蔡嘉哲林怜君Ling-Chun微生物免疫研究所2026-10-289999-12-312012https://handle.ncl.edu.tw/11296/ygu2y6https://scholars.csmu.edu.tw/handle/123456789/35747研究目的：痛風的成因是由於單鈉尿酸鹽（monosodium urate；MSU）沉積於體內關節及關節周邊所引起的發炎性疾病。在先前的文獻指出，在MSU所引起的小鼠腹腔發炎中，第二型轉麩胺脢(transglutaminase 2；TG2)會調節凋亡細胞的清除而抑制發炎。本研究主要研究TG2 在MSU引起之發炎反應之角色。研究方法及資料：蒐集痛風病人關節之滑膜組織以及關節液以分離單核球。分別利用組織免疫染色(Immunohistochemistry)與RT-PCR (Reverse Transcriptase-PCR)檢測TG2的表現量。接著利用MSU 引發小鼠巨嗜細胞株(RAW 264.7)吞噬並產生發炎反應。為了進一步探討TG2與MSU引起的發炎之關係，利用TG2 over expression (TG2 o/e) RAW264.7 cells、TG2 small interfering RNA (TG2 siRNA) knockdown RAW264.7 cells、及mouse embryonic fibroblasts deficient for TG2 (TG2 KO MEF cells)等細胞株來研究TG2在MSU引起細胞發炎的角色。最後加入ZDON ，一種TG2 cross-linking 功能抑制劑作進一步研究。研究結果：在痛風病人的關節液及滑膜當中發現TG2表現量增加。MSU刺激RAW 264.7細胞株之後IL-1β、TNF-α、TGF-β和TG2之 mRNA增加。在TG2 o/e RAW 264.7細胞株中，MSU所誘發之IL-1β及TNF-α之mRNA表現量會較野生型RAW 264.7細胞株減少；而TGF-β的mRNA表現會提高；相反的在TG2-siRNA RAW 264.7細胞株及TG2 KO MEF細胞株中，IL-1β的mRNA和細胞激素表現量提高且TGF-β的mRNA和細胞激素表現量降低。在MSU刺激WT MEF cells後加入anti- TGF-β Antibody培養之後IL-1βmRNA表現量會大幅上升，而加入ZDON可減少MSU所引起之TGF-β mRNA表現量。結論：本研究發現在痛風病人關節液和滑膜中及受MSU刺激之RAW264.7細胞株皆發現TG2增加。MSU刺激TG2 o/e RAW 264.7細胞株後，可經由TGF-β大量表現而降低IL-1β表現量以緩解發炎現象。Objective：Gouty arthritis is an inflammatory disease induced by accumulation of monosodium urate (MSU) crystals in joints. Previous studies show that transglutaminase 2 (TG2) affects the clearance of apoptotic neutrophils in murine peritoneal acute gout-like inflammation. We studied the role of TG2 in the inflammation induced by MSU.Methods and Materials： The synovium tissue and synovial fluid mononuclear cells (SFMCs) collected from patient with gouty arthritis (GA) for the TG2 expression by immunohistochemistry and reverse transcriptase-PCR (RT-PCR). MSU crystal stimulated RAW264.7 mouse macrophages were analysed for mRNA expression by RT-PCR. TG2 overexpression (TG2 o/e) RAW 264.7 cells and TG2 small interfering RNA (TG2 siRNA)-mediated knockdown RAW264.7 cells and mouse embryonic fibroblasts deficient for TG2 (TG2 KO MEFcells) were used to investigate the role of TG2 in resolving MSU induced inflammation. ZDON (TG2 cross-linking inhibitor) was used to do further investigation.Results：TG2 expression was up-regulated in the synovium tissue and SFMCs from patient with GA. The expressions mRNA of IL-1β, TNF-α, TGF-β and TG2 were increased in MSU-stimulated RAW 264.7 cells. IL-1β and TNF-αbut not TGF-β decreased in MSU-induced TG2 o/e RAW264.7 cells. On the contrary, IL-1β and TNF-αbut not TGF-β increased in TG2 siRNA-RAW264.7 cells and TG2 KO MEF cells., IL-1β mRNA was significant increased after anti-TGF-β antibody treatment in MSU-induced WT MEF cells. TGF-β mRNA was decreased after ZDON treatment.Conclusions： TG2 increased in synovium tissue and SFMCs from patient with GA and in MSU-stimulated RAW264.7 cells. In TG2 o/e RAW 264.7 cells, both TG2 and TGF-βwere increased and IL-1β was decreased with MSU stimulation.TG2 may play a role in resolution of MSU-induced inflammation.thesis