Chlorella vulgaris 가 장내 균총 변화 및 면역 조절에 미치는 영향
- 발행기관 강릉원주대학교 일반대학원
- 발행년도 2021
- 학위수여년월 2021. 8
- 학위명 석사
- 학과 및 전공 도움말 일반대학원 KIST강릉분원학.연협동과정
- 실제URI http://www.dcollection.net/handler/kangnung/000000010940
- UCI I804:42001-000000010940
- 본문언어 한국어
초록/요약 도움말
장내 균총은 전체 60% 항원을 제시할 만큼 면역시스템에 밀접한 영향을 미치고 자가면역질환 예방의 주요 인자로 알려지면서, 최근 장내 균총에 유익한 영향을 미치는 물질을 이용한 면역질환 치료제 연구가 대두되고 있다. 클로렐라 불가리스(Chlorella vulgaris)는 단세포 녹조류로 단백질, 지방, 식이섬유, 무기염류, 다당류, 엽록소, Carotenoid와 같은 장내 균총에 유익한 성분을 풍부하게 함유하고 있다. 그러나 클로렐라 불가리스가 장내 균총 및 면역 조절에 미치는 영향에 관한 연구는 미흡한 수준이다. 따라서 본 연구에서는 클로렐라 불가리스가 장내 균총의 변화 및 면역 조절에 미치는 영향을 평가하였으며, DSS(dextran sulfate sodium)로 유도된 염증성 장 질환(Inflammatory Bowel Disease) 모델에서 클로렐라 불가리스의 예방효과를 조사하였다. 실험군은 정상군(CON), 클로렐라 불가리스 열수 추출물 경구 투여군(CV), DSS 대조군(DSS), 클로렐라 열수 추출물 경구 투여 후 DSS 처리군 (DSS+CV)까지 4군으로 구성하였다. 정상군(CON)은 일반 식수를 투여하고, 클로렐라 불가리스 열수 추출물 투여군(CV, DSS+CV)에는 매일 2mg/g 용량으로 3주간 마우스에 경구 투여한 후, 면역 조절을 평가하기 위해 비장과 장간막 림프절의 면역세포를 유세포 분석기로 분석하였다. 실험 결과, CV 군에서 정상군과 비교하면 항염증 세포인 조절 T세포의 발현량이 유의성 있게 증가한 것을 관찰하였다. 마우스의 맹장에서 DNA를 추출하여 16S rRNA gene sequencing을 수행하여 미생물 군집을 분석한 결과, CV군에서 정상군과 비교하여 장내 균총의 다양성 및 Bifidobacterium과 Lactobacilli와 같은 프로바이오틱스 균이 증가함을 확인하였다. Gas chromatography(GC-FID)로 장내 균총의 대사 산물인 단쇄지방산(SCFA)을 분석한 결과, CV 군에서 정상군과 비교하여 acetate와 propionate 생산이 증가함을 확인하였다. 다음으로 염증성 장 질환을 유발하기 위해 DSS 대조군(DSS)과 클로렐라 열수 추출물 경구 투여 후 DSS 처리군(DSS+CV)에 DSS를 2.5%로 7일간 투여하였다. DSS 공급 후 체중과 대장 길이의 감소가 현저하였으나, DSS+CV 군은 DSS 군에 비교하여 체중과 대장 길이의 감소가 적었다. 또한, 대장 염증 지표인 DAI 점수와 조직 병리학 평가에서 DSS+CV 군이 DSS 군과 비교하여 현저히 낮은 점수를 나타냈으며, 항염증 기능을 갖는 조절 T세포의 빈도와 개체 증가가 관찰되었다. 이상의 결과로 볼 때, 클로렐라 불가리스 추출물은 장내 균총 다양성 및 프로바이오틱스 균의 증가를 유도하고, 조절 T세포 증가를 촉진하여 염증반응 억제 효과를 나타냄을 알수 있었다.
more목차 도움말
Abstract··························································································06
List of Figures················································································08
List of Tables·················································································10
국문 초록·························································································11
Ⅰ. 서론(Introduction)······································································13
II. 재료 및 방법 (Materials & Methods)············································18
2.1. 시료·····················································································18
2.2. 실험동물 및 동물군의 분류·····················································18
2.3. 염증성 장 질환 평가······························································18
2.4. 표본 채취 ············································································19
2.5. 대장점막의 조직병리학 분석····················································20
2.6. 세포 분리·············································································22
2.7. 유세포 분석기·······································································22
2.8. DNA추출··············································································22
2.9. 16S rRNA gene sequencing·················································23
2.10. GC-FID분석········································································23
2.11 세포배양··············································································24
2.12. 상피전위 전기저항(transepithelial electrical resistance, TEER)측정····················································································24
2.13 통계분석··············································································25
Ⅲ. 결과(Results)··············································································26
3.1. 클로렐라 불가리스 경구 투여 마우스의 조절 T세포의 발현 평가·····························································································26
3.2. 클로렐라 불가리스 경구 투여 마우스의 장내 균총의 다양성과 군집 평가·····················································································29
3.3. 클로렐라 불가리스 경구 투여 마우스의 분변 내 장내 균총의 상대적인 차이 분포 평가······························································31
3.4. 클로렐라 불가리스 경구 투여 마우스의 분변 내 문 수준에서 장내 균총 분류············································································33
3.5. 클로렐라 불가리스 경구 투여 마우스의 분변 내 종 수준에서 장내 균총 분류············································································35
3.6. 클로렐라 불가리스 경구 투여 마우스의 분변 내 단쇄지방산 평가························································································38
3.7. DSS 2.5% 유도 클로렐라 불가리스 경구 투여 마우스의 병리학적 염증성 장 질환 평가······························································41
3.8. DSS 2.5% 유도 클로렐라 불가리스 경구 투여 마우스의 비장과 맹장의 무게 평가······································································44
3.9. DSS 2.5% 유도 클로렐라 불가리스 경구 투여 마우스의 조직 병리학 평가··········································································47
3.10. DSS 2.5% 유도 클로렐라 불가리스 경구 투여 마우스의 조절 T세포의 발현 평가······································································51
3.11. DSS 1% 유도 클로렐라 불가리스 경구 투여 마우스의 병리학적 염증성 장 질환 평가 ·····························································55
3.12. DSS 1% 유도 클로렐라 불가리스 경구 투여 마우스의 비장과 림프노드 무게 평가···································································59
3.13. 분화된 장상피세포에서 클로렐라 불가리스의 염증 억제 평가································································································63
Ⅳ. 결론 및 고찰(Conclusion and Discussion)···································66
Ⅴ. 참고 문헌(References)································································71
