臺灣博碩士論文加值系統

發炎性腸道疾病如今在其他新興工業化地區的發病率症在上升，台灣也不例外。IBD患者的睡眠品質與他們的發炎狀態緊密相連。目前有許多研究探討了睡眠對IBD的影響，但尚且缺乏針對不同程度的睡眠間斷對IBD病情的量化研究。因此，本研究將專注於探討這一關聯。
本研究使用40隻10週大的C57BL雄性小鼠。以2%，36-50kDa的葡聚糖硫酸鈉（DSS）誘發小鼠潰瘍性結腸炎。實驗分成五組，分別是對照組、高頻率睡眠剝奪組、DSS組、低頻率睡眠剝奪加DSS組與高頻率睡眠剝奪加DSS組。本研究發現單純高強度睡眠剝奪會導致發炎與腸道屏障受損。結腸炎的情況下，再加上睡眠剝奪會使疾病的嚴重程度加劇，然而嚴重程度與睡眠剝奪的強度卻不是線性關係。在小鼠體重、腸道長度與腸道屏障上，高頻率睡眠剝奪加DSS組影響較為劇烈；而在組織學分數、腸道菌叢與短鏈脂肪酸上，低頻率睡眠剝奪加DSS組損害較為嚴重。然而，還需要後續研究來探討背後的機轉與原因。

The incidence of inflammatory bowel disease （IBD） is rising in newly industrialized regions, including Taiwan. IBD patients' sleep quality is closely linked to their inflammatory state. Although many studies have explored sleep's impact on IBD, research on the effects of varying levels of sleep disruption is lacking. This study addresses this gap.
Using 40 male C57BL mice aged 10 weeks, we induced ulcerative colitis with 2% dextran sulfate sodium （DSS）. The mice were divided into five groups: control, high-frequency sleep deprivation, DSS, low-frequency sleep deprivation plus DSS, and high-frequency sleep deprivation plus DSS. Results showed that high-intensity sleep deprivation alone causes inflammation and intestinal barrier damage. When combined with colitis, sleep deprivation worsens the disease, but not in a linear relationship to its intensity. The high-frequency sleep deprivation plus DSS group had more severe effects on body weight, intestinal length, and barrier integrity. In contrast, the low-frequency sleep deprivation plus DSS group showed greater damage in histological scores, gut microbiota, and short-chain fatty acids. Further research is needed to understand the underlying mechanisms.

目 錄
中文摘要 i
Abstract ii
目 錄 iii
圖 目 錄 v
表 目 錄 vii
第一章 緒論 1
1-1發炎性腸炎之介紹 1
1-2 IBD與睡眠的雙向關係—發炎性腸道疾病造成睡眠問題 3
1-3 IBD與睡眠的雙向關係—睡眠問題加重發炎性腸道疾病 5
1-4 IBD、睡眠與腸道微生物 7
1-5腸腦軸 9
1-6不同層次的睡眠剝奪對IBD之影響 10
第二章 研究目的 13
第三章 實驗方法 15
3-1實驗動物 15
3-2睡眠剝奪裝置及光照/黑暗時間 16
3-3潰瘍性結腸炎的模型 17
3-4實驗分組 18
3-5實驗結果評估 19
3-6統計分析 25
第四章 結果 26
4-1 生理變化與臨床指標 26
4-2 腸道巨觀變化 33
4-3 腸道組織學變化 34
4-4 腸道免疫化學染色 35
4-5生化標記物 37
4-6 流式細胞儀分析 45
4-7 腸道微生物與短鏈脂肪酸分析 46
第五章 討論 63
第六章 結論 73
參考資料 74
 
圖 目 錄
圖一：小鼠睡眠碎片化裝置 17
圖二：實驗流程 19
圖三：小鼠體重變化表 26
圖四：小鼠每日攝食量 28
圖五：小鼠每日飲水量 30
圖六：小鼠疾病活動指數變化 32
圖七：小鼠腸道長度 34
圖八：小鼠組織學分數與腸道切片 35
圖九：小鼠的腸道屏障指標 36
圖十：小鼠腸道之血管活性肽指標 37
圖十一：小鼠之CRP、GM-CSF與G-CSF含量 39
圖十二：小鼠Eotaxin、KC和MCP-1之含量 40
圖十三：小鼠IL-1α、IL-1β、IL-6與IL-17之含量 42
圖十四：小鼠IL-12（70）、IFN-γ之含量 43
圖十五：小鼠IL-4、IL-9之含量 44
圖十六：流式細胞儀分析結果 46
圖十七：腸道微生物α diversity 48
圖十八：腸道菌稀釋曲線 49
圖十九：腸道微生物排序階梯圖 51
圖二十：腸道微生物UpSet plot分析圖 52
圖二十一：Weighted UniFrac 53
圖二十二：β diversity中的PCA圖 55
圖二十三：NMDS分析圖 56
圖二十四：科階層上的物種相對豐度柱狀圖 57
圖二十五：各組小鼠腸道內厚壁菌門與擬桿菌門豐富度比例 58
圖二十六：各組小鼠腸道內物種群落的顯著性差異 60
圖二十七：進化分支圖及 LDA 值分布柱狀圖 61
圖二十八：各組小鼠腸道短鏈脂肪酸的含量 62

表 目 錄
表一：體重減輕評分表 20
表二：糞便稠度評分表 20
表三：組織發炎評分表 22

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