Human Endodermal Stem Cells Improve Liver Failure In A Model Of Acute Liver Failure By IL-10

Part 1:The therapeutical effect of endoderm stem cells transplantation on acute liver failure.Objective:To probe the therapeutical effect of endoderm stem cell transplantation on acute liver failure.Method:We induced the acute liver failure of rats by intraperitoneal injection of D-galactosamine(500mg/kg)and LPS(50ug/kg).All rats were divided randomly into four groups:normal control group(n=20);ALF group(n=20):D-galactosamine(500mg/kg)and LPS(50ug/kg)were injected intraperitoneally;sham-operated group(n=20),1 ml saline were injected via protal vein under the condition of asepsis after injection of D-galactosamine and LPS;EPC treatment group,1�10^7 EPCs were injected via protal vein under the condition of asepsis after injection of D-galactosamine and LPS.10 rats from each group were picked up randomly for praxiology and survival analysis.At different time points after intervention,rats in each group were sacrificed with an overdose of isoflurane.ALT,AST,TBil,blood ammonia,PT,hematoxylin-eosin staining of liver,TUNEL staining for apoptosis and Immunohistochemistry ki67 staining for the proliferation of hepatocytes were detected.Results:The praxiology,survival,liver function,blood ammonia,PT,hematoxylin-eosin staining,TUNEL staining,Immunohistochemistry ki67 of sham-operated group are similar to ALF group.In comparison to the ALF group and sham-operated group,the EPC treatment group had a better praxiology,a higher survival rate and a significantly better liver function,blood ammonia and PT,.The liver HE staining showed that the liver necrosis and inflammatory cell infiltration was significantly relieved.Immunohistochemical Ki67 staining showed EPC treatment group have higher levels of liver cell proliferation.TUNEL staining showed that the level of liver cell apoptosis in EPC treatment group was lower.Conclusions:There was a significant therapeutical effect of endoderm stem cell transplantation via protal vein on acute liver failure.Part 2:The research of the mechanism of endoderm stem cells on the treatment of acute liver failure.Obiective:To explore the mechanism of EPCs on the treatment of acute liver failure.Method:We first tested the level of rat IL-10 and human IL-10 in the rat liver after EPCs transplantation by ELISA.Then we put the serum of ALF rats into EPC culture,and tested the level of human IL-10 in the supernatant by ELISA after 24h or 48h.60 rats were divided randomly into three groups:normal control group(n=20);ALF group(n=20),D-galactosamine(500mg/kg)and LPS(50ug/kg)were injected intraperitoneally;IL-10 treatment group,mouse IL-10(30ug/kg)were injected via tail vein after injection of D-galactosamine and LPS.To compare treatment effects by detecting survival,IL-10 of livers,ALT,AST,TBiL,blood ammonia,PT and histopathology.80 rats were divided randomly into four groups:normal control group(n=20);sham-operated group(n=20),1 ml saline were injected via protal vein under the condition of asepsis after injection of D-galactosamine(500mg/kg)and LPS(50ug/kg);EPC treatment group,1�10^7 EPCs were injected via protal vein under the condition of asepsis after injection of D-galactosamine and LPS;anti-IL-10 group,anti-IL-10 antibody(20mg/kg)was injected after injected of D-galactosamine,LPS and EPCs.To compare treatment effects by detecting survival,ALT,AST,TBiL,blood ammonia,PT and histopathology.Results:The result of ELISA showed the levers of rat IL-10 in ALF group and sham-operated group are nearly the same.In comparison to the ALF group and sham-operated group,the level of the rat IL-10 of the EPC treatment group was significantly higher.But the levels of the human IL-10 of all groups ware very low and not statistically significant,and a similar result was got in the supernatant.The injection of rat IL-10(10ug/kg)also had a therapeutic effect,reduced the mortality,improved liver function,reduced blood ammonia and PT.The liver HE staining showed that the liver necrosis and inflammatory cell infiltration was significantly relieved.When anti-IL-10 antibody was injected on the ALF rats with treatment of EPCs,the levels of ALT,AST,TBiL,blood ammonia,PT were rised obviously.The liver HE staining showed that the liver necrosis and inflammatory cell infiltration was significantly aggravated.Conclusions:EPCs played a therapeutic role by stimulating ALF rats secreting IL-10.