TXNIP介导的NLRP3炎症小体激活在心肌微血管内皮细胞缺氧/复氧损伤中的作用被引量：6

Effect of TXNIP mediated NLRP3 inflammasome activation on cardiac microvascular endothelial cells during hypoxia / reoxygenation injury

下载PDF

导出

摘要目的研究硫氧还蛋白结合蛋白(TXNIP)介导的NLRP3炎症小体激活在心肌微血管内皮细胞(CMECs)缺氧/复氧(H/R)损伤中的作用。方法分离培养C57BL/6J小鼠CMECs,随机分为对照组、H/R损伤组、H/R+Scrambled siRNA组和H/R+TXNIP siRNA组。各组处理后使用ELISA试剂盒检测细胞IL-1β水平,采用免疫共沉淀的方法检测TXNIP和NLRP3的相互作用,Western blot检测TXNIP和NLRP3的表达水平,乳酸脱氢酶试剂盒检测细胞培养上清LDH释放情况和细胞Caspase-3活性。结果与对照组相比,H/R后CMECs的NLRP3表达水平明显升高(P<0.05),NLRP3炎症小体激活(IL-1β水平升高,P<0.01),且TXNIP和NLRP3的结合效果明显增强(P<0.01)。运用TXNIP siRNA抑制TXNIP表达后行H/R处理,与H/R+Scrambled siRNA组相比,NLRP3炎症小体激活水平明显下降(IL-1β水平降低),细胞Caspase-3活力和LDH释放水平均显著降低(均P<0.05)。结论在CMECs发生H/R损伤时,NLRP3炎症小体激活,TXNIP和NLRP3的相互作用增强;抑制TXNIP表达后,NLRP3炎症小体激活水平降低,内皮损伤减轻。说明TXNIP介导的NLRP3炎症小体激活参与CMECs的H/R损伤,可能成为CMECs的H/R损伤的新机制。 AIM To determine the effect of thioredoxin-interacting protein （TXNIP） mediated NLRP3 inflammasome activation on cardiac microvascular endothelial cells （CMECs） during hypoxia/reoxygenation （H/R） injury. METHODS CMECs isolated from the hearts of adult mice were randomized into control group, H/R group, H/R ＋ scrambled siRNA group and H/R ＋ TXNIP siRNA group. Levels of IL-IL were detected by ELISA kits. The interaction between TXNIP and NLRP3 was determined by immunoprecipitation. Expressions of TXNIP and NLRP3 were analyzed by Western blot, and caspase-3 activity and LDH release were assessed with test kits. RESULTS H/R increased NLRP3 expression （ P 〈 0. 05 ）, IL-1 [3 level （ P 〈 0. 01 ） and the interaction between TXNIP and NLRP3 （ P 〈 0. 01 ） in CMECs compared with control group. Blocking TXNIP signaling with siRNA significantly inhibited NLRP3 activation （P 〈 0. 05 ） and alleviated endothelial injury evidenced by increased caspase-3 activity LDH release （P 〈 0.05 ）. CONCLUSION H/R injury promotes NLRP3 activation and increases the interaction between TXNIP and NLRP3. Blocking TXNIP signaling reduces NLRP3 activation and endothelial injury. TXNIP mediated NLRP3 activation may be a novel mechanism in I-L/R injury of CMECs.

作者许倡涛刘毅朱迪辛超杜志超穆楠陶凌

机构地区第四军医大学西京医院心血管内科第四军医大学药学院药物基因组学教研室第四军医大学药学院生物制药学教研室

出处《心脏杂志》 CAS 2015年第5期510-513,519,共5页 Chinese Heart Journal

基金国家自然科学基金项目资助(81170186)

关键词 NLRP3 TXNIP 心肌微血管内皮细胞缺氧/复氧损伤 NLRP3 thioredoxin-interacting protein cardiac mierovascular endothelial cells hypoxia/ reoxygenation injury

分类号 R322.1 [医药卫生—人体解剖和组织胚胎学]

引文网络
相关文献

参考文献16

1Iyer SS, Pulskens WP, Sadler JJ, et al. Necrotic ceils trigger a ster- ile inflammatory response through the Nlrp3 inflammasome[ J]. Proc Natl Acad Sci U S A, 2009, 106(48) :20388 -20393.
2Martinon F, Bums K, Tschopp J. The inflammasome: a molecular platform triggering activation of inflammatory caspases and processing of prolL-beta [ J ]. Mol Cell, 2002, 10 (2) :417 - 426.
3Stutz A, Golenhock DT, Latz E. Inflammasomes: too big to miss [J]. J Clin Invest, 2009, 119(12) :3502 -3511.
4Sandanger O, Ranheim T, Vinge LE, et al. The NLRP3 inflamma- some is up-regulated in cardiac fibmblasts and mediates myocardial ischaemia-reperfusion injury [ J ]. Cardiovasc Res, 2013, 99 ( 1 ) : 164 - 174.
5Nishiyama A, Matsui M, Iwata S, et al. Identification of thioredox- in-binding protein-2/vitamin D( 3 ) up-regulated protein 1 as a nega- tive regulator of thioredoxin function and expression [ J ]. J Biol Chem, 1999, 274(31 ) :21645 -21650.
6Yoshioka J, Chutkow WA, Lee S, et al. Deletion of thioredoxin- interacting protein in mice impairs mitochondrial function but pro- tects the myocardium from ischemia-reperfusion injury [ J ]. J Clin Invest, 2012, 122( 1 ) :267 -279.
7Zhou R, Tardivel A, Thorens B, et al. Thioredoxin-interaeting protein links oxidative stress to inflammasome activation [ J ]. Nat Immunol, 2010, 11(2) :136 -140.
8Mezzarema E, Toldo S, Farkas D, et al. The inflammasome pro- motes adverse cardiac remodeling following acute myocardial infarc- tion in the mouse[ J]. Proc Natl Acad Scl U S A, 2011, 108 (49) : 19725 - 19730.
9Pober JS, Sessa WC. Evolving functions of endothelial cells in inflammation [ J ]. Nat Rev Immunol, 2007, 7 (10) : 803 - 815.
10Nishida M, Carley WW, Gerritsen ME, et al. Isolation and charac- terization of human and rat cardiac microvascular endothelial cells [ J ]. Am J Physiol, 1993, 264 (2 Pt 2) : H639 - H652.