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標題: | 探討腺苷酸激酶-4在傳統活化巨噬細胞的角色 The Role of Adenylate Kinase 4 in Classically Activated Macrophages |
作者: | Chi-Ying He 何季盈 |
指導教授: | 繆希椿(Shi-Chuen Miaw) |
關鍵字: | 核甘酸代謝,傳統活化巨噬細胞, nucleotide metabolism,classically activated macrophage, |
出版年 : | 2019 |
學位: | 碩士 |
摘要: | 巨噬細胞在先天性免疫扮演重要角色。巨噬細胞依據基因信號表達、表面受體及分泌的細胞激素分為傳統活化巨噬細胞(M1)和替代性活化巨噬細胞(M2)兩個子集。不同的刺激可以使骨髓源性巨噬細胞(M0)極化成為M1及M2細胞。此外,由脂多糖及γ型干擾素極化的M1巨噬細胞表達較高量的CD80、Nos2及Tnfa,而由介白素-4和介白素-13極化的M2巨噬細胞表達較高量的CD206、Arg1及Egr2。代謝對於巨噬細胞的功能及型態是重要的。相較於M0細胞和M2細胞,我們發現在粒線體基質中負責ATP/ADP/AMP平衡的腺苷酸激酶-4顯著地上調於M1細胞。為了調查Ak4基因在M1細胞的角色,Ak4基因在M0細胞中被小分子干擾核糖核酸及小髮夾核醣核酸敲落並用脂多糖及γ型干擾素刺激M0細胞。Ak4基因敲落後並不影響體外骨髓源性巨噬細胞的成熟,但Nos2、Hif1a及Il1b信使核糖核酸表達減少。再者,iNOS和HIF1α的蛋白表現量下降且IL-1β和TNF-α的細胞激素產量變少。此外,Arg1、CD80、CD86、CD206和MHC II在Ak4敲落的M1細胞及對照組M1細胞並無差異。總結來說,我們假說Ak4基因對於M1巨噬細胞的功能是重要的。 Macrophage has an important role in innate immunity. Macrophages are classified into two subsets, classically activated macrophages (M1) and alternatively activated macrophages (M2), based on their gene expression signatures, surface receptors, and cytokine productions. Different stimuli can skew bone marrow-derived macrophages (BMDM; M0) into M1 and M2. Furthermore, M1 macrophages skewed by LPS/IFN-γ exhibit higher expression of CD80, Nos2 and Tnfa, whereas M2 macrophages skewed by IL-4/IL-13 show higher expression of CD206, Arg1and Egr2. Metabolism is crucial for functions and phenotypes of macrophages. We found that Ak4 responsible for ATP/ADP/AMP homeostasis and located in mitochondria matrix, was significantly upregulated in M1 compared to M0 and M2. To investigate the role of Ak4 in M1, Ak4 was knockdowned by Ak4 siRNA or Ak4 shRNA treatment in M0 macrophages which were further stimulated with LPS/IFN-γ. After Ak4 knockdown (KD), BMDM maturation remained unchanged by Ak4 siRNA and Ak4 shRNA KD in vitro. The expressions of Nos2, Hif1a and Il1b mRNAs were decreased by Ak4 shRNA KD. Moreover, protein expressions of iNOS and HIF1α were declined by Ak4 shRNA KD. Furthermore, cytokine productions of IL-1β and TNF-α were decreased by Ak4 shRNA KD. However, the expressions of Arg1, CD80, CD86, CD206 and MHC class II were comparable between scramble and Ak4 siRNA KD M1 macrophages. Taken together, we hypothesized that Ak4 is critical for the functions of M1 macrophages. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/78615 |
DOI: | 10.6342/NTU201903349 |
全文授權: | 有償授權 |
顯示於系所單位: | 免疫學研究所 |
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