| Background and Aims: Acute liver failure(ALF)is defined as the rapid development of hepatocellular necrosis caused by a consequence of various etiologies,with a progressive serious clinical syndrome with high mortality.Currently,liver transplantation(LT)has been considered the most effective therapy for ALF.Yet,the management for ALF is limited by a shortage of available donor organs and the operation cost is expensive.Recently,many studies reported that transplantation of bone marrow mesenchymal stem cells(BMSCs)has emerged as a promising novel therapy for acute liver failure(ALF),but with limited homing efficacy of BMSCs to injured sites.Thus,the urgent need is to enhance the homing property,then improve the therapeutic efficacy of BMSCs for ALF.Currently,it has been demonstrated that the concentration of hepatocyte growth factor(HGF)in the injured liver apparently increased.Moreover,HGF/c-met signaling pathway play a critical role in the BMSCs migrated to the injured liver.Based on the present study,we established the BMSCs engineered by recombinant c-met lentivirus(c-met-BMSCs)over-expressing c-met would promote the homing efficacy of BMSCs to the injured liver,with the high level of HGF.Methods: 1.Constructing c-met lentiviral vector for permanent transfected bone marrow mesenchymal stem cells,Thereby,the cell line of c-met-BMSCs was established(c-met-BMSCs).Then the migration of c-met-BMSCs under different concentrations of HGF was measuredby transwell system in vitro.2.A total of thirty-six SD rats were randomly divided into three groups:c-met-BMSCs group,BMSCs group and the control group(12 in each group).The model of ALF was induced by co-injection of D-Gal N and LPS.After 24 hours,the transfusion(1.0×107 cells/kg)of the c-met-BMSCs and BMSCs were injected via the vena caudalis suspended in 1ml normal saline respectively,while the control group was injected with 1ml of normal saline(NS).The survival rates of three groups were documented daily.Meanwhile,peripheral blood of all rats were collected at 0,24,48,and 72 h post co-injection of D-Gal N/LPSfor testing liver function,and liver tissues were collected for HE stainingat 24,48,and 72 h post co-injection of D-Gal N/LPS.Then,the pathological changes of liver tissue were observed and assessed by HAI scores.3.c-met-BMSCs and BMSCs were dyed by Di R with amount of 1×106 cells were transplanted into the liver through vena caudalis.After 24 hours,the cells migration to the injured liver were examined via vivo imaging system,thenthe radiant efficiency was measured.Results: 1.c-met lentiviral vector was constructed successfully via RT-PCR and the titer was 2×108 TU/ml.c-met lentiviral vector was used to transfected into BMSCs(c-met-BMSCs).With puromycinthe screening,transduction efficiency was99%.The transgene was continuously present after integration and structurally stable via Western blot,which demonstrate BMSCs transduced by the lentiviral vector carried the c-met gene,as compared to the BMSCs.Moreover,transwell chamber in vitro showed that the migration of c-met-BMSCs relied on HGF was significantly increased,compared with BMSCs.2.After co-injection of D-Gal N and LPS,the concentration of HGF from liver lysates significantly increased at 24 h and 48 h.The survival rates of ALF rats increased by50% and 83.3% respectively after transplantation of BMSCs and c-met-BMSCs.Compared with the transplantation of BMSCs and the control group,transplanted c-met-BMSCs remarkably improved the liver function and HAI scores obviously decreased.3.The radiant efficiency of transplanted c-met-BMSCs was 20 times higher than of transplanted BMSCs via vivo imaging system.It showed that the efficacy of c-met-BMSCs migrating to injured liver significantly further increased.Conclusions: c-met lentiviral vector was constructed successfully,and the transgenic c-met-BMSC was continuously present after integration and structurally stable.c-met-BMSC targeted homing tothe injured liver effectively promoted the repair of liver in ALF. |
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