臺灣博碩士論文加值系統

系統性紅斑性狼瘡 (SLE) 是一種多器官遭受破壞的自體免疫性疾病，除了免疫系統的調控失常外，並且伴隨著多發性的自體免疫抗體產生。DHEA 是一種腎上腺皮質所產生的類固醇，具有少許雄性素的活性。有文獻報告 DHEA 可以改善 NZB/W F1 小鼠在腎炎上的症狀，改變細胞激素的分泌，並延長生命。而細胞激素在紅斑性狼瘡的致病過程中，具有重要的角色。在本實驗中，我們以 DHEA 餵食 NZB/W F1 小鼠後，觀察在血液中、腎臟和脾臟內表現細胞激素的細胞。小鼠餵食 DHEA 後，在周邊血液單核細胞所表現的細胞激素 mRNA 會受到改變，鼠齡 8 個月大時，DHEA 組的 IL-6、IL-10囥M IFN-γ較控制組有明顯的降低。在鼠齡 3個月至 6個月大期間，細胞激素受到 DHEA 的影響不大。在鼠齡 10 個月，腎臟的蘇木紫伊紅染色結果，可在腎小動脈發現免疫細胞的浸潤情況和腎絲球體萎縮的現象。若以冷凍切片，免疫組織染色觀察組織表現的細胞激素 IL-10、TNF-α和 IFN-γ，DHEA 組在腎臟的浸潤細胞表現 IL-10、TNF-α 數目減少，IFN-γ出現的時間提前。在脾臟的染色結果，鼠齡 8、10、12 個月大時，控制組都有表現 IL-10、TNF-α 和 IFN-γ，DHEA 組表現這三種的細胞激素數目較少。用抗 TCR β chain 和 IgG 抗體區分 T 細胞和 B 細胞，後再用染色法觀察這些細胞所表現的三種細胞激素。在腎臟內 IgG+ B 細胞會表現 IL-10 和 TNF-α，部份 T 細胞會產生 TNF-α。在脾臟的一些 T 細胞會表現 IL-10、TNF-α和 IFN-γ，IgG+ B 細胞會表現 IL-10。另外，以時間來看，在脾臟表現細胞激素的細胞出現時間較在腎臟的細胞早。在血液中的細胞，IgM+ B 細胞會產生IL-10，T 細胞產生 IFN-γ，TNF-α 可能由單核球所產生。綜合以上的結果，在 NZB/W F1 小鼠的脾臟和腎臟產生 IL-10 的細胞以 B 細胞為多數，產生 TNF-α 和 IFN-γ的細胞，有一部份是由 T 細胞產生而來。DHEA 可提早浸潤細胞製造 IFN-γ出現的時間，能降低腎臟和脾臟表現 IL-10 和 TNF-α 的細胞數目。而這些細胞對 SLE 的致病原因應佔有重要的角色。

Systemic lupus erythematosus (SLE) is a multisystem disease characterized by disturbances in the immune system associated with the production of autoantibodies to self-antigens. NZB/W F1 mice, which spontaneously develop a lupus-like syndrome characterized by an increased level of autoantibodies in old mice, is a mode for SLE. Dehydroepiandrosterone (DHEA), an abundant adrenal steroid with limited intrinsic androgenic activity, was shown to ameliorate nephritis in NZB/W F1 mice. Cytokines might play an important roles in the development of SLE. IL-10 and IL-6 are produced at high levels of patients with SLE. In this study, we investigated cytokine-producing cells in kidney, spleen and blood lymphocytes of NZB/W F1 mice after DHEA treatment.
Infiltrating cells in the renal arteriole and around sites showing glomerulus atrophy were observed by H&E staining in kidney at 10 month-old. Some of these cells expressed IL-10, TNF-a or IFN-g. DHEA could decrease the expression number of IL-10 and TNF-a in infiltrating cells, early expression of IFN-g in kidney as detected by immunohistochemical staining. IL-10-, TNF-a- and IFN-g- producing cells in spleen were found in control mice at 8, 10, 12 month-old. These cytokines expressing cells in mice received DHEA are lower than that of control group. Double staining experiments revealed some of IgG+ B cells produced IL-10 and TNF-a, part of TCR+ T cells produced TNF-a in kidney. Some TCR+ T cells produced IL-10, TNF-a and IFN-g. Some IgG+ B cells produced IL-10. In view of time, cytokine producing cells appeared in spleen earlier than kidney. In peripheral blood, IgM+ B cells produce IL-10 and T cells produce IFN-g.
In conclusion, IL-10 producing cells mostly were B cells in kidney and spleen of NZB/W F1 mice. Some of TNF-a and IFN-g producing cells were T cells. DHEA could promote the expression of IFN-g in kidney and decrease the number of IL-10 - and TNF-a - producing cells in kidney and spleen. The emergence of cytokine producing cells in kidney and spleen may play a role in the pathogenesis of SLE.

中文摘要I
英文摘要II
誌謝III
目錄IV
符號及縮寫VI
第一章 序論1-1
第二章 實驗材料與方法
第一節實驗動物餵食2-1
第二節抽取周邊血液單核細胞全量RNA2-1
第三節反轉錄a聚合a連鎖反應2-2
第四節DNA 電泳分析2-4
第五節蛋白尿偵測2-5
第六節石蠟切片2-6
第七節冷凍切片2-6
第八節分離血液中白血球2-7
第九節免疫組織化學染色2-9
第十節蘇木紫伊紅染色2-11
第三章 實驗結果
第一節餵食 DHEA 對體重的影響3-1
第二節NZB/W F1 小鼠周邊血液單核細胞激素的影響3-2
第三節DHEA 對蛋白尿的影響3-2
第四節DHEA 對腎臟浸潤細胞的影響3-3
第五節在腎臟浸潤細胞所分泌的細胞激素3-4
第六節在脾臟細胞所分泌的細胞激素3-6
第七節在周邊血液單核細胞所分泌的細胞激素3-7
第四章 討論4-1
第五章 結論5-1
參考文獻R-1
附錄一 圖表A-1
附錄二 藥品儀器A-21
附錄三 試劑配方A-27

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