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MB Sample ID: SA237439
| Local Sample ID: | LPS_ATP_2 |
| Subject ID: | SU002468 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
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Collection:
| Collection ID: | CO002461 |
| Collection Summary: | Mouse bone marrow was cultured in low glucose DMEM supplemented with 30% L929 cell-conditioned medium, 20% FBS, 2 mM L-glutamine, 1 mM sodium pyruvate, 100 U/ml penicillin, and 100 µg/ml streptomycin for 6-7 days until the cells reached confluence. BMDMs were then reseeded in culture dishes overnight in RPMI-1640 medium containing 1% Nutridoma-SP medium (Sigma-Aldrich). LPS-primed BMDMs were stimulated with 5 mM ATP in the presence or absence of 10 uM JX06 for 30 min. Unstimulated macrophages were used as control.13C tracing started by replacing medium with glucose-free medium supplemented with 2.06 g/L [U-13C]-glucose (Cayman Chemical) for 90 min. Cells were washed with ice-cold saline and intracellular metabolites were extracted using cold methanol and H2O (80:20; HPLC Grade; Sigma-Aldrich). Here, we show that pharmacological or genetic inhibition of pyruvate dehydrogenase kinase (PDHK) significantly attenuates NLRP3 inflammasome activation in murine and human macrophages and septic mice by lowering caspase-1 cleavage and IL-1beta secretion. Inhibiting PDHK reverses NLRP3 inflammasome-induced metabolic reprogramming, enhances autophagy, promotes mitochondrial fusion over fission, preserves cristae ultrastructure, and attenuates mitochondrial ROS production. The suppressive effect of PDHK inhibition on the NLRP3 inflammasome is independent of its canonical role as a pyruvate dehydrogenase regulator. We suggest that PDHK inhibition improves mitochondrial fitness by reversing NLRP3 inflammasome activation in acutely inflamed macrophages. |
| Sample Type: | Macrophages |
| Collection Method: | Cells were washed with ice-cold saline and intracellular metabolites were extracted using cold methanol and H2O (80:20; HPLC Grade; Sigma-Aldrich). The samples were pre-dissolved into 40 µL solution (LC-water, methanol, Acetonitrile 2:1:1) before injection. |
| Storage Conditions: | -80℃ |
