Identification of Fibrinogen as a Key Anti-Apoptotic Factor in Human Fresh Frozen Plasma for Protecting Endothelial Cells In Vitro

Abstract

Resuscitation with human fresh frozen plasma (FFP) in hemorrhagic shock (HS) patients is associated with improved clinical outcomes. Our group has demonstrated that the beneficial effect of FFP is due to its blockade on endothelial hyperpermeability, thereby improving vascular barrier function. The current study aimed to investigate HS-induced endothelial cell apoptosis, a potential major contributor to the endothelial hyperpermeability, and to determine the effect and the key components/factors of FFP on protecting endothelial cells from apoptosis. We first measured and demonstrated an increase in apoptotic endothelial microparticles (CD146+AnnexinV+) in patients in shock compared to normal subjects, indicating the induction of endothelial cell activation and apoptosis in shock patients. We then transfused HS rats with FFP and showed that FFP blocked HS-induced endothelial cell apoptosis in gut tissue. To identify the anti-apoptotic factors in FFP, we utilized high-performance liquid chromatography, fractionated FFP, and screened the fractions in vitro for the anti-apoptotic effects. We selected the most effective fractions, performed mass spectrometry, and identified fibrinogen as a potent anti-apoptotic factor. Taken together, our findings suggest that HS-induced endothelial apoptosis may constitute a major mechanism underlying the vascular hyperpermeability. Furthermore, the identified anti-apoptotic factor fibrinogen may contribute to the beneficial effects of FFP resuscitation, and therefore, may have therapeutic potential for HS.

Cryoprecipitate attenuates the endotheliopathy of trauma in mice subjected to hemorrhagic shock and trauma

“…34 Fibrinogen appears to stabilize EC surface syndecan-1, 34,35 which is a key component of the endothelial glycocalyx, and may protect against HS-induced EC apoptosis. 52 Moreover, there have been hundreds of other proteins identified in FFP outside of clotting factors, many of which have important biologic functions and could potentially mediate some of these protective effects. 53 It is possible that many of these proteins are also present in cryoprecipitate, and we hypothesize that the similar effects we find between FFP and cryoprecipitate are due to these EOT-modulatory proteins common to both products.…”

Endothelial Dysfunction is Associated With Increased Incidence, Worsened Severity, and Prolonged Duration of Acute Kidney Injury After Severe Trauma

“…Fresh frozen plasma is one of the leading clinically available treatments that has been shown to be effective at stabilizing the endothelium after severe trauma and in experimental hemorrhage models (33,34). Both fibrinogen and antithrombin III have been recognized for their roles in this observed effect (35)(36)(37). The benefit of these individual factors on AKI specifically has yet to be evaluated.…”

Spray‐dried plasma: A post‐traumatic blood “bridge” for life‐saving resuscitation

“…23,42,43 Further investigation has identified fibrinogen as a key component of plasma protection against endothelial apoptosis during hemorrhagic shock. 44 Fibrinogen can protect against endothelial barrier dysfunction by maintaining cell surface syndecan-1 in vitro 45 and may exert its protective effect directly on endothelial cells by its binding to syndecan-1 and activation of the P21 activated kinase 1 (PAK1) to reduce cytoskeletal stress fiber formation and endothelial permeability. 46 Additional plasma proteins, including antithrombin III and adiponectin have been identified as also contributing to endothelial barrier maintenance during hemorrhagic shock.…”

“…In preclinical studies, plasma can restore and/or prevent endotheliopathy by preserving endothelial glycocalyx and preventing endothelial barrier disruption during resuscitation of hemorrhagic shock 23, 42, 43 . Further investigation has identified fibrinogen as a key component of plasma protection against endothelial apoptosis during hemorrhagic shock 44 . Fibrinogen can protect against endothelial barrier dysfunction by maintaining cell surface syndecan‐1 in vitro 45 and may exert its protective effect directly on endothelial cells by its binding to syndecan‐1 and activation of the P21 activated kinase 1 (PAK1) to reduce cytoskeletal stress fiber formation and endothelial permeability 46 .…”