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首页> 外文期刊>Peritoneal dialysis international: Journal of the International Society for Peritoneal Dialysis >Nitric oxide production in peritoneal macrophages from peritoneal dialysis patients with bacterial peritonitis.
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Nitric oxide production in peritoneal macrophages from peritoneal dialysis patients with bacterial peritonitis.

机译：细菌性腹膜炎腹膜透析患者腹膜巨噬细胞中一氧化氮的产生。

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摘要

Nitric oxide (NO) is produced by various cell types, and it is an important mediator in many biological processes, including macrophage-mediated cellular host defense. The relevance and amount of NO production in peritonitis during peritoneal dialysis (PD) treatment is still not clear. We studied whether human peritoneal macrophages (PMphi) isolated from healthy PD patients or PD patients with peritonitis showed different spontaneous or lipopolysaccharide (LPS)/interferon gamma (IFN-gamma)-induced NO production (LPS, 1 ng/mL-10 microg/mL; IFN-gamma, 10-1000 U/mL; incubation between 6-48 hours; measured by Griess reagent). Results were compared with human blood monocytes (HBM) isolated from buffy coats. Inducible nitric oxide synthetase (iNOS) mRNA expression was looked for in PMphi by reverse transcriptase polymerase chain reaction (RT-PCR). Furthermore, plasma (P) and peritoneal dialysate effluent (D) nitrite concentrations were measured in vivo. The dialysate-to-plasma ratio (D/P) of nitrite concentration was inverse in the case of peritonitis compared to infection-free patients (peritonitis D/P = 1.3, non peritonitis D/P = 0.4; p < 0.01). PMphi from peritonitis patients produced higher amounts of NO than did those from infection-free patients (0.040+/-0.044 nmol per microgram cell protein versus 0.018+/-0.015 nmol per microgram cell protein, p < 0.05). NO release could not be further enhanced by stimulation with LPS plus IFN-gamma (1 ng/mL, 250 U/mL, respectively). However, NO production in PMphi from infection-free patients increased during in vitro stimulation (0.044+/-0.031 nmol per microgram cell protein versus 0.018+/-0.015 nmol per microgram cell protein, p < 0.01). An increase of iNOS mRNA expression could be demonstrated by RT-PCR. Blood monocytes from healthy donors also increased NO release during cytokine stimulation (0.032+/-0.015 nmol per microgram cell protein versus 0.019+/-0.009 nmol per microgram cell protein, p < 0.05). Our results indicate that significant amounts of NO are released intraperitoneally in the case of bacterial peritonitis. PMphi represent a site of NO production, though the absolute amounts released in vitro are only moderate. NO production can be induced in PMphi and HBM by LPS/IFN-gamma stimulation in vitro.
机译：一氧化氮（NO）由多种细胞类型产生，在许多生物过程中都是重要的介质，包括巨噬细胞介导的细胞宿主防御。腹膜透析（PD）治疗期间腹膜炎中NO产生的相关性和数量尚不清楚。我们研究了从健康的PD患者或腹膜炎PD患者中分离出的人类腹膜巨噬细胞（PMphi）是否表现出不同的自发性或脂多糖（LPS）/干扰素γ（IFN-γ）诱导的NO产生（LPS，1 ng / mL-10 microg / mL;IFN-γ，10-1000 U / mL;孵育6-48小时;通过Griess试剂测量）。将结果与从血沉棕黄层分离的人血单核细胞（HBM）进行比较。通过逆转录酶聚合酶链反应（RT-PCR）在PMphi中寻找诱导型一氧化氮合成酶（iNOS）mRNA表达。此外，在体内测量血浆（P）和腹膜透析液流出物（D）的亚硝酸盐浓度。与无感染的患者相比，腹膜炎患者中亚硝酸盐浓度的透析液与血浆的比率（D / P）是相反的（腹膜炎D / P = 1.3，非腹膜炎D / P = 0.4; p <0.01）。腹膜炎患者的PMphi产生的NO量高于无感染患者的PMphi（每微克细胞蛋白0.040 +/- 0.044nmol对每微克细胞蛋白0.018 +/- 0.015nmol，p <0.05）。用LPS加IFN-γ（分别为1 ng / mL，250 U / mL）刺激不能进一步增强NO的释放。但是，无感染患者的PMphi中NO的产生在体外刺激期间增加（每微克细胞蛋白0.044 +/- 0.031nmol与每微克细胞蛋白0.018 +/- 0.015nmol，p <0.01）。 RT-PCR证实iNOS mRNA表达增加。在细胞因子刺激期间，来自健康供体的血液单核细胞也增加了NO的释放（每微克细胞蛋白0.032 +/- 0.015nmol与每微克细胞蛋白0.019 +/- 0.009nmol，p <0.05）。我们的结果表明，在细菌性腹膜炎的情况下，腹膜内会释放大量的NO。尽管在体外释放的绝对量只有中等水平，但PMphi代表NO的产生位置。体外LPS /IFN-γ刺激可在PMphi和HBM中诱导NO的产生。

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