白细胞介素-18与深静脉血栓疾病的相关性研究被引量：3

Correlation between interleukin-18 and deep venous thrombosis disease

下载PDF

导出

摘要目的探讨白细胞介素-18(IL-18)的表达变化与深静脉血栓疾病(DVT)发生、发展的相关性及临床意义。方法收集临床DVT病例(DVT组,n=40)、实验对照组(n=40)、健康对照组(n=20)的血液,以ELISA法检测其中IL-18的表达并分析差异;培养原代培养的人脐静脉内皮细胞(HUVECs)行目标蛋白免疫荧光检测明确其定位;构建人IL-18过表达和干扰病毒载体(IL-18-pCDH-GFP、IL-18-LMP-shRNAmir1)感染HUVECs并进行基因表达谱芯片检测和KEGG Pathway等生物信息技术,分析IL-18与疾病的相关性。结果人血ELISA检测发现在DVT组和两对照组中IL-18的表达差异均有统计学意义(F=11.248,P<0.01);实验对照组与健康对照组之间IL-18的表达差异无统计学意义(P>0.05);免疫荧光检测显示目标蛋白定位于细胞质;人IL-18过表达和干扰载体经病毒转染后感染HUVECs;基因芯片检测发现与正常细胞比较,过表达载体IL-18-pCDH-GFP感染细胞有17条信号通路下调、16条上调;干扰载体IL-18-LMP-shRNAmir1感染细胞有23条信号通路下调、9条上调。结论 IL-18对HUVECs有确切影响,同时与DVT疾病密切相关,有希望作为深静脉血栓疾病临床预测诊断的候选分子标记物。 Objective To investigate the correlation between IL-18 and deep venous thrombosis disease and its clinical signification.Methods To detect the expression of IL-18 by ELISA,we collected the blood samples of DVT patients as the experimental group（n=40）compared to the control group（n=40）and normal group（n=20）.IL-18 over expression/interference vectors were constructed and transfected human vein endothelial cells,analyzed by microarray and KEGG Pathway as biology information technology.Then discuss the association between IL-18 and DVT.Results Results of ELISA showed that compared with control group and normal group,the expression of IL-18 gene in DVT patient were up-regulated（F=11.248,P〈0.01）.Compared with normal group,the IL-18 expression in control group have not been significantly up-regulated（P〉0.05）.Immunofluorescence detected IL-18 gene expression in cytoplasm of human umbilical vein endothelial cells（HUVECs）.According to the microarray analysis we found in the IL18-pCDH-GFP transfected cells 17 signaling pathways were down-expressed while 16 signaling pathways were upexpressed.Compared with normal group cells,in the IL18-LMP-shRNAmir1 transfected cells 23 signaling pathways were down-expressed and 9signaling pathways were up-expressed.Conclusion Based on the above experimental data,it is very clear that IL-18 influenced HUVECs and plays an important role in DVT,it is possible to predict the diagnosis of DVT and act as candidate molecular markers.

作者李光第宋恩赵学凌白云城彭志周如丹

机构地区昆明医科大学第一附属医院骨科

出处《重庆医学》 CAS 北大核心 2015年第5期600-604,共5页 Chongqing medicine

关键词静脉血栓形成白细胞介素18 分子标记物人脐静脉内皮细胞 deep venous thrombosis interleukin-18 biomarker human umbilical vein endothelial cells

分类号 R543.6 [医药卫生—心血管疾病]

引文网络
相关文献

参考文献20

1Levi M, van der Poll T, Schultz M. Infection and inflam-mation as risk factors for thrombosis and atherosclerosis[J]. Semin Thromb Hemost,2012,38(5) :506-514.
2Okamura H,Nagata K,Komatsu T,et al. A novel costim-ulatory factor for gamma interferon induction found in thelivers of mice causes endotoxic shock [J]. Infect Immun,1995,63(10) :3966-3972.
3Ushio S’Namba M, Okura T, et al. Cloning of the cDNAfor human IFN-gamma-inducing factor, expression inEscherichia coli ? and studies on the biologic activities ofthe protein[J]. J Immunol, 1996 ,156(11) : 4274-4279.
4Goon PK,Watson T,Shantsila E,et al. Standardization ofcirculating endothelial cell enumeration by the use of hu-man umbilical vein endothelial cells[J]. J Thromb Haem-ost,2007 ?5(4):870-872.
5孙慧琳,湛奕,刘珍珍,李筠,蔡德鸿.人脐静脉内皮细胞的原代培养及鉴定[J].广东医学,2012,33(6):744-746. 被引量：6
6Strukova S. Blood coagulation-dependent inflammation.Coagulation-dependent inflammation and inflammation-dependent thrombosis[J]. Front Biosci, 2006?11 (2) j 59-80.
7Roumen-Klappe EM,Janssen MC, Van Rossum J, et al.Inflammation in deep vein thrombosis and the develop-ment of post-thrombotic syndrome: a prospective study[J]. J Thromb Haemost,2009 ,7(4) : 582-587.
8Jezovnik MK, Poredos P. Idiopathic venous thrombosis isrelated to systemic inflammatory response and to in-creased levels of circulating markers of endothelial dys-function[J]. Int Angiol,2010,29(3) :226-231.
9Lin PH, Barshes NR, Annambhotla S, et al. Advances inendovascular interventions for deep vein thrombosis [J].Expert Rev Med Devices,2008,5(2) : 153-166.
10Samnegard A?Hulthe J,Silveira A, et al. Gender specificassociations between matrix metalloproteinases and in-flammatory markers in post myocardial infarction patients[J]. Atherosclerosis, 2009 ?202 (2) : 550-556.

二级参考文献7

1REICHLIN T,WILD A,DURRENBERGER M,et al.Investiga-ting native coronary artery endothelium in situ and in cell cultureby scanning force microscopy[J].J Struct Biol,2005,152(1):52-63.
2GOON P K,WASTON T,SHANTSILA E,et al.Standardizationof circulating endothelial cell enumeration by the use of human um-bilical vein endothelial cells[J].J Thromb Haemost,2007,5(4):870-872.
3VERMA S,BUCHANAN M R,ANDERSON T J.Endothelialfunction testing as a biomarker of vascular disease[J].Circula-tion,2003,108(17):2054-2069.
4LI X,XING S,ZHANG L,et al.High glucose promotes there-lease and expression of novel vasoactive peptide,coupling factor 6,in human umbilical vein endothelial cells[J].Peptides,2007,28(3):683-690.
5MARIN V,KAPLANSKI G,GRS S,et al.Endothelial cell cul-ture:protocol to obtain and cultivate human umbilical endothelialcells[J].J Immunol Methods,2001,254(1/2):183-190.
6司徒镇强吴军正.细胞培养[M].西安:世界图书版出版公司,2004.78-202.
7刘长乐,郑心田,李广平,孟恒星,邱录贵.体外人脐静脉内皮细胞的培养鉴定及抗原表达分析[J].山东医药,2011,51(36):25-26. 被引量：8