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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林璧鳳 | |
| dc.contributor.author | Hsueh-Yun Lu | en |
| dc.contributor.author | 呂學耘 | zh_TW |
| dc.date.accessioned | 2021-05-20T21:44:18Z | - |
| dc.date.available | 2015-08-19 | |
| dc.date.available | 2021-05-20T21:44:18Z | - |
| dc.date.copyright | 2010-08-19 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-08-09 | |
| dc.identifier.citation | &吳繼恆（2007）具影響干擾素γ分泌的食材對OVA致敏BALB/c小鼠免疫反應的影響 國立臺灣大學微生物與生化學研究所 碩士論文
洪永瀚（2008）苜蓿芽乙酸乙酯萃取物改善MRL-lpr/lpr自體免疫鼠病程發展之探討 國立臺灣大學微生物與生化學研究所 博士論文 莊佳穎（2004）山苦瓜活化過氧化體增殖劑活化受器 PPARs 之成分分離與鑑定 國立臺灣大學微生物與生化學研究所 碩士論文 陳妙齡（2006）松杉靈芝對過敏免疫反應之調節研究 國立臺灣大學微生物與生化學研究所 博士論文 陳永如（2007）數種植物雌激素食材萃物對PPAR、脂肪細胞生成及雌激素活性特質鑑定 國立臺灣大學微生物與生化學研究所 碩士論文 康智雄（2009）影響3T3-L1細胞株分泌細胞激素之食材對高油飲食小鼠的免疫調節作用 國立臺灣大學微生物與生化學研究所 碩士論文 羅文音（2006）以卵蛋白致敏小鼠模式探討攝食山藥對過敏性氣喘的影響 國立臺灣大學微生物與生化學研究所 碩士論文 Agrawal, A., Lingappa, J., Leppla, S. H., Agrawal, S., Jabbar, A., Quinn, C., et al. (2003). Impairment of dendritic cells and adaptive immunity by anthrax lethal toxin. Nature, 424(6946), 329-334. Agrawal, S., Agrawal, A., Doughty, B., Gerwitz, A., Blenis, J., Van Dyke, T., et al. (2003). Cutting edge: different Toll-like receptor agonists instruct dendritic cells to induce distinct Th responses via differential modulation of extracellular signal-regulated kinase-mitogen-activated protein kinase and c-Fos. J Immunol, 171(10), 4984-4989. Akbari, O., Freeman, G. J., Meyer, E. H., Greenfield, E. A., Chang, T. T., Sharpe, A. 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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/10619 | - |
| dc.description.abstract | 已有研究指出山苦瓜的降血糖功效與其乙酸乙酯萃物含共軛次亞麻油酸 (9c, 11t, 13t-conjugated linolenic acid, CLN)活性成份，但對於免疫調節的影響，尚未有研究發表。本研究首先以細胞培養探討花蓮四號山苦瓜正己烷(BGP-Hex)、乙酸乙酯(BGP-EA)、酒精(BGP-EtOH)與水萃物(BGP-H2O)和CLN，對巨噬細胞株RAW264.7、小鼠T細胞株EL4、小鼠初代細胞分泌細胞激素的作用。結果發現BGP-EA、BGP-Hex和BGP-EtOH能顯著的降低RAW264.7的TNFα及IL-6分泌，也顯著的抑制初代腹腔細胞IL-6的分泌。CLN能降低初代腹腔細胞TNFα及IL-6的分泌，也能顯著減少小鼠初代脾臟細胞IFNγ與IL-4。在體外實驗初步觀察到山苦瓜萃物與CLN具有抗發炎的潛力及調節免疫的效果。本研究接續探討山苦瓜與所含的CLN對氣喘模式小鼠發炎與過敏免疫反應的影響，將 BALB/c 小鼠8週大時以卵蛋白（ovalbumin, OVA）加佐劑氫氧化鋁，隔週腹腔致敏以誘發過敏免疫傾向。三次致敏之後，將以小鼠分為五組，分別為OVA-oil控制組 、飼料含5%山苦瓜凍乾粉的OVA-BGP組、每天管餵35 mg共軛次亞麻油酸OVA-CLN組、每天管餵35 mg共軛亞麻油酸 (conjugated linoleic acid, CLA) 的OVA-CLA組、和固醇類藥物prednisolone正對照OVA-Pred組 ，另有一組為未致敏的PBS-oil組。餵食期間以5%霧化之OVA給予致敏小鼠呼吸道刺激，誘發過敏性呼吸道發炎，而PBS-oil組給予霧化PBS。小鼠餵食13天後，以methacholine (Mch)刺激呼吸道，測定呼吸道過度反應(airway hyperresponsiveness, AHR)時的呼吸阻力。小鼠餵食16天後犧牲，採血液、肺氣管沖洗液、分離脾臟及腹腔細胞、取肺組織進行切片觀察與抽取RNA分析基因表現。結果顯示，OVA-BGP組與OVA-CLN組在Mch各濃度下呼吸道阻力係數Penh值顯著低於OVA-oil控制組，OVA-CLA組在大於25 mg/mL Mch刺激下也顯著較低，表示山苦瓜、CLN和CLA可減緩過敏性氣喘。與OVA-oil控制組相較， OVA-BGP組顯著減少肺氣管沖洗液中的總細胞、嗜伊紅性白血球及嗜中性白血球的數目、也降低Th2細胞激素IL-4、IL-5、IL-13與促發炎介質IL-6、PGE2和eotaxin含量，肺組織中的COX-2 mRNA表現也顯著較低，且顯著的回復肺組織中的PPARα基因表現量，顯示山苦瓜抑制發炎反應與Th2免疫反應。並且，OVA-BGP組的脾臟細胞在ConA刺激之下，IFN-γ分泌量顯著高於OVA-oil組，在特異性OVA刺激下的脾臟細胞IL-13分泌量顯著較低，血清總IgE生成量也顯著較低，顯示山苦瓜對調節傾免疫向Th1反應。 OVA-CLN組則是顯著降低肺氣管沖洗液中嗜伊紅性白血球的數目和IL-5含量，回復肺組織PPARγ mRNA的表現量，顯示CLN的抗發炎作用。而OVA-CLA組顯著降低脾臟細胞在ConA刺激之下IL-13的分泌，肺組織IL-12與PPARα mRNA表現量也顯著較高，故也測得顯著低的血清總IgE生成量，顯示CLA具調節免疫傾向Th1的作用。因此，由誘發致敏小鼠產生過敏性氣喘的動物模式顯示，山苦瓜具抑制過敏的免疫調節與抗發炎作用，所含成份的CLN則以降低呼吸道發炎作用較顯著，山苦瓜能減少過敏性抗體IgE的生成，應有助於減緩過敏免疫反應，值得推廣。 | zh_TW |
| dc.description.abstract | Studies show bitter gourd (Momordica charantia) exerts hypoglycemic effect and 9c, 11t, 13t-conjugated linolenic acid (CLN) from its ethyl acetate extract is the bioactive compounds. Only few reports on immunomodulatory effect of bitter guard have been published. First, the effect of hexane extract (BGP-Hex), ethyl acetate extract (BGP-EA), ethanol extract (BGP-EtOH), water extract (BGP-H2O) and CLN on cytokines secretion by RAW264.7 cell, EL4 cell and primary peritoneal cells or splenocytes from BALB/c mice was investigated. The results showed that BGP-EA, BGP-Hex and BGP-EtOH significantly inhibited TNFα and interleukin (IL)-6 secretion in LPS-stimulated RAW264.7. Furthermore, BGP-EA and BGP-EtOH significantly reduced IL-6 secretion and CLN could decrease the TNFα and IL-6 production in LPS-stimulated primary peritoneal cells. Treatment with CLN inhibited IFNγ and IL-4 secretion in ConA-stimulated splenocytes. Therefore, bitter gourd and CLN may have an anti-inflammatory and immunoregulatory effect. Subsequently, we investigated effect of bitter gourd and 9c, 11t, 13t-CLN may on airway inflammation and immune response using a murine model of allergic asthma. Eight-week-old BALB/c mice sensitized intraperitoneally and challenged with ovalbumin (OVA) were fed a diet either supplemented with 5 % (wt/diet) bitter gourd powder (OVA-BGP group), or tube-feeding 35 mg/day CLN (OVA-CLN group), 35 mg/day 9c, 11t-conjugated linoleic acid (CLA) (OVA-CLA group), or oil as control (OVA-oil group) for 16 days. The OVA-sensitized mice treated with prednisolone were the positive control (OVA-Pred group). Mice without sensitization were the negative control (PBS-oil group). The hyperresponsiveness (AHR) challenged by methacholine (Mch), airway inflammation, serum antibody levels, cytokine secretions from splenocytes and lung mRNA expression were measured. BGP, CLN and CLA supplementation significantly suppressed the AHR, suggesting the alleviative effects on allergy asthma. Compared to the OVA-oil group, the OVA-BGP group had significantly lower total cells, eosinophil and neutrophil infiltration and Th2 cytokines IL-4, IL-5 and IL-13, and inflammatory mediators IL-6, PGE2 and eotaxin in bronchoalveolar lavage fluid (BALF), and pulmonary COX-2 mRNA expression, suggesting anti-inflammatory effect of BGP. The OVA-BGP group also had lower OVA-specific IL-13 secretion and IFN-γ secretion from OVA- or ConA-stimulated splenocytes, and lower serum total immunoglobulin (Ig) E level, suggesting immunomodulatory effect of BGP. The OVA-CLN group showed significantly lower eosinophil infiltration and IL-5 in BALF, restored PPARγ mRNA expression that was down-regulated by sensitization, suggesting anti-inflammatory effect of CLN. The CLA group had significantly lower IL-13 secretion from Con-A stimulated splenocytes, higher pulmonary IL-12 and PPARα mRNA expression, lower total IgE level in serum, suggesting the immunoregulatory effect of CLA toward Th1 responses. In conclusion, Bitter gourd possessed both anti-inflammatory and immunoregulatory effects, while CLN exerted mainly reduced airway inflammation in a murine model of allergic asthma. Our study suggests that bitter guard suppressing allergic immune responses including serum IgE level may be beneficial for alleviation of allergic responses and worthy of dietary promotion. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-20T21:44:18Z (GMT). No. of bitstreams: 1 ntu-99-R97b47308-1.pdf: 13272176 bytes, checksum: 1670c82e0810ac6bd967c5a80b0d52b5 (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | 中文摘要 Ⅰ
英文摘要 Ⅲ 目錄 Ⅴ 表目錄 Ⅸ 圖目錄 Ⅹ 第一章、 序論 第一節 文獻回顧 1 一、過敏性氣喘 1 （一）簡介與定義 1 （二）過敏性氣喘發生機制 2 （三）樹突細胞與過敏性氣喘 4 （四）T輔助細胞與過敏性氣喘 4 二、PPAR 7 （一）PPAR之種類及分佈 7 （二）PPAR活化劑與 ligands 7 （三）PPAR與發炎反應之相關 8 三、苦瓜 11 （一）苦瓜之介紹 11 （二）山苦瓜之相關研究 11 四、共軛脂肪酸 12 （一）共軛次亞麻酸之介紹與相關研究 12 （二）共軛亞麻油酸之介紹與相關研究 13 第二節 研究動機與架構 一、研究動機 15 二、研究架構 16 第二章、以體外模式評估山苦瓜與共軛次亞麻油酸對發炎與免疫反應的影響 第一節 前言 17 第二節 材料與方法 18 一、山苦瓜萃取物製備 18 二、CLN與CLA樣品製備 19 三、RAW 264.7 小鼠巨噬細胞株 19 四、EL4 小鼠T細胞株 19 五、腹腔細胞的取得與培養 20 六、脾臟細胞之取得與培養 21 七、樹突細胞之取得與培養 22 八、細胞激素含量測定 23 九、MTT染色法測定細胞存活率 26 十、統計方法 26 第三節 結果 27 一、山苦瓜各種萃物與CLN對RAW264.7小鼠巨噬細胞株的影響 27 二、山苦瓜各種萃物與CLN對初代腹腔細胞的影響 27 三、山苦瓜各種萃物與CLN對EL4小鼠T細胞株的影響 27 四、山苦瓜各種萃物與CLN對初代脾臟細胞的影響. 28 五、山苦瓜各種萃物與CLN對骨髓細胞衍生之樹突細胞的影響 29 第四節 討論 37 一、山苦瓜與CLN可能藉由活化PPARα與PPARγ而具有抗發炎的效果 37 二、各樣品的免疫調節作用 38 三、山苦瓜萃物中的CLN 38 第三章、山苦瓜與共軛三烯次亞麻油酸對氣喘模式小鼠發炎與過敏免疫反應的影響 第一節 前言 40 第二節 材料與方法 41 一、樣品配製 41 二、動物實驗設計 41 （一）動物飼養 41 （二）致敏方法 41 （三）實驗流程及分組 42 （四）呼吸道過度反應測定 44 三、動物犧牲及實驗材料收集與分析 44 （ㄧ）血液樣品的收集 44 （二）血清特異性抗體IgE、IgG1、IgG2a測定 45 （三）血清總IgE、IgG、IgA、IgM抗體含量測定 46 （四）肺氣管沖洗液細胞之取得與處理 47 （五）脾臟細胞之取得與培養 47 （六）脾臟細胞增生測定 48 （七）腹腔細胞之取得與培養 48 （八）細胞激素含量測定 49 （九） Real-time PCR 法分析分析肺部組織相關基因 51 （十） 吞噬細胞活性測定 53 （十一）自然殺手細胞活性分析 54 （十二）肺部組織切片 55 （十三）統計方法 55 第三節 結果 一、生長情形與攝食狀況 56 二、器官重量與相對重量 56 三、呼吸道過度反應 (AHR) 56 四、肺沖洗液 (BALF) 中細胞激素含量 57 五、肺沖洗液中各種類細胞數目 57 六、肺組織形態與mRNA基因表現 58 七、脾臟細胞細胞激素分泌量 58 八、脾臟細胞增生反應 59 九、血清中OVA特異性抗體含量 59 十、血清中總抗體含量 60 十一、腹腔細胞細胞激素分泌量 60 十二、循環血液中吞噬細胞之吞噬能力 60 十三、脾臟細胞中自然殺手細胞活性 61 第四節 討論 76 一、餵食BGP、CLN或CLA能降低氣喘模式小鼠呼吸道發炎 76 二、餵食BGP、CLN或CLA對過敏性小鼠全身性免疫反應的調節作用 78 三、 餵食BGP與CLA能降低氣喘模式小鼠血清中過敏性抗體的含量 79 四、餵食BGP與CLA能回復氣喘模式小鼠肺組織ppar-α基因表現量 80 五、餵食CLN能回復氣喘模式小鼠肺組織ppar-γ基因表現量 81 六、Th17在氣喘模式小鼠中扮演的角色 81 第四章 綜合討論與總結 第一節 綜合討論 83 一、細胞模式與氣喘小鼠模式之比較 83 二、山苦瓜減緩過敏性氣喘的可能有效成份 84 三、CLN與CLA之比較 85 四、活化PPAR的食材具有減緩過敏性氣喘症狀之潛力 86 第二節 總結 88 參考文獻 89 表目錄 表2.1 山苦瓜各萃物、CLN與CLA對發炎與免疫反應的影響 39 表3.1a 飼料配方 43 表3.1b 各組管餵內容物與飼料 43 表3.2 餵食山苦瓜、CLN與CLA對OVA致敏小鼠生長情形與攝食的影響 62 表3.3 餵食BGP、CLN與CLA對OVA致敏小鼠組織器官重量與相對組織器官重量的影響 63 表3.4 餵食BGP、CLN與CLA對OVA致敏小鼠BALF中細胞激素與化學趨化激素的影響 65 表3.5 餵食BGP、CLN與CLA對OVA致敏小鼠脾臟細胞在OVA刺激下細胞激素影響 69 表3.6 餵食BGP、CLN與CLA對OVA致敏小鼠脾臟細胞在conA刺激下細胞激素分泌的影響 70 表3.7 餵食BGP、CLN與CLA對OVA致敏小鼠腹腔細胞分泌細胞激素影響 74 表3.8 餵食BGP、CLN與CLA對OVA致敏小鼠吞噬細胞能力的影響 75 表3.9 餵食BGP、CLN與CLA對OVA致敏小鼠NK細胞能力的影響 75 表4.1 餵食BGP、CLN與CLA對氣喘模式小鼠過敏免疫反應的影響 87 圖目錄 圖1.1 各種細胞與相關細胞激素或蛋白質在過敏性氣喘中的作用 6 圖1.2 PPARs合成與天然之ligands 8 圖2.1 山苦瓜各萃物、CLN與CLA對RAW264.7細胞株存活率與細胞激素的影響 30 圖2.2 山苦瓜各萃物、CLN與CLA對初代腹腔細胞存活率與細胞激素分泌的影響 31 圖2.3 山苦瓜各萃物、CLN與CLA對EL4細胞存活率與細胞激素分泌的影響 32 圖2.4 山苦瓜各萃物、CLN與CLA對初代脾臟細胞存活率的影響 33 圖2.5 山苦瓜各萃物、CLN與CLA對小鼠初代脾臟細胞細胞激素分泌的影響 34 圖2.6 山苦瓜各萃物、CLN與CLA對初代脾臟細胞分泌細胞激素Th1/Th2比例的影響 35 圖2.7 山苦瓜各萃物、CLN與CLA對樹突細胞存活率與細胞激素分泌的影響 36 圖3.1 氣喘模式小鼠致敏與餵食飼養流程 42 圖3.2 餵食BGP、CLN與CLA對OVA致敏小鼠呼吸道過度反應的影響 64 圖3.3 餵食BGP、CLN與CLA對OVA致敏小鼠BALF中各種類細胞數目的影響 66 圖3.4 餵食BGP、CLN與CLA的OVA致敏小鼠肺切片圖 67 圖3.5 餵食BGP、CLN與CLA對OVA致敏小鼠肺部(a) ppar-α(b)ppar-γ(c) il-12p35 (d) cox-2 mRNA表現量的影響 68 圖3.6 餵食BGP、CLN與CLA對OVA致敏小鼠脾臟細胞增生能力的影響 71 圖3.7 餵食BGP、CLN與CLA對OVA致敏小鼠血清中OVA特異性抗體含量的影響 72 圖3.8 餵食BGP、CLN與CLA對OVA致敏小鼠血清中總抗體含量的影響 73 | |
| dc.language.iso | zh-TW | |
| dc.title | 山苦瓜與共軛次亞麻油酸對氣喘模式小鼠發炎與過敏免疫反應的影響 | zh_TW |
| dc.title | Effects of bitter gourd and 9c, 11t, 13t-conjugated linolenic acids on airway inflammation and immune responses in murine allergic asthma model | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 黃青真,江伯倫,吳文勉,趙蓓敏 | |
| dc.subject.keyword | 氣喘,山苦瓜,共軛次亞麻油酸,IgE,呼吸道發炎,共軛亞麻油酸, | zh_TW |
| dc.subject.keyword | asthma,bitter gourd,conjugated linolenic acid,IgE,airway inflammation,conjugated linoleic acid, | en |
| dc.relation.page | 101 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2010-08-09 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 微生物與生化學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
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