Treatment Of Liver Injury By Interleukin-35 Modified Mesenchymal Stem Cells In Mice

Objective:This study was designed to investigate the result of interleukin-35(IL-35)modified mesenchymal stem cells as treatment of acute liver injury in mice;We established a stable model of acute liver injury model used Con A in mice through intravenous injection of IL-35 gene-modified mesenchymal stem cells,to observe its treatment of acute liver injury in mice.Methods:Adipose-derived MSCs was obtained using type I collagenase from mouse fat.IL-35 gene was integrated to working plasmid of lentivirus,and co-transfadecl with packing plasmcols into packaging too alls produce lestvira which preoprecis IL-35.This paper through the isolation and culture of mesenchymal stem cells from mouse adipose tissue,IL-35 gene inserted into the lentiviral packaging plasmid transfected into packaging cells overexpressing IL-35 lentiviral particles,IL-35overexpression lentivirus vector(IL-35-LV)was made.The mice were administrated with IL-35 gene modified MSCs,wild type MSCs or control saline solution intravenously after constructing acute liver injury model.The effects of the treatment were evaluated by determination of the survival of the state,the degree of liver injury and other physiological and pathological changes.Result:1.Isolation adipose-derived MSCs successfly.The primary cells need culture7-10 days,then the passage cells only need culture 2 days,the best activity for 3-6passage cells.2.After infection of IL-35-LV,MSCs can secrete IL-35,it can be obtained at the beginning of liquid after 48 hours since transfection with the virus infection function.3.15mg/g Con A can induce liver injury of mice successfully.4.1000ul/2500ul the virus liquid can keep the rate of MSC infection being 25.4%,the best number of cells injected is 5×10~5 cells.5.To observe the state of life in the model all the mice appear listlessness,lazy,cold,shivering.ALT in 6 hours bigeng to increase,and 24 h reached a peak,after 48 hours was restored.There 18h results show that con a group 2664.63+28.795,MSC group948.67+13.788,IL-35-MSC group 477.41+185.71;24h results show that con a group 3536.62+117.94,MSC group 2388.53+671.38,IL-35-MSC group 709.69+212.72.The level of ALT in the treatment group significantly lower than the other two groups,P<0.05.After 72 hours dissected the survive mice in each group to find that in IL-35-MSC group liver ischemic injury was lighter than the other two groups;HE staining and pathological changes of IL-35-MSC group was significantly less than the other two groups.Conclusion:1.The key point of adipose derived mesenchymal stem cells is the primary and culture.Cells provce to variation after too much passage.We used the cells after 3-6 activity decreased.2.Lentivirus can be used as effective carrier of IL-35 gene modification on MSC.3.Results there were obvious differences in IL-35-MSC treatment group comperde to other two groups,has the role of immunological factors in treatment of IL-35-MSC,by regulating liver injury mice body immune cell activity,IL-35-MSC are significantly reduce the seram ALT level and attanuase the live dying dgree,apoptosis,thereby effectively inhibit liver failure.