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• W4377029385 abstract "<b>Abstract ID 29339</b> <b>Poster Board 86</b> Each year in the United States, there are 190,600 cases of acute lung injury (ALI), associated with over 74,000 deaths. Data shows acyl-coenzyme A acetyltransferase-1 (ACAT-1) inhibition improves pulmonary inflammation in an <i>in vivo</i> murine model of ALI. We hypothesize that ACAT-1 inhibition has anti-inflammatory effects beyond its intended use to reduce cholesterol esterification. The purpose of this study is to establish an <i>in vitro</i> bone marrow-derived macrophage (BMDM) model to investigate the effect of ACAT-1 inhibition on macrophage activation. Monocytes were harvested from the bone marrow of 6-8 week old C57BL/6J (wild-type) mice. Cells were stimulated with M-CSF on d0, 3, and 7 to induce macrophage differentiation. To examine if ACAT-1 inhibition limits macrophage activation, K-604 was co-administered with M-CSF in the wild-type derived cells. Cells were then treated with LPS on d7 and harvested after 24h. Nitrite, Nos2 expression, and iNOS protein density were determined through nitrite colorimetric measurement, RT-qPCR, and western blot, respectively. Glycolytic function and oxygen consumption rate were determined through Seahorse^TM Glycolysis and Mito Stress Tests. Nitrite (μm/protein) was observed to significantly increase (*p<0.05) in LPS-stimulated cells compared to control (8±1.5* v. 0.9±0.1), which was reduced with K-604 treatment (3.2±1.0). LPS-induced Nos2 fold change (11,709±2,955* v. 1.0±0.1) was reduced with K-604 conjunctive treatment compared to LPS alone (2,235±586). These data suggest that LPS-induced NO production was hindered due to a lack of the iNOS protein, which may have implications for macrophage activation. This aligns with reported <i>in vivo</i> data, where K-604 reduced pulmonary inflammation in a rodent model of ALI. Metabolic and signaling implications of ACAT-1 inhibition in this model will be discussed." @default.
• W4377029385 created "2023-05-19" @default.
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• W4377029385 date "2023-05-18" @default.
• W4377029385 modified "2023-09-29" @default.
• W4377029385 title "ACAT-1 Inhibition Limits iNOS in an<i>In Vitro</i>Model of Macrophage Activation" @default.
• W4377029385 doi "https://doi.org/10.1124/jpet.122.293390" @default.
• W4377029385 hasPublicationYear "2023" @default.
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