| ObjectiveTo investigate Human Liver-Derived Stem Cells Protective Effect on AFL and ALI induced by Concanavalin A in mice and its Protective Mechanisms of ALI.Methods1.Eighty-eight mice were randomly divided into three groups.Control group,mice were given PBS without ConA or Experimental group.According to different injection time of ConA after the HYX1 pretreatment,model group and experimental group can be divided into3,6,12,24,48 hours groups;mice were injected with 200 ul PBS via tail vein in control group.mice were treated with human liver stem cells(HYX1)via intraperitoneal before challenge with ConA in experimental group.mice were treated with PBS 200 ul via intraperitoneal before challenge with ConA in model group.The levels of alanine aminotransferase(ALT),aspartate transaminase(AST)and total bilirubin(TBIL)in peripheral blood were detected after 24 hours.Mice liver tissues of each group were estimated by pathological examination and Ishak scoring.2.Total of male C57BL/6 mice were randomly divided into 0,2,6,12 and 24 hours groups according to the different time points after injection of ConA,The four groups(2?6?12?24 hours)were further divided into model groups and human liver-derived stem cells treated groups except for 0 hour group which were only treated by human liver-derived stem cells.Each group has 8 mice.The levels of alanine aminotransferase(ALT)?aspartate transaminase(AST)?TNF-? inperipheral blood were measured.Liver tissue sections were used to observe pathological changes and detect the apoptosis of hepatocytes;The expression of TNF-??IL-6 mRNA from the liver tissue were determined by real-time fluorescent quantitative PCR.3.Thirty male mice were randomly divided into blank group(N),model group(M)and stem cell treatment group(S).The mice in group N were fed with a normal diet,and those in the other groups were fed with Lieber-DeCarli alcohol liquid diet,and,at the same time,mice in group S were received intraperitoneal transplantation of human liver-derived stem cells twice per week.Four weeks after intervention,the body weight,liver index,ALT,AST,total bilirubin(TBIL),total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C)in mice were studied.Besides,the levels of TG and free fatty acids(NEFA)in the liver were measured and liver pathological examination was performed.Results1.the levels of AST and ALT were significantly higher in model groups 3hours,6hours,12 hours,24hours than in experimental groups 3hours?6hours?12hours?24hours(P<0.05),while there was no statistically significant difference between model group 48 hours and experimental group 48hours(P> 0.05).And the levels of TBIL were higher in groups 6hours,12 hours,24hours than in groups 6hours,12 hours,24hours(P<0.05).The levels of ALT?AST?TBIL were higher in model groups than in control group(P<0.01).The levels of ALT?AST?were higher in groups(3hours?24hours?48hours)than in control group(P<0.01).Ishak inflammation scores decreased significantly in in group experiment as compared with the group model(P < 0.05).2.The levels of alanine aminotransferase(ALT)aspartate transaminase(AST)were significantly decreased in the human liver-derived stem cells treated 12 h group and 24 h group in peripheral blood compared to their model groups,and the pathological sections of liver tissue of 12 h group and 24 h group showed that compared with their model groups had significant reductions in the degree of necrosis and Ishak score(P<0.05 or P<0.01).2h ? 6h ? 12 h human liver-derived stem cells treated group showed a significantly decreasing of serum TNF-? and TNF-??IL-6 mRNA in liver tissue(P<0.05 or P<0.01).The hepatocyte apoptosis decreased significantly in the 12 h human liver-derived stem cells treatedgroup.3.Compared with group M,serum ALT?AST?TG in group S were significantly decreased(P <0.05).Though the levels of serum TBIL,TC and HDL-C were also decreased,but the differences were not statistically significant(P > 0.05).And the levels of TG and NEFA of liver tissues were significantly decreased(P <0.05).Liver HE staining and oil red O staining showed that the degree of liver steatosis in group S was significantly less than that in group M.Conclusion1.Intraperitoneal transplantation of human liver stem cells,especially group 6 hours and group 12 hours,can obviously protect against ConA-induced acute liver injury in mice2.human liver-derived stem cells protects against ConA-induced liver injury,which may be mediated by its anti-liver cell apoptosis and decreasing TNF-? ? IL-6 level in both peripheral blood and liver tissue.3.Intraperitoneal transplantation of human liver stem cells can have obviously protective effect on alcohol liver injury in mice. |
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