title: NLRP3 inflammasome activation in dialyzed chronic kidney disease patients.
abstract: 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 P2X7 receptor 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.
authors: Renal Unit, Department of Medicine, University-Hospital of Verona, Verona, Italy.
authors: Simona
authors: S
authors: Granata
authors: Renal Unit, Department of Medicine, University-Hospital of Verona, Verona, Italy.
authors: Valentina
authors: V
authors: Masola
authors: Interdepartmental Laboratory for Medical Research (LURM), University of Verona, Verona, Italy.
authors: Elisa
authors: E
authors: Zoratti
authors: Interdepartmental Laboratory for Medical Research (LURM), University of Verona, Verona, Italy.
authors: Maria Teresa
authors: MT
authors: Scupoli
authors: Department of Pathology and Diagnostics, Section of General Pathology, University of Verona, Verona, Italy.
authors: Anna
authors: A
authors: Baruzzi
authors: Renal Unit, Department of Medicine, University-Hospital of Verona, Verona, Italy.
authors: Michele
authors: M
authors: Messa
authors: Department of Emergency and Organ Transplantation-Nephrology, Dialysis and Transplantation Unit, University of Bari, Bari, Italy.
authors: Fabio
authors: F
authors: Sallustio
authors: Department of Emergency and Organ Transplantation-Nephrology, Dialysis and Transplantation Unit, University of Bari, Bari, Italy.
authors: Loreto
authors: L
authors: Gesualdo
authors: Renal Unit, Department of Medicine, University-Hospital of Verona, Verona, Italy.
authors: Antonio
authors: A
authors: Lupo
authors: Renal Unit, Department of Medicine, University-Hospital of Verona, Verona, Italy.
authors: Gianluigi
authors: G
authors: Zaza
mesh_terms: D000328:Adult; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D016022:Case-Control Studies; D020170:Caspase 1 / Q000378:metabolism; D003497:Cyclic N-Oxides / Q000378:metabolism; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D058847:Inflammasomes / Q000235:genetics / Q000378:metabolism*; D018836:Inflammation Mediators / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D008928:Mitochondria / Q000378:metabolism; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; D021381:Protein Transport; D017382:Reactive Oxygen Species / Q000378:metabolism; D006435:Renal Dialysis; D051436:Renal Insufficiency, Chronic / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism* / Q000628:therapy
chemical_list: D002352:Carrier Proteins; D003497:Cyclic N-Oxides; D016207:Cytokines; D058847:Inflammasomes; D018836:Inflammation Mediators; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C453881:NLRP3 protein, human; D017382:Reactive Oxygen Species; D020170:Caspase 1; C003959:TEMPO