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Abstract Title:

Ascorbic acid attenuates endothelial permeability triggered by cell-free hemoglobin.

Abstract Source:

Biochem Biophys Res Commun. 2018 01 1 ;495(1):433-437. Epub 2017 Nov 9. PMID: 29129689

Abstract Author(s):

Jamie L Kuck, Julie A Bastarache, Ciara M Shaver, Joshua P Fessel, Sergey I Dikalov, James M May, Lorraine B Ware

Article Affiliation:

Jamie L Kuck

Abstract:

BACKGROUND: Increased endothelial permeability is central to shock and organ dysfunction in sepsis but therapeutics targeted to known mediators of increased endothelial permeability have been unsuccessful in patient studies. We previously reported that cell-free hemoglobin (CFH) is elevated in the majority of patients with sepsis and is associated with organ dysfunction, poor clinical outcomes and elevated markers of oxidant injury. Others have shown that Vitamin C (ascorbate) may have endothelial protective effects in sepsis. In this study, we tested the hypothesis that high levels of CFH, as seen in the circulation of patients with sepsis, disrupt endothelial barrier integrity.

METHODS: Human umbilical vein endothelial cells (HUVEC) were grown to confluence and treated with CFH with or without ascorbate. Monolayer permeability was measured by Electric Cell-substrate Impedance Sensing (ECIS) or transfer ofC-inulin. Viability was measured by trypan blue exclusion. Intracellular ascorbate was measured by HPLC.

RESULTS: CFH increased permeability in a dose- and time-dependent manner with 1 mg/ml of CFH increasing inulin transfer by 50% without affecting cell viability. CFH (1 mg/ml) also caused a dramatic reduction in intracellular ascorbate in the same time frame (1.4 mM without CFH, 0.23 mM 18 h after 1 mg/ml CFH, p < 0.05). Pre-treatment of HUVECs with ascorbate attenuated CFH induced permeability.

CONCLUSIONS: CFH increases endothelial permeability in part through depletion of intracellular ascorbate. Supplementation of ascorbate can attenuate increases in permeability mediated by CFH suggesting a possible therapeutic approach in sepsis.

Study Type : In Vitro Study

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Sayer Ji
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