Clopidogrel Protects Endothelium by Hindering TNFα-Induced VCAM-1 Expression through CaMKKβ/AMPK/Nrf2 Pathway

Abstract

Clopidogrel (INN), an oral antiplatelet drug, has been revealed to have a number of biological properties, for instance, anti-inflammation and antioxidation. Oxidative stress plays an imperative role in inflammation, diabetes mellitus, atherosclerosis, and cancer. In the present study, human aortic endothelial cells (HAECs) were employed to explore the anti-inflammatory activity of INN. INN reduced TNFα-induced reactive oxygen species (ROS) generation and time-dependently prompted the expression and activity of heme oxygenase 1 (HO-1). Cellular glutathione (GSH) levels were augmented by INN. shHO-1 blocked the INN suppression of TNFα-induced HL-60 cell adhesion. The CaMKKβ/AMPK pathway and Nrf2 transcriptional factor were implicated in the induction of HO-1 by INN. Additionally, TNFα dramatically augmented VCAM-1 expression at protein and mRNA levels. INN treatment strikingly repressed TNFα-induced expression of VCAM-1 and HL-60 cell adhesion. Compound C, an AMPK inhibitor, and shNrf2 abolished TNFα-induced expression of VCAM-1 and HL-60 cell adhesion. Our data suggest that INN diminishes TNFα-stimulated VCAM-1 expression at least in part via HO-1 induction, which is CaMKKβ/AMPK pathway-dependent.

Impact of CYP2C19 Variants on Clinical Efficacy of Clopidogrel and 1-Year Clinical Outcomes in Coronary Heart Patients Undergoing Percutaneous Coronary Intervention

“…Clopidogrel exerts not only anti-inflammatory activity but also protective effects on the vascular endothelium (Liu et al, 2012; Willoughby et al, 2014). The drug protects the endothelium by hindering the expression of vascular cell adhesion molecule-1 (VCAM-1) that is induced by a variety of cytokines (Yang et al, 2016). E-selectin, a cell adhesion molecule located on the vascular endothelial cell surface, is the first to be expressed in the inflammatory reaction, in turn causing aggravation of endothelial damage, enhanced inflammation, plaque rupture and thrombosis (Smadja et al, 2012).…”

Effects of clopidogrel on inflammatory cytokines and adhesion molecules in human endothelial cells: Role of nitric oxide mediating pleiotropic effects

“…For the first time, we found that clopidogrel reduces ICAM‐1 expression in the surface of endothelial cells induced by TNF‐α and that this reduction is dependent on NO bioavailability. A recent work also using an in vitro model of endothelial cells, particularly human aortic endothelial cells (HAECs), showed that clopidogrel also reduced the surface expression of the vascular cellular adhesion molecule (VCAM‐1) . VCAM‐1 is an adhesion molecule which acts together with ICAM‐1 in the stable adhesion of leukocytes, allowing further recruitment of inflammatory cells to the intima of arteries .…”

“…A recent work also using an in vitro model of endothelial cells, particularly human aortic endothelial cells (HAECs), showed that clopidogrel also reduced the surface expression of the vascular cellular adhesion molecule (VCAM-1). 29 VCAM-1 is an adhesion molecule which acts together with ICAM-1 in the stable adhesion of leukocytes, allowing further recruitment of inflammatory cells to the intima of arteries. 1,4 Therefore, clopidogrel has a beneficial effect by preventing instauration and progression of atherosclerotic lesions by a NO-dependent mechanism, which is also independent of platelet aggregation inhibition, as suggested by the direct effect in an in vitro model of vascular endothelium.…”

Catalpol attenuates oxidative stress and promotes autophagy in TNF-α-exposed HAECs by up-regulating AMPK

“…35,36 In AS, inhibition of AMPK activity has been reported in multiple cell types. Decreased AMPK activity promotes smooth muscle cell proliferation, monocyte-to-macrophage differentiation, 37 accelerates endothelium inammation 14,38 and so on. As catalpol has been reported to play a relieving role in diabetes through regulating glucose and lipid metabolism, 39 and AMPK is a critical enzyme in energy metabolism, we posited that the protective effect of catalpol on TNF-a-treated HAECs was depending on activating AMPK.…”

“…Adenosine 5 cells from AS through decreasing adhesion molecule expression, attenuating endothelial inammation 13,14 and reducing oxidative stress. Thus, augmentation of AMPK activity is posited to be a potential target for therapeutic interventions in AS.…”