Effects of factor Xa on the expression of proteins in femoral arteries from type 2 diabetic patients

Abstract

AIMFurther to its pivotal role in haemostasis, factor Xa (FXa) promotes effects on the vascular wall. The purpose of the study was to evaluate if FXa modifies the expression level of energy metabolism and oxidative stress-related proteins in femoral arteries obtained from type 2 diabetic patients with end-stage vasculopathy. METHODSFemoral arteries were obtained from 12 type 2 diabetic patients who underwent leg amputation. Segments from the femoral arteries were incubated in vitro alone and in the presence of 25 nmol l −1 FXa and 25 nmol l −1 FXa + 50 nmol l −1 rivaroxaban. RESULTSIn the femoral arteries, FXa increased triosephosphate isomerase and glyceraldehyde-3-phosphate dehydrogenase isotype 1 expression but decreased pyruvate dehydrogenase expression. These facts were accompanied by an increased content of acetyl-CoA. Aconitase activity was reduced in FXa-incubated femoral arteries as compared with control. Moreover, FXa increased the protein expression level of oxidative stress-related proteins which was accompanied by an increased malonyldialdehyde arterial content. The FXa inhibitor, rivaroxaban, failed to prevent the reduced expression of pyruvate dehydrogenase induced by FXa but reduced acetyl-CoA content and reverted the decreased aconitase activity observed with FXa alone. Rivaroxaban + FXa but not FXa alone increased the expression level of carnitine palmitoyltransferase I and II, two mitochondrial long chain fatty acid transporters. Rivaroxaban also prevented the increased expression of oxidative stress-related proteins induced by FXa alone. CONCLUSIONSIn femoral isolated arteries from type 2 diabetic patients with end-stage vasculopathy, FXa promoted disruption of the aerobic mitochondrial metabolism. Rivaroxaban prevented such effects and even seemed to favour long chain fatty acid transport into mitochondria. WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT• Factor Xa (FXa) modifies the vascular wall which suggests that FXa is a modulator of vascular function.• Patients with type 2 diabetes mellitus have an increased risk of cardiovascular events associated with thrombotic and pro-coagulant conditions.• In diabetes, abnormalities in vascular energy metabolism contribute to vascular dysfunction and higher risk of cardiovascular events WHAT THIS STUDY ADDS• FXa stimulates the glycolytic pathway in diabetic arteries but not to pyruvate anaerobic catabolism.• There is no involvement of long chain fatty acid beta oxidation as alternative source to increase acetyl-CoA in FXa-incubated arteries. • FXa changes the expression level of glucose oxidation-related proteins suggesting disruption of mitochondrial metabolism.

Coagulation Factor Xa and Protease-Activated Receptor 2 as Novel Therapeutic Targets for Diabetic Nephropathy

“…40 In our studies, Noxs mRNA expressions were not altered among Figure IV in the online-only Data Supplement), which is consistent with our previous finding that exacerbation of DN in the absence of eNOS is not associated with oxidative stress marker (reduced and oxidized glutathiones). 6 NADPH oxidases or oxidative stress dose not cause injurious effects in our model.…”

FXa inhibition by rivaroxaban modifies mechanisms associated with the pathogenesis of human abdominal aortic aneurysms

“…The fact that in supernatants from AAA explants, IL-10 production was not modified with respect to control while in rivaroxaban-incubated AAA was significantly increased might suggest an independent effect of rivaroxaban on endogenous FXa. In this regard, in a previous study performed in femoral arteries of diabetic patients, we have also observed an effect of rivaroxaban probably unrelated to FXa on the expression of long mitochondrial fatty acid transporters [25]. As we mentioned in that report, it is known the existence of rivaroxaban-derived metabolites and, therefore, we cannot rule out that some of the apparently FXa-independent effects of rivaroxaban could be attributed to such metabolites [48].…”

“…At the AAA site, the FXa inhibitor, rivaroxaban, significantly reduced the expression level of the gp67-and 91-phox NADPH oxidase isotypes, suggesting the involvement of FXa in such increased expression. Accordingly, in femoral arteries obtained from diabetic patients we have also demonstrated that rivaroxaban reduced the expression of oxidative stressrelated proteins [25] . Interestingly, it was suggested that NOS2, whose expression was significantly reduced by rivaroxaban in the AAA site, might be link between inflammation and oxidative stress in AAA since NOS2 was also demonstrated as primary sources of superoxide anion in AAA [50].…”

“…In this regard, it was demonstrated that FXa increased oxidative stress in human vascular smooth muscle cells from saphenous vein [24]. Moreover, in femoral arteries from diabetic patients, FXa modified the expression level of proteins associated with oxidative stress and energetic metabolism, effects that were prevented by rivaroxaban [25]. However, to our knowledge, possible effects of endogenous FXa have not been examined on proteins associated with inflammation and/or oxidative stress in AAA.…”

Effects of Oral Anticoagulant Therapy on Gene Expression in Crosstalk between Osteogenic Progenitor Cells and Endothelial Cells

“…Thrombin-PAR-1 interaction activates the signaling axis involved in actinic cytoskeleton modifications of endothelial cells, thus contributing to endothelial barrier disruption and increased permeability [5]. In patients suffering from vascular disease, factor Xa modifies the expression levels of oxidative stress-induced proteins, thereby affecting aerobic mitochondrial metabolism [6]. Through the stimulation of PAR-1 and -2 and the activation of insulin-like growth factor binding protein 5 (IGFBP5) and p53, factor Xa induces a senescent phenotype and the expression of pro-inflammatory molecules in the smooth muscle cells of the vascular wall [7].…”