- 學位論文
探討腺苷酸激酶-4於傳統及替代性活化巨噬細胞的功能
The Function of Adenylate Kinase 4 in Classically and Alternatively Activated Macrophages
摘要
巨噬細胞（也稱為Mφ，MΦ或MP）是由Ilya Metchnikoff於19世紀首次發現的，根據明顯的表現型和功能將其分為經典活化巨噬細胞（M1）和交替激活巨噬細胞（M2），在免疫系統和免疫反應中扮演不可缺少的角色。經典活化的巨噬細胞（M1）由脂多醣（LPS）及干擾素-γ（IFN-γ）誘導，並表達高水平的促發炎細胞因子，例如介白素-1（IL-1），介白素-6（IL-6），腫瘤壞死因子-α（TNF-α）。被介白素-4（IL-4）及介白素-13（IL-13）激活的巨噬細胞稱為交替激活巨噬細胞（AAM）或M2巨噬細胞。 M2巨噬細胞產生介白素-10（IL-10），負責組織修復與維持和抗發炎反應。腺苷酸激酶4（Ak4）是腺苷酸激酶家族的成員，在粒線體的基質中表達。它參與細胞腺嘌呤核苷酸的能量代謝和衡定。它們催化γ-磷酸基團從ATP（或GTP）可逆地轉移到AMP，釋放ADP（或GDP）和ADP。但是Ak4在巨噬細胞的功能尚未釐清。我們發現與其他巨噬細胞亞型(包括M0和M2)相比，Ak4的表達在M1中被高度誘導。為了進一步研究Ak4在M1巨噬細胞中的功能，在M0巨噬細胞中以Ak4 shRNA處理，敲低（KD）了Ak4的表現，這些巨噬細胞進一步被LPS及IFN-γ刺激而極化成M1。M1在敲低（KD）Ak4下，Nos2，Hifla，Il1b，Il6和Tnfa的mRNA表達降低，同時也會降低M1細胞的殺菌能力。Ak4的敲除（KO）在體內不影響骨髓細胞群，包括髓樣和淋巴樣細胞。Ak4的敲除（KO）亦不會影響BMDM的成熟以及M1和M2的極化。但是，在M1巨噬細胞中缺乏Ak4時，炎症基因的表達包括 Il1b、Il6、Tnfa、Nos2、Nox2 和 Hif1a表達下降。同樣， M1巨噬細胞的殺菌能力也會降低。因此，我們的研究顯示了將巨噬細胞的能量消耗與發炎連結的潛在機制。
並列摘要
Macrophages (abbreviated as Mφ, MΦ, or MP) were first discovered in the 19th century by Ilya Metchnikoff, are classified into classically activated macrophages (M1) and alternatively activated macrophages (M2) based on distinct phenotype and functions. Macrophages play an indispensable role in immunity and immune responses. Classically activated macrophages (M1) are induced by lipopolysaccharides (LPS) and interferon-γ (IFN-γ) and express a high level of pro-inflammatory cytokines, such as interleukin-1 (IL-1), IL-6, tumor necrosis factor-α (TNF-α). Macrophages that are activated by IL-4 and IL-13 are called alternatively activated macrophages (AAM) or M2 macrophages. M2 macrophages are responsible for tissue repair, maintenance, and anti-inflammatory responses, by producing IL-10. Adenylate kinase 4 (Ak4) is a member of the Ak family, expressed in the matrix of mitochondria. It is involved in energy metabolism and homeostasis of cellular adenine nucleotide composition. Ak4 catalyzes reversibly transfer the γ-phosphate group from ATP (or GTP) to AMP, releasing ADP (or GDP) and ADP. However, the role of Ak4 in regulating the function of macrophages remains elusive. Here we report that the expression of Ak4 is induced in M1 compare with other macrophages subsets (M0 and M2). To further investigate the role of Ak4 in M1, Ak4 was knockdown (KD) by Ak4 shRNA transduction in M0 macrophages which were further polarized with LPS and IFN-γ into the M1 subset. After Ak4 KD, the mRNA expressions of Nos2, Hifla, Illb, Il6, and Tnfa were decreased. Moreover, suppressing the expression of Ak4 in M1 macrophages with shRNA decreases bactericidal ability in M1 cells. Knockout (KO) Ak4 does not affect the population of bone marrow cells, including myeloid and lymphoid cells. Furthermore, Ak4 deficiency does not affect the maturation of BMDMs and the polarization of M1 and M2. However, the expression of inflammation genes, including Il1b, Il6, Tnfa, Nos2, Nox2, and Hif1a and the bactericidal ability are reduced in M1 macrophages. Thus, our data depict a potential mechanism linking energy consumption and inflammation in macrophages
並列關鍵字
Ak4 ； Hif1α ； classically activated macrophage ； alternatively activated macrophages ； shRNA ； siRNA ； knockdown ； knockout參考文獻
延伸閱讀
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