Expression of anti and pro‐inflammatory genes in human endothelial cells activated by 25‐hydroxycholesterol: A comparison of rivaroxaban and dabigatran

Abstract

Atherosclerosis is associated with a haemostatic imbalance characterized by excessive activation of pro‐inflammatory and pro‐coagulant pathways. Non‐vitamin K antagonists oral anticoagulant (NOACs) may reduce the incidence of cardiovascular events, cerebral ischemia, thromboembolic events and atherosclerosis. Chronic inflammation, vascular proliferation and the development of atherosclerosis is also influenced by 25‐hydroxycholesterol (25‐OHC). The aim of the study was to assess the effect of rivaroxaban and dabigatran on the messenger RNA (mRNA) expression of anti‐inflammatory cytokines transforming growth factor β (TGF‐β), interleukin (IL)‐37, IL‐35 as well as of pro‐inflammatory cytokines IL‐18 and IL‐23, in endothelial cells damaged by 25‐OHC. Human umbilical vascular endothelial cells (HUVECs) were treated with 25‐OHC (10 μg/mL), rivaroxaban (100, 500 ng/mL), dabigatran (100, 500 ng/mL), 25‐OHC + rivaroxaban, and 25‐OHC + dabigatran. The mRNA expression of TGF‐β, IL‐37, IL‐35 subunits EBI3 and p35, IL‐18, and IL‐23 was analysed using real‐time polymerase chain reaction (PCR). The results showed that 25‐OHC decreased TGF‐β and IL‐37 mRNA expression and increased EBI3, p35, IL‐18, IL‐23 mRNA expression in endothelial cell as compared to an untreated control (P < .05). Messenger RNA expression of TGF‐β and IL‐37 significantly increased following stimulation with rivaroxaban and dabigatran as compared to an untreated control (P < .01). In HUVECs pre‐treated with oxysterol, rivaroxaban and dabigatran increased mRNA expression of TGF‐β, IL‐37 and decreased mRNA expression of EBI3, p35, IL‐23 and IL‐18 as compared to 25‐OHC (P < .01). Our finding suggests that both rivaroxaban and dabigatran inhibit the inflammatory activation caused by oxysterol in vitro.

Distinct pleiotropic effects of direct oral anticoagulants on cultured endothelial cells: a comprehensive review

“…Following the examples of statins, metformin, and several other widely used cardiovascular drugs, a search for additional (pleiotropic) effects is currently underway in several preclinical and clinical studies. As it collectively derives from the studies reviewed herein, DOACs have several pleiotropic actions in ECs; it appears that apart from anticoagulation, they exert several additional beneficial actions on the endothelium (depicted in Figure 2 ), including anti-oxidant ( Torramade-Moix et al, 2021 ), anti-inflammatory ( Álvarez et al, 2018 ), atheroprotective ( Gorzelak-Pabiś et al, 2022b ), anti-senescence ( Sanada et al, 2016 ), and anti-fibrotic effects ( Imano et al, 2021 ). Moreover, they preserve endothelial integrity ( Benelhaj et al, 2019 ; Gorzelak-Pabiś et al, 2022a ; Zolotoff et al, 2022 ) and reduce thrombogenicity ( Torramade-Moix et al, 2021 ).…”

“… Gorzelak-Pabiś et al (2022b) also evaluated the effects of the same DOACs in a HUVECs inflammation model. Stimulation of HUVECs with 25-OHC caused downregulation of transforming growth factor beta 1 ( TGFB1 ) and IL-37, and upregulation of IL-18, IL-23, and IL-35, which were counterbalanced when ECs were pre-incubated with rivaroxaban or dabigatran.…”

“…Stimulation of HUVECs with 25-OHC caused downregulation of transforming growth factor beta 1 ( TGFB1 ) and IL-37, and upregulation of IL-18, IL-23, and IL-35, which were counterbalanced when ECs were pre-incubated with rivaroxaban or dabigatran. Thus, both drugs exert anti-inflammatory properties ( Gorzelak-Pabiś et al, 2022b ). The same team also looked into the effects of dabigatran and rivaroxaban on DNA oxidative damage in HUVECs induced by 25-OHC.…”

Pleiotropic Effects of Direct Oral Anticoagulants in Chronic Heart Failure and Atrial Fibrillation: Machine Learning Analysis

“…As mentioned above, preclinical studies showed anti-inflammatory, antioxidant and anti-fibrotic effects on endothelial cells. Importantly, several studies highlighted DOAC-mediated effects on endothelial cells in terms of improvement in endothelial function and integrity and the inhibition of neo-angiogenesis [ 22 , 39 , 40 ]. Although peculiar effects of a single molecule have not yet been determined, it is generally accepted that factor Xa inhibitors produce anti-angiogenic and anti-fibrotic effects beyond the anti-inflammatory action and stabilization of endothelial cells.…”

Endothelial cell dysfunction: Implications for the pathogenesis of peripheral artery disease

“…Forearm blood flow was also improved in diabetic patients administered rivaroxaban for 20 weeks, although treatment was associated with higher bleeding events ( 150 ). In vitro , rivaroxaban reduced ROS (reactive oxygen species), improved DNA repair ( 151 ) and reduced inflammatory gene expression in ECs exposed to hydroxycholesterol ( 152 , 153 ). Rivaroxaban also stimulated blood flow and increased capillary density in a mouse model of diabetic PAD ( 154 ).…”