颈动脉粥样硬化患者外周血单核细胞NLRP3炎性小体表达的研究

谢雯丽; 石昕; 孔维玮; 陈东; 曲鹏

doi:10.13201/j.issn.1001-1439.2015.12.014

颈动脉粥样硬化患者外周血单核细胞NLRP3炎性小体表达的研究

- 1 大连医科大学附属第二医院心内科(大连, 116023);
- 2 大连市第五人民医院心内科;
- 3 大连市中心医院综合神经外科
基金项目:
辽宁省自然科学基金(No:2013023025)

详细信息

通讯作者: 曲鹏:E-mail:qupeng777@aliyun.com

中图分类号: R543.1

收稿日期: 2015-05-06

Expression of NLRP3 inflammasome in peripheral blood mononuclear cells of patients with carotid atherosclerosis

- 1 Department of Cardiology, Second Affiliated Hospital of Dalian Medical University, Dalian, China, 116023;
- 2 Department of Cardiology, The Fifth People Hospital, Dalian;
- 3 Department of Neurosurgery, Dalian Centre Hospital

More Information

Corresponding author: QU Peng, E-mail: qupeng777@aliyun.com

Received Date: 06 May 2015

摘要

摘要: 目的:检测重度颈动脉粥样硬化患者外周血单核细胞NLRP3炎性小体表达水平,探讨NLRP3炎性小体在颈动脉粥样硬化形成中的作用。方法:收集重度颈动脉粥样硬化行颈动脉内膜剥脱术(CEA)患者30例为观察组,无颈动脉粥样硬化形成的健康者20例为对照组。采静脉血20 ml,采用Real-time PCR检测外周血单核细胞NLRP3炎性小体各组分、IL-1β、IL-18 mRNA表达,Western-blot法检测其蛋白表达,酶联免疫吸附试验(ELISA)检测血浆IL-1β和IL-18水平。应用多元线性逐步回归分析外周血单核细胞NLRP3炎性小体各组分、IL-1β、IL-18 mRNA及血浆IL-1β、IL-18的危险因素,并采用Pearson相关分析其与颈动脉mean-IMT、max-IMT的关系。结果:观察组外周血单核细胞NLRP3 mRNA(5.51±2.52)、Caspase-1 mRNA(3 467.84±1 057.99)、IL-1β mRNA(1 852.04±1 503.22)、IL-18 mRNA(5.67±3.15)表达量,血浆IL-1β(31 701.63±1 930.16)pg/L、IL-18(332.70±35.61)ng/L水平均显著高于对照组(均P<0.001)。观察组外周血单核细胞ASC mRNA表达量(84.17±25.78)与对照组(73.54±10.30)差异无统计学意义。多元线性逐步回归分析显示,高血压、LDL-C、TG、UA是激活NLRP3炎性小体表达及血浆IL-1β、IL-18水平的重要危险因素。观察组颈部超声 mean-IMT与外周血单核细胞NLRP3 mRNA(r=0.380,P=0.038)、Caspase-1 mRNA(r=0.381,P=0.038)、血浆IL-1β(r=0.432,P=0.017)、血浆IL-18(r=0.441,P=0.015)呈正相关。观察组颈部超声 max-IMT与血浆IL-1β(r=0.419,P=0.021)、血浆IL-18(r=0.382,P=0.037)呈正相关。结论:重度颈动脉粥样硬化患者外周血单核细胞NLRP3炎性小体活化,并与颈动脉粥样硬化的严重程度相关,提示NLRP3炎性小体可能参与了颈动脉粥样硬化发生发展,LDL-C、TG、UA、高血压可能是激活NLRP3炎性小体表达的重要危险因素。
- NLRP3炎性小体 /
- 颈动脉粥样硬化 /
- 血脂 /
- 血尿酸 /
- 内膜中膜厚度
Abstract: Objective: To test the expression level of the NLRP3 inflammasome mRNA and protein in peripheral blood mononuclear cells from patients with severe carotid atherosclerosis and to explore the effect of NLRP3 inflammasome in carotid atherosclerosis.Method: The research group consisted of 30 patients who were categorized as having severe carotid atherosclerosis waiting for CEA. And another 20 patients with no carotid stenosis and coronary artery stenosis were categorized as the control group.We collected 20 ml venous blood to get the mononuclear cells and plasma.Real time PCR was used to analyze the expression level of the NLRP3 inflammasome, IL-1β, IL-18 mRNA in PBMCs and Western-blot was used to investigate the expression levels of them. Enzyme linked immunosorbent assay (ELISA) was used to explore the IL-1β and IL-18 in plasma. Then t-test was applied to compare the differences of these indicators in each group. Multiple linear stepwise regression was used to analyze the risk factors of the expression of NLRP3 inflammasome, IL-1β, IL-18 mRNA in PBMCs, and plasma IL-1β, IL-18. Pearson correlation was used to evaluate the relationship between NLRP3 inflammasome, IL-1β, IL-18 mRNA in PBMCs and mean-IMT,max-IMT. Result: The expression of NLRP3 mRNA (5.51±2.52), Caspase-1 mRNA(3 467.84±1 057.99), IL-1βmRNA(1 852.04±1 503.22), IL-18 mRNA(5.67±3.15)in PBMCs and plasma IL-1β level (3 1701.63±1 930.16 pg/l), IL-18 (332.70±35.61) ng/l were significantly higher in CEA group than those in the control group (P<0.001), while there was no difference of ASC mRNA between CEA group(84.17±25.78) and the control group(73.54±10.30). Multiple linear stepwise regression analysis showed that hypertension, LDL-C, TG, UA were the influencing factors of the expression of NLRP3 inflammasome and the level of plasma IL-1β, IL-18. In CEA patients, Pearson correlation analysis indicated significant correlation between mean- IMT and NLRP3 mRNA(r=0.380,P=0.038), Caspase-1 mRNA(r=0.381,P=0.038), plasma IL-1β(r=0.432,P=0.017), plasma IL-18(r=0.441,P=0.015). And then there were significant correlations between max-IMT and plasma IL-1β(r=0.41,P=0.021), plasma IL-18(r=0.382,P=0.037) as well. Conclusion: NLRP3 inflammasome in PBMCs is activated in patients with severe carotid atherosclerosis. Meanwhile, there is a close relationship between the expression of NLRP3 inflammasome and severity of carotid atherosclerosis. NLRP3 inflammasome in PBMCs may play a critical role in the process of atherosclerosis, LDL-C, TG, UA and hypertension are important risk factors which could activate NLRP3 inflammasome.
- NLRP3 mtlammasome /
- carotid atherosclerosis /
- lipid /
- uric acid /
- mtima media thinckness

HTML全文

参考文献(20)

[1]	GARG N J.Inflammasomes in cardiovascular diseases[J].Am J Cardiovasc Dis,2011,1:244-254.
[2]	ZHENG F,XING S,GONG Z,et al.NLRP3 inflammasomes show high expression in aorta of patients with atherosclerosis[J].Heart Lung Circ,2013,22:746-750.
[3]	WANG L,QU P,ZHAO J,et al.NLRP3 and downstream cytokine expression elevated in the monocytes of patients with coronary artery disease[J].Arch Med Sci,2014,10:791-800.
[4]	中华医学会外科学分会血管外科学组.颅外段颈动脉狭窄治疗指南[J].中国实用外科杂志,2008,28(11):913-915.
[5]	European Carotid Surgery Trialists'Collaborative Group.Randomised trial of endarterectomy for recently symptomatic carotid stenosis:final results of the MRC European Carotid Surgery Trial(ECST)[J].Lancet,1998,351:1379-1387.
[6]	MORITO N,INOUE Y,URATA M,et al.Increased carotid artery plaque score is an independent predictor of the presence and severity of coronary artery disease[J].J Cardiol,2008,51:25-32.
[7]	CHEUK B L,CHENG S W.Annexin A1 expression in atherosclerotic carotid plaques and its relationship with plaque characteristics[J].Eur J Vasc Endovasc Surg,2011,41:364-371.
[8]	LIU H,LI W,CHEN Z,et al.Expression of the NLRP3 inflammasome in cerebral cortex after traumatic brain injury in a rat model[J].Neurochem Res,2013,38:2072-2083.
[9]	HOFFMAN H M,BRYDGES S D.Genetic and molecular basis of inflammasome-mediated disease[J].J Biol Chem,2011,286:10889-10896.
[10]	KIRII H,NIWA T,YAMADA Y,et al.Lack of interleukin-1beta decreases the severity of atherosclerosis in ApoE-deficient mice[J].Arterioscler Thromb Vase Biol,2003,23:656-660.
[11]	ELHAGE J,RUDLING M.Reduced atherosclerosis in interleukin-18 deficient apolipoprotein E-knockout mice[J].Cardiovasc Res,2003,59:234-240.
[12]	RAJAMAKI K,LAPPALAINEN J,OORNI K,et al.Cholesterol crystals activate the NLRP3 inflammasome in human macrophages:a novel link between cholesterol metabolism and inflammation[J].Plos One,2010,5:e11765.doi:10.1371/journal.pone.0011765.
[13]	DUEWELL P,KONO H,RAYNER K,et al.NLRP3 inflammasomes are required for atherogenesis and activated by cholesterol crystals[J].Nature,2010,464:1357-1361.
[14]	WANG L,QU P.Role of NLRP3 inflammasome and downstream cytokines in atherosclerosis[D].Dalian:Dalian Medical University,2013.
[15]	PENG K,LIU L,QU P,et al.P2X7R is involved in the progression of atherosclerosis by promoting NLRP3 inflammasome activation[J].Int J Mol Med,2015,35:1179-1188.
[16]	STEHLIK C,FIORENTINO L,DORFLEUTNER A,et al.The PAAD/PYRIN-family protein ASC is a dual regulator of a conserved step in nuclear factor kappa B activation pathways[J].J Exp Med,2002,196:1605-1615.
[17]	CASSEL S L,JOLY S,SUTTERWALA F S,et al.The NLRP3 inflammasome:a sensor of immune danger signals[J].Semin Immunol,2009,21:194-198.
[18]	ABELA G S,AZIZ K,VEDRE A,et al.Effect of cholesterol crystals on plaques and intima in arteries of patients with acute coronary and cerebrovascular syndromes[J].Am J Cardiol,2009,103:959-968.
[19]	LIPPI G,MONTAGNANA M,FRANCHINI M,et al.The paradoxical relationship between serum uric acid and cardiovascular disease[J].Clin Chim Acta,2008,392:1-7.
[20]	HOKANSON J E.Hypertriglyceridemia and risk of coronary heart disease[J].Curr Cardiol Rep,2002,4:488-493.