Atherosclerosis 202 (2009) 41–47 Anthocyanin attenuates CD40-mediated endothelial cell activation and apoptosis by inhibiting CD40-induced MAPK activation Min Xia, Wenhua Ling ∗ , Huilian Zhu, Jing Ma, Qing Wang, Mengjun Hou, Zhihong Tang, Honghui Guo, Chi Liu, Qingyuan Ye Department of Nutrition, School of Public Health, Sun Yat-Sen University (Northern Campus) 74 Zhongshan Road 2, Guangzhou 510080, Guangdong Province, PR China Received 9 April 2007; received in revised form 30 March 2008; accepted 3 April 2008 Available online 12 April 2008 Abstract CD40-mediated inflammatory signaling is a potent activator of endothelial cells (ECs) and effective in triggering the pathogenesis of atherosclerosis, a chronic inflammatory disease. Anthocyanin is considered to exert potent cardiovascular-protective effect partially through its anti-inflammatory property, however, the precise mechanism is still unknown. Here we chose cultured human umbilical vein endothelial cells (HUVECs) to explore the influence of anthocyanin on CD40-mediated endothelial activation and apoptosis and the underlying mechanism. Stimulation of human primary HUVECs by CD40 with its physiological ligand CD40L not only augmented MMP-1, -9 secretion and promoted MMP-1, -9 activities, but also induced endothelial cell apoptosis and death. Treatment of ECs with anthocyanins cyanidin-3- O--glucoside (Cy-3-g) and peonidin-3-O--glucoside (Pn-3-g) prevents CD40-induced endothelial activation by inhibiting production of proinflammatory cytokines and matrix metalloproteinases (MMPs). In addition, exposure to anthocyanins inhibits CD40-induced endothelial apoptosis. Anthocyanins also decreased activation of JNK and p38 induced by CD40. Collectively, our findings suggested that the inhibition of JNK and p38 activation interrupts CD40 induced endothelial cell activation and apoptosis, which thereby may represent a mechanism that would explain the anti-inflammatory response of anthocyanin and its athero-protective function. © 2008 Elsevier Ireland Ltd. All rights reserved. Keywords: Atherosclerosis; CD40; Endothelium; Adhesion molecules; Matrix metalloproteinases 1. Introduction Nowadays, a large body of evidence supports the view of atherosclerosis as a chronic inflammatory disease and implicates components of the immune system in atheroge- nesis [1–3]. Recently, many reports have documented that the immune mediator CD40 and its counterpart CD40 ligand (CD40L) mediated inflammation are potent activators in the pathogenesis of this disease [1–3]. CD40 and CD40L are members of the tumor necrosis fac- tor (TNF) and TNF-receptor (TNFR) family [3]. Although the initial role of the CD40–CD40L system was thought to be predominantly restricted to T- and B-lymphocyte inter- actions, it is now known that this dyad, also expressed by ∗ Corresponding author. Tel.: +86 20 87331597; fax: +86 20 87330446. E-mail address: lingwh@mail.sysu.edu.cn (W. Ling). a variety of nonimmune cells, including vascular cells like endothelial cells (ECs) exerted a wide range of functions [3]. CD40 ligation by CD40L induces production of various inflammatory cytokines and chemokines such as interleukins, monocyte chemoattractant protein-1 (MCP-1) and adhesion molecules which may activate the pro-atherogenic pathways [4,5]. Beside these, CD40–CD40L interactions are also pos- tulated to play an important role in the development of plaque rupture. Many studies have found that CD40 and CD40L abundantly presented in the shoulder region of rupture prone lesions, and stimulation of their signaling pathway resulted in the upregulation of the expression of matrix metallopro- teinases (MMPs) [6,7]. Interference with CD40 signaling is now considered to be a promising target for chronic inflam- matory response therapies such as atherosclerosis. Anthocyanins are naturally occurring compounds that impart color to fruits, vegetables, and plants. Apart from 0021-9150/$ – see front matter © 2008 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.atherosclerosis.2008.04.005