천년초에서 분리한 점질다당의 면역자극 활성

국문 초록 (Abstract)

천년초(Opuntia humifusa)에 존재하는 점질다당의 새로운 이용방안을 모색하기 위해 천년초로부터 다당을 분리하여 면역자극활성을 측정하였다. 천년초의 주요 다당 CNC-E는 상업용 효소처리, 물 추출 및 ethanol 침전을 거쳐 조제하고 이들의 특성을 검토하였다. CNC-E는 분자량 약 700 kDa의 다당체로, 구성당 조성을 확인한 결과, arabinose, galactose 및 xylose를 높은 비율로 함유하고 있었으며 rhamnose와 fucose를 미량 함유하고 있었다. 한편 CNC-E는 비장세포 유래 림프구에 대해서는 비교적 높은 증식활성을 농도 의존적으로 나타냈다. CNC-E로 자극한 macrophage는 IL-6, IL-12, IL-10 및 TNF-α와 같은 cytokine의 생산을 농도 의존적으로 증진시켰으며 CNC-E를 mouse에 정맥 주사한 후 얻어진 NK세포는 Yac-1 종양 세포주에 대한 높은 치사작용을 보였다. 특히 CNC-E를 mouse 당 100μg 처리한 후 얻어진 NK세포는 미처리 대조군에 비해 3배 높은 치사활성을 나타냈다. CNC-E는 인체 초기 면역반응에 있어 중요한 역할을 담당하는 보체계에 대하여 농도 의존적인 활성을 나타냈으며, anti-human C3를 이용한 면역전 기영동실험과 Ca2+이온이 제거된 상태에서의 항보체 활성실험을 통해 천년초 다당 CNC-E는 고전경로와 부경로를 모두 경유하여 활성을 나타내고 있음을 확인하였다. 이상의 결과로부터 한국산 선인장 천년초에는 인체에 유익한 면역 활성 다당체를 다량 함유함을 알 수 있었다.

다국어 초록 (Multilingual Abstract)

To examine the new practical utilization of mucilages in Opuntia humifusa, the Korean pear cactus, the polysaccharides were isolated from O. humifusa, and immuno-stimulating activities were assayed. The main polysaccharide, CNC-E, was prepared by a commercial enzyme treatment, water extraction, and ethanol precipitation. The molecular mass of CNC-E was estimated to be about 700 kDa, and it consisted mainly of arabinose, galactose and xylose in addition to two minor sugars such as rhamnose and fucose. On the other hand, CNC-E showed considerably high splenocyte proliferation activity in a dose-dependent manner. Peritoneal macrophages stimulated with CNC-E produced cytokines such as IL-6, IL-12, IL-10, and TNF-α. The intravenous administration of CNC-E significantly augmented the cytotoxicity of natural killer (NK) cells against Yac-1 tumor cells. Especially, NK cells obtained from the mice treated with 100 μg of CNC-E showed threefold higher cytolytic activity than those of untreated mice. CNC-E also showed potent anti-complementary activity in a dose-dependent fashion. Identification of C3 activation products by the crossed immunoelectrophoresis using anti-human C3 and the anti-complementary activity of CNC-E in a Ca2+-free condition suggested complement activations by CNC-E that occur via both alternative and classical pathways. These results indicate that Korean pear cactus contains selected polysaccharides that provide immuno-stimulating activities beneficial to human health.

목차 (Table of Contents)

• Abstract
• 서론
• 재료 및 방법
• 결과 및 고찰
• 요약
• 감사의 글
• 문헌

참고문헌 (Reference)

• 1 박민경, "천년초(Opuntia humifusa) 추물물의 사염화탄소를 처치한흰쥐에서의 간보호 효과" 한국식품과학회 37 (37): 822-826, 2005
• 2 이경석, "천년초 선인장 추출물의 항산화 효과" 한국식품과학회 37 (37): 474-478, 2005
• 3 김경태, "천년초 선인장 분말을 첨가한 우리밀 식빵의 품질특성" 한국식품조리과학회 23 (23): 461-468, 2007
• 4 송영주, "손바닥선인장이 고지방식이 흰쥐의 유산소 운동능력에 미치는 영향" 한국운동영양학회 9 (9): 183-188, 2005
• 5 이경석, "병원성 식중독 미생물에 대한 천년초 선인장 추출물의 항균 활성" 한국식품영양과학회 33 (33): 1268-1272, 2004
• 6 Hudson L, "Two dimensional or crossed immunoelectrophoresis. In Practical Immunology" Blackwell Scientific Publications 244-246, 1989
• 7 Whaley K, "The complement system, In Complement in Health and Disease" MTP Press 1-35, 1986
• 8 Kim SY, "Studies on the separation of antioxidative and anti-microbial compounds of Korean perennial cactus Cheonnyuncho" Hoseo University 2003
• 9 Iino K, "Structure-function relationship of antitumor β- 1,3-glucan obtained from matted mycelium of cultured Grifora frondosa" 33 : 4950-4956, 1985
• 10 Yadomae T, "Structure and biological activities of fungal β-1,3 glucans" 120 : 413-431, 2000
• 11 Shimura K, "Screening of host- mediated antitumor polysaccharides by crossed immunoelectrophoresis using fresh human serum" 33 : 403-408, 1983
• 12 Ruoslahti E, "Proteoglycans in cell regulation" 264 : 13369-13372, 1989
• 13 Saito H, "PSK, a polysaccharide from Coriolus versicolor, enhances oxygen metabolism of murine peritoneal macrophages and the host resistance to listerial infection" 134 : 1029-1035, 1988
• 14 Blumenkrantz N, "New method for quantitative determination of uronic acid" 54 : 484-489, 1973
• 15 Keller R, "Mechanism of macrophage-mediated tumor cell killing: a comparative analysis of the roles of reactive nitrogen intermediates and tumor necrosis factor" 46 : 682-686, 1990
• 16 Hikino H, "Mechanism of hypoglycemic activity of aconitan A, a glycan of Aconitum carmichaeli roots" 25 : 295-304, 1989
• 17 김장현, "Lactobacillus plantarum 균체 중 항보체 활성물질의 특성과 작용양식" 한국식품과학회 34 (34): 290-295, 2002
• 18 Zhu H, "Isolation and characterization of an anti-complementary protein-bound polysaccharide from the stem barks of Eucommia ulmoides" 8 : 1220-1230, 2008
• 19 Wang HX, "Immunomodulatory and antitumor activities of a polysaccharide-peptide complex from mycelia culture of Tricholoma sp., a local edible mushroom" 57 : 269-281, 1995
• 20 Hunter CA, "IL-1 beta is required for IL-12 to induce production of IFN-gamma by NK cells. A role for IL-1 beta in the T cell-independent mechanism of resistance against intracellular pathogens" 155 : 4347-4354, 1995
• 21 Paulson JC, "Glycoprotein: What are the sugar chains for?" 14 : 272-276, 1989
• 22 Young-Chul Lee, "Food Functionality of Opuntia ficus-indica var. Cultivated in Jeju Island" 한국식품영양과학회 10 (10): 103-110, 2005
• 23 Shida K, "Essential roles of monocytes in stimulating human peripheral blood mononuclear cells with Lactobacillus casei to produce cytokines and augment natural killer cell activity" 13 : 997-1003, 2006
• 24 Nathan CF, "Current concepts: the macrophage as an effector cell" 303 : 662-665, 1980
• 25 Kabat EA, "Complement and complement fixation, In Experimental Immunochemistry. 2nd ed" Thormas Publisher 133-240, 1971
• 26 Dubois M, "Colorimetric method for determination of sugars and related substances" 28 : 350-356, 1956
• 27 Kwon MH, "Characteristics of immune response by polysaccharides with complement system activity" 30 : 30-43, 1997
• 28 Male D, "Cells, tissues, and organs of the immune system. In Immunology. 7th ed" Mosby 19-58, 2006
• 29 Wang H, "Asian and Siberian ginseng as a potential modulator of immune function: an in vitro cytokine study using mouse macrophages" 327 : 123-128, 2003
• 30 Lasek W, "Antitumor effects of the combination immunotherapy with interleukin-12 and tumor necrosis factor alpha in mice" 45 : 100-108, 1997
• 31 Lee SJ, "Antimetastatic and immunomodulating properties of a new herbal prescription, Bojung-bangam-tang" 3 : 147-157, 2003
• 32 Bradford MM, "A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding" 72 : 248-254, 1976
• 33 Kantakamalakul W, "A novel enhanced green fluorescent protein (EGFP)-K562 flow cytometric method for measuring natural killer (NK) cell cytotoxic activity" 272 : 189-197, 2003
• 34 Jones TM, "A gas chromatography method for the determination of aldose and uronic acid constituents of plant cell wall polysaccharide" 49 : 926-936, 1972