TY - JOUR
T1 - Suppression of the pro-inflammatory NLRP3/interleukin-1β pathway in macrophages by the thioredoxin reductase inhibitor auranofin
AU - Isakov, Elina
AU - Weisman-Shomer, Pnina
AU - Benhar, Moran
N1 - Funding Information: We thank Dr. Liat Linde and Dr. Nili Avidan from the Genomics Core Facility at the Rappaport Faculty of Medicine & Research Institute in the Technion for BeadChips processing and data analysis. We thank Dr. Yuval Shaked and Mr. Dror Alishekevitz from the Rappaport Faculty of Medicine & Research Institute in the Technion for providing primary macrophages. This work was supported by grants from the Israel Science Foundation (grant no. 1336/10 ), the Israel Cancer Association ( 20120121 ) and the FP7 European Commission (Marie Curie) grant program ( Pirg06-GA-2009-256438 ) (to M.B.).
PY - 2014/10
Y1 - 2014/10
N2 - Background The thioredoxin/thioredoxin reductase system, which is best known for its essential role in antioxidant defense and redox homeostasis, is increasingly implicated in the regulation of multiple cellular signaling pathways. In the present study, we asked if the thioredoxin system in macrophages might regulate toll-like receptor 4 (TLR4)-dependent gene expression and consequent responses. Methods Using microarray analysis we analyzed the effect of auranofin, a highly potent and specific inhibitor of thioredoxin reductase, on the transcriptional program activated in J774 macrophages by the TLR4 agonist, lipopolysaccharide (LPS). We used quantitative real-time PCR (qPCR), Western blotting, ELISA and cytotoxicity assays to confirm and extend the microarray results. Results Global transcriptional profiling revealed that macrophage treatment with auranofin exerted a selective effect on LPS-induced gene expression, suppressing the induction of a small number of genes. Interestingly, among these suppressed genes were three members of the interleukin-1 (IL-1) family of genes, among which IL-1β was most affected. qPCR analyses confirmed the repressive effects of auranofin on IL-1 genes. In addition, qPCR and Western blot analyses showed that auranofin impaired TLR4-dependent induction of the inflammasome receptor NLRP3, which plays a critical role in IL-1β processing. Consistent with these findings, inflammasome-dependent release of IL-1β from stimulated macrophages was suppressed by auranofin as was inflammasome-mediated cell death. Conclusions Our findings suggest a regulatory role for the thioredoxin system in macrophage inflammatory signaling. Inhibition of the thioredoxin system in macrophages exerts an anti-inflammatory effect by repressing the activation of the NLRP3/IL-1β pathway.
AB - Background The thioredoxin/thioredoxin reductase system, which is best known for its essential role in antioxidant defense and redox homeostasis, is increasingly implicated in the regulation of multiple cellular signaling pathways. In the present study, we asked if the thioredoxin system in macrophages might regulate toll-like receptor 4 (TLR4)-dependent gene expression and consequent responses. Methods Using microarray analysis we analyzed the effect of auranofin, a highly potent and specific inhibitor of thioredoxin reductase, on the transcriptional program activated in J774 macrophages by the TLR4 agonist, lipopolysaccharide (LPS). We used quantitative real-time PCR (qPCR), Western blotting, ELISA and cytotoxicity assays to confirm and extend the microarray results. Results Global transcriptional profiling revealed that macrophage treatment with auranofin exerted a selective effect on LPS-induced gene expression, suppressing the induction of a small number of genes. Interestingly, among these suppressed genes were three members of the interleukin-1 (IL-1) family of genes, among which IL-1β was most affected. qPCR analyses confirmed the repressive effects of auranofin on IL-1 genes. In addition, qPCR and Western blot analyses showed that auranofin impaired TLR4-dependent induction of the inflammasome receptor NLRP3, which plays a critical role in IL-1β processing. Consistent with these findings, inflammasome-dependent release of IL-1β from stimulated macrophages was suppressed by auranofin as was inflammasome-mediated cell death. Conclusions Our findings suggest a regulatory role for the thioredoxin system in macrophage inflammatory signaling. Inhibition of the thioredoxin system in macrophages exerts an anti-inflammatory effect by repressing the activation of the NLRP3/IL-1β pathway.
KW - Inflammation
KW - Interleukin-1β
KW - Macrophage
KW - NLRP3
KW - Thioredoxin reductase
KW - Toll-like receptor
UR - http://www.scopus.com/inward/record.url?scp=84907880426&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.bbagen.2014.07.012
DO - https://doi.org/10.1016/j.bbagen.2014.07.012
M3 - مقالة
SN - 0304-4165
VL - 1840
SP - 3153
EP - 3161
JO - Biochimica et Biophysica Acta - General Subjects
JF - Biochimica et Biophysica Acta - General Subjects
IS - 10
ER -