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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 黃青真 | |
dc.contributor.author | Li-Hui Wang | en |
dc.contributor.author | 王麗卉 | zh_TW |
dc.date.accessioned | 2021-05-16T16:18:55Z | - |
dc.date.available | 2015-08-28 | |
dc.date.available | 2021-05-16T16:18:55Z | - |
dc.date.copyright | 2013-08-28 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-08-13 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/5971 | - |
dc.description.abstract | 共軛脂肪酸為一群特殊脂肪酸。共軛亞麻油酸 (conjugated linoleic acid,CLA) 混合物 (9c,11t-CLA與10t,12c-CLA ) 或10t,12c-CLA可活化PPARα、降低體脂肪,但造成脂肪肝及胰島素抗性。本實驗室曾發現自山苦瓜分離之共軛次亞麻油酸9c,11t,13t-conjugated linolenic acid (CLN) 可活化PPAR α及PPAR γ。 本研究目的為探討CLA及CLN對小鼠脂質代謝之影響 (in vivo)。
C57BL/6J 雌鼠以實驗飼料分成四組並飼養五週: 基礎飼料 (B組,控制組)、1 % CLA 混合物 (9c,11t-CLA: 10t,12c-CLA = 1:1) (A組)、 1% crude 9c,11t,13t-CLN (純度76 %) (N組) 或1% pure 9c,11t,13t-CLN (純度99 %) (H組)。 與B組小鼠相比,A組肝臟重量增加、肝臟TG及CHOL含量較高 (p < 0.05),但腎周圍脂肪組織(RWAT) TG含量顯著較低。N組與H組RWAT重量較大,但只有N組RWAT之TG含量較高 (p < 0.05)。 H組肝臟TG濃度及含量皆顯著低於B組 (p < 0.05)。新陳代謝儀分析顯示H組具較高呼吸商 (RQ)。N組禁食血清之葡萄糖、TG濃度顯著較高 (p < 0.05)。 A組非禁食血清胰島素濃度及禁食血清CHOL濃度顯著較高,但禁食血清TG濃度顯著較低 (p < 0.05)。 肝臟、RWAT、血清、肌肉以GC-FID分析脂肪酸組成。於四種組織中N、H組皆測得9c,11t,13t-CLN及 9c,11t-CLA,A組皆測得9c,11t-CLA 及10t,12c-CLA。與B組相較,A組肝臟MUFA含量較高、n-6 PUFA含量較低。A組肝臟delta-5-desaturase (D5D)、delta-6-desaturase (D6D) mRNA表現量與D5D index (C20:4 n-6/C20:3n-6)、D6D index (C18:3n-6/C18:2n-6)皆顯著低於B組 (p < 0.05)。 N組肝臟SCD1 (delta-9-desaturase, D9D) mRNA表現量顯著較低 (p < 0.05),但D9D index (C16:1n-7/C16:0)則否。四組小鼠肝臟組織中,與脂質代謝相關基因之表現量皆無顯著差異。 肝、血清、RWAT、尿液經LC-MS分析並預測代謝物。經標準品確認A、H組Phe於肝臟含量降低、於血清含量較高 (p < 0.05)。 本實驗結果顯示,CLA混合物對脂肪組織重量無顯著差異,但造成脂肪肝;CLN增加脂肪組織重量,但無肝臟脂質堆積。CLA混合物降低肝臟D5D及D6D酵素活性,但CLN則否。高純度CLN能降低肝脂並提高RQ,低純度則否。兩組CLN餵食結果之差異,究與劑量或低純度CLN樣品含有其他活性成分有關,尚待進一步研究。 | zh_TW |
dc.description.abstract | Conjugated linoleic acid (CLA) (a mixture of 9c,11t-CLA and 10t,12c-CLA or 10t,12c-CLA) were shown to activate peroxisome proliferator activated receptor (PPAR) alpha decrease body fat mass (BFM), but lead to fatty liver and insulin resistance. Previous study of our Lab demonstrated that 9c,11t,13t-conjugated linolenic acid (CLN) isolated from bitter gourd activated both PPAR alpha and gamma. This study aimed to examine effects of CLA and CLN on the lipid metabolism (in vivo).
Four groups of C57BL/6J female mice were respectively fed: a basal diet (control group, B group), basal diet supplemented with 1% CLA mixture (9c,11t-CLA: 10t,12c-CLA = 1:1) (A group), 1% crude CLN (76 % pure) (N group) or 1% pure CLN (99 % purity) (H group) for 5 weeks. Compared to the B group, the A group of mice had significantly higher liver weight and its TG and cholesterol content, but lower triglyceride (TG) concentration in the perirenal white adipose tissue (RWAT) (p < 0.05). The N and H groups had higher adipose weight but only the N group had higher RWAT TG content (p < 0.05). The H group showed significantly lower TG concentration and content in the liver (p < 0.05). Indirect calorimetry analysis showed that the H group had significantly higher Respiratory quotient (p < 0.05). The N group had higher fasting serum glucose and TG, while the A group had higher non-fasting serum insulin and fasting serum cholesterol, but lower fasting serum TG (p < 0.05). Fatty acid compositions in liver, RWAT, serum and muscle were analyzed by GC-FID. Both 9c,11t,13t-CLN and 9c,11t-CLA were detected in serum and all 3 tissues of the H and N groups, while both 9c,11t-CLA and 10t,12c-CLA were detected in the A group. Compared to the B group, the A group had higher MUFA and lower n-6 PUFA in the liver, while the N group had lower n-6 PUFA in the liver. The hepatic mRNA expressions of delta-5-desaturase (D5D), delta-6-desaturase (D6D), fatty acids D5D index (C20:4 n-6/C20:3n-6) and D6D index (C18:3n-6/C18:2n-6) were significantly lower in the A group (p < 0.05). The N group had lower SCD1 (delta-9-desaturase, D9D) hepatic mRNA (p < 0.05) but not lower D9D index (C16:1n-7/C16:0). No significant differences were observed in the mRNA of remaining lipid metabolism related genes in the liver among the 4 groups. Metabolites in liver, serum, RWAT, and urine of the 4 groups of mice were analyzed by using LC-MS and the metabolomics approach. Confirmed by standard compounds, phenylalanine was lower in the liver, but higher in the serum in both of A and H groups (p < 0.05). In this study, the CLA mixture led to fatty liver without a reduction in the adipose mass. CLN increased adipose mass but not liver lipids. The CLA mixture also decreased liver D5D and D6D desaturation of fatty acids, but not CLN. Reduced liver lipids and elevated respiration quotient were only observed in mice fed pure CLN. Further study is needed to elucidate whether the dose or other constituents in the crude CLN sample might contribute to the differences between the crude and pure CLN fed mice. | en |
dc.description.provenance | Made available in DSpace on 2021-05-16T16:18:55Z (GMT). No. of bitstreams: 1 ntu-102-R00b22039-1.pdf: 4178159 bytes, checksum: c0915beae160e5c34f5ee44ce9418043 (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 中文摘要---------------------------------------------------------------------------------------- i
Abstract------------------------------------------------------------------------------------------ iii 第一章 緒論-----------------------------------------------------------------------------------1 第一節 研究動機與目的-----------------------------------------------------------------1 第二節 文獻回顧--------------------------------------------------------------------------1 一、肥胖--------------------------------------------------------------------------------1 二、糖尿病-----------------------------------------------------------------------------2 三、肝臟之醣類調控及脂質堆積-------------------------------------------------3 四、PPARs------------------------------------------------------------------------------4 五、共軛脂肪酸-----------------------------------------------------------------------6 (一) 共軛亞麻油酸(CLA) ------------------------------------------------------6 1.介紹-------------------------------------------------------------------------6 2. CLA抗肥胖 (antiobesity) ----------------------------------------------8 (1) 降低攝食量----------------------------------------------------------8 (2) 增加能量消耗-------------------------------------------------------8 (3) 抑制脂肪細胞生合成----------------------------------------------8 3. 促進發炎反應------------------------------------------------------------10 4. 調控脂質代謝------------------------------------------------------------11 (1) 抑制脂質合成 (lipogenesis) --------------------------------------11 (2) 促進脂肪細胞三酸甘油酯分解----------------------------------11 5. 造成胰島素抗性---------------------------------------------------------12 6. 促進 (pre)adipocyte凋亡(apoptosis) --------------------------------13 (二) 共軛次亞麻油酸(CLN) ---------------------------------------------------14 1.介紹-------------------------------------------------------------------------14 2. 共軛次亞麻油酸之功效------------------------------------------------15 3. 共軛次亞麻油酸 (CLN) vs共軛亞麻油酸 (CLA) ---------------15 4. 苦瓜中共軛次亞麻油酸之相關研究---------------------------------16 六、代謝體學--------------------------------------------------------------------------16 七、CLA及CLN文獻整理----------------------------------------------------------18 第三節 實驗假說與設計-----------------------------------------------------------------29 一、實驗假說-------------------------------------------------------------------------29 二、研究內容--------------------------------------------------------------------------29 第二章 前言-----------------------------------------------------------------------------------30 第三章 材料與方法--------------------------------------------------------------------------31 一、實驗大綱--------------------------------------------------------------------------31 二、動物飼養--------------------------------------------------------------------------32 三、飼料--------------------------------------------------------------------------------32 (一) Basal diet----------------------------------------------------------------------32 (二) CLA diet-----------------------------------------------------------------------32 (三) cis-9, trans-11, trans-13 conjugated linolenic acid (Crude, 76 % purity)--------------------------------------------------------------------------------32 (四) cis-9, trans-11, trans-13 conjugated linolenic acid (high purity CLN, 99 % purity)-----------------------------------------------------------------32 四、糞便、尿液收集----------------------------------------------------------------35 五、氧氣消耗量、二氧化碳排出量、呼吸商分析----------------------------35 六、飽食血清收集-------------------------------------------------------------------35 七、動物犧牲-------------------------------------------------------------------------35 (一) 器具準備---------------------------------------------------------------------35 (二) 實驗流程---------------------------------------------------------------------36 七、血醣濃度分析-------------------------------------------------------------------36 八、胰島素濃度分析----------------------------------------------------------------36 九、Folch method組織萃取--------------------------------------------------------37 (一) 內標配製---------------------------------------------------------------------37 (二) 萃取前器具處理------------------------------------------------------------37 (三) 肝臟、腎周圍脂肪組織、肌肉、血清萃取---------------------------37 (四) 水層再萃取 (去除蛋白) --------------------------------------------------38 十、三酸甘油酯含量分析----------------------------------------------------------38 (一) 血清 (犧牲當天操作) -----------------------------------------------------38 (二) 組織---------------------------------------------------------------------------39 十一、膽固醇含量分析-------------------------------------------------------------39 (一) 血清 (犧牲當天操作) -----------------------------------------------------39 (二) 組織---------------------------------------------------------------------------39 十二、GC-FID分析組織脂肪酸組成--------------------------------------------40 (一) 無水甲醇製備--------------------------------------------------------------40 (二) 組織有機層萃取液甲基酯化---------------------------------------------40 (三) GC-FID脂肪酸分析--------------------------------------------------------40 十三、尿液creatinine定量--------------------------------------------------------41 十四、LC-MS分析組織代謝物---------------------------------------------------41 十五、RNA抽取---------------------------------------------------------------------42 十六、RNA轉錄成cDNA---------------------------------------------------------42 十七、Real-time qPCR分析PPARs及其下游基因表現---------------------43 十八、統計分析----------------------------------------------------------------------43 第四章 實驗結果-----------------------------------------------------------------------------44 一、體重變化、攝食量、攝食利用率、能量攝取及能量利用率----------44 二、器官及組織絕對重量與相對重量-------------------------------------------44 三、氧氣消耗量及二氧化碳排出量分析----------------------------------------45 四、血清分析結果-------------------------------------------------------------------45 (一) 血清葡萄糖及胰島素分析------------------------------------------------45 (二) 血清三酸甘油酯及膽固醇分析------------------------------------------46 五、組織三酸甘油酯及膽固醇含量分析----------------------------------------46 (一) 肝臟組織---------------------------------------------------------------------46 (二) 腎臟週圍脂肪組織---------------------------------------------------------46 (三) 肌肉組織---------------------------------------------------------------------46 六、 組織脂肪酸組成--------------------------------------------------------------47 (一) 肝臟脂肪酸組成(%)--------------------------------------------------------47 (二) 腎周圍脂肪組織 (RWAT) 脂肪酸組成-------------------------------47 (三) 血清脂肪酸組成------------------------------------------------------------48 (四) 肌肉脂肪酸組成------------------------------------------------------------49 七、 肝臟real-time PCR基因表現----------------------------------------------50 八、 子宮周圍脂肪 (UWAT) 組織real-time PCR基因表現--------------50 九、組織代謝物與B組具顯著差異之代謝物/代謝途徑預測--------------50 (一) 肝臟組織代謝物預測------------------------------------------------------50 (二) 腎周圍脂肪組織代謝物預測---------------------------------------------51 (三) 血清代謝物預測------------------------------------------------------------51 (四) 尿液代謝物預測------------------------------------------------------------51 十、 A、N、H組與B組具顯著差異之組織代謝途徑預測---------------51 (一) 肝臟組織代謝途徑預測---------------------------------------------------51 (二) 腎週圍脂肪組織代謝途徑預測------------------------------------------52 (三) 血清代謝途徑預測---------------------------------------------------------52 (四) 尿液代謝途徑預測---------------------------------------------------------52 第五章 討論-----------------------------------------------------------------------------------87 一、共軛脂肪酸影響組織脂質含量----------------------------------------------87 (一) 10t,12c-CLA造成肝臟脂質堆積------------------------------------------87 (二) 9c,11t,13t-CLN未造成肝臟脂質堆積-----------------------------------88 二、CLA與胰島素抗性------------------------------------------------------------88 三、高純度c9t11t13 CLN增高呼吸商 (RQ) ----------------------------------89 四、共軛脂肪酸改變組織脂肪酸組成-------------------------------------------89 (一) 9c,11t,13t-CLN轉變成c9t11 CLA---------------------------------------89 (二) 共軛脂肪酸影響脂肪酸代謝---------------------------------------------90 (三) t10c12 CLA活化PPARα------------------------------------------------91 (四) 共軛脂肪酸之代謝---------------------------------------------------------91 1. 9c,11t-CLA與10t,12c-CLA代謝--------------------------------------91 2. 共軛脂肪酸藉由競爭抑制脂肪酸合成------------------------------92 3. 共軛脂肪酸調控desaturase活性及表現量-------------------------92 五、共軛脂肪酸對肝臟及脂肪組織基因之調控-------------------------------93 六、代謝體學結果分析-------------------------------------------------------------95 第六章 結論-----------------------------------------------------------------------------------97 第七章 參考文獻-----------------------------------------------------------------------------98 | |
dc.language.iso | zh-TW | |
dc.title | 共軛脂肪酸對 C57BL/6J 雌鼠脂質代謝之影響 | zh_TW |
dc.title | Effects of conjugated fatty acid on lipid metabolism in C57BL/6J female mice | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 蘇慧敏,林璧鳳,呂紹俊,趙蓓敏 | |
dc.subject.keyword | 共軛亞麻油酸(CLA),共軛次亞麻油酸(CLN),脂肪酸組成,脂質代謝, | zh_TW |
dc.subject.keyword | conjugated linoleic acid (CLA),conjugated linolenic acid (CLN),fatty acid composition,lipid metabolism, | en |
dc.relation.page | 105 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2013-08-13 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科技學系 | zh_TW |
顯示於系所單位: | 生化科技學系 |
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