Release of salusin-β from human monocytes/macrophages
Release of salusin-β from human monocytes/macrophages
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DOI：
10.1016/j.regpep.2010.02.010
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发表时间：
2010-06-08
影响因子：
--
通讯作者：
Shichiri, Masayoshi
中科院分区：
文献类型：
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作者：
Sato, Kengo;Fujimoto, Kazumi;Shichiri, Masayoshi
Salusin-α和Salusin-beta是由相同的前体prosalusin生物合成的相关生物活性多肽。尽管salusin-beta具有强大的血流动力学和动脉粥样硬化活性，但由于其独特的物理化学特性(如与聚丙烯和聚苯乙烯的显著粘附性)相关的技术困难，其确切分布和生物学功能在很大程度上仍未确定。通过绕过这些问题，我们最近建立了一种检测免疫反应的人Salusin-beta的特异性放射免疫分析方法。在目前的研究中，我们证实了人单核细胞白血病细胞系THP-1和U937释放Salusin-beta。两种细胞提取的条件培养液的稀释曲线与标准人Salusin-β的放射免疫分析结果一致。反相高效液相色谱结合放射免疫分析检测培养上清液，发现了与正品Salusin-beta共洗脱的主要免疫反应成分。两种细胞株在无血清培养上清液中均可分泌Salusin-β样免疫反应性(LI)，且呈时间依赖性(每24 h分别分泌1234.3+/-122.7和186.7+/-9.1fmol/10(5)个细胞)。当THP-1和11937细胞在2-O-十四酰佛波醇-13-乙酸酯孵育后分化为巨噬细胞时，它们向培养上清液中分泌更多的Salusin-β-Li(每24 h分泌3351.9+/-899.3和1545.8+/-183.3 fmol/10(5)个细胞)。TPA处理加速了原杨柳素的裂解，提示THP-1诱导的巨噬细胞分泌增加的原杨柳素-β-Li是由于原杨柳素的细胞内加工增强所致。在炎性细胞因子肿瘤坏死因子-α(TNF-α)和脂多糖(LPS)的刺激下，Salusin-β的分泌增加，而不诱导前Prosalusin基因的表达，提示INF-α和LPS刺激Salusin-β的释放。这些数据表明，诱导巨噬细胞泡沫细胞形成的Salusin-beta是以其真实形式从人单核/巨噬细胞分泌的。(C)2010爱思唯尔B.V.保留所有权利。
Salusin-alpha and salusin-beta are related bioactive peptides biosynthesized from the same precursor, prosalusin. Despite the potent hemodynamic and proatherosclerotic activities of salusin-beta, its exact distribution and biological functions remain largely undetermined because of technical difficulties associated with its unique physicochemical characteristics, such as marked adhesiveness to polypropylene and polystyrene. By circumventing these problems, we recently established a specific radioimmunoassay for detecting immunoreactive human salusin-beta. In the current study, we demonstrated the release of salusin-beta from the human monoblastic leukemia cell lines, THP-1 and U937. Dilution curves of extracted conditioned media from both cells were parallel with those of standard human salusin-beta by radioimmunoassay. Reverse-phase high performance liquid chromatography coupled with radioimmunoassay detection of the culture supernatants revealed a major immunoreactive component that co-eluted with authentic salusin-beta. Both cell lines secreted salusin-beta-like immunoreactivity (LI) into serum-free media as a function of time (1234.3 +/- 122.7 and 186.7 +/- 9.1 fmol/10(5) cells per 24 h). When THP-1 and 11937 cells differentiated into macrophages after incubation with 2-O-tetradecanoylphorbol-13-acetate (TPA), they secreted far greater amounts of salusin-beta-LI into the culture supernatant (3351.9 +/- 899.3 and 1545.8 +/- 183.3 fmol/10(5) cells per 24 h). TPA treatment accelerated the processing of prosalusin into its cleaved fragments, suggesting that the increased secretion of salusin-beta-LI in THP-1-derived macrophages was caused by the enhanced intracellular processing of prosalusin. Stimulation with the inflammatory cytokines, tumor necrosis factor alpha (TNF-alpha) and lipopolysaccharide (LPS), resulted in increased secretion of salusin-beta without inducing expression of the gene for preprosalusin, suggesting that INF-alpha and LPS stimulated the release of salusin-beta. These data demonstrate that salusin-beta, which induces macrophage foam cell formation, is secreted in its authentic form from human monocytes/macrophages. (C) 2010 Elsevier B.V. All rights reserved.