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研究生(外文):Jing-Bo Chou
論文名稱(外文):Effects of Lipopolysaccharide on The Receptor for Advanced Glycation End Products in The Folliculo-Stellate Like Cell
指導教授(外文):Sai-Koong Tan
外文關鍵詞:Folliculostellate cellpituitary glandLipopolysaccharidethe receptor for advanced glycation end products/ RAGE
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高醣化終產物受體(The receptor for advanced glycation end products,RAGE)與發炎反應之產生有關,近年來發現阻斷RAGE的訊息傳遞可降低及改善與糖尿病之發炎相關的併發症,故RAGE已成為藥物開發之重要標的物。腦下腺前葉之濾泡星狀細胞(Folliculo-stellate cell,FS cell),其不分泌荷爾蒙,但可經由LPS刺激後而產生許多細胞激素及生長因子,故推測其可能扮演似吞噬細胞的角色。利用似濾泡星狀細胞之細胞株TtT/GF,我們發現其也會表現RAGE,且經由LPS刺激後RAGE mRNA及蛋白質的表現量都會上升,但對於RAGE之相關配體(ligand)S100B並無影響,同時利用免疫螢光染色,我們發現LPS也會改變RAGE在細胞膜上的分布,使其產生聚集的現象,而LPS也能促使細胞骨架蛋白actin產生actin bundle的細胞數目增加。此外,我們意外發現LPS可促使α-tubulin的蛋白質表現增加,並觀察到α-tubulin會從細胞質往細胞核內聚集。由以上結果顯示LPS所誘發RAGE的表現量上升可能進一步活化濾泡星狀細胞以參與免疫反應,而LPS可能經由影響S100B以外之其他配體來改變RAGE膜蛋白的分布,並可能與細胞的移動有關。目前已完成RAGE siRNA(small interfering RNA)表現質體之建構,未來將可進一步利用其探討此些現象的生物效應。
The receptor for advanced glycation end products(RAGE)is a member of the immunoglobulin super family of cell surface molecules, and the biology of RAGE driven by its ligands is associated with inflammatory responses. And it has been defined as a target of drug design to reduce the inflammatory disorders of diabetes, recently. Folliculo-stellate cell(FS cell), the non-hormone secreting cell of anterior pituitary gland, could produce many cytokines or growth factors which could influence the hormone production and modulate the immunoendocrine connections. Since the biological function of RAGE of FS cell had not been discussed, we investigate if RAGE participates in the inflammatory responses of FS cell. By using a FS-like cell line, TtT/GF, as a model, we first found that TtT/GF cell expressed RAGE, which was up-regulated by LPS at both mRNA and protein level. Horever, a ligand of RAGE, S100B, which normally expressed on FS cells, was not regulated by LPS at both mRNA and protein level. We also found the distribution of RAGE on the membrane was clustered after LPS stimulation. LPS could stimulate the actin bundling and the assembling of free form α-tubulin in the cytoplasm toward to the nucleus. Protein expression of α-tubulin was also up-regulated by LPS. We suggest that the expression of RAGE up-regulated by LPS may couple to inflammatory responses of FS cell. The distribution of RAGE altered by LPS may drive by its ligands except S100B, and may involve in the mobility of FS cell with cytoskeletons. We have build the siRNA construct of RAGE, and it may be useful to investigate the biological significant of these changes in the future.
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