Laboratory of Molecular Genetics of Hematology, Graduate School of Health Care Sciences, Tokyo Medical and Dental University, Tokyo, Japan ; Department of Endocrinology, Diabetes and Metabolism, Kitasato University School of Medicine, Kanagawa, Japan.
PLoS One. 2013 Oct 3;8(10):e76714. doi: 10.1371/journal.pone.0076714. eCollection 2013.
Using bioinformatics analysis, we previously identified salusin-β, an endogenous bioactive peptide with diverse physiological activities. Salusin-β is abundantly expressed in the neuroendocrine system and in systemic endocrine cells/macrophages. Salusin-β acutely regulates hemodynamics and chronically induces atherosclerosis, but its unique physicochemical characteristics to tightly adhere to all types of plastic and glassware have prevented elucidation of its precise pathophysiological role. To quantitate plasma total salusin-β concentrations, we produced rabbit and chicken polyclonal antibodies against the C- and N-terminal end sequences, circumvented its sticky nature, and successfully established a sandwich enzyme-linked immunosorbent assay (ELISA). Salusin-β was abundantly present in the plasma of healthy volunteers, ranging from 1.9 to 6.6 nmol/L. Reverse phase-high performance liquid chromatography analysis showed that a single immunoreactive salusin-β peak coincided with synthetic authentic salusin-β. Plasma salusin-β concentrations were unaffected by postural changes and by potent vasopressin release stimuli, such as hypertonic saline infusion or smoking. However, salusin-β concentrations showed significant circadian variation; concentrations were high during the daytime and reached the lowest concentrations in the early morning. Plasma salusin-β levels in subjects with diabetes mellitus, coronary artery disease, and cerebrovascular disease showed distinctly higher levels than healthy controls. Patients with panhypopituitarism combined with complete central diabetes insipidus also showed significantly higher plasma salusin-β levels. Therefore, the ELISA system developed in this study will be useful for evaluating circulating total salusin-β levels and for confirming the presence of authentic salusin-β in human plasma. The obtained results suggest a limited contribution of the neuroendocrine system to peripheral total salusin-β concentrations and a role for plasma total salusin-β concentrations as an indicator of systemic vascular diseases.
使用生物信息学分析,我们之前鉴定出了 salusin-β,这是一种具有多种生理活性的内源性生物活性肽。Salusin-β 在神经内分泌系统和全身内分泌细胞/巨噬细胞中大量表达。Salusin-β 可急性调节血液动力学,慢性诱导动脉粥样硬化,但由于其独特的理化特性,使其紧密附着于所有类型的塑料和玻璃器皿,从而阻止了其精确病理生理学作用的阐明。为了定量检测血浆总 salusin-β 浓度,我们针对 C 端和 N 端序列产生了兔和鸡的多克隆抗体,避开了其粘性,成功建立了夹心酶联免疫吸附测定(ELISA)。健康志愿者的血浆中存在大量的 salusin-β,范围为 1.9 至 6.6 nmol/L。反相高效液相色谱分析表明,单一免疫反应性 salusin-β 峰与合成的天然 salusin-β 相吻合。体位变化和强效血管加压素释放刺激(如高渗盐水输注或吸烟)不会影响血浆 salusin-β 浓度。然而,salusin-β 浓度存在明显的昼夜节律变化;白天浓度较高,清晨达到最低浓度。糖尿病、冠状动脉疾病和脑血管疾病患者的血浆 salusin-β 水平明显高于健康对照组。伴有全垂体功能减退症和完全中枢性尿崩症的患者的血浆 salusin-β 水平也显著升高。因此,本研究中开发的 ELISA 系统将有助于评估循环总 salusin-β 水平,并确认人血浆中天然 salusin-β 的存在。研究结果表明,神经内分泌系统对周围总 salusin-β 浓度的贡献有限,而血浆总 salusin-β 浓度可作为全身血管疾病的指标。