보라성게(Anthocidaris crassispina)의 알과 껍질 추출물의 항균활성 및 항산화 효과와 껍질의 열수추출물이 넙치의 비특이적 면역반응에 미치는 영향 = Antibacterial, antioxidant effect of sea-urchin (Anthocidaris crassispina) roe and shell extract and the effect of the hot water extract of shell on the non-specific immune responses in Paralichthys olivaceus

다국어 초록 (Multilingual Abstract)

Side effects of synthetic antibiotics and antioxidants, natural antibacterial substances and antioxidants present in natural products, and their human impact are the major research topics conducted to date. Recently, a study on natural antibacterial substances and antioxidants are concentrated in bio-active compounds of marine source has produced a unique metabolism and specificity. Sea-urchin called as “haedam” in oriental medicine, known for its excellent physiological activity.
In this study, we use purple sea-urchin (Anthocidaris crassispina)'s shell and egg, was used to measured antimicrobial activity against human and fish bacterial pathogens, and its antioxidant activity. Also, we investigate effect of hot water extract of sea-urchin shell in the immune response of olive flounder and to check it's usage as feed additives.
Hence for the study, 8.3955 g of A. crassispina egg hot water extract, 2.009 g of A. crassispina shell hot water extract, 3.1344 g of A. crassispina egg hexane layer, 0.3199 g of A. crassispina egg methylene chloride (CH2Cl2) layer, 0.2998 g of A. crassispina egg ethyl acetate layer, 2.5713 g of A. crassispina egg aqueous layer, 0.517 g of A. crassispina shell hexane layer, 0.0085g of A. crassispina shell CH2Cl2 layer, 0.0153 g of A. crassispina shell ethyl acetate layer and 0.8838 g of A. crassispina shell aqueous layer were extracted.
Antibacterial activity of A. crassispina egg extracts were compared with that of A. crassispina shell extracts, which reveals that egg extracts were more effect than shell extracts. In DPPH radical scavenging activity, both egg and shell extracts show similar reducing activity but higher than that of control BHA and BHT. Especially, aqueous layer of egg extracts, hot water extract and ethyl acetate layer of shell’s antioxidant activities were excellent in low concentration. In hydroxyl radical scavenging activity, also show similar results with DPPH radical scavenging assay. However, hydroxyl radical scavenging activity of egg extracts is slightly higher than that of shell extracts.
As a result in in vivo test, the hot water extract of A. crassispina shell was showed that do not effect on the non-specific immune responses of the olive flounder. The reason for this result, the feed uptake of olive flounder is presumably due to low during the experiment period. Further studies are need to detailed analyses of sea-urchin shell extracts and to encouraging the isolating biologically more potential substance.

목차 (Table of Contents)

• Ⅰ. 서 론 1
• Ⅱ. 재료 및 방법 4
• 2.1. 시료의 구입 및 추출 4
• 2.1.1. 시료의 구입 4
• 2.1.2. 성게알 및 껍질의 분말 제조 4
• 2.1.3. 성게알 및 성게껍질의 열수추출물 제조 4
• 2.1.4. 성게알 및 성게껍질의 유기용매 추출 및 분획물 제조 5
• 2.2. 성게알 및 성게껍질 추출물의 항균활성 확인 6
• 2.2.1. 항균실험 사용균주 6
• 2.2.2. 추출물의 항균활성 탐색 7
• 2.3. 성게알 및 성게껍질 추출물의 항산화 효과 확인 8
• 2.3.1. DPPH radical 소거활성 측정 8
• 2.3.2. Hydroxyl radical 소거활성 측정 9
• 2.4. 성게껍질 열수추출물의 첨가에 따른 양식 넙치의 비특이적 면역 반응 10
• 2.4.1 실험어 사육관리 10
• 2.4.2. 실험사료 제작 및 투여 10
• 2.4.1 Respiratory burst activity 측정 11
• 2.4.2. 혈청 내 lysozyme activity 측정 11
• Ⅲ. 결과 및 고찰 12
• 3.1. 시료의 추출 12
• 3.1.2. 성게알 및 껍질의 열수추출물과 각 분획물의 수율 12
• 3.2. 성게알 및 성게껍질 추출물의 항균활성 확인 14
• 3.3. 성게알 및 성게껍질 추출물의 항산화 효과 확인 19
• 3.3.1. DPPH radical 소거활성 측정 19
• 3.3.2. Hydroxyl radical 소거활성 측정 23
• 3.4. 성게껍질 열수추출물의 첨가에 따른 양식 넙치의 비특이적 면역 반응 27
• 3.4.1 Respiratory burst activity 측정 27
• 3.4.2. 혈청 내 lysozyme activity 측정 29
• Ⅳ. 요 약 31
• Ⅴ. 참고 문헌 32
• Ⅵ. 감사의 글 38