Asmis R, Stevens J, Begley J G, Grimes B, Van Zant G, Fanti P
Division of Nephrology, 7703 Floyd Curl Drive, University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, USA.
Clin Nephrol. 2006 Apr;65(4):267-75. doi: 10.5414/cnp65267.
Whole blood and peripheral blood mononuclear cells from hemodialysis (HD) patients show increased production and secretion of inflammatory cytokines. We determined the contribution of blood monocytes to the production of inflammatory cytokines in whole blood from HD patients.
Whole blood and isolated mononuclear cells from HD patients and healthy control subjects were preincubated with the isoflavone genistein and stimulated with LPS. TNFalpha, IL-6 and IL-10 formation in the whole blood was measured with ELISA and intracellular cytokine formation in CD 14-positive monocytes was determined by flow cytometry.
Unstimulated blood levels of TNFalpha, IL-6 and IL-10 were significantly elevated in HD patients compared to controls, but intracellular monocyte content of these cytokines was identical between groups. LPS induced a robust TNFalpha response in both whole blood and monocytes, and TNFalpha formation was 2.3-fold higher in blood from HD patients compared to controls. A similar trend was observed in monocytes. Conversely, LPS stimulation increased IL-6 levels >1000-fold in whole blood, albeit without a noticeable difference between groups. Only minor increases in monocyte IL-6 content were observed. The isoflavone genistein did not inhibit IL-6 formation and did not alter basal TNFalpha levels, but genistein selectively blocked LPS-induced TNFalpha formation in whole blood and monocytes from both groups.
Intracellular levels of TNFalpha, IL-6 and IL-10 in monocytes are indistinguishable between HD patients and healthy controls. However, monocytes from HD patients are selectively primed for enhanced TNFalpha secretion in response to LPS. The selective inhibition of monocyte TNFalpha production by genistein may explain the anti-inflammatory action of this phytochemical observed in vivo.
血液透析（HD）患者的全血和外周血单个核细胞显示炎症细胞因子的产生和分泌增加。我们确定了血液单核细胞对HD患者全血中炎症细胞因子产生的贡献。
将HD患者和健康对照者的全血及分离的单个核细胞与异黄酮染料木黄酮预孵育，并用脂多糖刺激。用酶联免疫吸附测定法（ELISA）测量全血中肿瘤坏死因子α（TNFα）、白细胞介素6（IL-6）和白细胞介素10（IL-10）的生成，通过流式细胞术测定CD14阳性单核细胞内细胞因子的生成。
与对照组相比，HD患者未受刺激时血液中TNFα、IL-6和IL-10水平显著升高，但两组间这些细胞因子的细胞内单核细胞含量相同。脂多糖在全血和单核细胞中均诱导了强烈的TNFα反应，HD患者血液中TNFα的生成比对照组高2.3倍。在单核细胞中也观察到类似趋势。相反，脂多糖刺激使全血中IL-6水平升高>1000倍，尽管两组间无明显差异。仅观察到单核细胞IL-6含量有轻微增加。异黄酮染料木黄酮不抑制IL-6的生成，也不改变基础TNFα水平，但染料木黄酮选择性地阻断了两组全血和单核细胞中脂多糖诱导的TNFα生成。
HD患者和健康对照者单核细胞内TNFα、IL-6和IL-10水平无差异。然而，HD患者的单核细胞对脂多糖刺激有选择性的预激活，以增强TNFα分泌。染料木黄酮对单核细胞TNFα生成的选择性抑制可能解释了该植物化学物质在体内观察到的抗炎作用。
