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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 林璧鳳(Bi-Fong Lin) | |
dc.contributor.author | Yu-Ting Chen | en |
dc.contributor.author | 陳郁婷 | zh_TW |
dc.date.accessioned | 2021-05-20T00:49:24Z | - |
dc.date.available | 2025-08-20 | |
dc.date.available | 2021-05-20T00:49:24Z | - |
dc.date.copyright | 2020-09-17 | |
dc.date.issued | 2020 | |
dc.date.submitted | 2020-08-17 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/8157 | - |
dc.description.abstract | 台灣末期腎臟病盛行率多年來於全球排名中高居首位,近期的研究發現腎臟病與腸道共生菌的失衡有關。給予末期腎臟病人補充乳桿菌屬代田菌後,可降低病人血清的尿素含量。目前對於益生菌的補充是否可改善腎損傷尚未有定論,故本研究欲探討代田菌 (Lactobacillus casei Shirota, 簡稱LcS) 的給予對腎臟損傷及其免疫調控的影響。 實驗一為每天給予VHL基因缺失而自發性腎癌之Vhlhfl/fl小鼠1×10^9 CFU的LcS,結果得知生命期顯著延長。實驗二接著利用馬兜鈴酸 (aristolochic acid , 簡稱AA) 誘發Vhlhfl/+小鼠腎病變,探討代田菌的影響機轉。先給予8到12週齡小鼠1×10^9 CFU/d LcS 4週後,加AA (4 mg/kg BW/d)餵2週後犧牲分析指標。共四組小鼠:Con組、LcS組、AA組、與LcS-AA組。結果顯示LcS可顯著降低AA誘發之腎損傷程度、尿蛋白肌酸酐比值、尿液KIM-1及血清尿素氮濃度等腎功能指標,全身性發炎指標血清IL-6濃度及纖維化相關基因Lgals3與Fn1的mRNA表現量也有減少趨勢;腎臟的免疫分析觀察到,AA誘發M2巨噬細胞、Th細胞及Treg細胞浸潤至腎臟,同時,MCP-1及IL-10濃度上升且IL-1b、TNF-a及IL-4濃度下降。補充LcS顯著降低M1巨噬細胞的浸潤比例及M1/M2比例,M2巨噬細胞及Treg浸潤程度也有下降的趨勢,此外,TGF-b和 IL-4的濃度增加。腸道和脾臟細胞免疫指標顯示,AA促進IL-6、MLN和脾臟細胞TNF-a、PP細胞IL-10、MLN細胞TGF-b的分泌。補充LcS增加腸道固有層的樹突細胞比例,抑制PP免疫細胞分泌IL-17A,促進PP和脾臟免疫細胞分泌IL-10,MLN和脾臟免疫細胞分泌IFN-g,抑制脾臟免疫細胞分泌IL-6。綜合以上結果,補充LcS可能藉由影響調節型和發炎性的細胞激素平衡,能顯著促進PP進而脾臟細胞的IL-10分泌和MLN細胞與腎的IL-4分泌,能抑制PP細胞IL-17A及脾臟細胞IL-6的分泌進而降低血清IL-6濃度,影響腸道免疫進而調控全身性及腎臟的免疫反應,顯著降低AA造成的尿蛋白和KIM-1濃度、BUN和IL-6濃度增加,且延長自發性腎炎致腎癌小鼠的生命期,具有抑制發炎以達護腎之潛力。 | zh_TW |
dc.description.abstract | It has been noted that the prevalence and incidence rate of end-stage renal disease in Taiwan are the highest in the world. It is interesting to find out if probiotic could delay the progression of renal disease by ameliorating inflammatory response and reducing pathobionts which generate uremic toxin. Lactobacillus casei Shirota (LcS) has been demonstrated that to reduce serum urea level in III-IV stage chronic kidney disease patients. In this study, we investigated whether Lactobacillus casei Shirota could improve renal injury and exert the immunomodulation effect. The results demonstrated that supplementary of LcS (1×10^9 CFU/day) prolonged the lifespan of Vhlhfl/fl mice. Further, 8-12-week-old Vhlhfl/+ mice were fed AIN-93 diet with or without LcS for 4 weeks and then fed aristolochic acid added diets (AA; 4 mg/kg BW) for induction of renal injury for 2 weeks to investigate the effects. There are the Con, LcS, AA, LcS-AA groups. The results showed urinary protein and KIM-1, serum urea, M1 macrophages infiltration in kidney were decreased by LcS supplement. AA promoted renal MCP-1 and IL-10 level, decreased IL-1b, IL-6, TNF-a, and IL-4 level. LcS supplement increased IL-4 and TGF-b secretion under AA induction. LcS supplement increased dendritic cells infiltration in lamina propria, downregulated IL-17A and upregulated IL-10 secretion by Peyer’s patch. Levels of IFN-g and TGF-b were elevated in mesenteric lymph node. Splenocyte decreased IL-6 and promoted IL-2, IFN-g and IL-10 secretion. In conclusion, LcS supplement might have the nephroprotective effects by regulating the immune response and possibly maintaining the balance of gut microbiota. | en |
dc.description.provenance | Made available in DSpace on 2021-05-20T00:49:24Z (GMT). No. of bitstreams: 1 U0001-1708202016090300.pdf: 3984422 bytes, checksum: 01fabf5e37aefc04d2c17f170d9e6a20 (MD5) Previous issue date: 2020 | en |
dc.description.tableofcontents | 目錄 中文摘要 i Abstract ii 目錄 iii 圖目錄 vii 表目錄 ix 縮寫對照表 x 第一章 序論 1 第一節 文獻回顧 1 一、 腎臟疾病 1 (一) 腎臟簡介 1 (二) 急性腎臟疾病 1 (三) 腎臟疾病之免疫反應 2 (四) 腎臟纖維化 3 二、 馬兜鈴酸致腎病變 4 三、 von Hippel-Lindau (VHL) 基因缺失小鼠 5 四、 腸道免疫 6 (一) 簡介 6 (二) 腎臟疾病之腸道免疫 8 五、 益生菌 9 (一) 簡介 9 (二) 益生菌對腸道免疫之調控 10 (三) 益生菌對腎臟病之免疫調控機制 11 六、 代田菌 (Lactobacillus casei Shirota, LcS) 12 (一) 簡介 12 (二) 代田菌對免疫之調控 13 (三) 代田菌對腎臟病之影響 13 第二節 研究動機與目的 14 第二章 實驗設計與材料方法 15 第一節 實驗設計 15 第二節 代田菌培養與定量 16 一、 代田菌培養 16 二、 代田菌液gDNA萃取 16 三、 代田菌培養液菌數定量方法 16 第三節 動物繁殖 18 一、 繁殖基因剔除鼠 18 二、 基因鑑定 18 三、 動物飼養 19 第四節 實驗方法 20 一、 尿液及血清收集與檢測 20 (一) 尿液收集 20 (二) 血液收集 20 (三) 尿液kidney injury molecular (KIM)-1測定 20 (四) 尿蛋白檢測 20 (五) 尿液與血清肌酸酐檢測 21 1. 尿液肌酸酐檢測 21 2. 血液肌酸肝檢測 21 (六) 血清尿素氮檢測 21 二、 直腸及腎臟組織切片染色 21 三、 腎臟纖維化相關mRNA分析 22 (一) 腎臟RNA萃取 22 (二) RNA反轉錄 22 (三) Real-time PCR 分析mRNA表現量 23 四、 腸道菌群分析 23 (一) 糞便及盲腸收集 23 (二) 樣品gDNA萃取 23 (三) Real-time PCR分析腸道菌群 24 五、 淋巴細胞分離 25 (一) 腸道固有層淋巴細胞單離 25 (二) 腸繫膜淋巴結與皮耶氏體細胞單離 26 (三) 脾臟細胞單離 26 (四) 腎臟細胞單離 27 (五) 細胞計數 27 六、 免疫細胞表型分析 28 (一) 細胞前處理 29 (二) Surface staining (外染) 29 (三) Intracellular staining (內染) 30 七、 細胞激素含量分析 30 (一) 免疫細胞培養 30 (二) 腎臟組織測定前處理 30 (三) 細胞激素含量分析 31 八、 統計分析 32 第三章 結果 33 第一節 實驗一 補充代田菌對Vhlhfl/fl腎癌模式小鼠生命期之影響 33 第二節 實驗二 補充代田菌對馬兜鈴酸致腎病變之Vhlhfl/+小鼠腎病變之影響 34 一、 代田菌定殖確認 34 二、 Vhlhfl/+小鼠體重與攝食量變化 35 三、 Vhlhfl/+小鼠器官重量 36 四、 Vhlhfl/+小鼠腎臟損傷及纖維化程度 37 五、 Vhlhfl/+小鼠腎臟的細胞激素濃度 40 六、 Vhlhfl/+小鼠腎臟中巨噬細胞與調節型T細胞比例 41 七、 補充代田菌對Vhlhfl/+小鼠直腸組織損傷及免疫細胞之影響 43 第四章 討論與結論 48 第一節 討論 48 一、 補充代田菌對Vhlhfl/fl小鼠生命期之影響 48 二、 口服馬兜鈴酸致Vhlhfl/+小鼠腎病變模式探討 48 三、 補充代田菌對馬兜鈴酸致Vhlhfl/+小鼠腎病變之腎損傷影響 49 四、 補充代田菌對馬兜鈴酸致Vhlhfl/+小鼠腎病變之腸道免疫調節 51 五、 補充代田菌對馬兜鈴酸致Vhlhfl/+小鼠腎病變之全身性免疫調節 53 第二節 結論 56 附錄表 58 附錄圖 60 參考文獻 61 | |
dc.language.iso | zh-TW | |
dc.title | 益生菌Lactobacillus casei Shirota 對VHL基因缺失小鼠腎損傷及免疫調節的影響 | zh_TW |
dc.title | Renal protective effect of Lactobacillus casei Shirota on renal injury and immunomodulation in Vhlh gene-knockout mice | en |
dc.type | Thesis | |
dc.date.schoolyear | 108-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 鄭光成(Kuan-Chen cheng),江孟燦(Meng-Tsan Chiang),徐沺(Tien Hsu),林金源(Jin-Yuarn Lin) | |
dc.subject.keyword | Lactobacillus casei Shirota,馬兜鈴酸,腎病變,腸道免疫,抗發炎, | zh_TW |
dc.subject.keyword | Lactobacillus casei Shirota,aristolochic acid,nephropathy,gut-immune system,anti-inflammation, | en |
dc.relation.page | 79 | |
dc.identifier.doi | 10.6342/NTU202003787 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2020-08-18 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科技學系 | zh_TW |
dc.date.embargo-lift | 2025-08-20 | - |
顯示於系所單位: | 生化科技學系 |
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