Kupffer Cells Mediated Emulsified Isoflurane Precondition Against Hepatic Ischemia And Reperfusion Injury In The Rats And Its Functional Mechanism Analysis

Partâ… Kupffer cells mediated emulsified isoflurane preconditioning against hepatic ischemia and reperfusion injury in the rats in vivoObjective To investigate the protective effect of pretreatment with emulsified isoflurane on hepatic ischernia and reperfusion injury in the rats and this effect may mediated by Kupffer cells.Methods A total of 32 healthy adult male SD rats, weighing between 250 g to 300 g,were randomly divided into four groups: preconditioning with intralipid group(C,n = 8),inhibition of Kupffer cells and preconditioning with intralipid group(KC,n = 8),preconditioning with emulsified isoflurane group(IP,n = 8),inhibition of Kupffer cells and preconditioning with emulsified isoflurane group(IK,n = 8).Rats suffered from ischemia for 30 min and were followed by reperfusion for 2 hours.The animals were killed at 2 hours after reperfusion. Serum levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were determined.Hepatic cell homogenate levels of MDA and SOD were determined.The histopathologic changes in the liver were also observed.The protective effect of pretreatment with emulsified isoflurane was reversed after inhibition of Kupffer cells.Results Compared with those in C group,in IP group serum levels of ALT and AST decreased significantly after hepatic ischernia and reperfusion.Hepatic cell homogenate levels of MDA decreased and SOD increased.The injury degrees of hepatic cell histopathology in IP group were much less than that in C group.While serum levels of ALT and AST,hepatic cell homogenate levels of MDA and SOD and the histopathologic changes in the liver in KC and IK groups have no marked changes compared with those in C group.Conclusion Pretreatment with emulsified isoflurane can protect liver from hepatic ischemia and reperfusion injury in the rats and this effect may mediated by Kupffer cells.Partâ…¡Kupffer cells which depended on the levels of NO mediated emulsified isoflurane preconditioning against hepatic ischemia and reperfusion injury in the rats in vivoObjective To investigate the protective effect of pretreatment with emulsified isoflurane on hepatic ischemia and reperfusion injury in the rats and observe if this effect is related with Kupffer cells and the levels of NO.Methods A total of 48 healthy adult male SD rats,weighing between 180 g to 250 g,were randomly divided into six groups:preconditioning with intralipid group(C,n = 8),inhibition of Kupffer cells and preconditioning with intralipid group(KC,n = 8),preconditioning with emulsified isoflurane group(IP,n = 8),inhibition of Kupffer cells and preconditioning with emulsified isoflurane group(IK,n = 8);activation of NO and preconditioning with emulsified isoflurane group(LA,n = 8) inhibition of NO and preconditioning with emulsified isoflurane group(LN,n = 8).Rats suffered from ischemia for 30 rain and were followed by reperfusion for 2 hours.The animals were killed at 2 hours after reperfusion. Serum levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were determined.Hepatic cell homogenate levels of MDA,SOD,NO and iNOS were determined.The histopathologic changes in the liver were also observed.Western blot were performed to analysis the iNOS expression in liver tissue after hepatic I/R.Results Compared with those in C group,in IP group serum levels of ALT and AST decreased significantly after hepatic ischemia and repeffusion.Hepatic cell homogenate levels of MDA decreased and SOD increased.Hepatic cell homogenate levels of NO and iNOS increased.The injury degrees of hepatic cell histopathology in IP group were much less than that in C group.While serum levels of ALT and AST,hepatic cell homogenate levels of MDA and SOD and the histopathologic changes in the liver in KC,IK,LA and LN groups have no marked changes compared with those in C group.Hepatic cell homogenate levels of NO and iNOS significantly increased in group LA.Hepatic cell homogenate levels of NO and iNOS significantly decreased in group LN.Western blot analysis show that iNOS expression increased significantly in IP and LA group compare to C,KC,IK and LN group.Conclusion Kupffer cells mediated emulsified isoflurane preconditioning against hepatic ischemia and reperfusion injury in the rats and this effect may be related with the levels of NO and iNOS. Partâ…¢Primary cultured Kupffer cells from rat's liver mediated emulsified isoflurane preconditioning against hepatic ischemia and reperfusion injury in vitroObjective To explore a stable and effective method for the isolation and purification of Kupffer cells from rat liver,investigate the effect of oxidative stress and apoptosis of primary cultured Kupffer cells after pretreatment with emulsified isoflurane in vitro and verify the protective effect of pretreatment with emulsified isoflurane on hepatic ischemia and reperfusion injury in the rats.Methods To explore a stable and effective method for the isolation and purification and identification of Kupffer cells from rat liver. Primary cultured Kupffer ceils were randomly divided into five groups:Control group(C), no drug added;preconditioning with 500μmol/L H₂O₂ group(H₂O₂),preconditioning with 500μmol/L H₂O₂ group for 12 hours;pretreatment with 0.2%emulsified isoflurane group(0.2%IP),pretreatment with 0.2%emulsified isoflurane for 6 hours and preconditioning with 500μmol/L H₂O₂ group for 12 hours;pretreatment with 0.1% emulsified isoflurane group(0.1%IP),pretreatment with 0.1%emulsified isoflurane for 6 hours and preconditioning with 500μmol/L H₂O₂ group for 12 hours;pretreatment with 0.05%emulsified isoflurane group(0.05%IP),pretreatment with 0.05%emulsified isoflurane for 6 hours and preconditioning with 500μmol/L H₂O₂ group for 12 hours.The ROS levels in cells were observed with fluorometric method and apoptosis levels were also measured using a flow cytometry method(FCM) that measures Annexin V and propidium iodide(PI) staining.Results In 0.2%IP group,0.1%IP group and 0.05%IP group the ROS levels in cells with fluorometric method were lower than that in H₂O₂ group.In 0.2% IP group and 0.1%IP group the apoptosis levels using a flow cytometry method(FCM) were lower than that in H₂O₂ group.Conclusion Pretreatment with emulsified isoflurane can decrease the oxidative stress and inhibit the apoptosis of primary cultured Kupffer cells and this effect may be related with the protective effect of pretreatment with emulsified isoflurane on hepatic ischemia and reperfusion injury in the rats.