在体外培养的人脐静脉内皮细胞血管损伤模型中，芬戈莫德不能阻止Syndecan-4从内皮糖萼脱落。
Fingolimod does not prevent syndecan-4 shedding from the endothelial glycocalyx in a cultured human umbilical vein endothelial cell model of vascular injury.
作者信息
Milford Elissa M, Meital Lara, Kuballa Anna, Reade Michael C, Russell Fraser D
机构信息
Faculty of Medicine, University of Queensland, Herston, QLD, Australia.
Intensive Care Unit, Royal Brisbane and Women's Hospital, Butterfield St., Herston, QLD, Australia.
出版信息
Intensive Care Med Exp. 2022 Aug 18;10(1):34. doi: 10.1186/s40635-022-00462-7.
BACKGROUND
Shedding of the endothelial glycocalyx (EG) is associated with poor outcomes in a range of conditions including sepsis. Fresh frozen plasma (FFP) restores the damaged EG to baseline thickness, however the mechanism for this effect is unknown, and some components of FFP have adverse effects unrelated to the EG. There is some limited evidence that sphingosine-1-phosphate (S1P) within FFP restores the EG by activating the endothelial cell S1P receptor 1 (S1PR). However, there are disadvantages to using S1P clinically as an EG restorative therapy. A potential alternative is the S1PR agonist fingolimod (FTY720). The aim of this study was to assess whether FTY720 prevents EG shedding in injured cultured human umbilical vein endothelial cells.
METHODS
Shedding of the EG was induced in cultured human umbilical vein endothelial cells (HUVECs) by exposure to adrenaline, TNF-α and HO. The cells were then assigned to one of six conditions for 4 h: uninjured and untreated, injured and untreated, injured and treated with FTY720 with and without the S1PR inhibitor W146, and injured and treated with 25% FFP with and without W146. Syndecan-4, a component of the EG, was measured in cell supernatants, and syndecan-4 and thrombomodulin mRNA expression was quantitated in cell lysates.
RESULTS
The injury resulted in a 2.1-fold increase in syndecan-4 (p < 0.001), consistent with EG shedding. Syndecan-4 and thrombomodulin mRNA expression was increased (p < 0.001) and decreased (p < 0.05), respectively, by the injury. Syndecan-4 shedding was not affected by treatment with FTY720, whereas FFP attenuated syndecan-4 shedding back to baseline levels in the injured cells and this was unaffected by W146. Neither treatment affected syndecan-4 or thrombomodulin mRNA expression.
CONCLUSIONS
FTY720 did not prevent syndecan-4 shedding from the EG in the HUVEC model of endothelial injury, suggesting that activation of S1PR does not prevent EG damage. FFP prevented syndecan-4 shedding from the EG via a mechanism that was independent of S1PR and upregulation of SDC-4 production. Further studies to examine whether FTY720 or another S1PR agonist might have EG-protective effects under different conditions are warranted, as are investigations seeking the mechanism of EG protection conferred by FFP in this experimental model.
背景
在内皮糖萼（EG）脱落与包括脓毒症在内的一系列病症的不良预后相关。新鲜冷冻血浆（FFP）可将受损的EG恢复至基线厚度，然而这种作用的机制尚不清楚，并且FFP的某些成分具有与EG无关的不良影响。有一些有限的证据表明，FFP中的鞘氨醇-1-磷酸（S1P）通过激活内皮细胞S1P受体1（S1PR）来恢复EG。然而，临床上将S1P用作EG恢复疗法存在缺点。一种潜在的替代物是S1PR激动剂芬戈莫德（FTY720）。本研究的目的是评估FTY720是否能防止受损的人脐静脉内皮细胞培养物中EG的脱落。
方法
通过暴露于肾上腺素、TNF-α和HO诱导人脐静脉内皮细胞（HUVECs）培养物中EG的脱落。然后将细胞分配到六种条件之一，持续4小时：未受损且未处理、受损且未处理、受损并用FTY720处理（有无S1PR抑制剂W146），以及受损并用25% FFP处理（有无W146）。在细胞上清液中测量EG的一种成分Syndecan-4，并在细胞裂解物中定量Syndecan-4和血栓调节蛋白的mRNA表达。
结果
损伤导致Syndecan-4增加2.1倍（p < 0.001），与EG脱落一致。损伤分别使Syndecan-4和血栓调节蛋白的mRNA表达增加（p < 0.001）和降低（p < 0.05）。FTY720处理不影响Syndecan-4的脱落，而FFP可将受损细胞中Syndecan-4的脱落减弱至基线水平，且不受W146影响。两种处理均不影响Syndecan-4或血栓调节蛋白的mRNA表达。
结论
在HUVEC内皮损伤模型中，FTY720不能防止EG中Syndecan-4的脱落，这表明S1PR的激活不能防止EG损伤。FFP通过一种独立于S1PR和SDC-4产生上调的机制防止EG中Syndecan-4的脱落。有必要进一步研究FTY720或其他S1PR激动剂在不同条件下是否可能具有EG保护作用，以及在该实验模型中寻找FFP赋予EG保护作用的机制的研究。
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