de Guevara Yuliannis Lugones Ladrón, Hidalgo Odalys Blanco, Santos Sidnéia Sousa, Brunialti Milena Karina Colo, Faure Roberto, Salomao Reinaldo
Centro Nacional de Sanidad Agropecuaria (CENSA), San José de las Lajas, Mayabeque, Cuba.
Division of Infectious Diseases, Hospital São Paulo, Escola Paulista de Medicina, Universidade Federal de São Paulo, Brazil.
Int Immunopharmacol. 2014 Aug;21(2):369-74. doi: 10.1016/j.intimp.2014.05.020. Epub 2014 May 27.
Surfacen® is a clinical surfactant preparation of porcine origin. In the present study, we have evaluated the effect of Surfacen® in the modulation of oxidative burst in monocytes and neutrophils in human blood and pro-inflammatory cytokine production in peripheral blood mononuclear cells (PBMC). Reactive oxygen species (ROS) level was measured in monocytes and neutrophils by flow cytometry using 2,7-dichlorofluorescein diacetate (DCFH-DA) as substrate, while, tumor necrosis factor (TNF)-α and interleukin (IL)-6 levels were estimated in PBMC supernatant by enzyme-linked immunosorbent assays (ELISA). Our results show that Staphylococcus aureus-induced ROS level was slightly affected by Surfacen® added to whole blood monocytes and neutrophils. The time course experiments of pre-incubation with Surfacen® showed no significant increase of ROS level at 2h; however, the ROS level decreased when pre incubated for 4h and 6h with Surfacen®. Pre-incubation of PBMC cells with Surfacen® at 0.125 and 0.5mg/mL showed a dose-dependent suppression of TNF-α levels measured after 4h of S. aureus stimulation, an effect less impressive when cells were stimulated for 24h. A similar behavior was observed in IL-6 release. In summary, the present study provides experimental evidence supporting an anti-inflammatory role of Surfacen® in human monocytes and neutrophils in vitro.
Surfacen®是一种猪源性临床表面活性剂制剂。在本研究中,我们评估了Surfacen®对人血中单核细胞和中性粒细胞氧化爆发的调节作用以及对外周血单核细胞(PBMC)促炎细胞因子产生的影响。使用2,7-二氯荧光素二乙酸酯(DCFH-DA)作为底物,通过流式细胞术测定单核细胞和中性粒细胞中的活性氧(ROS)水平,同时,通过酶联免疫吸附测定(ELISA)评估PBMC上清液中的肿瘤坏死因子(TNF)-α和白细胞介素(IL)-6水平。我们的结果表明,添加到全血单核细胞和中性粒细胞中的Surfacen®对金黄色葡萄球菌诱导的ROS水平影响较小。用Surfacen®预孵育的时间进程实验表明,在2小时时ROS水平没有显著增加;然而,当与Surfacen®预孵育4小时和6小时时,ROS水平下降。用0.125和0.5mg/mL的Surfacen®预孵育PBMC细胞显示,在金黄色葡萄球菌刺激4小时后测量的TNF-α水平呈剂量依赖性抑制,当细胞刺激24小时时,这种效果不太明显。在IL-6释放中观察到类似的行为。总之,本研究提供了实验证据,支持Surfacen®在体外对人单核细胞和中性粒细胞具有抗炎作用。