Investigation Of The Protective Effect Of Mycophenolate Mofetil On Hepatic Warm Ischemia-reperfusion Injury In Mice

Objective To investigate the protective effects and newly possible mechanisms of mycophenolate mofetil(MMF) on the expression of Toll-like receptor 4, autophagy induction and inflammation in hepatic warm ischemia-reperfusion injury in mice.Methods The partial hepatic warm ischemia-reperfusion model was established in mice. A total of 96 healthy male Balb/c mice were divided randomly into 4 groups(24 mice each group): Group A, control group;Group B, mycophenolate mofetil(MMF) group; Group C, chloroquine(CQ) group; Group D, MMF plus CQ combination group. According to reperfusion time, every group was divided into four subgroups randomly:sham operation, 1 h, 6 h and 24 h after reperfusion. Histopathological analysis and serum ALT level were examined for assessing liver function and degree of hepatic damage. The expression levels of TNF-? and IL-6m RNA were detected by Real-time PCR while p-NF-?B by western blot.TLR4 and HMGB-1 expression were monitored by western blot and immunehistochemisty. Autophagy and apoptotic related markers,including LC3, p62, Beclin-1, Caspase-3 and Bcl-2, were determined by western blot analysis. And TUNEL analyse was also tested.Results Compared to control group, MMF and CQ decreased serum ALT level significantly and lessened hepatic damage with lower Suzuki's score(P<0.05). MMF could induce autophagy by increasing LC3 II/I ratio and the numbers of autophagosomes comparing with other three groups(P<0.05). At one hour after reperfusion, MMF increased the expression of TLR4 and HMGB-1(P<0.05). For hepatic inflammation, MMF and CQ decreased the transcription of p-NF-?B and the expressions of TNF-? and IL-6 m RNA comparing with control group. Also, MMF and CQ lessened hepatocytes' apoptosis by decreasing Caspase-3 expression and TUNEL positive cells' numbers.Conclusion MMF could protect liver from warm ischemia-reperfusion injury by induction of autophagy, decreasing hepatic inflammation and apoptosis. The mechanism of induction of autophagy could be achieved by activation of TLR4 signal.