C‐reactive protein promotes atherosclerosis by increasing LDL transcytosis across endothelial cells

• DOI: 10.1111/bph.12616
• Publication Date: 2014-02-12T12:24:21Z
• Authors: Fang Bian; Xiaoyan Yang; Fan Zhou; Pin‐Hui Wu; Shasha Xing; Gao Xu; Wenjing Li; Jiangyang Chi; Changhan Ouyang; Yonghui Zhang; Bin Xiong; Yongsheng Li; Tao Zheng; Dan Wu; Xiaoqian Chen; Si Jin
• Keywords: Male; Mice, Knockout; 0301 basic medicine; Time Factors; Endothelial Cells; Atherosclerosis; Antioxidants; Lipoproteins, LDL; Mice, Inbred C57BL; Disease Models, Animal; 03 medical and health sciences; Apolipoproteins E; C-Reactive Protein; Reducing Agents; Human Umbilical Vein Endothelial Cells; Animals; Humans; Reactive Oxygen Species; N-Ethylmaleimide-Sensitive Proteins; Protein Kinase Inhibitors; Cells, Cultured; Protein Kinase C

Background and PurposeThe retention of plasma low‐density lipoprotein (LDL) particles in subendothelial space following transcytosis across the endothelium is the initial step of atherosclerosis. Whether or not C‐reactive protein (CRP) can directly affect the transcytosis of LDL is not clear. Here we have examined the effect of CRP on transcytosis of LDL across endothelial cells and have explored the underlying mechanisms.Experimental ApproachEffects of CRP on transcytosis of FITC‐labelled LDL were examined with human umbilical vein endothelial cells and venous rings in vitro and, in vivo, ApoE‐/‐ mice. Laser scanning confocal microscopy, immunohistochemistry and Oil Red O staining were used to assay LDL.Key ResultsCRP increased transcytosis of LDL. An NADPH oxidase inhibitor, diphenylene iodonium, and the reducing agent, dithiothreitol partly or completely blocked CRP‐stimulated increase of LDL transcytosis. The PKC inhibitor, bisindolylmaleimide I and the Src kinase inhibitor, PP2, blocked the trafficking of the molecules responsible for transcytosis. Confocal imaging analysis revealed that CRP stimulated LDL uptake by endothelial cells and vessel walls. In ApoE‐/‐ mice, CRP significantly promoted early changes of atherosclerosis, which were blocked by inhibitors of transcytosis.Conclusions and ImplicationsCRP promoted atherosclerosis by directly increasing the transcytosis of LDL across endothelial cells and increasing LDL retention in vascular walls. These actions of CRP were associated with generation of reactive oxygen species, activation of PKC and Src, and translocation of caveolar or soluble forms of the N‐ethylmaleimide‐sensitive factor attachment protein.