Expression of NLRP3 inflammasome, cytokines and vascular mediators in the skin of systemic sclerosis patients.
BACKGROUND
The activated NLRP3 inflammasome is associated with the etiology of fibrotic diseases. The role of inflammasomes in SSc is still poorly understood.


OBJECTIVE
To determine the expression of NLRP3 (nucleotide-binding domain, leucine-rich-repeat-containing family, pyrin domain-containing 3) in the skin of patients with systemic sclerosis (SSc) and its relationship with pro-inflammatory cytokines and vascular mediators expression.


METHODS
Skin biopsies were taken from 42 patients with either limited or diffuse SSc (21 lcSSc and 21 dcSSc), and from 13 healthy individuals. Using real-time polymerase chain reaction (PCR), the relative expression of caspase-1, IL-1β, IL-18, IL-33, TGF-β, ET-1, iNOS and eNOS genes, were measured. The location of NLRP3 and IL-1β were also determined by immunohistochemistry. Clinical characteristics were evaluated.


RESULTS
The mean age of the patients was 49.3 ± 12.9 (lcSSc), 44.6 ± 1 3.8 (dcSSc), and 45 ± 14.1 (healthy individuals). Compared to healthy individuals, the skin of both subtypes of SSc showed a significant increase (P < 0.05) in NLRP3, caspase-1, IL-1β, IL-18 and ET-1. Samples of lcSSc also showed a significant increase of eNOS (P < 0.029), iNOS (P < 0.04) and TGF-β (P < 0.05). Dermal fibrosis evaluated by modified Rodnan skin score (MRSS) had significant correlation with NLRP3, IL-1β, IL-18, and ET-1. Immunohistochemical analysis showed stronger staining of NLRP3 and IL-1β cytoplasmic expression in the keratinizing squamous epithelium of skin from SSc patients compared to controls.


CONCLUSIONS
This study identified NLRP3 over-expression in skin of patients with SSc. Skin thickness correlates positively with the NLRP3 inflammasome gene expression and with the vascular mediator and pro-fibrotic ET-1, suggesting that NLRP3 inflammasome plays a role in the pathophysiology of skin fibrosis in human SSc.
Maria A
MA
Martínez-Godínez
Maria P
MP
Cruz-Domínguez
Luis J
LJ
Jara
Aarón
A
Domínguez-López
Rosa A
RA
Jarillo-Luna
Olga
O
Vera-Lastra
Daniel H
DH
Montes-Cortes
Rafael
R
Campos-Rodríguez
Dulce M
DM
López-Sánchez
Cesar M
CM
Mejía-Barradas
Enrique E
EE
Castelán-Chávez
Angel
A
Miliar-García
D000328:Adult; D001706:Biopsy; D002352:Carrier Proteins / Q000235:genetics*; D016207:Cytokines / Q000378:metabolism*; D019332:Endothelin-1 / Q000378:metabolism; D005260:Female; D005355:Fibrosis; D005786:Gene Expression Regulation; D006801:Humans; D058847:Inflammasomes / Q000378:metabolism*; D018836:Inflammation Mediators / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; D060888:Real-Time Polymerase Chain Reaction; D012595:Scleroderma, Systemic / Q000235:genetics / Q000276:immunology / Q000473:pathology*; D012867:Skin / Q000473:pathology*
D002352:Carrier Proteins; D016207:Cytokines; D019332:Endothelin-1; D058847:Inflammasomes; D018836:Inflammation Mediators; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C453881:NLRP3 protein, human