Kim Jin-Man, Kim Hyunsoo, Kwon Soon Bok, Lee Soo Young, Chung Sung-Chang, Jeong Dae-Won, Min Byung-Moo
Department of Oral Biochemistry and Craniomaxillofacial Reconstructive Sciences Major, Dental Research Institute, College of Dentistry, Seoul National University, Seoul, Republic of Korea.
Biochem Biophys Res Commun. 2004 Dec 3;325(1):101-8. doi: 10.1016/j.bbrc.2004.09.220.
Although a redox shift can regulate the development of cells, including proliferation, differentiation, and survival, the role of the glutathione (GSH) redox status in macrophage differentiation remains unclear. In order to elucidate the role of a redox shift, macrophage-like cells were differentiated from the bone marrow-derived monocytes that were treated with a macrophage colony stimulating factor (M-CSF or CSF-1) for 3 days. The macrophagic cells were characterized by a time-dependent increase in three major symptoms: the number of phagocytic cells, the number of adherent cells, and the mRNA expression of c-fms, a M-CSF receptor that is one of the macrophage-specific markers and mediates development signals. Upon M-CSF-driven macrophage differentiation, the GSH/GSSG ratio was significantly lower on day 1 than that observed on day 0 but was constant on days 1-3. To assess the effect of the GSH-depleted and -repleted status on the differentiation and phagocytosis of the macrophages, GSH depletion by BSO, a specific inhibitor of the de novo GSH synthesis, inhibited the formation of the adherent macrophagic cells by the down-regulation of c-fms, but did not affect the phagocytic activity of the macrophages. To the contrary, GSH repletion by the addition of NAC, which is a GSH precursor, or reduced GSH in media had no effect on macrophage differentiation, and led to a decrease in the phagocytic activity. Furthermore, we observed that there is checkpoint that is capable of releasing from the inhibition of the formation of the adherent macrophagic cells according to GSH depletion by BSO. Summarizing, these results indicate that the intracellular GSH status plays an important role in the differentiation and phagocytosis of macrophages.
尽管氧化还原状态的改变可以调节细胞的发育，包括增殖、分化和存活，但谷胱甘肽（GSH）氧化还原状态在巨噬细胞分化中的作用仍不清楚。为了阐明氧化还原状态改变的作用，从骨髓来源的单核细胞中诱导分化出巨噬样细胞，这些单核细胞用巨噬细胞集落刺激因子（M-CSF或CSF-1）处理3天。巨噬细胞的特征表现为三个主要指标随时间增加：吞噬细胞数量、贴壁细胞数量以及c-fms的mRNA表达，c-fms是M-CSF受体，是巨噬细胞特异性标志物之一，介导发育信号。在M-CSF驱动的巨噬细胞分化过程中，第1天的GSH/GSSG比值显著低于第0天，但在第1 - 3天保持恒定。为了评估GSH耗竭和补充状态对巨噬细胞分化和吞噬作用的影响，用BSO（一种从头合成GSH的特异性抑制剂）消耗GSH，通过下调c-fms抑制贴壁巨噬细胞的形成，但不影响巨噬细胞的吞噬活性。相反，添加NAC（一种GSH前体）或在培养基中添加还原型GSH来补充GSH，对巨噬细胞分化没有影响，并导致吞噬活性降低。此外，我们观察到存在一个检查点，它能够根据BSO导致的GSH耗竭解除对贴壁巨噬细胞形成的抑制。综上所述，这些结果表明细胞内GSH状态在巨噬细胞的分化和吞噬作用中起重要作用。
