Homocysteine induces the expression of C-reactive protein via NMDAr-ROS-MAPK-NF-κB signal pathway in rat vascular smooth muscle cells
Homocysteine induces the expression of C-reactive protein via NMDAr-ROS-MAPK-NF-κB signal pathway in rat vascular smooth muscle cells
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DOI：
10.1016/j.atherosclerosis.2014.06.021
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发表时间：
2014-09-01
期刊：
影响因子：
5.3
通讯作者：
Wu, Di
中科院分区：
文献类型：
--
作者：
Pang, Xiaoming;Liu, Juntian;Wu, Di
目的：同型半胱氨酸（Homocysteine，Hcy）是动脉粥样硬化的独立危险因素。C反应蛋白（CRP）直接参与动脉粥样硬化的发生和发展。然而，没有直接证据表明Hcy通过CRP对血管平滑肌细胞（VSMCs）的促炎作用。本研究采用Western blot和ELISA方法检测同型半胱氨酸对VSMCs CRP表达的影响，并探讨同型半胱氨酸对VSMCs CRP表达的影响机制。RT-PCR检测mRNA表达。用光泽精比色法测定超氧阴离子的含量，用荧光显微镜观察免疫荧光染色。结果表明，Hcy可显著诱导VSMCs CRP mRNA和蛋白的表达，抗IL-1 β或抗IL-6中和抗体单独或联合应用可部分抑制Hcy诱导的CRP表达。同型半胱氨酸可增加血管平滑肌细胞N-甲基-D-天冬氨酸受体（NMDAR）NR 1亚单位的表达，MK-801可减轻同型半胱氨酸诱导的CRP表达。进一步的研究表明Hcy刺激VSMCs产生超氧阴离子。然而，MK-801，TTFA和DPI预处理细胞显着减少Hcy刺激的超氧阴离子的产生，抗氧化剂NAC降低Hcy诱导的CRP在VSMCs的表达。此外，PD 98059、SB 205380或PDTC可拮抗Hcy诱导的CRP表达，MK-801、NAC、PD 98059或SB 205380可抑制Hcy激活的ERK 1/2和p38磷酸化。目前的研究表明，同型半胱氨酸能够通过刺激CRP的产生启动VSMCs的炎症反应，其通过NMDAr-ROS-ERK 1/2/p38-NF-κ B信号通路介导。这些发现为Hcy在动脉粥样硬化发病机制中的作用提供了新的证据。(C)2014爱思唯尔爱尔兰有限公司版权所有。
Objective: Homocysteine (Hcy) is known as an independent risk factor for atherosclerosis. C-reactive protein (CRP) directly participates in initiation and progression of atherosclerosis. However, there is no direct evidence to demonstrate pro-inflammatory effect of Hcy on vascular smooth muscle cells (VSMCs) through CRP. In the present study, we examined the effect of Hcy on CRP expression and investigated the related mechanism in VSMCs.Methods and results: Protein expression and secretion were detected by Western blot and ELISA, respectively. mRNA expression was detected by RT-PCR. Superoxide anion was detected by lucigenin chemiluminometry and the immunofluorescence staining was observed by a fluorescence microscope. The results revealed that Hcy significantly induced mRNA and protein expressions of CRP in VSMCs both in vitro and in vivo, and anti-IL-1 beta or anti-IL-6 neutralizing antibody alone or in combination partially reduced Hcy-induced CRP expression. Hcy increased the expression of NR1 subunit of N-methyl-D-aspartate receptor (NMDAr), and MK-801 alleviated Hcy-induced CRP expression in VSMCs. Further studies showed that Hcy-stimulated superoxide anion generation in VSMCs. Nevertheless, pretreatment of the cells with MK-801, TTFA and DPI significantly reduced Hcy-stimulated superoxide anion generation, and antioxidant NAC decreased Hcy-induced CRP expression in VSMCs. Additionally, PD98059, SB205380 or PDTC antagonized Hcy-induced CRP expression, and MK-801, NAC, PD98059 or SB205380 inhibited Hcy-activated phosphorylations of ERK1/2 and p38.Conclusion: The present study demonstrates that Hcy is able to initiate an inflammatory response in VSMCs by stimulating CRP production, which is mediated through NMDAr-ROS-ERK1/2/p38-NF-kappa B signal pathway. These findings provide new evidence for a role of Hcy in pathogenesis of atherosclerosis. (C) 2014 Elsevier Ireland Ltd. All rights reserved.