Objective　:　To　explore　the　role　of　Kupffer　cells　in　APAP-induced　liver　injury　by　using　the　Cre/iDTR　system　to　specifically　deplete　Kupffer　cells.　Methods　:　ClecAfCre/iDTR　mice　were　injected　intraperitoneally　with　1μg　D　T　(Diphtheria　Ttoxin,　diphtheria　toxin),　and　the　depletion　efficiency　of　Kupffer　cells　in　the　liver　was　detected　by　flow　cytometry.　Then,　ClecAfCre/iDTR　mice　were　divided　into　four　groups,　namely　control　group,　DT　group,　APAP　group,　DT　and　APAP　group.　300　mg/kg　APAP　was　injected　intraperitoneally　after　18　h　of　starvation　to　establish　an　acute　liver　injury　model.　ELISA　and　CBA　analysis　were　used　to　detect　the　release　of　ALT　and　AST　and　the　secretion　of　inflammatory　factors　in　serum.　H　&　E　staining　and　T　UNEL　staining　were　used　to　detect　the　necrosis　of　liver　tissue　and　the　apoptosis　of　hepatocytes.　Immunohistochemistry　was　used　to　detect　inflammatory　cell　infiltration　in　the　liver　and　immunoblotting　to　detect　activation　of　the　J　N　K　signaling　pathway.　qPCR　was　used　to　detect　the　transcriptional　expression　of　inflammatory　factors　in　the　liver.　Results　:　Hepatic　Kupffer　cells　in　ClecAfCre/iDTR　mice　were　almost　completely　depleted　at　12　and　24　h　after　injection　of　1　μg　DT.　Depletion　of　Kupffer　cells　potently　ameliorated　APAP-induced　liver　injury,　as　manifested　by　decreased　serum　ALT　and　AST　levels,　less　hepatic　necrotic　area　and　hepatocyte　apoptosis,　reduced　inflammatory　cell　infiltration　and　production　of　inflammatory　cytokines,　and　abrogated　activation　of　the　JNK　pathway.　Conclusion　:　This　study　demonstrated　that　removal　of　Kupffer　cells　significantly　attenuated　APAP-induced　liver　injury,　suggesting　an　essential　contribution　of　Kupffer　cells　to　APAP　hepatotoxicity.　©　2024　China　Biotechnology　Press.　All　rights　reserved.