NLRP3 inflammasome activation in dialyzed chronic kidney disease patients.
NLRP3 inflammasome activation in dialyzed chronic kidney disease patients.
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DOI：
10.1371/journal.pone.0122272
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Zaza G
中科院分区：
文献类型：
--
作者：
Granata S;Masola V;Zoratti E;Scupoli MT;Baruzzi A;Messa M;Sallustio F;Gesualdo L;Lupo A;Zaza G
为了评估NLR pyrin domain containing protein 3（NLRP 3）炎性体（一种介导半胱天冬酶-1（CASP-1）和促炎细胞因子IL-18和IL-1β活化的多蛋白复合物）是否可能参与在接受血液透析治疗（CKD-HD）的慢性肾病患者中观察到的慢性炎症状态，我们采用了几种生物分子技术，包括RT-PCR、蛋白质印迹、FACS分析，共聚焦显微术和微阵列。有趣的是，与15名健康受试者相比，15名CKD-HD患者的外周血单核细胞显示出更高的NLRP 3、CASP-1、ASC、IL-1β、IL-18和P2 X7受体mRNA水平。Western blotting分析证实了上述结果。特别是，与对照组相比，CKD-HD中CASP-1、IL 1-β和IL-18的活性形式显著上调。此外，线粒体ROS水平升高，CKD-HD患者外周血单核细胞中NLRP 3/ASC/线粒体共定位，以及在线粒体ROS产生抑制剂mitoTEMPO存在下用LPS/ATP刺激的CKD-HD患者免疫细胞中CASP-1、IL 1-β和IL-18蛋白水平下调，表明该细胞器在上述CKD相关炎性小体活化中可能发挥作用。然后，微阵列分析证实，在一个独立的微阵列研究队列中，NLRP 3和CASP-1，沿着其他炎性小体相关基因，在17名CKD-HD患者中上调，并且他们能够清楚地将这些患者与5名健康受试者区分开来。总之，这些数据首次表明，NLRP 3炎性体在接受透析治疗的尿毒症患者中被激活，并且他们表明这种非生理状况可能由线粒体功能障碍诱导。
To assess whether NLR pyrin domain-containing protein 3 (NLRP3) inflammasome, a multiprotein complex that mediates the activation of caspase-1 (CASP-1) and pro-inflammatory cytokines IL-18 and IL-1β, could be involved in the chronic inflammatory state observed in chronic kidney disease patients undergoing hemodialysis treatment (CKD-HD), we employed several biomolecular techniques including RT-PCR, western blot, FACS analysis, confocal microscopy and microarray. Interestingly, peripheral blood mononuclear cells from 15 CKD-HD patients showed higher mRNA levels of NLRP3, CASP-1, ASC, IL-1β, IL-18 and P2X7receptor compared to 15 healthy subjects. Western blotting analysis confirmed the above results. In particular, active forms of CASP-1, IL1-β and IL-18 resulted significantly up-regulated in CKD-HD versus controls. Additionally, elevated mitochondrial ROS level, colocalization of NLRP3/ASC/mitochondria in peripheral blood mononuclear cells from CKD-HD patients and down-regulation of CASP-1, IL1-β and IL-18 protein levels in immune-cells of CKD-HD patients stimulated with LPS/ATP in presence of mitoTEMPO, inhibitor of mitochondrial ROS production, suggested a possible role of this organelle in the aforementioned CKD-associated inflammasome activation. Then, microarray analysis confirmed, in an independent microarray study cohort, that NLRP3 and CASP-1, along with other inflammasome-related genes, were up-regulated in 17 CKD-HD patients and they were able to clearly discriminate these patients from 5 healthy subjects. All together these data showed, for the first time, that NLRP3 inflammasome was activated in uremic patients undergoing dialysis treatment and they suggested that this unphysiological condition could be possibly induced by mitochondrial dysfunction.
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DOI：
10.1179/096805105x76850
发表时间：
2005-12-01
期刊：
JOURNAL OF ENDOTOXIN RESEARCH
影响因子：
--
作者：
Fritz, JH;Girardin, SE
通讯作者：
Girardin, SE
影响因子：
4
作者：
Gyorgy Hajnoczky;Gyrogy Csordas;Yi, Muqing
通讯作者：
Yi, Muqing
影响因子：
19.6
作者：
Himmelfarb, J;Stenvinkel, P;Hakim, RM
通讯作者：
Hakim, RM
影响因子：
19.6
作者：
Handelman, GJ;Walter, MF;Blumberg, JB
通讯作者：
Blumberg, JB
影响因子：
13.2
作者：
Coresh, J;Astor, BC;Levey, AS
通讯作者：
Levey, AS