臺灣博碩士論文加值系統

發炎疾病是世界性之重大醫療衛生主題，勢必對國軍健康具潛在之威脅，進而危及我國的國防戰力，因而建立有效之預防及治療策略勢在必行。目前已知NLRP3發炎體主要參與先天性免疫反應過程。NLRP3發炎體由NLRP3、ASC以及procaspase-1三種蛋白所構成，活化後使發炎細胞激素包括IL-1及IL-18釋放，加重發炎反應。
LHK小分子是阿斯匹靈衍生物，可經由腸道吸收，並具有多種藥理特性，包含治療發燒、疼痛及風濕等作用。然而LHK小分子對發炎相關疾病是否具有明確療效，仍尚待研析。因此本研究利用 [1]細胞模式：J774A.1巨噬細胞； [2]動物模式：加速惡化型狼瘡腎炎小鼠模式，探討LHK小分子是否可以抑制NLRP3發炎體相關路徑，進而改善小鼠發炎疾病。
結果顯示： [1]細胞模式：LHK小分子有效抑制脂多醣 (Lipopolysaccharides, LPS)誘導巨噬細胞NLRP3發炎體活化之相關路徑及促進自噬作用相關路徑； [2]動物模式：LHK小分子有效降低狼瘡腎炎小鼠腎臟損傷，並降低相關發炎激素之表現。綜合以上，LHK小分子具有潛力成為抑制發炎之候選藥物，但需進一步探討其關鍵之生效機轉，希望未來發展為臨床治療策略或做為輔佐藥物之應用。

As the proverb says, the power of eyes is the power of country. Among the three armies, the Air Force has the most pressure-bearing capacity. In view of this, The Air Force has become the cornerstone of the Taiwan Strait stable and peace. Inflammatory diseases are a major topic theme all over the world. It is a potential threat to our military, and then endanger our national defense capability. Therefore, it is necessary to establish effective prevention and treatment strategies. In present study, we reported that LHK, a derivative from aspirin, proved to bear potent anti-inflammatory property to inflammatory-related renal disorder. The process of inflammation usually regulated by different molecular proteins and signal pathways. Macrophages play as an important role in the progression of inflammation, including antigen presentation, phagocytosis, immune modulation and cytokines production. It has been known that NLRP3 inflammasome initiates an inflammatory form of cell damages and mediates proinflammtory cytokines production, such as IL-1 and IL-18. Herein, we validated its therapeutic effects of LHK on the progression of an inflammatory-related renal injury, using an accelerated and severe LN (ASLN) mousse model and murine macrophage cell line. The results showed that LHK significantly increased induction of autophagy in activated murine macrophage cells treated with the compound. LHK substantially inhibited priming and activating signals of NLRP3 inflammasome in murine macrophage cells treated with the compound. Furthermore, LHK dramatically improved renal function and renal conditions in ASLN mice treated with LHK, including glomerular sclerosis, proliferation, renal mononuclear lymphocyte infiltration. These findings justify its potential of LHK as a drug candidate for the progression of LN.

目錄
正文目錄 Ⅰ
圖目錄 III
英文摘要 IV
中文摘要 V
第一章、緒論 5
一、 發炎之免疫角色與重要性 5
二、 巨噬細胞 (Macrophage) 6
三、 NLRP3發炎體 (NLR family pyrin domain containing 3 inflammasome, NLRP3 inflammasome ) 7
四、 自噬作用 (Autophagy) 9
五、 狼瘡腎炎 (Lupus nephritis, LN) 11
六、 LHK小分子 12
七、 發炎疾病與航太醫學之關聯重要性 13
第二章、研究動機與目的 14
第三章、材料與方法 15
一、 LHK小分子之取得 15
二、 細胞培養 15
三、 酵素連結免疫吸附分析法 （Enzyme-linked immunosorbent assay, ELISA） 15
四、 西方墨點法（Western blot analysis） 16
五、 動物模式：加速惡化型狼瘡腎炎小鼠模式 (Acceleration and Severe Lupus Nephritis, ASLN) 17
六、 腎臟病理組織型態學 17
七、 免疫組織化學染色 18
八、 免疫沉澱法 (Immunoprecipitation) 19
九、 統計分析 19
第四章、結果 20
壹、LHK小分子抑制巨噬細胞之NLRP3發炎體活化相關訊息路徑 20
(1) LHK小分子抑制巨噬細胞受脂多醣刺激後，細胞上清液IL-1表現 (圖三) 20
(2) LHK小分子抑制巨噬細胞受脂多醣刺激後，細胞上清液proIL-1 及 IL-1表現 (圖四) 20
(3) LHK小分子抑制巨噬細胞受脂多醣刺激後，細胞上清液procaspase-1及Caspase-1表現 (圖五) 21
(4) LHK小分子抑制巨噬細胞受脂多醣刺激後，細胞上清液NLRP3、ASC 及 IL-18蛋白表現 (圖六) 21
(5) LHK小分子抑制巨噬細胞MAPKs訊息路徑活化 (圖七) 22
(6) LHK小分子抑制巨噬細胞IB訊息路徑活化 (圖八) 23
(7) LHK小分子抑制巨噬細胞受脂多醣刺激後，NLRP3蛋白與ASC蛋白結合 (圖九) 23
貳、LHK小分子促進巨噬細胞之自噬作用活化相關訊息路徑 24
(1) LHK小分子促進巨噬細胞自噬作用的表現 (圖十) 24
(2) 以自噬作用抑制劑驗證LHK小分子促進巨噬細胞自噬作用 (圖十一) 25
參、LHK小分子改善加速惡化型狼瘡腎炎小鼠腎臟損傷 25
(1) LHK小分子改善加速惡化型狼瘡腎炎腎臟損傷 (圖十二) 25
(2) LHK小分子改善加速惡化型狼瘡腎炎小鼠腎臟CD3+T細胞浸潤 (圖十三) 26
(3) LHK小分子改善加速惡化型狼瘡腎炎小鼠腎臟F4/80+細胞浸潤(圖十四) 26
第五章、討論 27
壹、發炎疾病與軍陣醫學相關性 27
貳、LHK小分子與發炎疾病之應用 28
第六章、結論 31
第七章、參考文獻 32

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