Study On The Therapeutical Effect Of G-CSF On Mice With Acute Live Failure And Exploration Of The Possible Anti-apoptosis Mechanism

Objectives:To investigate the protective role of granulocyte colony-stimulatingfactor(G-CSF) on mice with acute liver failure induced by D-GalN/LPS,andexplore the possible mechanism of anti-apoptoticon liver.Methods:1、The acute liver failure mice model was induced by intraperitonealinjection of D-GalN/LPS.Three factors,incluing dosages of D-GalN,LPS anddilution rate of D-GalN/LPS mixture, which may influence the success ofmodeling were chosen for the experiment. Each factor was tested in4level inthe experiment. The orthogonal array L16(43) was selected to arrange the testprogram. The mortality rate of mice within24hours of the administration ofD-GalN/LPS was observed. Serum ALT level, histological changes in liver, andhepatocytes apoptosis were also detected.2、Male C57/BL6mice were divided into G-CSF treatment group, controlgroup and normal control group. The mice in the first two groups were given aintraperitoneal dose of D-GalN(350mg/kg body weight)/LPS(30μg/kg bodyweight)with and without subcutaneous treatment of G-CSF(250mg/kg bodywight) at96h,72h,48h,24h and0h before D-GalN/LPS administration.Survival rate of mice within24h after D-GalN/LPS administration was observed.Serum ALT was tested to assess liver injury. Apoptosis of liver tissue weredetected by TUNEL. Liver Bcl-2, Bcl-xl expression were detected on mRNAlevel by Real-time PCR. Western blot was used to analyze liver Bcl-2expressionin liver tissue and to furtherobserve the anti-apoptptic role of G-CSF.Results:1、An optimized administration was selected for the ideal model of acute liver failure in mice:350mg/kg D-GalN combined with LPS30μg/kg injectedintraperitoneally and the optimal dilution rate is3..By measuring serum ALTlevel, detecting liver tissue HE and TUNEL dying.The optimal dosage wasmake sure for the mice model with acute liver failure.2、G-CSF significantly improved the survival rate of mice within24h afterD-GalN/LPS administration（χ~2=3.989, P=0.046）. In treatment group,serumALT was lower than control group6h after D-GalN/LPSadministration(681.0±113.2VS1371±210.9t=2.882,P=0.044). Liverhistopathology changes in G-CSF group6h after D-GalN/LPS administrationmice were characterized by hepatocyte swelling, inflammatory infiltrationwithout mass necrosis, dissociation of hepatic cord andintralobular haemorrahge which were observed in the control group. Liver tissuefrom those G-CSF group showed significant lower TUNEL-positve cell countthan mice in control group6h after D-GalN/LPS administration(654±64.29VS1155±192.6t=2.467P=0.033). Bcl-2, Bcl-xl expression of livertissue were upragulated in G-CSF group(P=0.017, P=0.033inspectively).Conclusions: G-CSF may exerts its protective effects on mice model with acuteliver failure induced by D-GalN/LPS by suppressing hepatocyte apoptosis. Theanti-apoptotic effects of G-CSF may be take effect by upregulation ofBcl-2,Bcl-xl.