Suzuki Eriko, Sugiyama Chie, Umezawa Kazuo
Department of Applied Chemistry, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-0061, Japan.
Biomed Pharmacother. 2009 Jun;63(5):351-8. doi: 10.1016/j.biopha.2008.05.003. Epub 2008 Jun 20.
Previously, we designed and synthesized a new NF-kappaB inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ), and found that racemic DHMEQ inhibited cytokine secretion and phagocytosis by cells of the macrophage cell line RAW264.7. In the present research, we looked into the effect of optically active (-)-DHMEQ on the NO production, inflammatory cytokine secretion, and prostaglandin secretion in mouse bone marrow-derived macrophages (BMMs). We also studied the effect of (-)-DHMEQ on the differentiation of macrophages. DHMEQ inhibited lipopolysaccharide (LPS)-induced NF-kappaB activation. It also inhibited the expression of inducible NO synthase (iNOS) and NO production induced by LPS. Using enzyme-linked immunosorbent assays, we showed DHMEQ to inhibit LPS-induced secretion of IL-6 and TNF-alpha. It also inhibited COX-2 expression and prostaglandin E(2) production and secretion. It did not inhibit the phagocytosis of fluorescently labeled Escherichia coli by BMMs treated with LPS, unlike in the case of RAW264.7 cells. Next we examined the effect of the inhibitor on M-CSF-induced differentiation of bone marrow cells to macrophages. DHMEQ showed no effect on the differentiation in terms of reactive oxygen species production and F4/80 expression. However, although BMM incorporated oxidized LDL to give rise to foam cells, the (-)-DHMEQ-treated bone marrow cells did not take up oxidized LDL. Taken together, our data show that (-)-DHMEQ inhibited LPS-induced activation of BMM in terms of NO and cytokine secretion, but its effect on phagocytosis differed between BMMs and RAW264.7 cells. We also found that the functional differentiation into macrophages was inhibited by (-)-DHMEQ.
此前，我们设计并合成了一种新型核因子κB抑制剂——去羟甲基环氧喹霉素（DHMEQ），并发现外消旋DHMEQ可抑制巨噬细胞系RAW264.7细胞的细胞因子分泌和吞噬作用。在本研究中，我们研究了旋光性（-）-DHMEQ对小鼠骨髓来源巨噬细胞（BMM）中一氧化氮（NO）生成、炎性细胞因子分泌和前列腺素分泌的影响。我们还研究了（-）-DHMEQ对巨噬细胞分化的影响。DHMEQ可抑制脂多糖（LPS）诱导的核因子κB激活。它还可抑制LPS诱导的诱导型一氧化氮合酶（iNOS）表达和NO生成。通过酶联免疫吸附测定，我们发现DHMEQ可抑制LPS诱导的白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）分泌。它还可抑制环氧合酶-2（COX-2）表达以及前列腺素E2生成和分泌。与RAW264.7细胞不同，它不会抑制经LPS处理的BMM对荧光标记大肠杆菌的吞噬作用。接下来，我们研究了该抑制剂对巨噬细胞集落刺激因子（M-CSF）诱导的骨髓细胞向巨噬细胞分化的影响。就活性氧生成和F4/80表达而言，DHMEQ对分化没有影响。然而，尽管BMM摄取氧化型低密度脂蛋白（ox-LDL）产生泡沫细胞，但经（-）-DHMEQ处理的骨髓细胞不摄取ox-LDL。综上所述，我们的数据表明，（-）-DHMEQ在NO和细胞因子分泌方面可抑制LPS诱导的BMM激活，但其对吞噬作用的影响在BMM和RAW264.7细胞之间存在差异。我们还发现，（-）-DHMEQ可抑制巨噬细胞的功能分化。
