For in-depth investigation, we evaluated the cell viability of NAFLD in vitro model using CCK8 assay and found that CTCF knockdown increased cell viability, mitigated pyroptosis, reduced ROS content in NAFLD in vitro model compared to control AML12 cells, which resembled the effects induced by 50 μM gardenoside treatment. Here, CTCF is linked to metabolic dysfunction-associated steatotic liver disease.