摘要
目的研究C反应蛋白对大鼠骨髓内皮祖细胞参与血管形成及其功能活性的影响,探讨C反应蛋白在动脉粥样硬化病变发展中的可能作用机制。方法用密度梯度离心法分离大鼠骨髓内皮祖细胞,并用DiI标记的乙酰化低密度脂蛋白和FITC标记的植物凝集素双染,在共聚焦显微镜下观察。将细胞与不同浓度C反应蛋白(0、1、5及10 mg/L)共同培养,建立内皮祖细胞与大鼠心脏微血管内皮细胞共同培养的体外血管形成模型。分别用MTT比色法、Transwell小室、Matrigel检测C反应蛋白对内皮祖细胞增殖活性、迁移及管腔形成能力的影响。用Western blotting分别检测凋亡相关蛋白Bcl-2和Bax、整合素β2和内源性血管内皮生长因子的表达。结果随着浓度的增加,C反应蛋白明显减少内皮祖细胞掺入模型中心脏微血管内皮细胞管腔结构的数量,减弱内皮祖细胞的增殖活性,减少迁移的细胞数及形成的管腔数,上调Bax蛋白的表达,下调Bcl-2蛋白、整合素β2、内源性血管内皮生长因子的表达。结论 C反应蛋白减弱了内皮祖细胞参与血管形成的能力,这与其抑制内皮祖细胞的功能活性有关,从而可能促进了动脉粥样硬化等缺血性疾病的发展。
Aim To investigate the effect of C-reactive protein（CRP）on rat bone marrow-derived endothelial progenitor cells（EPC）involved into vasculogenesis and functional activity in vitro,and discuss possible mechanism of CRP during progression of atherosclerosis. Methods EPC were separated from bone marrow of rat with density gradient centrifugation,and characterized as DiI-ac-LDL/FITC-UEA-l double positive cells detected by laser confocal microscopy.EPC were cultured with CRP of different concentrations（0,1,5 and 10 mg/L） for 24 h.A co-culture model of EPC and rat cardiac microvascular endothelial cells（CMEC） was used to analyze the effect of CRP on EPC involved into lumen formation in vitro.Proliferation viability,migration,vasculogenesis,and protein expression of EPC were assayed by MTT,Transwell chamber,Matrigel,Western blotting respectively. Results CRP dose-dependently reduced the number of EPC involved into capillary-like structures of CMEC,weakened proliferation viability of EPC,reduced the number of migration and lumen formation,up-regulated the expression of Bax protein,and down-regulated the expression of Bcl-2 protein,integrin β2,endogenous vescular endothelial growth factor（VEGF）. Conclusion CRP may impair EPC-mediated neovascularization by weakening its functions,which leads to progression of atherosclerosis.
出处
《中国动脉硬化杂志》
CAS
CSCD
北大核心
2011年第2期105-109,共5页
Chinese Journal of Arteriosclerosis
关键词
C反应蛋白
内皮祖细胞
血管形成
动脉粥样硬化
C-Reactive Protein
Endothelial Progenitor Cells
Tube Formation
Atherosclerosis
