牛樟芝為台灣所公認之民間藥物，具有護肝與解毒療效，而目前牛樟芝之護肝與解毒機制未明。因此本實驗以油膜包覆之新劑型牛樟芝萃取物為實驗樣品，探討油膜包覆之新劑型，其對四氯化碳誘導 SD 大鼠肝臟急性損傷之影響，以進行測試牛樟芝小分子新劑型抗發炎之研究。
本研究主要是以四氯化碳 (CCl4；Carbon tetrachloride) 在雄性鼠急性肝臟損傷模式，探討油膜包覆之新劑型牛樟芝萃取物 (NAE, New Antrodia cinnamomea ) 保護肝臟所引發的大鼠急性肝損傷之影響。實驗 SD 雄性鼠，經口腔餵食 NAE (10 mg/kg；50 mg/kg)，再予以腹腔注射四氯化碳 40% (1 ml/kg) 逆境處理後，分別測試血清與與肝臟組織中的促發炎細胞激素與一氧化氮含量。依不同時間抽血測試、天門冬胺酸轉胺酶 (aspartate aminotransferase, AST, SGOT)、丙胺酸轉胺酶 (alanine aminotransferase, ALT, SGPT)、介白素-1 (interleukin-1, IL-1)、介白素-6 (interleukin-6, IL-6)、活性氧 (reactive oxygen species, ROS)、與一氧化氮 (nitric oxide, NO) 的含量，以探討牛樟芝小分子對於急性肝損傷的保護作用程度；並且進行大鼠肝臟的 MAPKs、Caspase-3與 COX-2 等訊號傳遞蛋白質表現量測定，以了解牛樟芝小分子對於急性肝損傷的保護作用機制。
實驗結果證實，在投予四氯化碳注射後，血清一氧化氮濃度 (NO) 明顯上升，實驗組投予油膜包覆之新劑型牛樟芝萃取物的 NO 濃度明顯降低，且具有顯著性差異 (P<0.05)，在血清活性氧化自由基物質 (Reactive oxygen species, ROS) 分析， SD 大鼠投予四氯化碳後，在血清中 ROS 明顯上升；在實驗組投予 NAE 的 SD 大鼠， ROS 值較逆境對照組為低且有顯著性差異 (P<0.05) 。肝臟組織經 (High Performance Liquid Chromatography；HPLC) 高效能液向層析法分析，在實驗組發 NAE 之脂溶性小分子，並能證實 NAE 能透過小腸吸收直接達大鼠的肝臟。
由實驗結果顯示，NAE 明顯降低大鼠血清中的促發炎因子：NO、ROS、、AST、ALT、IL-1 與 IL-6 含量與肝臟組織中誘導、第二型環氧化酶 (COX-2) 的蛋白質表現量。因此推斷NAE藉由抑制大鼠的肝臟 p-ERK、Caspase-3 與COX-2 表現量，進而降低肝臟發炎的程度，達到保護肝臟的功能。NAE 於四氯化碳誘導逆境實驗證實，對 SD 大鼠抗氧化與解毒代謝能力增強，並可降低肝臟氧化及發炎程度。以上結論，NAE 具有保護肝臟作用。

The Antrodia cinnamomea are recognized as a folk medicine in Taiwan, detoxification effect with liver, however its mechanisms of detoxification until not clear. Therefore, the present experiment that we test the small molecule isolation of the Antrodia cinnamomea extract with new formulations of film-coated capsule, analysis its liver anti-inflammatory effect of the acute injury SD rats subjected to carbon tetrachloride (CCl4).
The experimental results demonstrate that administration of carbon tetrachloride after injection, SD rats serum concentrations of nitric oxide (NO) and reactive oxygen species (ROS) are significantly increased on the CCl4 group but the NO and ROS concentration significantly reduced on the Antrodia cinnamomea group (P <0.05). We also find the small molecule isolation compound of the Antrodia cinnamomea extract from the liver tissues of the Antrodia cinnamomea group SD rats by HPLC method.
The small molecule isolation of the Antrodia cinnamomea with new formulations could decrease the liver damage of SD rat subjected to CCl4.
Taken together, these results indicate that the mechanism of new formulations the Antrodia cinnamomea extract involves the antioxidant metabolism and detoxification capacity through scavenging of NO and ROS in CCl4-stressed SD rat.

第一章 緒論 13
1.1 發炎反應 13
1.2 發炎反應相關轉錄因子 15
1.2.1 一氧化氮 15
1.2.2 介白素 17
1.2.3 前列腺素 19
1.2.4 MAPKs路徑 21
1.2.5 環氧化酶 23
1.3 四氯化碳 (CCl4；Carbon tetrachloride) 28
1.4 牛樟芝 (Antrodia cinnamomea) 30
1.5 牛樟芝小分子 (small molecules) 33
1.5.1 多醣體 ( Polysaccharides ) 33
1.5.2 三萜類 ( Triterpenoids ): 34
1.6 研究目的 36
第二章 實驗材料與方法 37
2.1 儀器與設備 37
2.2 實驗藥品與材料 39
2.3 實驗步驟 41
2.3.1 實驗動物 41
2.4 動物實驗研究架構 42
2.5 樣品採集與前處理 43
2.6 一氧化氮含量之測定 44
2.7 清除活性氧化物能力測定 45
2.8 血液中生化值測定 46
2.9 ALT測定原理 46
2.10 AST測定原理 48
2.11 促發炎之激素 49
2.12 介白素-1β；6含量之測定 50
2.13 前列腺素E2含量之測定 53
2.14 肝臟組織蛋白質之萃取 55
2.15 肝臟組織特異性蛋白質含量之測定 56
2.16 肝組織病理切片觀察 64
第三章 結果 66
3.1 AE 與 NAE 各組織生物活性分析 66
3.2 AE 與 NAE 對 CCl4 誘導 SD 大鼠血清產生 NO 之影響 67
3.3 AE 與 NAE 對 CCl4 誘導 SD 大鼠血清清除 ROS 能力之影響 69
3.4 AE 與 NAE 對 CCl4 誘導 SD 大鼠血清產生 ALT 活性檢測 71
3.5 AE 與 NAE 對 CCl4 誘導 SD 大鼠血清產生 AST 活性檢測 72
3.6 AE 與 NAE 對 CCl4 誘導 SD 大鼠血清產生 IL-1β 之影響 73
3.7 AE 與 NAE 對 CCl4 誘導 SD 大鼠血清產生 IL-6 之影響 75
3.8 AE 與 NAE 對 CCl4 誘導 SD 大鼠血清產生 PGE2 之影響 77
3.9 AE 與 NAE 對 CCl4 誘導 SD 大鼠下分子機制之路徑 79
3.10 AE 與 NAE 對 CCl4 誘導 SD 大鼠產生肝損傷之影響 82
第四章 討論 84
第五章 結論 89

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