Lund T, Mangsbo S M, Scholz H, Gjorstrup P, Tötterman T H, Korsgren O, Foss A
Division of Surgery, Section for Transplantation, Oslo University Hospital, Oslo, Norway.
Exp Clin Endocrinol Diabetes. 2010 Apr;118(4):237-44. doi: 10.1055/s-0029-1241825. Epub 2010 Jan 29.
In clinical islet transplantation, inflammatory responses initiated by the transplanted islets and by the host immune system cause acute and chronic graft loss. The resolution of acute inflammation is an active process mediated by specific signals and mediators such as resolvin E1 (RvE1). We investigated the effect of RvE1 on i) the inflammatory status of human pancreatic islets, ii) islet viability and apoptosis, and iii) the instant blood-mediated inflammatory reaction (IBMIR) IN VITRO.
Pro-inflammatory cytokines and tissue factor (TF) in isolated human islets were determined by real-time RT-qPCR (mRNA levels), CBA and Gyrolab bioaffy (protein levels) after lipopolysaccaride (LPS) stimulation. Islet viability was measured using insulin secretion in a dynamic model, ADP/ATP ratio and total ATP content. Apoptosis was measured using commercial kits after stimulation with proinflammatory cytokines. To assess effect on IBMIR, human islets were mixed with non-anticoagulated, RvE1 or vehicle pretreated ABO-compatible blood in heparin-coated tubing loops.
Treatment of human islets with RvE1 (500 nM) for 24 h reduced LPS-induced increase in mRNA and protein levels of selected pro-inflammatory markers (IL-8, MCP-1, and TF). RvE1 lowered the ADP/ATP ratio, but had no effect on insulin secretion. RvE1 reduced the apoptotic effect of proinflammatory cytokines. Additionally, RvE1 reduced platelet consumption and TAT complex formation during the first 5 min after islet-blood contact.
RvE1 suppresses proinflammatory markers and lowers the ADP/ATP ratio in human islets IN VITRO. RvE1 demonstrates anti-apoptotic effects in a proinflammatory milieu. Additionally, RvE1 has modest dampening effects on IBMIR. We conclude that RvE1 may have potential in clinical islet transplantation.
在临床胰岛移植中，移植胰岛和宿主免疫系统引发的炎症反应会导致急性和慢性移植物丢失。急性炎症的消退是一个由特定信号和介质（如消退素E1，RvE1）介导的活跃过程。我们研究了RvE1对以下方面的影响：i）人胰岛的炎症状态；ii）胰岛活力和凋亡；iii）体外即时血液介导的炎症反应（IBMIR）。
通过实时逆转录定量聚合酶链反应（mRNA水平）、细胞因子检测技术（CBA）和Gyrolab生物芯片（蛋白质水平），测定脂多糖（LPS）刺激后分离的人胰岛中促炎细胞因子和组织因子（TF）。在动态模型中使用胰岛素分泌、ADP/ATP比值和总ATP含量来测量胰岛活力。在用促炎细胞因子刺激后，使用商业试剂盒测量凋亡情况。为了评估对IBMIR的影响，将人胰岛与未抗凝、经RvE1或载体预处理的ABO血型相容血液在肝素包被的管路环中混合。
用RvE1（500 nM）处理人胰岛24小时可降低LPS诱导的所选促炎标志物（IL-8、MCP-1和TF）mRNA和蛋白质水平的升高。RvE1降低了ADP/ATP比值，但对胰岛素分泌没有影响。RvE1降低了促炎细胞因子的凋亡作用。此外，RvE1减少了胰岛与血液接触后最初5分钟内的血小板消耗和凝血酶 - 抗凝血酶复合物（TAT复合物）形成。
RvE1在体外可抑制人胰岛中的促炎标志物并降低ADP/ATP比值。RvE1在促炎环境中表现出抗凋亡作用。此外，RvE1对IBMIR有适度的抑制作用。我们得出结论，RvE1在临床胰岛移植中可能具有潜力。
