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dc.contributor.advisor김유리-
dc.contributor.author김경미-
dc.creator김경미-
dc.date.accessioned2016-08-26T04:08:41Z-
dc.date.available2016-08-26T04:08:41Z-
dc.date.issued2015-
dc.identifier.otherOAK-000000110981-
dc.identifier.urihttps://dspace.ewha.ac.kr/handle/2015.oak/212219-
dc.identifier.urihttp://dcollection.ewha.ac.kr/jsp/common/DcLoOrgPer.jsp?sItemId=000000110981-
dc.description.abstractInflammatory bowel disease (IBD), including Crohn’s disease and ulcerative colitis, involves chronic inflammation of the gastrointestinal tract. Sasa quelpaertensis leaves extract (SQE) have been shown to mediate anti-inflammatory and anti-carcinogenic effects, although it remains unclear whether Sasa leaves are able to attenuate inflammation-related intestinal diseases. Therefore, the aim of this study is to investigate whether SQE inhibits gut inflammation in vitro and in vivo. At the cellular level, a co-culture consisting of intestinal epithelial Caco-2 cells and RAW 264.7 macrophage cells were stimulated by lipopolysaccharide (LPS) and were induced inflammation. Treatment with SQE significantly suppressed the secretion of LPS-induced nitric oxide (NO), prostaglandin E_(2) (PGE_(2)), interleukin (IL)-6, and IL-1β in co-cultured RAW 264.7 macrophage cells. In addition, expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX-2), and tumor necrosis factor (TNF)-α were down-regulated in response to inhibition of IκBα phosphorylation by SQE. In the dextran sulfate sodium (DSS)-induced colitis mouse model, mice were pretreatment with SQE (100 mg/kg or 300 mg/kg body weight) by gavage for a 2-week period. Mice then received either SQE or sulfasalazine (100 mg/kg body weight) with 2.5% DSS in drinking water for 7 days twice daily and 7 days of tap water ad libitum between DSS treatment. Treatment with SQE was found to attenuate the severity of DSS-induced colitis, as assessed by disease activity index scores, shrinkage of colon length, and histopathologic changes. SQE reduced DSS-induced proliferation in distal colon tissues. It also significantly suppressed levels of TNF-α in serum and colon tissues, iNOS, COX-2, and levels of phosphorylated c-Jun N-terminal kinases, p38, extracellular-signal–regulated kinases 1/2, and IκBα in colon tissues. In conclusion, this study provides the first demonstrate that SQE can represent to ameliorate inflammation-related disease, including IBD by limiting excessive production of pro-inflammatory mediators.;염증성 장질환 (IBD)은 크론병 (Crohn’s Disease)과 궤양성 장질환 (ulcerative colitis, UC)을 포함하는 대장암 발병의 중요한 원인으로 알려져 있다. 이에 본 연구는 제주 조릿대 추출물이 대장의 건강에 미치는 영향을 알아보기 위하여, 조릿대를 이용하여 염증성 장질환 개선에 대하여 세포 실험과 동물실험을 수행하였으며, 염증성 장질환에 관한 조릿대의 항염 효능을 규명 하고자 하였다. 조릿대의 항염 기능성을 규명하기 위한 세포 실험에서는 대장상피세포로 분화시킨 Caco-2 세포와 대식세포인 RAW 264.7 세포를 이용하여 co-culture함으로써 실제 인체 내의 대장과 비슷한 환경을 조성하였고, lipopolysaccharide (LPS) 로 염증을 유도한 후, 조릿대를 처리하여 염증 사이토카인 nitric oxide (NO), prostaglandin E_(2) (PGE_(2)), interleukin (IL)-6, 그리고 IL-1β의 발생을 억제함을 확인하였다. 게다가 nitric oxide synthase (iNOS) 와 cyclooxygenase (COX)-2, tumor necrosis factor (TNF)-α 와 Nuclear factor κB (NF-κB) inhibitor인 IκB의 인산화가 억제됨으로 조릿대의 효과를 밝혀내었다. 조릿대의 항염 기능성 규명을 위한 동물실험에서는 만성염증 장질환의 대표적인 모델인 dextran sulfate sodium (DSS) 모델을 이용하였다. 조릿대 추출물 (100 mg/kg or 300 mg/kg body weight)을 2주간 경구투여 한 후, 2.5% DSS가 포함된 식수를 7일간 공급하며 조릿대 (100 mg/kg or 300 mg/kg body weight)와 항염제 sulfasalazine (100 mg/kg body weight) 를 투여하였고, 다시 7일은 일반 식수, 그 후 다시 7일간 2.5% DSS가 포함된 식수를 공급하는 cycle을 반복하여 만성염증 장질환을 유도하여 관찰하였다. 염증이 유발된 조릿대 투여군에서 임상적 염증척도 (disease activity index)와 대장의 길이, 조직형태학적인 분석에서 DSS염증 유도군에 비해 완화되는 것을 확인 하였다. 세포의 분열과 염증과정에 중요한 Proliferating cell nuclear antigen (PCNA)의 발현이 조릿대 추출물의 처리함으로 억제됨을 확인하였다. 조릿대 추출물 투여에 따른 동물 혈장에서의 염증성 매개 인자인 TNF-α와 대장조직에서의 TNF-α mRNA 발현이 억제 되었으며, iNOS, COX-2의 단백질 발현과 extracellular-signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38, IκBα 의 phpsphorylation 이 유의적으로 감소함을 밝혀내었다. 이와 같이 조릿대 추출물은 만성 대장염뿐만 아니라 다른 염증유도 만성질병에도 활용 될 수 있는 의미 있는 연구로 항염증 치료법 개발에 유용하게 활용될 수 있을 것이다.-
dc.description.tableofcontentsI.Introduction 1 A.Literature Review 1 B.Hypothesis 5 II.Material and Methods 6 A.Preparation of SQE and HPLC analysis 6 B.Cell culture 9 C.Cell viability 9 D.Co-culture system 10 E.Assay for NO production 12 F.Measurements of cytokine secretion form RAW 264.7 macrophages 12 G.Western Blot analysis 12 H.Reverse transcription-PCR and quantitative RT-PCR 13 I.Mice groups and induction of chronic colitis 14 J.Serologic assessment of liver toxicity 17 K.Disease activity index 17 L.Macroscopic and histopathologic evaluations 19 M.Immunohistochemistry for proliferation cell nuclear antigen 21 N.Measurement of TNF-α production in mouse serum 22 O.Statistical Analysis 22 III.Results 23 Part 1. SQE inhibits inflammation in co-cultured cells 23 A.SQE treatment did not affect the viability of RAW 264.7 macrophages 24 B.SQE, tricin, p-coumaric acid down-regulated the expression of cytokines in a co-culture system of Caco-2 and RAW 264.7 macrophages 26 C.SQE, tricin, and p-coumaric acid down-regulated expression of iNOS and COX-2 in LPS-activated and co-cultured Caco-2 and RAW 264.7 macrophages 30 D.SQE, tricin, and p-coumaric acid reduced IκBα phosphorylation and inhibited expression of TNF-α 32 Part 2. SQE suppresses DSS-induced colitis in mice 36 E.SQE does not induce hepatotoxicity in mice 37 F.SQE attenuates the progression of DSS-induced chronic colitis in mice 39 G.SQE treatment reduces histologic damage associated with colitis 41 H.SQE supplementation reduces colon cell proliferation in a DSS-induced chronic colitis model 43 I.SQE supplementation attenuates the expression of inflammatory mediators, including TNF-α, iNOS, and COX-2 45 J.SQE supplementation inhibits MAPK phosphorylation and NF-κB activation in DSS-induced chronic colitis 47 IV.Discussion 49 V.Conclusion 56 References 57 국문초록 63-
dc.formatapplication/pdf-
dc.format.extent1423967 bytes-
dc.languageeng-
dc.publisher이화여자대학교 대학원-
dc.subject.ddc600-
dc.titleThe effect of Sasa quelpaertensis leaf extract on chronic inflammatory bowel disease in vitro and in vivo-
dc.typeMaster's Thesis-
dc.title.translated조릿대의 만성염증 장질환에서의 건강기능성 및 항염기능과 기전규명-
dc.creator.othernameKim, Kyung Mi-
dc.format.pageviii, 67 p.-
dc.contributor.examiner고광석-
dc.contributor.examiner김정현-
dc.contributor.examiner김유리-
dc.identifier.thesisdegreeMaster-
dc.identifier.major대학원 식품영양학과-
dc.date.awarded2015. 2-
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