Heparin and LPS-induced COX-2 expression in airway cells: a link between its anti-inflammatory effects and GAG sulfation

Abstract

Purpose/Aim Previous studies have indicated that the sulfated polysaccharide heparin has anti-inflammatory effects. However, the mechanistic basis for these effects has not been fully elucidated. Materials and Methods NCI-H292 (mucoepidermoid) and HBE-1 (normal) human bronchial epithelial cells were treated with LPS alone or in the presence of high-molecular-weight (HMW) fully-sulfated heparin or desulfated HMW heparin. Cells were harvested to examine the phosphorylation levels of ERK1/2, p38, and NF-κB p65 and COX-2 protein expression by Western blot and gene expression of both COX-2 and CXCL-8 by TaqMan qRT-PCR. Results Heparin is known to exert an influence on receptor-mediated signaling through its ability to both potentiate and inhibit the receptor-ligand interaction, depending upon its concentration. In H292 cells, fully-sulfated HMW heparin significantly reduced LPS-induced gene expression of both COX-2 and CXCL-8 for up to 48 hours, while desulfated heparin had little to no significant suppressive effect on signaling or on COX-2 gene or protein expression. Desulfated heparin, initially effective at preventing LPS-induced CXCL8 up-regulation, reduced CXCL8 transcription at 24 hours. In contrast, in normal HBE-1 cells, fully-sulfated heparin significantly suppressed only ERK signaling, COX-2 gene expression at 12 hours, and CXCL-8 gene expression at 6 and 12 hours, while desulfated heparin had no significant effects on LPS-stimulated signaling or on gene or protein expression. Sulfation determines heparin’s influence and may reflect the moderating role of GAG sulfation in lung injury and health. Conclusions Heparin’s anti-inflammatory effects result from its non-specific suppression of signaling and gene expression and are determined by its sulfation.

Up‐regulation of microRNA‐574 attenuates lipopolysaccharide‐ or cecal ligation and puncture‐induced sepsis associated with acute lung injury

“…The dose of LPS was also used in other study. Certainly it might be a limitation not showing culture images . HBE cells were transfected with siC3 and NC or/and incubated with LPS.…”

Pulmonary immunity and extracellular matrix interactions

“…For example, AECs grown in the presence of de-sulfated chondroitin sulfate or de-sulfated heparin along with laminin proliferate in response to FGF-2 whereas sulfated heparin can inhibit this proliferation [93, 94] suggesting that the sulfation status of the basement membrane proteoglycans may be an important determinant of AEC responses to injury, repair and differentiation. Not surprisingly, the well-known anti-inflammatory effects of heparin have recently been determined to be mediated by the sulfation status of the molecule [95].…”

Role of Adaptor Protein Myeloid Differentiation 88 (MyD88) in Post-Subarachnoid Hemorrhage Inflammation: A Systematic Review

“…Nevertheless, advanced studies have proved that heparin has a diminutive effect on vasospastic damage, especially SAH-associated whereas it may still be effective in lowering inflammatory-related injuries that are related to SAH [67]. The underlying anti-inflammatory properties of heparin may stem from its ability to ameliorate both MAPK-p38 and NF-κB inflammatory pathways downstream of the MyD88 adaptor [68].…”