한국산 적송 (Pinus densiflora) 심재 에탄올 추출물의 지방간 개선 효과
- 발행기관 강릉원주대학교 일반대학원
- 발행년도 2021
- 학위수여년월 2022. 2
- 학위명 석사
- 학과 및 전공 도움말 일반대학원 웰니스바이오산업학과
- 실제URI http://www.dcollection.net/handler/kangnung/000000011118
- UCI I804:42001-000000011118
- 본문언어 한국어
초록/요약 도움말
간에 과도한 지질 축적은 비알코올성 지방간 질환 (NAFLD)에 기여하여 비알코올성 지방간 염(NASH)을 유발합니다. 한국의 최근 연구에 따르면 한국산 적송 심재 (RPH)의 항산화 함 량이 프랑스 바다 소나무 껍질에서 추출한 피크노제놀보다 높은 것으로 나타났다. 이를 바 탕으로 진행한 실험에서 국산 적송 심재는 정상 간세포에서 세포 증식 억제에 영향을 미치 지 않는 것으로 나타났다. 항산화 효능을 알아보기 위해 과산화지질과 환원글루타치온 함량 을 측정하여 국산 적송 심재의 항산화 효능을 확인하였다. 항염증 효능을 평가하기 위해 대 식세포와 인간 정상세포에서 산화질소와 사이토카인을 측정한 결과, RPH 스텔라 효과의 농 도를 관찰할 수 있었다. 또한 Western blot을 통해 에너지대사 관련 단백질 (pAMPK-)의 증가와 지질 축적에 관여하는 단백질 (SREBP-1, PPAR-)과 세포사멸 관여 단백질 (casp ase-8, caspase-3, PARP)을 억제하는 단백질을 확인할 수 있었다. 또한 동물실험용 마우스 인 C57BL/6J를 60kcal의 L-Amino Acid Diet with 60 kcal% Fat With 0.1% Methionine a nd No Added Choline (CDHFD)와 함께 고농도 (300mg/ml), 저농도 (100mg/ml)의 RPH로 8주간 경구투여하였다. 실험이 끝나면 혈액과 간 샘플을 수집하고 조직 병리학 및 생화학적 분석을 통해 RPH의 영향을 평가했습니다. 혈액 지표 분석 및 조직 지방 함량 결과, RPH 그룹은 CDHFD 다이어트 그룹에 비해 ALT 및 AST 수치와 지방증을 유의하게 감소시켰 다. 또한, 항산화 효과가 유의하게 나타나는 것을 알 수 있었다. 종합하면, 이러한 결과는 R PH가 NASH 유발 간 손상을 개선하는 데 도움이 될 수 있음을 나타낸다.
more목차 도움말
ABSTRACT ·········································································································································· 7
Ⅰ. 서론 ·················································································································································· 9
Ⅱ. 재료 및 방법 ······························································································································· 12
1. 시약 ············································································································································· 12
2. 시료 추출 ··································································································································· 12
3. 세포배양 및 세포 생장률 평가 ··························································································· 12
3.1 적송심재 에탄올추출물 및 실리마린, 케르세틴의 세포 독성 평가 ····················· 12
3.2 Palmitic acid에 의한 세포 생장율 측정 ····································································· 13
3.3 적송심재 에탄올 추출물 및 실리마린, 케르세틴의 세포 보호 효과 ··················· 13
4. 적송심재 에탄올추출물의 세포 단백질 발현 효과 ························································ 13
5. 적송심재 에탄올 추출물의 항산화 효과 ·········································································· 14
5.1 환원형 glutathion(GSH)함량 측정 ··············································································· 14
1)세포 샘플 ································································································································ 14
2)동물 샘플 ································································································································ 14
3)GSH/GSSG Ratio ···················································································································· 15
5.2 지질과산화물 측정 ··················································································································· 15
6. 적송심재 에탄올추출물의 항염증 효과 ······················································································ 15
6.1 산화질소 측정 ··························································································································· 15
6.2 사이토카인 측정 ······················································································································· 16
7. 적송심재 에탄올추출물(RPH)의 C57BL/6J mouse 비알코올성 지방간 개선 효과 ·············· 16
7.1 실험동물 사육 ··························································································································· 16
7.2 지방간 질환 유발 ····················································································································· 17
7.3 적송심재 에탄올 추출물(RPH)를 이용한 치료 ····································································· 17
7.4 적송심재 에탄올추출물(RPH)의 지방증 감소 효과 ······························································ 17
7.5 적송심재 에탄올추출물(RPH)의 생화학적 분석 ····································································· 18
1) Triglyceride 측정 ··············································································································· 18
2) AST(Aspartate Aminotransferase) 측정 ··································································· 18
3) ALT(Alanine Aminotransferase) 측정 ······································································· 18
목 차
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8. 통계처리 ····································································································································· 19
Ⅲ. 실험 결과 ··········································································································································· 20
1. 적송심재 에탄올추출물(RPH)의 palmitic acid에 의한 인간 정상 간세포 손상에 미치는 영향
···························································································································································· 20
2. 적송심재 에탄올추출물(RPH)의 palmitic aicd에 의한 지방증 및 Apoptosis 억제
효과 ············································································································································· 20
3. 적송심재 에탄올 추출물(RPH)의 항산화 효과 ······························································ 21
4. 적송심재 에탄올 추출물(RPH)의 항염증 효과 ······························································ 21
5. 비알코올성 지방간 질환 유발 마우스에 대한 적송심재 에탄올 추출물(RPH)의 간
개선 효과 ·································································································································· 22
Ⅳ. 고찰 ···················································································································································· 39
Ⅴ. 참고문헌 ········································································································································· 42
