C-反应蛋白抑制缺氧状态下人脐静脉内皮细胞表达低氧诱导因子-1α

摘要/Abstract

摘要： 目的研究C-反应蛋白(C-reactive protein,CRP)对低氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)表达的影响。方法利用COCl₂(浓度为100μmol/L)模拟缺氧环境,使其分别与4个试验组的CRP(浓度分别为5,10,50,100μg/ml)同时作用于脐静脉内皮细胞,以仅加CoCl₂(100μmol/L)组为对照;同时设空白对照组。刺激24h后提取蛋白。用Western-blotting半定量检测HIF-1α的蛋白表达量,数据用SPSS11.0软件进行统计分析。结果 CRP在5μg/ml水平即减少HIF-1α表达(P<0.001),在100μg/ml水平达到最大作用,其作用效果呈剂量反应关系(P<0.001)。结论 CRP抑制缺氧状态下人脐静脉内皮细胞表达HIF-1α。此结果证实CRP直接作用于血管,抑制缺氧组织血管新生。

Abstract: Objective To investigate the effect of C-reactive protein (CRP) on expression of hypoxia-inducible factor-1 (HIF-1). Methods Using COCl₂ (100 mol/L) to simulate hypoxic condition for culturing human umbilical vein endothelial cells (HUVECs), we examined the effects of CRP on expression of HIF-1α. The proteins were extracted from the cells after a 24-hour exposure of the cells to CRP of varied concentrations in the presence of COCl₂, and Western blotting was performed for quantification of HIF-1α expression, the results were analyzed statistically with SPSS software. Results CRP at the concentration of 5 μg/ml decreased the expression ofHIF-1α (P<0.001), producing the maximum inhibitory effect at the concentration of 100μg/ml, an effect exhibiting dose-dependence. Conclusion CRP inhibits the expression of HIF-1α in HUVECs subjected to hypoxic condition, which serves as an important evidence for the inhibitory effect of CRP on angiogenesis in hypoxic condition.

中图分类号:

R54
R-33

宋云峰, 吴平生, 赖文岩. C-反应蛋白抑制缺氧状态下人脐静脉内皮细胞表达低氧诱导因子-1α[J]. 南方医科大学学报, 2004, 24(07): 746-748.

SONG Yun-feng, WU Ping-sheng, LAI Wen-yan. C-reactive protein inhibits the expression of hypoxia-inducible factor-1α in human umbilical vein endothelial cells in hypoxic condition[J]. Journal of Southern Medical University, 2004, 24(07): 746-748.

参考文献

[1] Aronow WS. C-reactive protein:Should it be considered a coronary risk factor[J]? Geriatrics, 2003, 58(5):19-25.
[2] 迟东升,靳风霞,杨曙光,等.高脂血症患者C-反应蛋白含量的变化及氟伐他汀对它的影响[J].第一军医大学学报,2002,23(12):1109-11.Chi DS, Jin FX, Yang SG, et al. Effects of fluvastatin on the levels of C-reactive protein and lipids in patients with hyperli-pidemia [J].J First Mil Med Univ/Di Yi Jun Yi Da Xue Xue Bao, 2002, 23(12):1109-11.
[3] Lagrand WK, Niessen HW, Wolbink GJ, et al. C-reactive protein colocalizes with complement in human hearts during acute myocardial infarction[J]. Circulation, 1997, 95(1): 97-103.
[4] Verma S, Wang CH, Li SH, et al. A self-fufilling prophecy-C-reactive protein attenuates nitric oxide production and inhibits angiogenesis[J]. Circulation, 2002, 106(8): 913-9.
[5] Pugh CW, Ratcliffe RJ. Regulation ofangiogenesis by hypoxia: role of the HIF system[J]. Nat Med, 2003, 9(6): 677-84.
[6] 徐耕,金国栋,傅国胜,等.冠心病患者C反应蛋白水平及其1059G/C基因多态性的研究[J].中华心血管病杂志,2003,31(4):274-7.Xu G, Jin GD, Fu GS, et al. Plasma C reactive protein level and the 1059G/C polymorphism of CRP in patients with coronary artery disease[J]. ChinJ Cardiol, 2003, 31(4): 274-7.
[7] Shrive AK, Cheetham GMT, Holden D, et al. Three-dimensional structure of human C-reactive protein[J]. Nat Struct Biol, 1996, 3(4): 346-54.
[8] Danesh J, Muir J, Wong YK, et al. Risk factors for coronary heart disease and acute-phase proteins. A population-based study[J]. Eur Heart J, 1999, 20(13): 954-9.
[9] Zhang YX, Cliff WJ, Schoefi GI, et al. Coronary C-reactive protein distribution: its relation to development of atherosclerosis [J].Atherosclerosis, 1999, 145(2): 375-9.
[10] 付锐斌,吴平生,戴铁英,等.pcDNA3.1+-HIF-1α载体的构建和初步表达鉴定[J].第一军医大学学报,2003,23(11):1134-6.Fu RB, Wu PS, Dai TY, et al. Construction and expression analysis o f recombinant vector pcDNA3.1 +-HIF- 1 α [J]. J First Mil Med Univ/Di Yi Jun Yi Da Xue Xue Bao, 2003, 23(11): 1134-6.
[11] Yong-gun Yoo, Seung Hyun Oh, Eun Soo Park, et al. Hepatitis B virus X protein enhances transcriptional activity of hypoxiainducible factor-1α through activation of mitogen-activated protein kinase pathway[J]. J Biol Chem, 2003, 278(40): 39076-84.
[12] Yanaya K, Tsai AL, Kamitani T. Cobalt-and nickel-banding property ofcullin-2[J]. Biochem Biophys Res Commun, 2002, 290(1):294-299.
[13] Dulk J, Alica J. Regulation of endothelial growth factor synthesis by nitric oxide: Facts and controversies [J]. Antioxidants and redox signaling, 2003, 5(1): 123-32.
[14] Ridker PM. High-sensitivity C-reactive protein and cardiovascular risk: rational for screening and primary prevention[J]. Am J Cardiol,2003, 92(suppl): 17k-22k.
[15] Griselli M, Herbert J, Hutchinson WL, et al. C-reactive protein and complement are important mediators of tissue damage in acute myocardial infarction[J]. J Exp Med, 1999, 190:1733-9.
[16] Pepys MB, Hirschfield MG. C-reactive protein: a critical update [J].J Clin Invest, 2003, 111(12): 1805-12.