Native pentameric C-reactive protein displays more potent pro-atherogenic activities in human aortic endothelial cells than modified C-reactive protein
Native pentameric C-reactive protein displays more potent pro-atherogenic activities in human aortic endothelial cells than modified C-reactive protein
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DOI：
10.1016/j.atherosclerosis.2005.03.031
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发表时间：
2006-01-01
期刊：
影响因子：
5.3
通讯作者：
Jialal, I
中科院分区：
文献类型：
--
作者：
Devaraj, S;Venugopal, S;Jialal, I
炎症是动脉粥样硬化形成的关键。大量前瞻性研究表明，高敏C反应蛋白（CRP）水平可预测心血管事件。最近的研究表明，CRP可能是动脉粥样硬化血栓形成的介质。已显示CRP中五聚体对称性的丧失导致修饰的CRP（mCRP）的形成。本研究的主要目的是检查与人主动脉内皮细胞中的修饰形式相比，CRP的天然五聚体形式的生物学效应。纯化并使用来自两种不同来源（重组体和血清）的人五聚体天然CRP（n-CRP）。然后将其进行EDTA螯合和尿素处理以制备修饰的CRP（m-CRP）。通过凝胶电泳检查n-CRP和m-CRP制剂的纯度。将n-CRP和m-CRP与人主动脉内皮细胞（HAEC）孵育，通过测定白细胞介素-8（IL-8）、纤溶酶原激活物抑制剂-1（派-1）、环磷酸鸟苷（cGMP）和前列腺素F1-α（PGF 1-α）来检测其生物学活性。n-CRP在浓度>= 10 μ g/mL时显著上调IL-8，而m-CRP仅在浓度为50 μ g/mL时上调IL-8（p < 0.05）。与m-CRP相比，天然的派-1水平在更大程度上显著增加（p < 0.05）。虽然两者在50 μ g/mL浓度下均降低PGF 1-α，但天然CRP的作用更明显，在10 μ g/mL浓度下更明显（p < 0.05）。通过cGMP评估，在eNOS活性的抑制方面观察到最显著的差异，在10 μ g/ml天然CRP下观察到，而在50 μ g/ml m-CRP下仅观察到（天然CRP与mCRP：p < 0.001）。因此，与修饰的CRP相比，天然五聚体CRP在人主动脉内皮细胞中发挥更强的致动脉粥样硬化作用。(c)2005爱思唯尔爱尔兰有限公司保留所有权利。
Inflammation is pivotal in atherogenesis. Numerous prospective studies have shown that levels of high sensitive C-reactive protein (CRP) predict cardiovascular events. Recently, data suggests that CRP could be a mediator in atherothrombosis. Loss of pentameric symmetry in CRP has been shown to result in the formation of modified CRP (mCRP). The main aim of this study was to examine the biological effects of the native, pentameric form of CRP compared to a modified form in human aortic endothelial cells. Human pentameric native CRP (n-CRP) from two different sources (recombinant and serum) was purified and used. It was then subjected to EDTA chelation and urea treatment to prepare modified CRP (m-CRP). Purity of both n-CRP and m-CRP preparations was checked by gel electrophoresis. Both n-CRP and m-CRP were incubated with human aortic endothelial cells (HAEC) and biological activities was tested by assaying for interleukin-8 (IL-8), plasminogen activator inhibitor-1 (PAI-1), cyclic GMP and prostaglandin F1-alpha. n-CRP significantly upregulated IL-8 at concentrations >= 10 mu g/mL while m-CRP upregulated IL-8 only at concentrations of 50 mu g/mL (p < 0.05). PAI-1 levels were significantly increased to a greater extent with native compared to m-CRP (p < 0.05). While both decreased PGF1-alpha at concentrations of 50 mu g/mL, the effect of native CRP was more pronounced and was evident at 10 mu g/mL (p < 0.05). The most pronounced difference was observed with regard to inhibition of eNOS activity as assessed by cGMP which was observed at 10 mu g/ml of native CRP but only at 50 mu g/mL for m-CRP (native CRP versus mCRP: p < 0.001). Thus, native pentameric CRP compared to modified CRP exerts more potent atherogenic effects in human aortic endothelial cells. (c) 2005 Elsevier Ireland Ltd. All rights reserved.