COORDINATE REGULATION OF NITRIC-OXIDE AND 1,25-DIHYDROXYVITAMIN-D PRODUCTION IN THE AVIAN MYELOMONOCYTIC CELL-LINE HD-11
COORDINATE REGULATION OF NITRIC-OXIDE AND 1,25-DIHYDROXYVITAMIN-D PRODUCTION IN THE AVIAN MYELOMONOCYTIC CELL-LINE HD-11
复制标题
DOI：
10.1210/en.136.5.2262
复制
发表时间：
1995-05-01
期刊：
影响因子：
4.8
通讯作者：
GACAD, MA
中科院分区：
文献类型：
--
作者：
ADAMS, JS;REN, SY;GACAD, MA
单核细胞/巨噬细胞谱系的细胞能够分别通过表达诱导型一氧化氮合酶（iNOS）和推定的25-羟基维生素D（25-OHD）-1-羟化酶产生一氧化氮（NO）和1，25-二羟基维生素D [1，25-（OH）（2）D]。我们最近报道了鸡骨髓单核细胞系HD-11中1，25-（OH）（2）D的合成受到iNOS抑制的限制。在目前的一组实验中，测量亚硝酸盐，一种稳定的水溶性NO分泌代谢物作为iNOS活性的指标，并测量与200 nM 25-OHD 3孵育的细胞脂质提取物中的1，25-（OH）（2）D-3作为1-羟化酶活性的指标，我们证明HD-11细胞产生的NO和1，25-（OH）（2）D是时间相关的，由同种激活剂诱导，协同调控。HD-11细胞产生的NO和1，25-（OH）（2）D-3被巨噬细胞刺激剂γ-干扰素和脂多糖以及表观分子量约为10，000道尔顿的自体非脂质热不稳定因子刺激数倍。正如预期的那样，NO的合成1）依赖于细胞外培养基中L-精氨酸的存在，2）受到N-W-羟基-L-精氨酸（NO生物合成中L-精氨酸衍生的中间体）和硝普钠（非L-精氨酸）的显着刺激。依赖于细胞内NO的来源，3）受到iNOS竞争性抑制剂N-W-硝基-L-精氨酸甲酯的抑制。在高浓度脂多糖或硝普钠诱导的高NO生成速率下，1，25-（OH）（2）D-3合成明显下降，表明1-羟化酶对NO的功能依赖性，但在高水平的细胞内NO生成下最终抑制1，25-（OH）（2）D-3合成能力。根据这些结果，我们推测，巨噬细胞25-OHD-1-羟基化反应可能依赖于iNOS产生的NO作为可溶性电子源，并在自分泌模式下由巨噬细胞衍生的NO刺激因子和NO本身调节。
Cells of the monocyte/macrophage lineage are capable of both nitric oxide (NO) and 1,25-dihydroxyvitamin D [1, 25-(OH)(2)D] production through expression of inducible nitric oxide synthase (iNOS) and a putative 25-hydroxyvitamin D (25-OHD)-1-hydroxylase, respectively. We have recently reported that 1,25-(OH)(2)D synthesis in the chick myelomonocytic cell Line HD-11 is restricted by inhibition of iNOS. In the current set of experiments, measuring nitrite, a stable water-soluble secreted metabolite of NO as an index of iNOS activity and 1,25-(OH)(2)D-3 in lipid extracts of cells incubated with 200 nM 25-OHD3 as an index of 1-hydroxylase activity, we demonstrate that NO and 1,25-(OH)(2)D production by HD-11 cells are temporally related, induced by the same kinds of activating agents, and coordinately regulated. NO and 1,25-(OH)(2)D-3 production by HD-11 cells was stimulated severalfold by the macrophage stimulators interferon-gamma and lipopolysaccharide and by an autologous, nonlipid, heat-labile factor with an apparent molecular mass approximate to 10,000 daltons. As expected NO synthesis was 1) dependent upon the presence of L-arginine in the extracellular medium, 2) subject to significant stimulation by N-W-hydroxy-L-arginine, an L-arginine-derived intermediate in NO biosynthesis, and by sodium nitroprusside, a non-L-arginine-dependent source of intracellular NO, and 3) inhibited by N-W-nitro-L-arginine methyl ester, a competitive inhibitor of iNOS. At high NO production rates, induced either by high-dose lipopolysaccharide or by sodium nitroprusside exposure, there was an apparent downturn in 1,25(OH)(2)D-3 synthesis, suggesting functional dependence of the 1-hydroxylase on NO but ultimate inhibition of 1,25-(OH)(2)D-3 synthetic capacity at high levels of intracellular NO production. On the basis of these results we postulate that the macrophage 25-OHD-1-hydroxylation reaction may be dependent on iNOS-generated NO as a soluble source of electrons and regulated in an autocrine mode by a macrophage-derived NO stimulatory factor and NO itself.