Division of Neonatology, Pediatric Intensive Care and Neuropediatrics, Department of Pediatrics and Adolescent Medicine, Medical University of Vienna, Vienna, Austria.
Department of Surgery, Research Labs &Core Facility Flow Cytometry, Medical University of Vienna, Vienna, Austria.
Pediatr Res. 2017 Aug;82(2):215-225. doi: 10.1038/pr.2017.41. Epub 2017 May 24.
BackgroundPentoxifylline (PTX), a methylxanthine derivate with immunomodulating properties, has been used as adjunctive treatment in severe neonatal sepsis. The aim of the study was to investigate the anti-inflammatory effects of PTX on Lipopolysaccharides (LPS)-stimulated monocytes of preterm neonates in vitro compared with monocytes of term infants and adult controls.MethodsWhole cord blood samples and control adult blood samples were incubated with LPS and PTX. The expression of surface markers, phagocytosis, cytokine secretion, and Toll-like receptor (TLR)4 signaling of monocytes were assessed by flow cytometry. Changes of TLR4-messenger RNA (mRNA) levels were confirmed by reverse-transcriptase PCR.ResultsThe expression of CD14, CD11b, CD64, CD71, and CD80 was downregulated by PTX in a dose-dependent manner; the greatest effect was observed on CD14 and CD11b in preterm infants. PTX markedly downregulated LPS-induced tumor necrosis factor-α, interleukin (IL)-1β, and IL-6 levels in all age groups. Early IL-10 production was significantly downregulated by PTX in term and preterm neonates, while remaining unchanged in adults. Moreover, PTX downregulated TLR4 expression of monocytes on cellular and mRNA level, decreased signaling, and suppressed phagocytosis.ConclusionPTX downregulated TLR4 expression and signaling, thereby leading to strong anti-inflammatory properties in monocytes. Age-dependent differences were identified for CD14 and CD11b expression and IL-10 production.
己酮可可碱（PTX）是一种具有免疫调节特性的甲基黄嘌呤衍生物，已被用作严重新生儿败血症的辅助治疗。本研究的目的是研究 PTX 在体外对早产儿和成人对照单核细胞中脂多糖（LPS）刺激的单核细胞的抗炎作用。
用 LPS 和 PTX 孵育脐带血样本和成人对照血样本。通过流式细胞术评估单核细胞的表面标志物表达、吞噬作用、细胞因子分泌和 Toll 样受体（TLR）4 信号转导。通过逆转录-PCR 确认 TLR4-信使 RNA（mRNA）水平的变化。
PTX 以剂量依赖的方式下调 CD14、CD11b、CD64、CD71 和 CD80 的表达；在早产儿中，下调作用最明显的是 CD14 和 CD11b。PTX 显著下调所有年龄组中 LPS 诱导的肿瘤坏死因子-α、白细胞介素（IL）-1β 和 IL-6 水平。PTX 在足月和早产儿中显著下调 LPS 诱导的早期 IL-10 产生，而在成人中则保持不变。此外，PTX 下调单核细胞的 TLR4 表达在细胞和 mRNA 水平上，降低信号转导，并抑制吞噬作用。
PTX 下调 TLR4 的表达和信号转导，从而导致单核细胞具有强大的抗炎作用。在 CD14 和 CD11b 表达和 IL-10 产生方面，观察到了年龄依赖性差异。
