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首页> 外文期刊>The Journal of Nutritional Biochemistry >Comparison of intracellular zinc signals in nonadherent lymphocytes from young-adult and elderly donors: role of zinc transporters (Zip family) and proinflammatory cytokines
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Comparison of intracellular zinc signals in nonadherent lymphocytes from young-adult and elderly donors: role of zinc transporters (Zip family) and proinflammatory cytokines

机译：成年人和老年人供体非粘附淋巴细胞中细胞内锌信号的比较：锌转运蛋白（Zip家族）和促炎细胞因子的作用

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摘要

Intracellular zinc homeostasis is crucial in regulating the inflammatory/immune response at any age. It is tightly regulated by zinc transporters that control influx, efflux and compartmentalization of zinc within the cells. Specific methods for detecting the age-related differences in intracellular zinc signaling are poorly described. We report a novel assay induced after the in vitro zinc addition in peripheral blood mononuclear cells (PBMCs) and in lymphocytes from young and old donors in the absence/presence of in vitro zinc depletion (using EDTA). The intracellular labile zinc variations are monitored over time by flow cytometry using Fluozin-3 AM probe. The best curve fit of the data is calculated using a nonlinear regression model defined as follows: pr3/[1+Exp(-pr1-pr2*Xt)]. Pr1 depends on the initial free zinc value (time 0); pr2 describes the rate of the speed in reaching the maximum intracellular free zinc concentration; pr3 represents the maximum intracellular zinc increment (plateau curve); Xt is the time course. Age-related intracellular free zinc variations occur in PBMCs and lymphocytes incubated in EDTA-supplemented medium. The higher plateau of the curve (pr3) was observed in younger subjects. An up-regulation of Zip genes (Zip1, Zip2, Zip3), influencing zinc influx, is more pronounced in the young than old donors. Interleukin-6 and tumor necrosis factor- alpha overproduction was enhanced in old individuals, suggesting the presence of more marked zinc deficiency and chronic inflammation. In conclusion, the determination of intracellular zinc signals induced by in vitro zinc addition using logistic parameters may be useful to estimate the rate of intracellular zinc homeostasis and its role in inflammatory/immune response in aging
机译：细胞内锌稳态对于调节任何年龄的炎症/免疫反应都至关重要。它受锌转运蛋白的严格调控，锌转运蛋白控制锌在细胞内的流入，流出和分隔。很少描述检测细胞内锌信号转导的年龄相关差异的具体方法。我们报告了一种新颖的测定方法，该方法在体外锌缺乏后/存在体外锌消耗（使用EDTA）的情况下，在外周血单核细胞（PBMC）和来自年轻和老年供体的淋巴细胞中添加锌后诱导。使用Fluozin-3 AM探针，通过流式细胞仪监测细胞内不稳定锌的变化。使用如下定义的非线性回归模型计算数据的最佳曲线拟合：pr3 / [1 + Exp（-pr1-pr2 * Xt）]。 Pr1取决于初始游离锌值（时间0）; pr2描述达到最大细胞内游离锌浓度的速度速率; pr3代表最大的细胞内锌增量（平台曲线）; Xt是时间过程。年龄相关的细胞内游离锌变异发生在PBMC和在EDTA补充培养基中孵育的淋巴细胞中。在较年轻的受试者中观察到较高的曲线平台（pr3）。影响锌流入的Zip基因（Zip1，Zip2，Zip3）的上调在年轻供者中比在老年供者中更为明显。白细胞介素6和肿瘤坏死因子-α的过度生产在老年个体中得到增强，表明存在更多明显的锌缺乏和慢性炎症。总之，使用逻辑参数确定体外添加锌诱导的细胞内锌信号可能有助于评估细胞内锌稳态的速率及其在衰老中炎症/免疫反应中的作用

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