Aflatoxin B1 (AFB1) is extremely carcinogenic and can cause
liver cancer in humans and animals with continued ingestion. As a natural compound,
curcumin (Cur) exhibits excellent anti-inflammatory, and anti-
cancer properties with few side effects. In this study, a total of 60 male mice (6-week-olds, 15 per group). After one week of acclimatization feeding, the mice were divided into control group (Con), AFB1 group,
curcumin group (Cur), and AF+Cur group. The mice were gavaged with
curcumin (Cur, 100 mg/kg) and/or AFB1 (0.75 mg/kg). To identify a new therapeutic target for AFB1-induced pyroptosis, we performed proteomic profiling for
curcumin alleviating liver injury caused by AFB1 to further validate the targets through volcano plot analysis, Venn analysis, heatmap analysis, correlation, cluster analysis, GO and KEGG enrichment. AFB1 exposure resulted in the loss of hepatocyte membrane, swelling of the endoplasmic reticulum, and a significant increase in
transaminase (ALT and AST) contents, while
curcumin greatly improved these changes. We found that differentially expressed
proteins are enriched in the endoplasmic reticulum membrane and identified ITPR2 as a target of
curcumin that alleviates AFB1-induced liver injury by proteomics. Furthermore, ITPR2 expression was detected by immunofluorescence, and qRT-PCR for
mRNA expression of genes downstream of ITPR2 (calpain1, calpain2,
caspase-12,
caspase-3). ITPR2-activated endoplasmic reticulum stress-related
proteins (calpain1, calpaini2, bcl-2, BAX, cl-caspase-12, cl-caspase-3), apoptosis (PARP) and pyroptosis (
DFNA5) related
proteins were examined by western blotting. The analysis showed that it effectively prevents AFB1-induced pyroptosis by lowering endoplasmic reticulum stress via interfering with ITPR2 and its downstream
proteins (calpain1, calpain2, bcl-2, Bax) and inhibiting
caspase-12/
caspase-3 pathway. Conclusively, this study applied proteomic profiling to elucidate ITPR2 as a new target, which might give a new perspective on the mechanism of
curcumin alleviating AFB1-induced pyroptosis.