Xia Brent T, Beckmann Nadine, Winer Leah K, Pugh Amanda M, Pritts Timothy A, Nomellini Vanessa, Gulbins Erich, Caldwell Charles C
Division of Research, Department of Surgery, College of Medicine, University of Cincinnati, Cincinnati, OH, USA.
Department of Molecular Biology, University Hospital, University of Duisburg-Essen, Essen, Germany.
Cell Physiol Biochem. 2019;52(3):565-579. doi: 10.33594/000000040.
BACKGROUND/AIMS: During sepsis, an unchecked pro-inflammatory response can be detrimental to the host. We investigated the potential protective effect of amitriptyline (AT).
We used two murine models of sepsis: Cecal ligation and puncture and endotoxemia following LPS challenge. Aural temperatures were taken and cytokines quantified by cytometric bead assay. Lung injury was determined histologically and by protein determination in bronchoalveolar lavage fluid. Cell accumulation in the peritoneum was analyzed by flow cytometry, as well as cytokine production and p38-phosphorylation. Neutrophil chemotaxis was evaluated using an in vitro transwell assay.
Our findings demonstrate that AT-treated septic mice have improved survival and are protected from pulmonary edema. Treatment with AT significantly decreased serum levels of KC and monocyte chemoattractant protein-1, as well as the accumulation of neutrophils and monocytes in the peritoneum of septic mice. Peritoneal IL-10 levels in septic mice were increased upon AT treatment. Direct treatment of septic mice with IL-10 recapitulated the effects of AT. Endotoxemic mice also exhibited enhanced IL-10 production upon AT-administration and peritoneal macrophages were identified as the ATinfluenced producers of IL-10. Treatment of these cells with AT in vitro resulted in increased p38-phosphorylation and IL-10 generation, whereas ceramide and p38 inhibition had the opposite effect.
Altogether, AT treatment improved survival, increased IL-10 levels, and mitigated a pro-inflammatory response during sepsis. We conclude that AT is a promising therapeutic to temper inflammation during septic shock.
背景/目的:在脓毒症期间,不受控制的促炎反应可能对宿主有害。我们研究了阿米替林(AT)的潜在保护作用。
我们使用了两种脓毒症小鼠模型:盲肠结扎穿刺模型和脂多糖(LPS)攻击后的内毒素血症模型。测量耳温,并通过细胞计数珠分析法对细胞因子进行定量。通过组织学检查和支气管肺泡灌洗液中的蛋白质测定来确定肺损伤。通过流式细胞术分析腹膜中的细胞积聚情况,以及细胞因子的产生和p38磷酸化情况。使用体外Transwell试验评估中性粒细胞趋化性。
我们的研究结果表明,接受AT治疗的脓毒症小鼠存活率提高,且免受肺水肿影响。AT治疗显著降低了血清中KC和单核细胞趋化蛋白-1的水平,以及脓毒症小鼠腹膜中中性粒细胞和单核细胞的积聚。AT治疗后,脓毒症小鼠腹膜中的IL-10水平升高。用IL-10直接治疗脓毒症小鼠可重现AT的作用。给予AT后,内毒素血症小鼠也表现出IL-10产生增加,且腹膜巨噬细胞被确定为受AT影响的IL-10产生细胞。在体外对这些细胞进行AT处理导致p38磷酸化增加和IL-10生成增加,而神经酰胺和p38抑制则产生相反的效果。
总之,AT治疗提高了存活率,增加了IL-10水平,并减轻了脓毒症期间的促炎反应。我们得出结论,AT是一种在脓毒性休克期间调节炎症的有前景的治疗方法。