李瑤玲高良帆Liang-Fan Kao醫學檢驗暨生物技術學系碩士班2026-10-159999-12-312016https://handle.ncl.edu.tw/11296/s2yw8whttps://scholars.csmu.edu.tw/handle/123456789/22104細胞與組織在缺氧環境下是藉由缺氧誘導因子 (hypoxia-inducible factor, HIF)所調控，此轉錄因子在缺氧狀態下會大量表現並參與細胞氧的代謝過程。在缺氧環境下，免疫細胞中特別是巨噬細胞會引發慢性炎症的病變，使巨噬細胞具有極化的能力。本研究目的是探討缺氧誘導對於RAW264.7巨噬細胞M1或M2表現型的影響。為了確立氯化亞鈷誘導缺氧的細胞模式，在RAW264.7巨噬細胞處理不同濃度的氯化亞鈷 (CoCl2)；利用反轉錄聚合酶鏈反應與西方墨點法檢測HIF-1α mRNA與蛋白質的表現，結果發現HIF-1α mRNA與蛋白質表現量隨著氯化亞鈷暴露時間和濃度的增加而上升。另外也利用細胞存活、傷口癒合與細胞穿透等試驗，發現RAW264.7巨噬細胞在高濃度氯化亞鈷處理下，細胞的增生、遷移與侵襲能力都會增加。最後探討在缺氧誘導下RAW264.7巨噬細胞極化類型的轉換，在本研究顯示RAW 264.7巨噬細胞暴露在氯化亞鈷後，其Arginase I mRNA明顯降低而iNOS則顯著增加。綜合上述結果顯示RAW264.7巨噬細胞在缺氧的條件下，會刺激細胞的存活、遷移與穿透能力；細胞在極化轉換的過程中更偏向於M1型。顯示在缺氧的情況下，巨噬細胞活化的類型轉換中扮演著重要的角色。The hypoxic response in cells and tissues is mediated by the family ofhypoxia-inducible factor (HIF) transcription factors. HIFs are stress-responsive transcriptional regulators of cellular and physiological processes involved in oxygen metabolism. The decreased of oxygen tension due to the reduced in oxygen supply or the increased consumption of oxygen relative to the supply. Previous studies have reported that immune cells, particularly macrophages, lead to the pathological changes in chronic inflammatory and possess the ability of polarization under hypoxic conditions. The aim of this study is to investigate the effects of hypoxia-induction in the RAW 264.7 macrophage cell line will tended to M1 or M2 phenotype. In order to confirm the effect of CoCl2 induced hypoxia cell model, RAW 264.7 macrophage cells were treated with 100 µM CoCl2 for 0-24 hours, or with various concentrations of CoCl2 (0 - 125 µM) for 24 hours. RT-PCR and western blotting were used to examine the expression of HIF-1α mRNA and protein. Results show that the HIF-1α mRNA and protein expression were increased by CoCl2 in time-dependent and dose-dependent manner. Cell viability, migration and invasion were evaluated by MTT, wound healing and Transwell migration assay respectively. The data indicate that the cell survival, migration rate and invasion ability of RAW 264.7 macrophage were increased under the hypoxic condition. Finally, RAW 264.7 macrophages exposure to CoCl2 will significantly decrease the mRNA expression of Arginase I and increase that of HIF-1α and iNOS. Take in together, our data show that the proliferation, migration and invasion ability of RAW 264.7 cell will be stimulated under hypoxic condition. Hypoxia also leads the macrophage polarized into M1 phenotype and intend for the inflammatory response. HIF-1α plays an important role in the hypoxia-induced activation of macrophages.巨噬細胞缺氧缺氧誘導因子遷移增生MacrophageHypoxiaHIF-1αmigrationproliferationthesis