Kawase T, Orikasa M, Oguro A, Burns D M
Department of Pharmacology, Niigata University School of Dentistry, Niigata 951, Japan.
Calcif Tissue Int. 1998 Jul;63(1):27-35. doi: 10.1007/s002239900485.
In previous studies we found that the calciotropic hormone 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] augments the action of either prostaglandin E1 (PGE1) or NaF to induce differentiation of human promyelocytic HL-60 cells, a process that features increased generation of nitric oxide (NO) via up-regulation of inducible nitric oxide synthase (iNOS). We have now examined the short-term interaction of 1,25(OH)2D3 with phorbol 12-myristate 13-acetate (PMA) and dimethylsulfoxide (DMSO) in these cells. PMA (100 nM) alone generally up-regulated several classical indices of macrophagic differentiation and stimulated cellular production of interleukin (IL)-1alpha, IL-6, tumor-necrosis factor (TNF)-alpha, PGE2, and NO. Increased generation of NO primarily resulted from increased expression of cellular iNOS. When 1,25(OH)2D3 (10 nM) was added to PMA treatments, most PMA-induced changes, particularly its effects to up-regulate iNOS-dependent NO production and change cell morphology, were multiplicatively augmented. In contrast, DMSO (1.3%) alone, an inducer of granulocytic differentiation, increased cytokine production, but failed to stimulate NO production or induce iNOS. In contrast to its striking interaction with PMA, 1,25(OH)2D3 could not augment DMSO's differentiative effects. Changes in cellular cytokine production were eliminated as the driving force in HL-60 differentiation when specific neutralizing antibodies failed to produce any attenuation of iNOS up-regulation or of the shifts in cell morphology. However, indomethacin (30 microM) blocked the synergistic interaction between 1,25(OH)2D3 + PMA to shift cell morphology and stimulate NO production. Subsequently adding PGE2 (1 ng/ml) to indomethacin-treated cells restored the ability of 1, 25(OH)2D3 + PMA to interactively increase cellular NO production, but failed to fully replicate the strong shift in cell morphology typical of PMA + 1,25(OH)2D3 treatments. Our findings suggest that interaction between 1,25(OH)2D3 and PMA to induce macrophagic differentiation increases iNOS-dependent NO production by a mechanism involving a cyclooxygenase product(s), possibly PGE2.
在先前的研究中，我们发现钙调节激素1,25 - 二羟基维生素D3 [1,25(OH)2D3]可增强前列腺素E1（PGE1）或氟化钠诱导人早幼粒细胞HL - 60细胞分化的作用，这一过程的特征是通过诱导型一氧化氮合酶（iNOS）的上调增加一氧化氮（NO）的生成。我们现在研究了1,25(OH)2D3与佛波酯12 - 肉豆蔻酸酯13 - 乙酸酯（PMA）和二甲基亚砜（DMSO）在这些细胞中的短期相互作用。单独使用PMA（100 nM）通常会上调巨噬细胞分化的几个经典指标，并刺激细胞产生白细胞介素（IL）-1α、IL - 6、肿瘤坏死因子（TNF）-α、PGE2和NO。NO生成增加主要是由于细胞iNOS表达增加所致。当将1,25(OH)2D3（10 nM）添加到PMA处理中时，大多数PMA诱导的变化，特别是其上调iNOS依赖性NO生成和改变细胞形态的作用，会成倍增强。相比之下，单独使用DMSO（1.3%），一种粒细胞分化诱导剂，可增加细胞因子的产生，但未能刺激NO生成或诱导iNOS。与它与PMA的显著相互作用不同，1,25(OH)2D3不能增强DMSO的分化作用。当特异性中和抗体未能减弱iNOS上调或细胞形态变化时，细胞因子产生的变化不再是HL - 60分化的驱动力。然而，吲哚美辛（30 microM）阻断了1,25(OH)2D3 + PMA之间在改变细胞形态和刺激NO生成方面的协同相互作用。随后向吲哚美辛处理的细胞中添加PGE2（1 ng/ml）恢复了1,25(OH)2D3 + PMA相互作用增加细胞NO生成的能力，但未能完全重现PMA + 1,25(OH)2D3处理典型的强烈细胞形态变化。我们的研究结果表明，1,25(OH)2D3与PMA之间诱导巨噬细胞分化的相互作用通过一种涉及环氧化酶产物（可能是PGE2）的机制增加了iNOS依赖性NO的生成。
