Endogenous Glycosaminoglycans in Various Pathologic Plasma Samples as Measured by a Fluorescent Quenching Method

Abstract

Endogenous glycosaminoglycans (GAGs) with a similar structure to heparin are widely distributed in various tissues. A fluorescence probe, namely Heparin Red, can detect polyanionic GAGs in plasma samples. The purpose of this study is to measure endogenous GAGs in various plasma samples obtained from different pathologic states in comparison to healthy controls utilizing this method. Plasma samples were obtained from patient groups including atrial fibrillation (AF), end-stage-renal-disease (ESRD), diabetes mellitus (DM), sepsis, cancer, liver disease (LD), and pulmonary embolism (PE). Normal human plasma (NHP) was used as healthy controls. The Heparin Red kit from Red Probes (Münster, Germany) was used for the quantification of endogenous GAGs in each sample before and after heparinase I degradation. All results were compiled as group means ± SD for comparison. NHP was found to have relatively low levels of endogenous GAGs with a mean concentration of 0.06 μg/mL. The AF, ESRD, DM, and sepsis patient samples had a mean endogenous GAG concentration of 0.55, 0.72, 0.92, and 0.94 μg/mL, respectively. The levels of endogenous GAGs were highest in cancer, LD, and PE patient plasma samples with a mean concentration of 1.95, 2.78, and 2.83 μg/mL, respectively. Heparinase I degradation resulted in a decline in GAG levels in plasma samples. These results clearly show that detectable Heparin Red sensitive endogenous GAGs are present in circulating plasma at varying levels in various patient groups. Additional studies are necessary to understand this complex pathophysiology.

The Relevance of Thrombo-Inflammatory Biomarkers and Their Relationship with Circulating Glycosaminoglycans in End-Stage Renal Disease Patients

“…Although this method needs to be further evaluated in more well-designed, controlled studies in different pathological disease states, it is important to note that we were able show elevated levels of endogenous GAGs in various pathological plasma samples including ESRD in one of our recent studies. 51 Interestingly, while the levels of vWF were correlated with the levels of tPA, endogenous GAGs were correlated with the levels of PAI-1 antigen. This finding might be due to the fact that while the levels of vWF are significantly associated with endothelial cell damage, endogenous GAGs are significantly associated with glycocalyx degradation.…”

Chemokines, molecular drivers of thromboinflammation and immunothrombosis

“…Whether other abundant chemokines, such as CXCL7, would also result in heparin neutralization is, to our knowledge, unknown. Fluorescent probes enable the quantification of endogenous circulating GAG without differentiating the species or functional activity (52). In healthy human plasma, the concentration of circulating GAGs is in the range of 60 ng/ml.…”