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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.advisor | 林璧鳳(Bi-Fong Lin | bifong@ntu.edu.tw | ), | |
dc.contributor.author | Wei-Yin Chen | en |
dc.contributor.author | 陳維茵 | zh_TW |
dc.date.accessioned | 2023-03-19T22:32:10Z | - |
dc.date.copyright | 2022-08-30 | |
dc.date.issued | 2022 | |
dc.date.submitted | 2022-08-25 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/84909 | - |
dc.description.abstract | 根據台灣健保資料庫數據顯示,乾癬就診人數逐年攀升。乾癬 (psoriasis) 為一種免疫失衡的慢性皮膚疾病。近年研究顯示,乾癬患者的腸道菌相多樣性較低,益生菌數量顯著減少。已知乳酸菌代田株 (Lactobacillus casei Shirota, LcS) 具有調節腸道菌組成與免疫調節的功能,但對於乾癬免疫調節的影響尚未研究。故本研究欲探討補充代田菌是否能減緩Imiquimod (IMQ) 誘發乾癬小鼠的發炎症狀。實驗設計將8週齡小鼠分為Control組、IMQ組、LcS/IMQ組、正控制組。於LcS/IMQ組小鼠的飲用水每天加入1 x 109 CFU/ml LcS 4週後,分別於小鼠12、20、26週齡時,進行連續5天背部皮膚與耳朵塗抹IMQ誘發乾癬,Ctrl組塗抹凡士林。於第三次誘發乾癬後將小鼠犧牲,分析免疫相關指標。結果顯示,LcS/IMQ組第一、二次誘發乾癬背部皮膚的皮屑程度與促發炎細胞激素IL-22與IL-6含量顯著較IMQ組低,且調節型細胞激素IL-10含量顯著較IMQ組高,顯示,LcS可能具有減緩皮膚發炎反應的能力。LcS/IMQ組的MLN細胞分泌IL-17A、TNF-α,以及脾臟細胞分泌IL-17A顯著低於IMQ組,IFN-γ的分泌顯著高於IMQ組,顯示,LcS可能具有減緩乾癬腸道與全身性發炎反應的能力。LcS/IMQ組腸道的Bifidobacterium菌含量趨勢性高於IMQ組,顯示,LcS可促進腸道益生菌的生長。綜合以上結果,推測補充代田菌藉由促進腸道益生菌生長與降低腸道促發炎細胞激素,進而影響全身性的免疫反應,降低脾臟分泌促發炎細胞激素,促進IFN-γ分泌,使T細胞偏向發育Th1,降低Th17細胞激素的分泌,並降低皮膚中促發炎細胞激素的含量,從而減緩乾癬的症狀。 | zh_TW |
dc.description.abstract | The prevalence of psoriasis in Taiwan has increased year by year. Psoriasis is a T-cell-mediated chronic inflammatory skin disorder. Recently, accumulating evidence has supported the pathogenic role of the gut microbiome in autoimmune and inflammatory diseases, including psoriasis. It has been demonstrated that Lactobacillus casei Shirota (LcS) can prevent pathogen colonization of the gut and reduce the incidence or relieve the symptoms of various diseases caused by dysregulated immune responses. However, influence of LcS on psoriasis remains unknown. Therefore, this study was designed to investigate whether LcS could affect commensal microbiota and intestinal immunity on Imiquimod (IMQ)-induced psoriasis-like model. Eight weeks old mice were pretreated with probiotic LcS for 4 weeks, followed by receiving IMQ on the shaved back to induce psoriasis, and continuously given LcS until mice were sacrificed. The mice received a topical application of 62.5 mg IMQ cream or vaseline on the back skin daily for 5 consecutive days. The age induction of psoriasis was 12, 20, and 26 week-old, respectively. Here, the results showed that oral administration of LcS significantly decreased scaling lesions during the first and second induction of psoriasis compared to the IMQ group. It also reduced the cytokine production of proinflammatory cytokines, including IL-22 and IL-6, and increased IL-10 in the back skin of imiquimod-treated mice. LcS supplements significantly decreased gut proinflammatory cytokines, including IL-17A and TNF-α in IMQ-induced psoriasis-like mice. Meanwhile, cytokines IL-17A produced by ConA-stimulated splenocyte was reduced, increasing the secretion of IFN-γ. Furthermore, the gut microbiota in cecum contents from IMQ-induced psoriasis-like mice fed LcS increased the population of Bifidobacterium. In conclusion, oral administration of LcS could affect the immune responses and the gut microbiota compositions, which might be beneficial for amelioration progression of psoriasis. | en |
dc.description.provenance | Made available in DSpace on 2023-03-19T22:32:10Z (GMT). No. of bitstreams: 1 U0001-2508202212510900.pdf: 4860486 bytes, checksum: fb54ef9ac262675d18ec984a16708df4 (MD5) Previous issue date: 2022 | en |
dc.description.tableofcontents | 誌謝 i 摘 要 iii Abstract iv 目 錄 vi 圖目錄 x 表目錄 xi 縮寫對照表 xii 第一章 序論 1 第一節 文獻回顧 1 一、乾癬簡介 1 (一) 皮膚 1 (二) 乾癬 1 (三) 乾癬之免疫 4 A. 乾癬致病機制 4 B. 乾癬相關細胞激素 5 (四) Imiquimod誘發乾癬動物模型 6 二、腸道共生菌與免疫系統 7 (一) 腸道共生菌 7 (二) 腸道免疫系統 7 (三) 腸道菌與免疫恆定 9 (四) 腸道與自體免疫疾病 10 三、益生菌 11 (一) 簡介 11 (二) 益生菌之免疫調控 12 (三) 益生菌與皮膚之關係 13 四、代田菌 (乾酪乳酸菌代田株Lactobacillus casei Shirota, LcS) 14 (一) 簡介 14 (二) 代田菌調節免疫的機制 15 五、Dexamethasone 16 第二章 實驗設計與材料方法 18 第一節 乾癬動物模式建立 18 第二節 代田菌培養與定量 20 一、代田菌培養 20 二、菌種保存 20 三、代田菌gDNA萃取與菌數定量 21 第三節 實驗方法 22 一、Psoriasis Area and Severity Index (PASI) score 22 二、背部皮膚組織與耳朵切片染色與量化 23 三、背部皮膚發炎相關細胞激素分析 23 四、腸道菌群分析 24 (一) 樣品 g-DNA萃取 24 (二) Real-time PCR 法分析腸道菌群基因的表現量 25 (三) 計算基因表現量 26 五、淋巴細胞分離與培養 27 (一) 脾臟細胞分離 27 (二) 腸繫膜淋巴結與皮耶氏體細胞分離 28 六、免疫細胞表型分析 29 七、細胞激素含量測定 32 八、統計分析 33 第三章 實驗結果 34 一、代田菌定殖確認 34 二、IMQ對誘發乾癬小鼠攝食量、體重、器官絕對重量與相對重量的影響 35 (一) 攝食量 35 (二) 體重 36 (三) 器官絕對與相對重量 37 (四) 脾臟絕對與相對重量 38 三、補充代田菌對IMQ誘發乾癬小鼠小鼠腸道指標菌群的影響 39 四、補充代田菌對IMQ誘發乾癬小鼠之腸道免疫細胞分泌細胞激素的影響 40 (一) 補充代田菌對IMQ誘發乾癬小鼠之PP細胞分泌細胞激素的影響 40 (二) 補充代田菌對IMQ誘發乾癬小鼠之MLN細胞分泌細胞激素的影響 41 五、補充代田菌對IMQ誘發乾癬小鼠之小鼠脾臟免疫細胞族群的影響 42 六、補充代田菌對IMQ誘發乾癬小鼠之脾臟細胞分泌細胞激素的影響 43 七、補充代田菌對 IMQ 誘發乾癬小鼠之背部皮膚相關細胞激素含量的影響 44 (一) 主要促成乾癬發展之細胞激素 44 (二) 其他促發炎及調節型細胞激素 45 八、補充代田菌對IMQ誘發乾癬小鼠之耳朵與背部皮膚表皮層增厚的影響 46 九、補充代田菌對 IMQ 誘發乾癬小鼠之背部皮膚表徵的影響 48 (一) 第一次誘發乾癬 48 (二) 第二次誘發乾癬 48 (三) 第三次誘發乾癬 50 第四章 討論與結論 51 - 一、 IMQ誘發乾癬小鼠模式之探討 51 二、補充代田菌與重複三次塗抹IMQ誘發乾癬對背部皮膚表徵的影響 53 A. 第一次IMQ誘發 53 B. 第二次IMQ誘發 53 C. 第三次IMQ誘發 53 三、 補充代田菌與IMQ誘發乾癬對小鼠腸道的影響 55 (一) 腸道菌指標分析 55 (二) 腸道細胞激素分析 56 (三) 腸道菌指標與腸道免疫之相關性 57 (四) 腸道免疫與皮膚免疫反應之相關性 58 A. 腸道免疫與皮膚病徵之相關性 58 B. 腸道免疫與皮膚細胞激素之相關性 59 四、 補充代田菌與IMQ誘發乾癬對小鼠全身性免疫反應的影響 60 (一) 脾臟細胞分泌細胞激素之分析 60 (二) 腸道免疫與全身性免疫反應之相關性 61 (三) 脾臟細胞分泌細胞激素之相關性 62 (四) 脾臟細胞分泌細胞激素與皮膚免疫反應之相關性 63 A. 脾臟細胞分泌細胞激素與皮膚病癥之相關性 63 B. 脾臟細胞分泌細胞激素與皮膚細胞激素之相關性 64 五、補充代田菌與IMQ誘發乾癬對背部皮膚的影響 65 (一) 背部皮膚細胞激素含量分析 65 (二) 皮膚細胞激素含量之相關性 66 (三) 皮膚細胞激素含量與皮膚表皮層厚度之相關性 67 (四) 皮膚細胞激素含量與病徵的相關性 68 第二節 結論 69 附錄圖 70 參考文獻 73 圖1-1乾癬發病之免疫機制 5 圖1-2益生菌免疫調控機制 13 圖2-1實驗設計 18 圖2-2代田菌標準曲線 21 圖2-3紅腫與皮屑評分參考標準 22 圖3-1 IMQ誘發乾癬小鼠補充代田菌之盲腸代田菌含量 34 圖3-2補充代田菌對 IMQ 誘發乾癬小鼠之脾臟大小的影響 38 圖3-3補充代田菌對IMQ誘發乾癬小鼠小鼠腸道指標菌的影響 39 圖3-4補充代田菌對IMQ誘發乾癬小鼠之PP細胞分泌細胞激素能力的影響 40 圖3-5補充代田菌對IMQ誘發乾癬小鼠之MLN細胞分泌細胞激素的影響 41 圖3-6補充代田菌對IMQ誘發乾癬小鼠之脾臟中 (a) CD4+ T 細胞 42 圖3-7補充代田菌對IMQ誘發乾癬小鼠之脾臟細胞分泌細胞激素的影響 43 圖3-8補充代田菌對IMQ誘發乾癬小鼠之背部皮膚細胞激素IL-17A、IL-22與IL-23含量的影響 44 圖3- 9補充代田菌對IMQ誘發乾癬小鼠之背部皮膚細胞激素IL-6、TNF-α、IL-1β與IL-10含量的影響 45 圖3-10補充代田菌對IMQ誘發乾癬小鼠之耳朵與背部皮膚表皮增厚程度的影響 47 圖3-11補充代田菌對第1次IMQ誘發乾癬小鼠之背部皮膚紅腫與皮屑程度的影響 48 圖3-12補充代田菌對第2次IMQ誘發乾癬小鼠之背部皮膚紅腫與皮屑程度的影響 49 圖3-13補充代田菌對第3次IMQ誘發乾癬小鼠之背部皮膚紅腫與皮屑程度的影響 50 圖4-1代田菌藉由調節腸道菌相與免疫調節作用減緩IMQ誘發乾癬小鼠病情之可能機制圖 69 表2-1 AIN-93飼料成分 19 表2- 2 LLV培養基成分 20 表2- 3 本研究使用的引子對 25 表2- 4本研究所使用的引子對PCR反應條件 25 表3-1 IMQ誘發乾癬小鼠之攝食量 35 表3-2 IMQ誘發乾癬小鼠之體重變化 36 表3-3 補充代田菌對IMQ誘發乾癬小鼠之絕對與相對器官重量的影響 37 表4-1補充代田菌及IMQ誘發乾癬對小鼠PASI score的影響 54 表4-2補充代田菌及IMQ誘發三次乾癬之PASI score比較 54 表4-3補充代田菌及IMQ誘發乾癬對小鼠腸道菌相的影響 55 表4-4補充代田菌及IMQ誘發乾癬影響小鼠腸道免疫細胞分泌細胞激素之整理表 56 表4-5 IMQ誘發乾癬小鼠之腸道菌指標與MLN分泌細胞激素之相關性 57 表4-6 IMQ誘發乾癬小鼠之MLN分泌細胞激素與與乾癬病徵的相關性 58 表4-7 IMQ誘發乾癬小鼠之MLN細胞激素含量與皮膚細胞激素含量的相關性 59 表4-8補充代田菌及IMQ誘發乾癬影響小鼠脾臟細胞分泌細胞激素之整理表 60 表4-9 IMQ誘發乾癬小鼠之MLN細胞與脾臟細胞分泌細胞激素之相關性 61 表4-10 IMQ誘發乾癬小鼠之脾臟細胞分泌細胞激素之相關性 62 表4-11 IMQ誘發乾癬小鼠之脾臟細胞激素含量與乾癬病徵的相關性 63 表4-12 IMQ誘發乾癬小鼠之脾臟細胞激素含量與皮膚細胞激素的相關性 64 表4-13補充代田菌及IMQ誘發乾癬對小鼠背部皮膚細胞激素含量的影響 65 表4-14 IMQ誘發乾癬小鼠之皮膚細胞激素含量的相關性 66 表4-15 IMQ誘發乾癬小鼠之皮膚細胞激素含量與耳朵、背部皮膚表皮層厚度的相關性 67 表4-16 IMQ誘發乾癬小鼠之皮膚細胞激素含量與乾癬病徵的相關性 68 | |
dc.language.iso | zh-TW | |
dc.title | 益生菌 Lactobacillus casei Shirota 對 imiquimod 誘發乾癬小鼠之免疫調節的影響 | zh_TW |
dc.title | Effects of Lactobacillus casei Shirota on immunoregulation in imiquimod-induced psoriasis-like mice | en |
dc.type | Thesis | |
dc.date.schoolyear | 110-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 江伯倫(Bor-Luen Chiang),江孟燦(Meng-Tsan Chiang),陳俊任(Chun-Jen Chen),何佳安(Ja-An Ho) | |
dc.subject.keyword | 乾酪乳酸菌代田株,咪喹莫特,自體免疫疾病,腸道黏膜免疫, | zh_TW |
dc.subject.keyword | Lactobacillus casei Shirota,Imiquimod,autoimmune disorders,intestinal mucosal immune system, | en |
dc.relation.page | 82 | |
dc.identifier.doi | 10.6342/NTU202202799 | |
dc.rights.note | 同意授權(限校園內公開) | |
dc.date.accepted | 2022-08-25 | |
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 | 2022-08-30 | - |
顯示於系所單位: | 生化科技學系 |
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