Non-alcoholic fatty liver disease (
NAFLD)is accompanied by typical inflammatory damage and cell death. As a pro-inflammatory form of cell death, pyroptosis participates in important
pathological processes involved in
NAFLD. Regulatory roles of both
CCCTC-binding factor (CTCF) and dipeptidyl peptidase-4 (DPP4) have been reported in
NAFLD, but it is still unclear whether the mechanism of action of
gardenoside, a potential therapeutic for
NAFLD, can be driven via these
proteins. In this study, the direct interaction between CTCF and DPP4 was first confirmed by a dual-
luciferase reporter assay system. Then, a cell model of
NAFLD was established by induction with
palmitic acid (PA) and
lipopolysaccharide (LPS). A mouse
NAFLD model was established, and the effect of
gardenoside on both the cell and mouse models of
NAFLD was also investigated. Increased
lipid accumulation, NLRP3
inflammasome activation, and hepatocyte pyroptosis were recorded in
NAFLD in vitro and in vivo.
Gardenoside treatment effectively reduced the
lipid accumulation, increased cell viability, reduced
reactive oxygen species (ROS) generation, and attenuated pyroptosis and apoptosis in
NAFLD in the in vitro and in vivo models. Alterations in these biological processes were evidenced by the decreased expression levels of several pro-pyroptotic markers including the NLR family, pyrin domain-containing 3 (NLRP3), apoptosis-related speckle-like
protein (ASC), caspase-1 p20, Gasdermin D N-terminal domain (GSDMD-N), and IL-1β, along with simultaneously decreased CTCF and DPP4 levels. Importantly, CTCF silencing or DPP4 silencing exhibited effects similar to
gardenoside treatment, while CTCF overexpression counteracted this trend, which indicated that CTCF might be a target responsible for
gardenoside-induced alleviation of
NAFLD, such
therapeutic effects might be achieved through controlling the expression of the direct target of CTCF (DPP4) and several downstream molecules. In general, the current study provides a promising strategy for
NAFLD treatment.