Skip to Content
Merck
  • Targeting the TXNIP-NLRP3 interaction with PSSM1443 to suppress inflammation in sepsis-induced myocardial dysfunction.

Targeting the TXNIP-NLRP3 interaction with PSSM1443 to suppress inflammation in sepsis-induced myocardial dysfunction.

Journal of cellular physiology (2021-01-17)
Linhua Wang, Hongsheng Zhao, Huifen Xu, Xiangxin Liu, Xinlong Chen, Qingyun Peng, Mingbing Xiao
ABSTRACT

Sepsis-induced myocardial dysfunction (SIMD), a deadly symptom in sepsis patients, is mainly caused by cardiovascular inflammation. However, it remains unclear how systemic inflammation triggers and aggravates cardiovascular inflammation in the pathogenesis of SIMD. This study found that proinflammatory cytokines and H2 O2 concentrations were significantly induced in SIMD-mice. In particular, a microarray analysis of CD63+ exosomes isolated from sham- and SIMD-monocytes revealed a significant induction of thioredoxin-interacting protein (TXNIP) and NLR family pyrin domain-containing 3 (NLRP3). We proved that oxidative stress caused the disassociation of the TXNIP-TRX2 (thioredoxin 2) complex and the assembly of the TXNIP-NLRP3 complex. In addition, this finding showed that the latter complex could be embedded into CD63+ exosomes and traffic from monocytes to the resident heart macrophages, where it activated caspase-1 and cleaved inactive interleukin 1β (IL-1β) and IL-18. Furthermore, using an amplified luminescent proximity homogeneous assay (Alpha) with GST-TXNIP and His-NLRP3, we obtained a small molecule named PSSM1443 that could disrupt the TXNIP-NLRP3 interaction in vitro, impairing NLRP3 downstream events. Of note, after administering PSSM1443 to the SIMD-mice, we found the small molecule could significantly suppress the activation of caspase-1 and the cleavage of pro-IL-1β and pro-IL-18, reducing inflammation in the SIMD-mice. Collectively, our results reveal that monocyte-derived exosomes harbor the overexpressed TXNIP-NLRP3 complex, which traffics from circulating monocytes to local macrophages and promotes the cleavage of inactive IL-1β and IL-18 in the macrophages, aggravating cardiovascular inflammation. PSSM1443 functions as an inhibitor of the TXNIP-NLRP3 complex and its administration can decrease inflammation in SIMD-mice.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Lipopolysaccharides from Escherichia coli O55:B5, purified by ion-exchange chromatography, TLR ligand tested
Sigma-Aldrich
DL-Glyceraldehyde 3-phosphate solution, 45-55 mg/mL in H2O
Sigma-Aldrich
Dulbecco′s Modified Eagle′s Medium - high glucose, With 4500 mg/L glucose, L-glutamine, and sodium bicarbonate, without sodium pyruvate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Mouse Tumor Necrosis Factor α ELISA Kit, for serum, plasma and cell culture supernatant
Sigma-Aldrich
Mouse IL-1 β ELISA Kit, for serum, plasma and cell culture supernatant
Sigma-Aldrich
Penicillin-Streptomycin, Solution stabilized, with 10,000 units penicillin and 10 mg streptomycin/mL, 0.1 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
TWEEN® 20, for molecular biology, viscous liquid
ECL Western Blotting Reagents, Cytiva RPN2106
Sigma-Aldrich
Goat Anti-Rabbit IgG Antibody, Peroxidase Conjugated, 1 mg/mL (after reconstitution), Chemicon®
Sigma-Aldrich
Goat Anti-Mouse IgG Peroxidase Conjugate, lyophilized, Calbiochem®
Roche
cOmplete, Mini Protease Inhibitor Cocktail, Tablets provided in a glass vial
Sigma-Aldrich
Fetal Bovine Serum, USA origin, Heat Inactivated, sterile-filtered, suitable for cell culture, suitable for insect cell culture, suitable for hybridoma
Sigma-Aldrich
Mouse Il18 / Interleukin-18 ELISA Kit, for serum, plasma and cell culture supernatants
Millipore
Immobilon®-FL PVDF, 0.45 m, 8.5 cm x 10 m roll, 0.45 um ( 0.45 micron filter ) pore size Hydrophobic PVDF Transfer Membrane for western blotting using Fluorescent, Chemifluorescence, Chromogenic, or Chemiluminescence. Comes in a 8.5 cm x 10 m roll.
Sigma-Aldrich
Mouse IL-6 ELISA Kit, for serum, plasma and cell culture supernatant