2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38749The etiology and treatment of Systemic lupus erythematosus (SLE) are still unclear. However, the impaired clearance and accumulation of apoptotic bodies has been recognized as a major factor to induce inflammation, breakdown of immuno-tolerance, and production of autoantibodies. Cystamine is known as an inhibitor of transglutaminase 2 could inhibit the caspase activity and neuro-cell degeneration, which has been used in treatment with the animal model and human phase-I clinical trial of Huntington’s disease. Notably, our recent publications have demonstrated the beneficial effects of cystamine on SLE by reducing the brain and hepatic apoptosis. Since macrophages play important role in autoimmune diseases and apoptosis, we intend to investigate the mechanism of cystamine on macrophage in NZB/W F1 mice via proteomic analysis and develop the therapeutic targets for SLE. The lupus prone mice, NZB/W F1, will receive PBS or cystamine intravenously and the peritoneal macrophages will be isolated for 2-dimentional electrophoresis analysis and production of the master PAGE/image. The protein spots with different expression level will be picked, purified, and in-gel digested for further identification by western blotting, N-terminal sequencing, and peptide fingerprinting. Additionally, database research will be performed to explore the possible candidates for further therapeutic and new drug development.紅斑性狼瘡的致病原因目前仍未釐清，但最新研究相信是因為細胞凋亡的過程失衡﹐凋亡小體無法被正確辨認和清除﹐持續堆積在體內引起發炎反應﹐導致免疫耐受破壞而產生自體抗體。胱胺酸是已知的殼胺醯胺轉移脢抑制劑﹐可以抑制凋亡相關的酵素活性和神經細胞的降解(degradation)﹐目前已成功應用在許多神經缺失的動物上﹐特別在舞蹈症(Huntington's disease)的動物模式，而在人類舞蹈症的應用上也已進入第一期的臨床試驗。而胱胺酸在狼瘡動物的研究上，我們發現胱胺酸可顯著改善狼瘡小鼠臨床表徵之效果﹐包括改善狼瘡小鼠腦部及肝臟的緊迫與凋亡。此外﹐更能減緩狼瘡小鼠巨嗜細胞的凋亡。由於巨嗜細胞在自體免疫疾病中扮演極重要的角色﹐所以我們將利用蛋白質體技術去探討胱胺酸對於紅斑性狼瘡小鼠巨嗜細胞的作用機轉﹐以開發可能之治療標的﹐研發更有效之藥物﹐並與臨床對於狼瘡的治療相連結。 首先我們將腹腔或靜脈注射PBS或胱胺酸至發病後之狼瘡小鼠﹐兩週後將採集巨嗜細胞進行雙向電泳分析﹐除了建立主圖樣外，將比較狼瘡鼠巨嗜細胞處理胱胺酸前後表現相異之蛋白質點﹐挑選表現差異之蛋白質點鑑定﹐包括西方免疫吸漬法（Western blotting﹐蛋白質N-端胺基酸定序 (N-terminal sequencing)﹐蛋白質生肽指紋分析（peptide fingerprinting。最後將進行資料庫及文獻搜尋比對﹐以開發可能之治療標的﹐進一步研發更有效藥物﹐以期與臨床接軌。如能完成這些研究﹐相信對紅斑性狼瘡致病機轉的了解及治療的應用上﹐將能夠有重要的貢獻。系統性紅斑性狼瘡胱胺酸蛋白體學治療生物標的Systemic lupus erythematosuscystamineproteomictherapeutic bio-targetProject