臺灣博碩士論文加值系統

發炎性腸病是一種重複發生的慢性腸道發炎疾病。研究指出發炎性腸炎會顯著增加大腸癌的風險，目前已知發炎性腸炎與生活習慣以及飲食有密切的關係。研究指出牛樟芝具有抗氧化、抗發炎以及保肝功效。但牛樟芝對於發炎性腸炎的抗發炎功效尚未明朗。首先本研究利用皿培式牛樟芝子實體利用水萃 (ACP) 與乙醇萃取 (ACE) 的產物，針對 LPS 誘導產生發炎反應的巨噬細胞 RAW264.7 觀察抑制NO、TNF-、IL-6 以及 PGE2 的作用，IL-6 與 iNOS 的 mRNA 的表現也有因 ACP 與 ACE 而降低表現。動物實驗以發炎性腸炎的模式， C57BL/6 小鼠給予含硫酸葡聚醣鈉 (3% DSS) 的飲用水誘導十四天，在後七天管餵牛樟芝粉末，DSS 造成粘膜水腫和血便促使腸道縮短並增加腸重量與長度的比值以及增加發炎反應。牛樟芝能夠減緩發炎性腸炎藉由抑制發炎激素的產生。大腸癌模式中，腹腔注射致癌劑 azoxymethane (AOM) ，再以 2% DSS重複誘導腸道發炎，結果顯示牛樟芝具有防止腫瘤的生成。綜合上述結果牛樟芝可以減緩 DSS 誘導的傷害並且在發炎性腸炎與腸癌中可能具有治療的功用。

Inflammatory bowel diseases (IBD) are associated with chronic relapsing inflammation of the intestinal tract. IBD are at increased risk of developing CRC since it is now known that 90–95% of the cause is attributable to lifestyle and diet. Many studies have confirmed that Antrodia cinnamomea have antioxidant and anti-inflammatory and protect liver property. But, anti-inflammatory effects of AC in IBD have not been investigated. First,we use petri dish cultured AC fruiting body water extract (ACP) and ethanol extract (ACE) was assessed via inhibitory effect of nitric oxide (NO) and tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) and prostaglandin E2 (PGE2) in lipopolysaccharide (LPS) induced RAW 264.7 macrophage cells.The mRNA expression levels of interleukin-6 (IL-6) and inducible NO synthase (iNOS) were reduced by the addition of ACP and ACE in a dose-dependent manner. Second , we used dextran sulfate sodium (DSS) (3% W/V) in drinking water fourteen days to induce male C57BL/6 mice colitis, and oral AC powder to mice for last seven days. DSS produced shrinkage of colon length and increased the relative colon weight/length ratio and inducing inflammatory response. AC attenuated the severity of DSS-induced IBD,reducing pro-inflammatory cytokine. In the colitis-associated colorectal cancer (CRC) model, mice were treated with 3 cycle DSS (2% W/V) for colitis and intraperitoneal injection of azoxymethane (10 mg/kg AOM), AC could be in prevent of CRC in the AOM-DSS model. The result shows that AC mitigated the injurious effects of DSS and inhibited pro-inflammatory cytokine and reduced tumor number. These results suggest that AC could possibly have improve injurious effects of DSS and that it may have therapeutic value in the setting of IBD and CAC.

目錄
謝誌 I
中文摘要 III
英文摘要 IV
名詞縮寫 V
目錄 VI
圖表目錄 XI
壹、序論 1
貳、文獻回顧 3
一、大腸直腸癌 (Colorectal cancer) 3
(一)分佈與症狀 3
(二)成因與因子 4
二、發炎性腸炎與大腸直腸癌的關係 5
三、發炎性腸炎 (Inflammatory bowel disease) 6
(一)潰瘍性腸炎 (Ulcerative colitis) 6
(二)克隆氏症 (Crohn's disease) 6
四、發炎性腸癌與發炎反應 7
(一)發炎反應 (Inflammatory response) 7
(二)活性氧 (Reactive Oxygen Species;ROS) 傷害 8
(三)環氧合酶 (Cyclooxygenase;COX) 9
(四)一氧化氮合成酶 (Nitric oxide synthase;NOS) 10
(五)NF-B 11
(六)細胞激素 11
(一)TNF- 11
(二)IL-6 12
(三)IL-1 13
五、惡病質 13
(一)成因 13
(二)急性期反應 15
六、牛樟芝 15
(一)牛樟芝 15
(二)命名由來 16
(三)牛樟芝分類 17
(四)生理活性 17
1.抗氧化活性 17
2.抗病毒活性 19
3.抑制腫瘤 19
4.護肝活性 21
5.降血脂 22
6.增強免疫活性 23
参、實驗設計 25
(一)Inflammatory bowel disease model 27
(二)Colitis-associated colon cancer model 28
肆、材料方法 29
一、實驗材料 29
(一)細胞株 29
(二)樣品 29
(三)藥品 29
1.細胞培養 29
2.細胞存活率測定 30
3.胞內活性氧分析 30
4.吞噬球呼吸爆 30
5.反轉錄酶鏈反應分析(RT-PCR) 30
6.動物 31
7.飼料來源 31
8.冷凍切片染色 31
9.血液細胞激素測定 32
10.急性蛋白 32
11.脾臟細胞分離 32
12.脾臟細胞增殖反應 32
(四)儀器設備 32
二、實驗方法 34
(一)樣品製備 34
(二)抗發炎分析 34
1.RAW264.7培養 34
2.細胞存活率測定 35
3.RT-PCR分析 iNOS 和 IL-6 36
3.細胞激素測定 39
4. 吞噬球呼吸爆 (Respiratory burst) NBT還原分析 40
5.細胞內活性氧分析 41
6.前列腺素E2測定 42
(三)、牛樟芝減緩動物發炎性腸病及腸癌實驗 42
1. 實驗動物之飼養 42
2. 動物實驗組別設定 43
3. 取得老鼠下腔大動脈血及其他臟器 43
4. 疾病指數的評分 44
5. 小鼠脾臟細胞之取得 44
6. 小鼠腸組織之處理 44
7. 血清之脂質過氧化影響 45
8. C 反應蛋白測定 45
9. 淋巴球增生數量測定 (MTT Assay) 46
10. 一氧化氮測定 46
(四)、統計方法 46
伍、結果與討論 47
陸、結論 58
柒、參考文獻 60
捌、圖表 74
玖、附錄 104

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