目 錄
頁次
正文目錄 ................................................................................................. I
圖目錄 .................................................................................................... V
中文摘要 ............................................................................................... VI
英文摘要 ............................................................................................ VIII
第一章、緒言 ......................................................................................................... 1
第一節 慢性腎臟病（Chronic Kidney Disease, CKD） ....................................... 1
壹、慢性腎臟病之定義與免疫複合物所引起之腎炎 ............................................. 1
貳、IgA腎病變（IgA nephropathy, IgAN） ............................................................ 2
參、 狼瘡腎炎（Lupus nephritis, LN）....................................................................... 2
肆、 IgA腎病變及狼瘡腎炎之治療............................................................................ 3
第二節IgA腎病變及狼瘡腎炎小鼠模式................................................................... 4
壹、誘發型IgA 腎病變小鼠模式............................................................................... 4
貳、加速惡化型狼瘡腎炎（Accelerated and severe lupus nephritis, ASLN）及自發型狼瘡腎炎（Spontaneous LN, spLN）小鼠模式..................................................... 4
第三節NLRP3 發炎體（NLR family pyrin domain containing 3 inflammasome, NLRP3 inflammasome） ......................................................................................... 5
第四節 自噬作用（Autophagy） ........................................................................... 5
第五節Sirtuin (SIRT) 1 和SIRT3 .......................................................................... 5
第六節 Tris (dibenzylideneacetone) dipalladium（Tris DBA）………………... 6
第七節 研究動機與目的............................................................................................ 6
第二章、 材料與方法
第一節 Tris DBA 取得及製備 ................................................................................. 7
第二節 製備IgA 抗體及PnC 抗原 ........................................................................ 7
第三節 動物模式及實驗設計 ................................................................................... 8
壹、誘發型IgA 腎病變小鼠模式 ............................................................................ 8
貳、加速惡化型狼瘡腎炎（Accelerated and severe lupus nephritis, ASLN）及自發型狼瘡腎炎（Spontaneous LN, spLN）小鼠模式 ................................................... 8
第三節 腎功能及蛋白尿測定 ................................................................................... 9
第四節 腎臟病理學分析 ......................................................................................... 10
第五節 免疫組織化學染色及免疫螢光分析 ......................................................... 10
第六節 西方墨點轉漬法 ......................................................................................... 11
第七節 抽取腎臟組織核糖核酸及即時聚合酶鏈鎖反應 .............................................................................................................................. 12
第八節 流式細胞儀分析 ........................................................................................ 12
第九節 T 細胞增生測定 ........................................................................................ 13
第十節 調控型T細胞體外抑制試驗 .................................................................... 13
第十一節 LPS / ATP誘發的急性腹膜炎 .............................................................. 14
第十二節 細胞培養.................................................................................................. 15
壹、巨噬細胞株（J774A.1）.................................................................................. 15
貳、骨髓分化之樹突狀細胞（Bone marrow derived dendritic cells, BMDCs）
第十三節 活性氧化物測定 ................................................................................... 16
第十四節 小鼠血清或細胞上清液中IL-1β，TNF-α，IL-6和抗dsDNA自體抗體濃度之偵測 ......................................................................................................... 16
第十五節 數據分析 ............................................................................................... 16
第三章、結果 ......................................................................................................... 17
第一節 Tris DBA治療IgA腎病變小鼠模式 ...................................................... 17
壹、Tris DBA治療IgA腎病變小鼠之腎功能，蛋白尿和腎臟病理分析................................................................................................................................ 17
貳、Tris DBA抑制IgA腎病變小鼠腎臟組織中的NLRP3發炎體之活化並促進自噬作用 .......................................................................................................................17
參、Tris DBA抑制腎臟組織和巨噬細胞中的ROS產生 .................................... 18
肆、Tris DBA抑制了巨噬細胞中的NLRP3發炎體的活化 ............................... 19
伍、Tris DBA促進SIRT1和SIRT3所引發之自噬作用來抑制了NLRP3發炎體的活化............................................................................................................................ 20
第二節 Tris DBA治療加速惡化型狼瘡腎炎及自發型狼瘡腎炎小鼠模式 ...............................................................................................................................21
壹、Tris DBA治療加速惡化型狼瘡腎炎小鼠模式之腎功能，蛋白尿和腎臟病理分析 ...............................................................................................................................21
貳、Tris DBA條控加速惡化型狼瘡腎炎小鼠模式之T細胞及B細胞活化 .... 22
參、Tris DBA抑制了BMDC的活化以及BMDC所導致的輔助性T細胞增殖/分化............................................................................................................................... 22
肆、在ASLN小鼠中Tris DBA透過促進自噬作用來抑制的NLRP3發炎體活化 ............................................................................................................................. 23
伍、Tris DBA抑制ASLN小鼠的ROS生成並增強其抗氧化活性 ............................................................................................................................ 24
陸、Tris DBA抑制MAPK（ERK，JNK）所導引的NLRP3發炎體的第一訊型路徑活化...................................................................................................................... 25
柒、自噬作用與NLRP3發炎路徑的交互作用.................................................... 25
捌、Tris DBA治療自發型狼瘡腎炎（Spontaneous LN, spLN）小鼠模式....... 26
第四章、討論 ....................................................................................................... 28
第一節 Tris DBA 於IgA 腎病變的治療效果 .................................................. 28
第二節 Tris DBA 於加速惡化型及自發型狼瘡腎炎的治療效果 ...................28
第五章、結論 ..................................................................................................... 29
第六章、參考文獻 ............................................................................................. 30
圖 ........................................................................................................................ 36
附錄 .................................................................................................................... 49
 
圖目錄
頁次
圖 1、Tris DBA顯著改善IgA腎病變之腎功能、腎臟組織形態學及巨噬細胞和T細胞浸潤................................................................................................................. 36
圖2、Tris DBA促進IgA腎病變腎臟自噬作用並抑制NLRP3發炎體的活化..37
圖3、Tris DBA 抑制IgA腎病變之氧化壓力並促進抗氧化路經的活化………38
圖4、Tris DBA 抑制巨噬細胞中MAPK (ERK, JNK)所導引之NLRP3第一訊息路徑之活化................................................................................................................. 39
圖5、Tris DBA 經由自噬作用抑制NLRP3發炎體的活化................................. 40
圖6、Tris DBA 透過SIRT1和 SIRT3所誘發之自噬作用來抑制巨噬細胞中NLRP3發炎體之活化............................................................................................... 41
圖7、Tris DBA顯著改善加速惡化行狼瘡腎炎之腎功能、腎臟組織形態學....... 42
圖8、Tris DBA抑制T cell功能及骨髓所分化之樹突狀細胞活化和樹突狀細胞所導引之輔助型T細胞之增生及分化......................................................................... 43
圖9、Tris DBA促進加速惡化型狼瘡腎炎腎臟自噬作用並抑制NLRP3發炎體的活化..............................................................................................................................44
圖10、 Tris DBA 抑制加速惡化行狼瘡腎炎腎臟氧化壓力並促進抗氧化路徑的活化..............................................................................................................................45
圖11、Tris DBA 抑制巨噬細胞中MAPK (ERK, JNK)所導引之NLRP3發炎體第一訊息路徑之活化..................................................................................................46
圖12、Tris DBA 調控巨噬細胞中自噬作用與NLRP3發炎體之交互作用........47
圖13、Tris DBA顯著改善自發性NZB / W F1小鼠，狼瘡腎炎疾病進展/嚴重程度..................................................................................................................................48

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