Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most common chronic liver disease, posing public health risks from potential irreversible liver damage. Curcumin (Cur), a polyphenolic compound from Zingiberaceae and Araceae, exhibits antihyperlipidemic and insulin-sensitizing effects. However, its targets and mechanisms in MASLD remain unclear. This study aimed to identify the potential targets and mechanisms by which Cur ameliorates MASLD. Cur markedly improved metabolic disorders and inflammation in rats and reduced intracellular lipid accumulation in HepG2 cells, primary hepatocytes, and AML12 cells. TKFC was identified as a direct target of Cur which binds to TKFC at Val136 and Glu538, thereby activating it. Gene microarray screening showed that Cur down-regulated the mRNA expression of Gpat3. TKFC knockdown attenuated the down-regulation of GPAT3 by Cur, whereas TKFC overexpression reversed this effect. In Tkfc-deficient mice, the therapeutic effects of Cur were attenuated and the down-regulation of GPAT3 and related proteins was hindered, thereby promoting triglyceride production in the liver. Further mechanistic studies revealed that Cur targets TKFC to inhibit GPAT3 expression by modulating the AMPK-STAT3 axis. Our study highlights TKFC as a novel and promising target for MASLD and offers new insights into the molecular mechanisms through which Cur ameliorates MASLD.

Curcumin  /  MASLD  /  Chemical proteomics  /  TKFC  /  GPAT3  /  Lipid metabolism  /  Activity-based protein profiling  /  Triacylglycerol