37 ± 43.31 pg / ml in the
면역억제제(Cyclophosphamide)를 통해 면역력을 저하시킨 ICR mouse를 통해 면역 증진 효과를 실험한 결과를 요약하면 다음과 같다.Immunosuppressive effect of immunosuppressive agent (Cyclophosphamide) through the ICR mice decreased immunity.
우선, 체중 및 식이섭취량 변화를 측정한 결과, 체중 및 식이섭취량은 SC2 200, 400 투여군에서 정상군과 대조군에 비해 차이가 나타나지 않았고, 간 및 비장 무게 변화를 측정한 결과, 간 및 비장 무게는 SC2 200, 400 투여군에서 정상군과 대조군에 비해 차이가 나타나지 않았다.The liver and spleen weights of liver and spleen weights were measured by SC2 and 400 in the
또한, 간 기능을 측정한 결과, AST, ALT는 SC2 200, 400 투여군에서 대조군에 비해 차이가 나타나지 않았고, 신 기능을 측정한 결과, BUN, Creatinine은 SC2 200, 400 투여군에서 정상군과 대조군에 비해 차이가 나타나지 않았다.As a result of measuring liver function, AST and ALT did not show any difference compared to the control group in the
또한, 혈액 내 면역세포 수를 측정한 결과, 백혈구와 림프구는 SC2 200, 400 투여군에서 증가가 나타났으나 SC2 400 투여군에서 유의성이 나타났고, 단핵구는 SC2 200, 400 투여군에서 유의성있는 감소가 나타났다. 또한 호중구와 호산구는 SC2 200, 400 투여군에서 증가가 나타났으나 SC2 400 투여군에서 유의성이 나타났고, 호염기구는 SC2 400 투여군에서 유의성 있는 증가가 나타났다.In addition, the number of white blood cells and lymphocytes was increased in the
또한, 혈청 내 면역글로불린 생성량을 측정한 결과 IgA는 SC2 200, 400 투여군에서 증가가 나타났으나 SC2 400 투여군에서 유의성이 나타났고, IgG, IgM은 SC2 200, 400 투여군에서 유의성 있는 증가가 나타났다.In addition, serum IgG production was increased in the
따라서, SC2는 면역억제제 (Cyclophosphamide)를 통해 면역력을 저하시킨 ICR mouse에서 체중 변화량의 감소와 식이섭취량의 증가가 나타났으며, 간 및 신 기능이 대조군에 비해 차이가 나타나지 않아 안전성이 확보되었다. 또한 면역세포 수, 면역글로불린의 증가 효능이 실험적으로 규명되었다. 다만 염증과정에서 대식세포로 분화하는 단핵구의 감소는 다른 면역세포들의 증가에 의해 비율이 감소된 것이라 사료된다. 또한 면역질환의 이상이 발생하면 최초로 반응하는 IgM과 혈액 및 세포조직에서 발견되며 2차적으로 반응하는 IgG, 타액같은 외분비액에 존재하여 감염방어에 관여하는 IgA의 증가는 면역증진에 효능이 있어 증가된 것이라 사료된다. Therefore, SC2 decreased the body weight change and increased the dietary intake in ICR mice, which had decreased immunity through immunosuppressive agents (cyclophosphamide), and the liver and the renal function were not different compared to the control group, and the safety was secured. In addition, the number of immune cells and the increasing effect of immunoglobulin were experimentally determined. However, the decrease of monocyte differentiated into macrophages during inflammation seems to be reduced by the increase of other immune cells. In addition, IgM that reacts first when an abnormality of the immune disease occurs is present in the secretory fluid such as IgG and saliva which are found in blood and cell tissue and reacts secondarily, and the increase of IgA involved in infection defense is effective for immunity enhancement .
따라서 본 발명에 따른 조성물은 급성 염증의 개선에 효과뿐만 아니라 면역 증진의 효과도 동시에 나타내는 것으로 파악되었다.Therefore, the composition according to the present invention was found to exhibit not only the improvement of acute inflammation but also the effect of immunity enhancement.
본 발명의 권리는 위에서 설명된 실시예에 한정되지 않고 청구범위에 기재된 바에 의해 정의되며, 본 발명의 분야에서 통상의 지식을 가진 자가 청구범위에 기재된 권리범위 내에서 다양한 변형과 개작을 할 수 있다는 것은 자명하다.It is to be understood that the invention is not limited to the disclosed embodiment, but is capable of many modifications and variations within the scope of the appended claims. It is self-evident.
Claims (5)
상기 녹용 분말은 녹용을 추출한 후 균주를 투여하여 배양하고 이를 동결건조하여 분말화한 발효 녹용 분말인 것을 특징으로 하는 급성 염증 개선 및 면역 증진용 조성물.
Antler powder, and a natural medicinal powder composed of Gakyung, McDonald, Omija and Ginseng at a weight ratio of 2: 1 to 1: 2,
Wherein the deer antler powder is a fermented deer antler powder obtained by extracting antler antler, culturing the antler antler extract, and lyophilizing the powder to obtain a fermented deer antler powder.
상기 천연 약재 분말은 길경, 맥문동, 오미자, 인삼을 환류 추출하고 감압 농축하여 얻어진 용액을 동결 건조하여 얻어진 분말인 것을 특징으로 하는 급성 염증 개선 및 면역 증진용 조성물.
The method according to claim 1,
Wherein the natural medicinal ingredient powder is a powder obtained by lyophilizing a solution obtained by refluxing and extracting Gakyung, McMundong, Omija, and Ginseng under reduced pressure, and then lyophilizing the obtained solution.
상기 조성물은 경구투여용인 것을 특징으로 하는 급성 염증 개선 및 면역 증진용 조성물.
The method according to claim 1,
The composition for acute inflammation improvement and immunity enhancement is characterized in that the composition is for oral administration.
상기 조성물은 식품 조성물인 것을 특징으로 하는 급성 염증 개선 및 면역 증진용 조성물.The method according to claim 1,
The composition for improving acute inflammation and immunity, wherein the composition is a food composition.
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| KR100532633B1 (en) | 2003-09-29 | 2005-12-02 | 코스맥스 주식회사 | Cosmetic composition having anti-inflammatory, skin-protecting, skin-elastic effects which comprise mixed plants extract |
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