Sulodexide up-regulates glutathione S-transferase P1 by enhancing Nrf2 expression and translocation in human umbilical vein endothelial cells injured by oxygen glucose deprivation

Abstract

Introduction: Sulodexide (SDX) is used for the treatment of many vascular disorders due to its anticoagulant, anti-inflammatory and anti-atherosclerotic properties. However, the detailed molecular mechanism of its endothelioprotective action is still not completely understood. There is increasing evidence suggesting that antioxidant enzymes play an important role in anti-ischemic properties of SDX. We postulate that up-regulation of glutathione-S-transferase P1 (GSTP1) mediated by the transcription factor Nrf2 could be associated with the antioxidant effect of SDX on vascular endothelial cells. Material and methods: In the present study, we investigated whether SDX affects GSTP1 and Nrf2 in oxygen glucose deprivation (OGD) treated human umbilical vein endothelial cells (HUVECs). The cells treated with/without SDX (0.5 LRU/ml) were subjected to OGD for 1-6 h. To study the influence of SDX on the Nrf2 nucleus accumulation, the cells were incubated with 0.5 LRU/ml SDX in OGD for 1 h. Results: We found that after short-term OGD (1-3 h), the drug increased the expression of both GSTP1 and Nrf2 mRNA/protein in HUVECs (p < 0.05), as determined by real-time PCR and enzyme-linked immunosorbent assay (ELISA). SDX treatment also enhanced the nuclear accumulation of Nrf2 in HUVECs after 1 h of OGD (p < 0.05). Conclusions: SDX induces a rapid onset of the antioxidant response by up-regulating the expression of GSTP1 and Nrf2 in endothelial cells subjected to in vitro simulated ischemia.

Sulodexide increases mRNA expression of glutathione-related genes in human umbilical endothelial cells exposed to oxygen-glucose deprivation

“…It is known that Nrf2 is an important regulator of GSH-related gene expression [8]. We recently reported that Nrf2 and glutathione S-transferase p1 (GSTP1) are up-regulated in response to SDX in HUVECs exposed to OGD [3]. Here we found that SDX induces very early and transient transcription of de novo GSH synthesis genes, including GCLc and xCT (Figures 1 A, B).…”

“…HUVECs (Lonza, USA) were cultured in EGM-2 with EGM-2 BulletKit as previously described [3]. SDX (0.5 LRU/ml) was added to glucose-free DMEM and cells were harvested in a hypoxic chamber (3% O 2 , 5% CO 2 , 92% N 2 at 37°C) after different OGD times (1-6 h).…”

“…SDX possesses endothelial protective effects due to its antioxidant, anti-inflammatory and antiproteolytic properties [1]. In endothelium, SDX interacts with nuclear factor-erythroid-2-related factor (Nrf2)-antioxidant response element (ARE) and nuclear factor-kB (NF-kB) signaling pathways [1,3]. Nrf2 regulates the expression of enzymes involved in glutathione (GSH) synthesis and utilization such as g-glutamylcysteine ligase (GCL), cysteine-glutamate exchanger (xCT), glutathione synthase (GS), glutathione peroxidase (GPx) and glutathione reductase (GR).…”

Overexpression of PGC-1α reduces inflammation and protects against focal cerebral ischaemia/reperfusion injury

“…Nrf-2 is mainly associated with Keap1 (Kelch-like ECH associated protein 1) protein in the cytoplasmic compartment [28,29]. Under oxidative stress, Nrf-2 translocates into the nucleus and binds to antioxidant response elements (ARE) to activate cytoprotective enzymes (GSH, heme oxygenase 1, NQO-1, GPX, GST) [30][31][32]. The present results showed that PGC-1α regulates apoptosis through the Nrf-2 pathway.…”

Glycocalyx sialic acids regulate Nrf2-mediated signaling by fluid shear stress in human endothelial cells

“…Notably, experimental restoration of SIA has been shown to be efficacious against atherosclerosis [ 99 ], obesity-related hypertension [ 100 ] and age-related renal microvascular dysfunction [ 101 ]. Furthermore, it was recently reported that GCX enhancement by the GAG supplement sulodexide activates Nrf2 signaling to confer cytoprotection against ischaemia-reperfusion injury [ 102 ]. Further studies are thus warranted to further elucidate interactions between GCX components in coordinating Nrf2-regulated antioxidant defences.…”