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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳明汝(Ming-Ju Chen) | |
dc.contributor.author | Shih-Hung Wei | en |
dc.contributor.author | 魏士弘 | zh_TW |
dc.date.accessioned | 2021-06-08T01:23:32Z | - |
dc.date.copyright | 2014-08-08 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-08-04 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18749 | - |
dc.description.abstract | 第一型糖尿病(type 1 diabetes, T1D)是一種自體免疫疾病,由於胰臟中β細胞逐漸失去功能並死亡,導致胰島素分泌不足及高血糖症狀。類升糖素胜肽-1(glucagon-like peptide-1, GLP-1)是由腸道內泌細胞所分泌之腸泌素,可調節胰臟β細胞之生長、增加胰島素分泌量以調節血糖之恆定。因此本試驗旨在篩選可刺激腸道內泌細胞分泌GLP-1之乳酸菌株。
試驗選用六十株乳酸菌分別與上皮內泌細胞株STC-1以1:100之比例共培養,測定上清液中之GLP-1濃度,並挑選與對照組相比可顯著提高GLP-1分泌量(p< 0.05)之兩株乳酸菌進行定序,分別為Lactobacillus kefiranofaciens 與 Lactobacillus kefiri。為進一步探討此二篩選菌株於動物模型中是否具有刺激GLP-1分泌能力及調節血糖之機能性,動物試驗選用4週齡C57BL/6小鼠進行試驗,以每公斤體重40 mg之鏈脲佐菌素連續腹腔注射4天以誘導T1D,成功誘導T1D之小鼠隨機分組,分別給予磷酸緩衝液之對照組,及每天每隻小鼠給予108 CFU之篩選菌株,一共8週。結果顯示,Lb. kefiranofaciens 與 Lb. kefiri皆能顯著降低T1D小鼠之空腹血糖值,而Lb. kefiri組別經口服糖耐量測定後也顯著改善了小鼠恆定血糖能力。藉由胰臟切片之免疫組織化學染色發現給予Lb. kefiranofaciens 與 Lb. kefiri後,胰臟β細胞對胰島之面積比例顯著高於STZ對照組,同時也觀察到篩選菌株具有調節細胞激素含量及提高血清GLP-1含量之能力。然而在非肥胖糖尿病(non obese diabetes, NOD)小鼠試驗中,給予Lb. kefiri無法顯著預防T1D之發生。本試驗嘗試探討篩選菌株降血糖功效之可能機制,然而結果顯示其降血糖功效似乎與類鐸受體2(Toll-like receptor 2)之訊息傳遞及短鏈脂肪酸(short chain fatty acid)之含量並無關聯。 綜觀上述,本試驗篩選之乳酸菌株Lb. kefiranofaciens與Lb. kefiri於體外試驗及動物試驗皆能刺激GLP-1分泌,且有助於STZ誘導T1D小鼠維持血糖恆定,此二篩選菌株或許具有高度潛力可開發成為具降血糖機能性之益生菌產品,並應用於糖尿病之輔助療法之一。然而其詳細降血糖機制仍有待釐清。 | zh_TW |
dc.description.abstract | Type 1 diabetes (T1D) is an autoimmune disease, which characterized by progressive dysfunction of pancreatic β cell and loss in β cell mass. The disease generally results in insulin deficiency and hyperglycemia. Glucagon-like peptide-1 (GLP-1) is a gut hormone which secreted by enteroendocrine cell. It has been well documented that GLP-1 could regulate β cell growth and increase insulin secretion to maintain glucose homeostasis. Therefore, our aim was to screen and select lactic acid bacteria (LAB) with abilities to stimulate intestinal GLP-1 secretion.
The murine enteroendocrine STC-1 cells were co-cultured with sixty LABs at a ratio of 1: 100 and GLP-1 productions in supernatant were measured. We selected and identified top two strains, Lactobacillus kefiranofaciens and Lactobacillus kefiri, significantly elevating GLP-1 secretion when compared with control group (p< 0.05). We next investigated GLP-1 secretion abilities and improvement of hyperglycemia in vivo. Four-week-old male C57BL/6 mice were induced diabetes by intraperitoneal injection of streptozotocin(STZ) for 4 consecutive days. Diabetic mice were randomly assigned to different groups and were administered PBS or selected two strains 108 CFU per mouse daily for eight weeks. The results exhibited that Lb. kefiranofaciens and Lb. kefiri could significantly lower fasting blood glucose than STZ group after eight week administration. Lb. kefiri also significantly improve glucose homeostasis after oral glucose challenge when compared with the STZ group. We found that Lb. kefiranofaciens and Lb. kefiri group showed significantly higher insulin/pancreas ratio by pancreas immunohistochemistry staining. Modulation effects on cytokines level and an elevation of serum GLP-1 levels were also observed. However, administration of Lb. kefiri did not show significant effect to delay the onset of diabetes in non obese diabetic (NOD) mice. We further tried to address possible mechanism and found that toll-like receptor 2 and short chain fatty acid production might be not involved in blood glucose lowering effect. Taken together, our present study concluded that Lb. kefiranofaciens and Lb. kefiri could stimulate GLP-1 secretion both in vitro and in vivo. Oral administration of Lb. kefiranofaciens and Lb. kefiri provided a benefit to blood glucose homeostasis in STZ-induced T1D model. It suggested that the selected strains might be a potential therapeutic supplementation to T1D. However, the mechanisms need more clarification. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T01:23:32Z (GMT). No. of bitstreams: 1 ntu-103-R01626012-1.pdf: 60456364 bytes, checksum: 3c3ea44a53d7cb7610cc20a0ecc00a65 (MD5) Previous issue date: 2014 | en |
dc.description.tableofcontents | 壹、文獻探討 1
第一節:糖尿病(diabetes) 1 一、糖尿病之定義 1 二、第一型糖尿病之病因 2 (一)遺傳基因之易致病性(predisposition) 2 (二)環境因子 8 三、糖尿病之治療 11 (一)胰島素注射 11 (二)胰島移植(islet transplantation) 11 (三)免疫抑制藥物 11 (四)類升糖素胜肽-1(glucagon-like peptide-1, GLP-1) 12 第二節:GLP-1 14 一、GLP-1之生理功能 14 二、GLP-1之分泌 17 (一)脂肪酸與GLP-1之分泌 17 (二)腸道菌相與GLP-1之分泌 17 第三節:乳酸菌與糖尿病 19 一、乳酸菌與腸道上皮完整性 19 二、乳酸菌與腸道菌相 20 貳、材料與方法 22 第一節:藉由體外試驗篩選可刺激GLP-1分泌之菌株 22 一、實驗材料 22 (一)試驗菌株與STC-1腸道上皮細胞 22 二、實驗方法 22 (一)菌株之活化與保存 22 (二)細胞之活化與保存 23 (三)菌株與STC-1細胞之共培養 24 (四)GLP-1之測定 24 (五)篩選菌株之定序 24 (六)篩選菌株之模擬胃腸液耐受性測試 28 (七)經模擬胃腸液消化之篩選菌株與STC-1細胞共培養 28 (八)共培養STC-1細胞存活率測定 29 第二節:由動物試驗探討篩選菌株之調節血糖能力 30 一、化學藥物誘導第一型糖尿病之動物模式 30 (一)試驗菌株 30 (二)實驗動物 30 二、自發性第一型糖尿病之動物模式 34 (一)試驗菌株 34 (二)實驗動物 34 第三節:探討調節血糖之機制 35 一、抗體阻斷(blocking)試驗 35 二、糞便之短鏈脂肪酸分析 35 三、糞便之菌群分析 38 統計分析 41 參、結果與討論 42 第一節:篩選可刺激腸道L細胞分泌GLP-1之乳酸菌株 42 一、菌株之篩選與定序結果 42 二、篩選菌株之模擬胃腸液耐受性 43 第二節:篩選乳酸菌株減緩STZ誘導小鼠糖尿病機能性評估 50 一、糖尿病臨床症狀評估 50 二、STZ誘導T1D小鼠之血糖變化 54 三、STZ誘導T1D小鼠之OGTT 56 四、血清中胰島素與GLP-1含量 59 五、胰臟組織切片之評估 61 六、胰臟、腎臟及脾臟之細胞激素含量 64 七、NOD小鼠 67 第三節:乳酸菌降血糖可能機制探討 72 一、TLR2路徑 72 二、糞便SCFA之含量 74 三、糞便微生物菌門比例 76 肆、結論 78 參考文獻 79 作者小傳 93 | |
dc.language.iso | zh-TW | |
dc.title | 篩選可刺激類升糖素胜肽-1分泌之益生菌及探討其減緩高血糖症之研究 | zh_TW |
dc.title | Selecting of probiotics with GLP-1 enhancing ability to attenuate hyperglycemia in vitro and in vivo | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 林慶文(Chin-Wen Lin),丁詩同(Shih-Torng Ding),潘子明(Tzu-Ming Pan),廖俊旺(Jiunn-Wang Liao) | |
dc.subject.keyword | 益生菌,類升糖素胜?-1,高血糖,糖尿病, | zh_TW |
dc.subject.keyword | Probiotic,Glucagon-like peptide-1,Hyperglycemia,Diabetes, | en |
dc.relation.page | 93 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2014-08-05 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 動物科學技術學研究所 | zh_TW |
顯示於系所單位: | 動物科學技術學系 |
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