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AIM: To investigate the importance of direct contact betweenKupffer cells (KCs) and hepatocytes (HCs) during hepaticinflammatory responses,and the effect of leflunomide’s activemetabolite,A_(771726),on cytokines in KCs,HCs and KC cocultures(DC cocultures).METHODS: KCs and HCs in liver were isolated by digestionwith pronase and collagenase.Lipopolysaccharide (LPS)-induced inflammatory response in monocultures of rat HCsand KCs was compared with that in DC cocultures.Tumornecrosis factor-α (TNF-α) and interleukin-1 (IL-1)concentrations in different culture supernatants weremeasured with ELISA.TNF-α mRNA in KCs of inflammatoryliver injury was analyzed with reverse transcriptasepolymerase chain reaction (RT-PCR).RESULTS: DC cocultures strongly exhibited the productionof TNF-α and IL-1 compared with other cultures,and thesecytokines were mainly produced by KCs,especially byactivated KCs.Time course studies revealed an increasedproduction of TNF-α preceding the IL-1 production,suggesting that increased TNF-α levels could be involvedin the increase of IL-1 production.Lefiunomide’s activemetabolite,A_(771726),had significantly inhibitory effect on TNF-αand IL-1 at protein and transcription levels,and the reducedproduction of IL-1 by A_(771726) was associated with theinhibitory action of A_(771726) on TNF-α.CONCLUSION: Leflunomide can inhibit hepatocyte damageby inhibiting proinflammatory cytokine release from KCs.
AIM: To investigate the importance of direct contact between Kupffer cells (KCs) and hepatocytes (HCs) during hepatic inflammatory response, and the effect of leflunomide’s active metabolite, A 771726, on cytokines in KCs, HCs and KC cocultures (DC cocultures). METHODS : KCs and HCs in liver were isolated by digestionwith pronase and collagenase. Lipopolysaccharide (LPS) -induced inflammatory response in monocultures of rat HCs and KCs was compared with that in DC cocultures. Tumornecrosis factor-α (TNF-α) and interleukin-1 ( IL-1) concentrations in different cultures supernatants weremeasured with ELISA.TNF-α mRNA was analyzed in KCs of inflammatoryliver injury with reverse transcriptase polymerase chain reaction (RT-PCR) .RESULTS: DC cocultures strongly exhibited the production of TNF-α and IL-1 compared with other cultures, and these cytokines were mainly produced by KCs, especially by activated KCs.Time course studies revealed an increased production of TNF-α preceding the IL-1 production, suggestin g that increased TNF-α levels could be involved in the increase of IL-1 production. Lefiunomide’s active metabolite, A 771726, had significant inhibitory effect on TNF-α and IL-1 at protein and transcription levels, and the reduced production of IL-1 by A_ (771726) was associated with the inhibitory action of A_ (771726) on TNF-α. CONCLUSION: Leflunomide can inhibit hepatocyte damage by inhibiting proinflammatory cytokine release from KCs.