남석현 서석진 2006-08 1556 https://aurora.ajou.ac.kr/handle/2018.oak/7012 학위논문(석사)--아주대학교 교육대학원 :생물교육,2006. 8 본 연구는 애기똥풀 추출물의 항산화 효과를 알아보기 위하여 세포를 손상시키는 자유라디칼을 어느 정도 제거할 수 있는지 DPPH 자유라디칼의 억제 능력을 in vitro에서 알아보고, 과산화라디칼에 대한 소거활성이 있는지 HL-60 cells을 대상으로 조사하였다. 애기똥풀 추출물의 DPPH 자유라디칼 소거능력은 줄기의 경우 8월과 고산지역에서 78.7%, 76.6%, 뿌리는 10월과 고산지역에서 87.6%, 84.6%로 나타났으며, 과산화라디칼 소거활성 능력은 줄기 8월이 84.9%, 고산지역은 82.1%로 나타나났다. 그리고 인터페론-γ나 LPS로 유도된 활성화된 대식세포에서 NO가 얼마나 생성되는지 조사해보고, 곽향 추출물을 처리하였을 때 사이토카인처럼 NO를 생성하는 상승효과가 있는지 알아보았다. 또한 사이토카인이나 곽향 추출물에 의해 활성화된 대식세포가 TNF-α를 분비하는지, 그리고 대식세포 내 NO생성효소가 NO 생성억제제에 어느정도 영향을 받는지도 알아보았다. 따라서 신체 외부로부터 감염된 세포는 활성화되어 NO를 생성하고 NO의 작용에 의해 염증회복 작용이 일어나는데, 그 과정에서 식물 추출물이 더욱 높은 수준의 NO를 생성하도록 유도하므로 대식세포의 내재면역성 기작에 중요한 역할을 하는 NO의 생성을 유도하는 효과는 면역증진을 통한 감염 저항성의 획득이나 염증반응의 완화와 같은 활성 성분이 될 수 있으므로 질병에 대한 인체 보호 역할을 할 수 있을 것으로 판단된다. 국문요약 Ⅰ. 서론 1. 연구필요성 및 목적 ····························································· 1 2. 연구내용 ·········································································· 2 Ⅱ. 이론적 배경 1. 천연물의 효과 ···································································· 3 2. 항산화 작용 ······································································ 4 3. Nitric oxide의 특징과 염증관련 기작·································· 6 Ⅲ. 연구 방법 1. 시약 ·············································································· 12 2. 시료추출 및 제조 ······························································· 12 3. DPPH 자유라디칼 소거활성 측정 ······································· 15 4. HL-60 cells의 분화유도와 하이드록시라디칼 소거활성측정················15 5. 마우스 복강대식세포의 배양과 NO 농도 측정 ·················· 16 6. MTT assay에 의한 세포독성 검정 ···································· 17 7. 시료내 endotoxin 혼입여부 측정 ········································· 18 8. ELISA에 의한 TNF-α의 측정 ··············································· 18 9. 통계분석 ········································································ 19 Ⅳ. 연구 결과 및 논의 1. 애기똥풀의 항산화효과 가. 애기똥풀 추출물의 DPPH 자유라디칼 소거활성 ············ 20 나. HL-60 세포에서 애기똥풀 추출물의 과산화라디칼 소거활성측정 ··························································································· 23 2. Nitric oxide 생성에 관한 연구 가. 복강대식세포내 Endotoxin이 NO 생성에 미치는 영향조사················26 나. 복강대식세포에서 애기똥풀 및 곽향 추출물의 NO 생성유도효과 ····························································································27 다. 곽향에 의한 NO 생성유도 기작 추정 ······························· 31 라. 복강대식세포에서 세포독성 검정 ····································· 37 마. 곽향의 TNF-α 생성 효과 ··················································· 39 Ⅴ. 요약 및 결론 ····································································· 40 참고문헌 kor The Graduate School, Ajou University 아주대학교 논문은 저작권에 의해 보호받습니다. 애기똥풀(Chelidonium majus L.) 및 곽향(Herba Agastache) 열수추출물의 항산화효과와 Nitric Oxide 생성 유도효과 SEO SEOK JIN Thesis 아주대학교 교육대학원 SEO SEOK JIN 교육대학원 생물교육 2006. 8 Master http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000001556 항산화 대식세포 nitric oxide