(a) Alignment of LXR responsive element (LXRE) on
Fasn and SCD-1 promoters.
(b) LXR–driven transcriptional activity was
determined from various macrophage lines upon stimulation with a synthetic
T0901317 (10 µM) or endogenous 25-hydroxycholesterol (10
µM) LXR ligand (luciferase activity is reported after
normalizing to transfection efficiency). (c) Relative
LXR–α and LXR–β protein
levels in various macrophage lines were examined by Western blotting
(d) Relative SCD-1 and Fasn mRNA levels
from aP2−/− macrophage
treated with a scrambled (−) or Nr1h3-specific
siRNA were examined by qRT-PCR. (e) Lysates from various
macrophages treated with scrambled (−) siRNA or a specific siRNA
against SCD-1 or Nr1h3 and stressed with
or without PA (500 µM) were examined for P-PERK and
P-eIF2–α by Western blotting. (f)
Lysates from peritoneal macrophages from
aP2−/−Nr1h3−/−
or WT mice stressed with or without PA (500 µM) were examined
for P-PERK, P-eIF2–α and cleaved PARP by Western
blotting. (g)
A cellular lipotoxicity model: Toxic levels of lipids are
sensed by the ER through an aP2-dependent pathway and induce the UPR and
lead to macrophage apoptosis. The absence of aP2 serves to reactivate
macrophage de novo lipogenesis pathways and promotes
desaturation, particularly through LXRα-mediated activation of
SCD-1, leading to increased production of bioactive lipids and resistance to
ER stress. Our findings indicate that alleviation of macrophage ER stress,
either through aP2 inhibition or enhancing ER function, is protective
against atherosclerosis.