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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 潘子明(Tzu-Ming Pan) | |
| dc.contributor.author | Meng-Chun Cheng | en |
| dc.contributor.author | 鄭孟純 | zh_TW |
| dc.date.accessioned | 2021-06-16T09:40:23Z | - |
| dc.date.available | 2022-02-17 | |
| dc.date.copyright | 2017-02-17 | |
| dc.date.issued | 2017 | |
| dc.date.submitted | 2017-02-07 | |
| dc.identifier.citation | Aderem, A., Underhill, D.M. 1999. Mechanisms of phagocytosis in macrophages. Annu. Rev. Immunol. 17: 593-623.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/59834 | - |
| dc.description.abstract | 腦血管疾病為一種嚴重的神經性疾病，亦為導致死亡與成年人長期殘疾的主要原因。腦血管疾病發生期間，氧氣和葡萄糖供應不足造成細胞損傷與大量自由基產生，最終導致神經細胞死亡及造成智力減退，由腦血管疾病引起之認知障礙及失智症統稱為血管性失智症。近年來研究指出萃取自蔬果與飲品中之天然化合物，藉由抗氧化、抗發炎及抑制微膠細胞活化等機制達到神經保護之效果。本研究室先前研究結果顯示，Lactobacillus paracasei subsp. paracasei NTU 101 (NTU 101) 發酵產物可改善數種與氧化壓力及/或發炎反應相關之疾病，亦能降低腦血管疾病之危險因子如：高血壓、粥狀動脈硬化、高血脂及肥胖。推論 NTU 101 發酵產物應具抗氧化及抗發炎等生理活性，故具預防及治療腦血管疾病之潛力。首先，以菌數、pH 值與乳酸變化作為指標，初步篩選出適合 NTU 101 發酵之基質為 25% 牛奶添加 5% 葡萄糖 (25% SM + 5% Glc)。25% SM + 5% Glc 發酵液之乙醇萃取物，於濃度 1 mg/mL 下，可預防由氧氣與葡萄糖剝奪及再灌注 (oxygen glucose deprivation-reoxygenation, OGD/R) 所造成之細胞死亡，與 OGD/R 相比，顯著回復細胞存活率達 42% (p < 0.05，回復率公式:(樣品組細胞存活率- OGD/R 組細胞存活率)/OGD/R 組細胞存活率 × 100%)。進一步探討 25% SM + 5% Glc 中具神經保護之生物活性物質，25% SM + 5% Glc 之 NTU 101 發酵液乙醇萃取物中的區分物 2-7-4 (F-2-7-4) 於濃度 10 μg/mL 下與 OGD/R 相比顯著增加細胞存活率達 40% (p < 0.05)。F-2-7-4 由超高效液相層析串聯質譜儀 (ultra-performance liquid chromatography-mass spectrometry) 得其偽離子峰為 m/z 551.5033 ([M-H2O+H]+)，利用核磁共振 (nuclear magnetic resonance) 儀鑑定其分子式為 C35H68O5、分子量為 568 之化合物 glyceryl 1,3-dipalmitate (GD)。動物試驗以含有 GD 之 25% SM + 5% Glc 之 NTU 101 發酵液乙醇萃取物 (NTU101F) 來評估其對於醋酸去氧皮質酮與鹽溶液 (deoxycorticosterone acetate and salt solution, DOCA-S) 高血壓誘導血管性失智症之改善效果，實驗結果顯示，NTU101F (11、22 或 110 mg/kg) 可改善弓動脈彈性蛋白排列散亂及增加內皮型一氧化氮合成酶 (endothelial nitric oxide synthase) 表現量，促使一氧化氮生成，達到血管舒張之效果，亦可降低腦部水分滯留及腦組織中基質金屬蛋白酶 9 (matrix metalloproteinases-9)、氧化壓力及發炎之情形，進而改善學習記憶之能力。以上結果顯示，NTU101F 可透過抗氧化及抗發炎機制，預防血管性失智症之效果，具有潛力應用於腦血管疾病。進一步利用人類神經纖維瘤母細胞 (SH-SY5Y) 探討 GD 之神經保護機制，GD 可減緩 OGD/R 引起胞內活性氧 (reactive oxygen species) 之生成，提升細胞核中過氧化體增生劑活化接受器 γ (peroxisome proliferator-activated receptor gamma) 與核因子-紅血球之 2 相關因子-2 (nuclear factor erythroid 2-related factor 2, Nrf2) 之蛋白表現量，進而促使 Nrf2 下游抗氧化酵素-第一型血紅素氧化酶 (heme oxygenase-1, HO-1) 之表現，因而提升神經細胞抗氧化之能力，進而降低氧化壓力所造成之損傷。GD 可抑制核因子活化 B 細胞 κ 輕鏈增強子 (nuclear factor kappa-light-chain-enhancer of activated B cells) 之表現量，降低發炎反應。亦可藉由提升鈣離子通道蛋白 (plasma membrane Ca2+ ATPase) 之表現量，調控細胞內鈣離子之含量，達到減緩神經細胞損傷之效果。我們的研究結果顯示 GD 具預防或治療缺血性腦損傷及神經退行性疾病之潛力。由反應曲面法所得之最適 GD 生成條件為牛奶濃度 25.53%、葡萄糖濃度 44.32 g/L 及培養溫度 39.52oC，於此最適培養條件下，可生成 71.51 ng/g 之GD。以葡萄糖與牛奶為替代培養基可降低生產成本，提升其應用於商業之可行性。 | zh_TW |
| dc.description.abstract | Cerebrovascular disease is a common and serious neurological disease, which is the leading cause of death and long-term disability in adults. During the development of cerebrovascular disease, glucose and oxygen deficiencies induce subcellular damage and produce large amounts of free radicals, finally leading to neuronal death and intelligence deterioration. Cerebrovascular disease causes cognitive impairment and vascular dementia. Recent studies have focused on the potential preventative effects of natural fruit and vegetable extracts and beverages against certain age-related neurological disorders. The underlying mechanisms include antioxidation, anti-inflammation, and inhibition of microglia activation. Therefore, natural products are attracting increasing attention in the field of drug discovery. Our research group found that the fermentation metabolites of Lactobacillus paracasei subsp. paracasei NTU 101 (NTU 101) have antioxidant and anti-inflammation effects. In addition, they reduce the risk factors of cerebrovascular diseases such as hypertension, atherosclerosis, hyperlipidemia, and obesity. NTU 101-fermented metabolites have the potential to prevent and treat cerebrovascular diseases. The addition of 5% glucose to a 25% milk solution (25% SM + 5% Glc) exerted a significant effect not only on the number of bacterial cells but also on their acid-producing ability. A more statistically significant increase in cell viability was observed in the group treated with the 1 mg/mL ethanol NTU 101 extract-fermented 25% SM + 5% Glc (NTU101F) than was observed in the control oxygen-glucose deprivation-reoxygenation (OGD/R) pre-treatment groups (42%, p < 0.05, recovery rate: (cell viability of sample-treatment group - cell viability of OGD/R group)/cell viability of OGD/R group × 100%). Therefore, we investigated the neuroprotective compounds in the NTU101F. At a concentration of 10 μg/mL, fraction 2-7-4 (F-2-7-4) significantly enhanced cell viability by 40% (p < 0.05). The molecular formula of F-2-7-4 was identified as glyceryl 1,3-dipalmitate (GD, C35H68O5) using nuclear magnetic resonance and ultra-performance liquid chromatography-mass spectrometry (m/z 551.5033, [M-H2O+H]+). The hypertension-induced vascular dementia rats were orally administered the GD-containing-NTU101F (11, 22, or 110 mg/kg), which improved the elastin arrangement and upregulated the expression of endothelial nitric oxide (NO) synthase (eNOS). The upregulated eNOS stimulated NO synthesis and, thereby, improved hypertension. NTU101F also improved the learning and memory ability, and decreased brain edema, matrix metalloproteinase (MMP)-9 and oxidative activity, and inflammation in the hippocampus. Our results indicate that NTU101F effectively inhibited oxidative stress- and inflammation-related mechanisms in the brain, thus providing neuroprotection and preventing hypertension-associated loss of cognitive abilities. Therefore, we further investigated the neuroprotective effect of GD in human neuroblastoma SH-SY5Y cells. GD ameliorated OGD/R-induced apoptosis by elevating the expression of nuclear peroxisome proliferator-activated receptor gamma and nuclear factor erythroid 2-related factor 2 (Nrf2) proteins and its downstream heme oxygenase-1, which attenuated reactive oxygen species generation. Pre-treatment with GD reduced the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) expression, which attenuated the induction of pro-inflammatory mediators. GD also increased the plasma membrane Ca2+ adenosine triphosphatase (ATPase, PMCA) level and, thereby, reduced the levels of cytosolic Ca2+, which also correlated with the reduced cell death. Our findings indicate the potential of the therapeutic application of GD in the prevention and treatment of cerebral ischemic and neurodegenerative diseases. The response surface plot predicted a maximum GD concentration (71.51 ng/g) at an incubation temperature of 39.52°C using the substrate containing 25.53% milk and 44.32 g/L glucose. The milk powder and glucose were used as an alternative medium which could decrease the production cost, with the potential prospects for application. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T09:40:23Z (GMT). No. of bitstreams: 1 ntu-106-D02b22004-1.pdf: 11715635 bytes, checksum: 20e529e3d6c04e468d532f8c4680dd02 (MD5) Previous issue date: 2017 | en |
| dc.description.tableofcontents | 中文摘要………………………………………………………………………………… II
英文摘要………………………………………………………………………………… IV 縮寫表…………………………………………………………………………………… VII 目錄……………………………………………………………………………………… IX 圖目錄…………………………………………………………………………………… XII 表目錄…………………………………………………………………………………… XV 第壹章 前言與文獻回顧…………………………………………………………….…. 1 ㄧ、腦血管疾病介紹…………………………………………………………………… 1 二、缺血型腦血管疾病造成細胞損傷的機轉……………………………………….…. 4 (一) 細胞酸中毒 (acidosis) 與鈉離子超載 (sodium overload)…………………… 4 (二) 興奮性毒性物質釋放與鈣離子超載 (calcium overload)……………………... 4 (三) 蛋白質降解 (protein degradation)………………………………..……. 5 (四) 自由基及氧化傷害…………………….……………………………………….. 5 (五) 發炎反應……………………………………….....…………………………….. 9 三、缺血型腦血管疾病治療方式……………………………………………….………. 9 (一) 血栓溶解劑……………………………………………………………………... 9 (二) 神經保護……………………………………………………………………….. 10 四、血管性失智症 (vascular dementia, VaD)…………………………………………... 12 (一) 血管性失智症危險因子……………………..…………………………………. 13 (二) 血管性失智症的動物模式………………………………………….………... 15 五、乳酸菌………………….……………………………………………………………. 18 (一) 改善腸道菌相…………………………………………………………...……… 18 (二) 調節免疫系統…...…………………………………………………….………... 18 (三) 調節血脂………………………………………………………………………... 19 (四) 預防動脈粥狀硬化…...………………………………………………………… 20 (五) 調節血壓…………………...…………………………………………………… 20 (六) 預防胃黏膜損傷…………………………...…………………………………… 20 (七) 骨質保健………..……………………………………………………...……….. 21 (八) 不易形成體脂肪………………………………………………………….…….. 21 (九) 預防齲齒………………………………………………..………………………. 22 (十) 抑制癌細胞增生…………………………………………………….……..…… 22 第貳章 研究動機與目的……………………………………………………………….. 23 第參章 材料與方法…………………………………………………………………….. 25 ㄧ、實驗材料…………………………………………………………………………… 25 (一) 藥品試劑………………………………………………………………………... 25 (二) 細胞株…………………………………………………………………………... 25 (三) 儀器設備………………………………………………………………………... 25 二、實驗方法…………………………………………………………………………… 27 第一部分 初步發酵條件篩選………………………………………………………….. 27 第二部分 具保護神經細胞之活性物質分離、純化與鑑定…………29 第三部分 探討 25% SM + 5% Glc 之 NTU 101 發酵液乙醇萃取物對血管性失智 症之改善效果……………………………………………………………34 第四部分 探討 NTU 101 發酵液乙醇粗萃取物中的活性物質 (glyceryl 1,3-dipalmitate, GD) 對人類神經纖維瘤母細胞之保護效果……………44 第五部分 以反應曲面法 (response surface methodology, RSM) 探討 NTU 101 發酵液乙醇粗萃取物中的活性物質最適培養條件組合…48 第肆章 結果與討論…………………………………………………………………….. 50 一、NTU 101 初步發酵條件篩選………………………………………………………50 二、具保護神經細胞活性物質之分離、純化與鑑定………………………52 三、25% SM + 5% Glc 之 NTU 101 發酵液乙醇萃取物對高血壓誘發血管性失智症大鼠之改善效果……………………………...…………………………….…71 四、探討 NTU 101 發酵液乙醇粗萃取物中的活性物質 (glyceryl 1,3-dipalmitate, GD) 對人類神經纖維瘤母細胞 (SH-SY5Y) 之保護效果……………………87 五、以反應曲面法 (response surface methodology, RSM) 探討 NTU 101 發酵液中的活性物質 (GD) 最適培養條件組合…………100 第伍章 結論…………………………………………………………………………….. 106 第陸章 參考文獻……………………………………………………………………….. 108 第柒章 附錄…………………………………………………………………………….. 135 | |
| dc.language.iso | zh-TW | |
| dc.subject | 腦血管疾病 | zh_TW |
| dc.subject | 3-dipalmitate | zh_TW |
| dc.subject | glyceryl 1 | zh_TW |
| dc.subject | 發酵產物 | zh_TW |
| dc.subject | NTU 101 | zh_TW |
| dc.subject | 氧氣與葡萄糖剝奪 | zh_TW |
| dc.subject | NTU 101 | en |
| dc.subject | cerebrovascular diseases | en |
| dc.subject | oxygen-glucose deprivation | en |
| dc.subject | 3-dipalmitate | en |
| dc.subject | glyceryl 1 | en |
| dc.subject | fermented product | en |
| dc.title | Lactobacillus paracasei subsp. paracasei NTU 101 發酵產物對改善腦血管疾病之研究 | zh_TW |
| dc.title | Study on the Lactobacillus paracasei subsp. paracasei NTU 101-fermented products for improvement in cerebrovascular diseases | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 105-1 | |
| dc.description.degree | 博士 | |
| dc.contributor.coadvisor | 陳俊任(Chun-Jen Chen) | |
| dc.contributor.oralexamcommittee | 周正俊(Cheng-chun Chou),廖啟成(Chii-Cherng Liao),蔡宗佑(Tsung-Yu Tsai),蔣慎思(Shen-Shih Chiang),呂彥禮(Yann-Lii Leu) | |
| dc.subject.keyword | 腦血管疾病,氧氣與葡萄糖剝奪,NTU 101,發酵產物,glyceryl 1,3-dipalmitate, | zh_TW |
| dc.subject.keyword | cerebrovascular diseases,oxygen-glucose deprivation,NTU 101,fermented product,glyceryl 1,3-dipalmitate, | en |
| dc.relation.page | 135 | |
| dc.identifier.doi | 10.6342/NTU201700388 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2017-02-08 | |
| 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 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-106-1.pdf 未授權公開取用 | 11.44 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。