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

• Published in: British journal of pharmacology 2014-05, Vol.171 (10), p.2671-2684
• Main Authors: Bian, Fang, Yang, Xiaoyan, Zhou, Fan, Wu, Pin‐Hui, Xing, Shasha, Xu, Gao, Li, Wenjing, Chi, Jiangyang, Ouyang, Changhan, Zhang, Yonghui, Xiong, Bin, Li, Yongsheng, Zheng, Tao, Wu, Dan, Chen, Xiaoqian, Jin, Si
• Format: Article
• Language: English
• Subjects: Animals; Antioxidants - pharmacology; Apolipoproteins E - deficiency; Apolipoproteins E - genetics; Atherosclerosis; Atherosclerosis - genetics; Atherosclerosis - metabolism; C-Reactive Protein - administration & dosage; C-Reactive Protein - metabolism; caveolae; Cells, Cultured; Cholesterol; C‐reactive protein; Disease Models, Animal; endothelial cells; Endothelial Cells - drug effects; Endothelial Cells - metabolism; Human Umbilical Vein Endothelial Cells - metabolism; Humans; Kinases; LDL; Lipoproteins, LDL - metabolism; Male; Mice, Inbred C57BL; Mice, Knockout; N-Ethylmaleimide-Sensitive Proteins - metabolism; PKC; Protein Kinase C - antagonists & inhibitors; Protein Kinase C - metabolism; Protein Kinase Inhibitors - pharmacology; Proteins; Reactive Oxygen Species - metabolism; Reducing Agents - pharmacology; Research Papers; ROS; Src; src-Family Kinases - antagonists & inhibitors; src-Family Kinases - metabolism; Time Factors; transcytosis; Transcytosis - drug effects
• ISSN: 0007-1188; 1476-5381
• DOI: 10.1111/bph.12616

Background and Purpose The 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 Approach Effects 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 Results CRP 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 Implications CRP 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.